EP4676554A1

EP4676554A1 - Porous and solid elastomeric bioactive polymer-ceramic composites for bone defect filling and bone tissue regeneration

Info

Publication number: EP4676554A1
Application number: EP23764737.5A
Authority: EP
Inventors: Malgorzata GAZINSKA; Sonia ZIELINSKA; Aleksandra Korbut; Konrad SZUSTKIEWICZ; Pawel PISZKO; Bartlomiej KRYSZAK; Katarzyna SIEJA; Andrzej Trochimczuk; Karolina Rudnicka; Przemyslaw Plocinski; Aleksandra SZWED-GRORGIOU; Agnieszka Krupa; Marcin WLODARCZYK; Agnieszka Sobczak-Kupiec; Dagmara SLOTA; Wioletta PLORKIEWICZ; Agnieszka Tomala; Karina PIETAK; Bozena TYLISZCZAK; Monika Biernat
Original assignee: Siec Badawcza Lukasiewicz Instytut Ceramiki I Materialow Budowlanych; Uniwersytet Lodzki; Politechnika Wroclawska; Politechnika Krakowska
Current assignee: Siec Badawcza Lukasiewicz Instytut Ceramiki I Materialow Budowlanych; Uniwersytet Lodzki; Politechnika Wroclawska; Politechnika Krakowska
Priority date: 2023-02-12
Filing date: 2023-06-30
Publication date: 2026-01-14
Also published as: WO2024167424A1

Abstract

The invention regards polymer-ceramic biocomposites in two embodiments based on poly(glycerol sebacate) (PGS) containing calcium phosphate particles, produced in the form of flexible porous materials for use as implant material for filling bone defects. Also an object of the invention is a method for producing biocomposites in the form of flexible porous and solid materials.

Description

Porous and solid elastomeric bioactive polymer-ceramic composites for bone defect filling and bone tissue regeneration

Application of PGS in soft tissue engineering is known. Patent application number US2021401580A1 describes the use of poly(glycerol sebacate) in the implantation procedure of sacroiliac joints. The composition described in the application allows bioactive compounds such as bone morphogenic proteins or selected antibiotics to be included in the polymer. The described polymeric filler is to be used as a stabilizing agent for the intercondylar spaces in the sacroiliac joint area. The next invention, known from the patent description WO2022146236A1, relates to artificial blood vessels for implant applications in the form of a cell-free extracellular matrix. The described blood vessel implant comprises an outer layer and an inner layer. Poly(glycerol sebacate) is used as the material simulating the extracellular matrix. In some described cases, a biologically active agent is added to the matrix, including growth factors (TGF-a, TGF-b, FGF-2, VEGF and others). The description allows the use of mammalian tissues devoid of cells as one of the implant layers. Another patent application (under patent number CN109289093A) describes a method for producing a bilayer material with a cylindrical shape and reticular structure, for blood flow targeting or other applications in cardiovascular regenerative medicine.

In the topic of bone tissue engineering, there are known solutions for the use in PGS-based systems. For example, patent application EP3420020A2 describes a thermosetting composite material. Both the filler and the matrix can be the product of a condensation reaction between dicarboxylic acid and a polyhydroxy alcohol (e.g. poly(glycerol sebacate), PGS). The polymeric matrix as well as the polymeric filler differ in their physico-chemical properties to allow any formulation of the material. The application allows doping with additional polymer fillers such as PCL, PGLA, PGA or PLA. The patent application describes a process for routing the PGS matrix to the filler by cryomilling (grinding at reduced temperature). The application also describes the potential use of the above-mentioned polymer-polymer composite in orthopedics when mixed with hydroxyapatite (finished paste mixture). Another known invention from patent description US2016158408A1 relates to a water-resistant composition for bone tissue repair made of bio-glass and poly(glycerin sebacate). The composition described in the application can have both thin and thick consistencies. Due to the mineral composition, the described bone filling promotes osteointegration. The resulting poly(glycerol sebacate) has a Young's modulus in the range of 0.05-1.5 MPa and can be obtained with selected degradation kinetics. The application allows doping with different bio-glass subtypes containing different elements. On the other hand, poly(glycerol sebacate) composites in which the filler and matrix are made of the same material to provide a homogeneous polymer composition and ternary composites with the addition of hydroxyapatite or tricalcium phosphate are known from international patent description US10525140B2. Also an object of the described invention is a method of obtaining a PGS as well as method of forming porous scaffolds using a salt leaching technique. Another formulation, relating to bone tissue engineering, is that of patent application WO2019152582A1. This is a formulation of an elastomeric and osteostimulating bone filling. The aforementioned recapture comprises microparticles of an elastomeric material and a filler. At least one of the elastomeric materials contains thermosetting poly(glycerol sebacate). The elastomeric matrix transfers the load thus promoting the bone regeneration process. The application also describes the possibility of forming the matrix from poly(glycerol sebacate) and producing its porous structure. In the description of the application one can find PGS-based compounds indicated as matrix compounds, e.g. PGSA, PGSU, PGS-salicylic acid as well as porophores such as calcium phosphates and hydrogen phosphates. Magnesium oxide, hydroxyapatite, silicon oxide and bio-glass, among others, are listed as fillers. The mixture of elastomer, filler and porophore is temperature cross-linked over a time range of 24-96h and then decrosslinked with a solvent depending on the porogen used. Composite materials based on poly(glycerol sebacate) in both solid and porous forms, dedicated to various applications, are known in the literature.

Referring to solid form composites, some of the more commonly studied are PGS- based materials with the addition of multi-walled carbon nanotubes. Akhilesh K. Gaharwa in his paper (Akhilesh K. Gaharwar et al in Biomaterials Science 3 (2015): 46 - 58, doi: 10.1039/c4bm00222a) produced a range of PGS composites with carbon nanotubes at 1- 10% by weight by solvent route. In vitro studies using human mesenchymal stem cells demonstrated the biocompatibility of the composites and the enhanced cell differentiation potential of PGS. Other work on PGS composites with multi-walled carbon nanotubes is also available. Yi Yan and his team (Yi Yan et al. in Journal of Colloid and Interface Science 521 (2018): 24-32, doi: org/10.1016/j.jcis.2018.03.015) produced composites with the addition of carbon nanotubes in amounts of 1-3 wt%, resulting in an increase in the mechanical performance of the material without loss of flexibility. Bioassays on an adult mouse hypothalamic neuron cell line, A59, demonstrated the potential of the fabricated materials to regenerate neural tissue. In another publication (Yi Yan et al. in Composites Science and Technology 142 (2017): 163-170, doi: 10.1016/j.compscitech.2017.02.007) the possibility of using such composites for piezoresistive sensors in biomedical engineering applications was suggested.

Existing research also indicates the potential of silica materials as additives for PGS. In a paper (C. Talla Ferrer et al. in International Journal of Polymeric Materials and Polymeric Biomaterials 69:12 (2020): 761-772, doi: 10.1080/00914037.2019.1616197) the potential of PGS composites with the addition of 5 wt% of two types of silica nanoparticles for tissue engineering in the dental field was shown. Studies conducted using mouse fibroblasts showed high bioactivity and high cell viability. Another silica- based additive used for PGS is silica glass (Xin Zhao et al. in Journal of Materials Chemistry B 3 (2015): 3222, doi: 10.1039/c4tb01693a). The composites showed the potential to regenerate bone tissue, supported by in vitro studies using cells from the MC3T3 osteoblast lineage. Another example is a PGS-based composite with the addition of micrometric bio-glass at 0-15 wt% dedicated to soft tissue engineering applications (Shu- Ling Liang et al. in Biomaterials 31 (2010): 8516-8529, doi: 10.1016/j. biomaterials.2010.07.105). PGSU (PGS enriched with HDI crosslinking compound) composites with aluminosilicates are also known (Mohammad Monem et al in Journal of Polymer research 29:25 (2022), doi: https://doi.org/10.1007/sl0965-021- 02866-7).

The solid materials based on chemically cross-linked poly(glycerol sebacate) (via HDI) enriched with curcumin and hydroxyapatite dedicated for hard tissue regenerative medicine are also known (Vafa Fakhri et al. in Polymer Chemistry 12 (2021): 6263, doi: 10.1039/dlpy01040a). The introduction of the additives increased the elastic modulus of the material, improved the proliferation and metabolic activity of cells from the mouse fibroblast L 929 line and improved the antibacterial properties of the material.

Several papers can be found in the literature focusing on the development of porous PGS-based materials for bone tissue engineering applications. In a paper (Marina Trevelin Souza et al. in Materials 10(83) (2017): 1-14, doi: 10.3390/mal0010083), using a salt leaching method, porous materials with 5 and 10 wt% bioactive glass fibers were produced, dedicated to cartilage tissue regeneration. The use of this filler improved the materials' mechanical properties, bioactivity and better control of hydrolytic degradation kinetics.

In contrast, the most commonly used fillers for bone tissue engineering applications include calcium phosphate-based compounds, including hydroxyapatite. In a publication (Yaozong Wang et al. in Scientific Reports 9 (2019): 7960, doi: 10.1038/s41598-019-44478-8) the porous composites of maleic anhydride-grafted PGS with 40-60 wt% hydroxyapatite are presented. A combination of salt leaching and freeze- drying techniques were used to produce them. The addition of the filler increased the viability of cells from a human adipose-derived stem cell (hADSC) line, as well as the expression of osteogenic genes. Another paper confirming the osteoconductive properties of PGS composites with hydroxyapatite produced by a similar technique is also known (Pawel Piszko et al. in International Journal of Molecular Sciences 22 (2021): 8587, doi: 10.3390/ijms22168587).

There are also literature reports presenting more complex composite systems, where, in addition to PGS, other polymers are introduced into the system as a blend or copolymer with PGS. Examples include materials dedicated to electrospinning fabrication, where PGS is usually present in a blend with polycaprolactone (PCL). Several PGS/PCL composite systems made by the electrospinning technique with various functional additives, including hydroxyapatite, are currently known - materials designed for neural tissue engineering applications (Ahmad Saudi et al. in Materials Chemistry and Physics 275 (2022): 125224, doi: doi.org/10.1016/j.matchemphys.2021.125224) and bone tissue engineering (Abdelrahman I. Rezk et al. in Polymers 12 (2020): 2667, doi: 10.3390/polyml2112667), with carbon quantum dots - for cardiac tissue engineering (Sara Rastegar et al. in Materials Chemistry and Physics 266 (2021): 124543, doi: 10.1016/j.matchemphys.2021.124543), and with graphene - for neural tissue engineering applications (Aref Fakhrali et al in Applied Polymer Science 138 (2021): 51177, doi: 10.1002/app.51177).

Composite blends of PGS with gelatine along with various additives are also known. An example is the work of Mahshid Kharaz , in which he demonstrated the feasibility of producing a PGS/gelatin blend with the addition of carbon nanotubes in mat form using an electrospinning technique (Mahshid Kharaziha et al. in Biomaterials 35 (2014): 7346 - 7354, doi: 10.1016/j.biomaterials.2014.05.014).

At present, several approaches to cross-link poly(glycerol sebacate) by chemical means are known in the scientific literature.

The most common is the use of crosslinking compounds from the isocyanate group. A precursor to this approach was the work by Maria Jose Nunes Pereira et al (Maria Jose Nunes Pereira, et al in Advanced Materials 25 (2013): 1209 - 12015, doi: 10.1002/adma.201203824), which used hexamethylene diisocyanate (HDI) as the crosslinking agent, forming a urethane bond between the isocyanate groups and the hydroxyl groups of the polymer chain. A similar approach was used by Martin Frydrych (Martin Frydrych et al. in Polymer 122 (2017): 159 - 168, doi: http://dx.doi.Org/10.1016/j.polymer.2017.06.064), using HDI as a chemical crosslinking agent for the production of PGS cell scaffolds for soft tissue engineering. HDI was also used to crosslink PGS in the production of composite materials with the addition of fillers: zinc oxide particles (Hooman Golbaten-Mofrad et al. in European Polymer Journal 159 (2021): 110749, doi: 10.1016/j.eurpolymj.2021.110749) and aluminosilicate nanoparticles (Mohammad Monem et al in Journal of Polymer research 29:25 (2022), doi: 10.1007/S10965-021-02866-7).

Another compound from the isocyanate group used to crosslink PGS reported in the literature is diphenylmethane diisocyanate (MDI) with a similar crosslinking mechanism to HDL Xinda Li (Xinda Li et al. in Biomacromolecules 16 (2015): 1525 - 1533, doi: 10.1021/acs.biomac.5b00018) presented the validity of its use, demonstrating that its addition to PGS at 7.8 wt% contributes to improving the mechanical performance of the final product to an extent that is not achievable by standard thermal crosslinking. MDI has also been used by Runcy Wilson (Runcy Wilson et al. in ACS Omega 3 (2018): 118714 - 18723, doi: 10.1021/acsomega.8b02451) to crosslink PGS and PGS block copolymers with 0 - 4 wt% of PTMO (poly(tetramethylene oxide)).

Several other approaches to PGS-based chemical crosslinking of materials have also been presented in the literature to date. Akhilesh K. Gaharwar (Akhilesh K. Gaharwar et al. in Biomaterials Science 3 (2015): 46 - 58, doi: 10.1039/c4bm00222a) proposed the use of carbon nanotubes as a crosslinking agent. Carbon nanotubes become attached to a polymer by forming ester bonds between the carboxyl groups present in their wall structure and the hydroxyl groups present in the polymer backbone. An example of another chemical crosslinking agent reported in the scientific literature used for PGS is gelatine, which increases crosslinking efficiency by forming additional ester and amide linkages to the prepolymer (Sungkwon Yoon et al. in Polymer Chemistry 9 (2018): 3727 - 3740, doi: 10.1039/c8py00544c).

The inconveniences of known polymer-ceramic biocomposites for use as bone defect fillers and for bone regeneration are: lack of effective osteoinductive effect, lack of pro adhesive properties towards bone cells, stiffness and brittleness, lack of shape-matching tolerance to the dimensions of the bone defect.

The aim of the invention was therefore to develop such biocomposites for use as bone defect fillers and for bone tissue regeneration, so that they have the following properties: they have proven osteoinductive properties, are biocompatible at the cellular level in vitro and in vivo, and are elastomeric materials and thus characterized by elasticity and deformation reversibility, exhibiting ease of adaptation to the dimensions of the bone defect.

These effects were achieved by developing composites based on chemically crosslinked poly(glycerol sebacate) using one cross-linking agent selected from three: ethyl ester of L-lysine diisocyanate "LDI" or isophorone diisocyanate "IDI" or 3- (triethoxysilyl)propyl isocyanate "ICPTES" and calcium phosphate particles of one type from four: hydroxyapatite in the form of nanometric particles "HAP_B" or nanometric hydroxyapatite particles covalently functionalized with L-lysine "HAP_B-Lys" or nanometric particles of biphasic calcium phosphate "nanoHAP1200" or hydroxyapatite in the form of nanometric particles with high surface development covalently modified with L-lysine "nanoHAP-Lys". The abbreviations for these components used hereafter are indicated in quotation marks.

Calcium phosphate particles covalently functionalized with L-lysine, HAP_B-Lys and nanoHAP-Lys were used because L-lysine promotes osteoblast adhesion and proliferation (Yoshikawa Masataka et al. in Journal of Biomedical Science and Engineering 8 (2015): 389-398 doi:10.4236/jbise.2015.86037, Liuyun Jang et al. in Journal of Biomaterials Applications 30 (2016): 750-758 doi: 10.1177/0885328215584491) and enhances the osteogenic potential of bone stem cells (Yoshikawa Masataka et al. in Journal of Biomedical Science and Engineering s (2012): 587-592, doi:10.4236/jbise.2012.510072).

A. COMPOSITION OF THE BIOCOMPOSITE

The essence of the invention is new osteoinductive elastomeric polymer-ceramic composites with proven cytocompatibility with normative and bone cells, based on chemically crosslinked poly(glycerol sebacate) using one crosslinking agent selected from three: LDI or IDI or ICPTES and calcium phosphate particles of one type selected from four: HAP_B or HAP_B-Lys or nanoHAP1200 or nanoHAP-Lys for use as an implant material for bone defect filling and bone tissue regeneration. The composite has medical applications as an osteoinductive material for filling bone defects and regenerating bone tissue. As for the amount of individual components in the porous biocomposite: poly(glycerol sebacate) chemically cross-linked using one cross-linking agent selected from three: LDI or IDI or ICPTES, is between 70 wt.% and 85 wt.%. relative to the total weight of the binary composite, calcium phosphate particles (one type selected from four: HAP_B or HAP_B-Lys or nanoHAP1200 or nanoHAP-Lys), is in an amount of 15-30 wt% relative to the total weight of the binary composite. In the solid composite, the amounts of the individual components in the biocomposite: poly(glycerol sebacate) chemically crosslinked using one crosslinking agent selected from two: LDI or IDI, is between 70 wt% and 80 wt% relative to the total weight of the binary composite, calcium phosphate particles (HAP_B or HAP_B-Lys or nanoHAP1200 or nanoHAP-Lys), is between 20-30 wt% relative to the total weight of the binary composite.

A two-component composite with osteoinductive properties for filling bone defects and regenerating bone tissue preferably contains, as a filler, non-surface-modified calcium phosphate particles selected from the two aforementioned: HAP_B or nanoHAP1200; or calcium phosphate particles surface-modified with L-lysine, i.e. HAP_B- Lys or nanoHAP-Lys with an L-lysine content of 1 to 10 wt.% relative to the total filler amount.

Advantageously, the content of calcium phosphate particles of one of the four types, HAP_B or HAP_B-Lys or nanoHAP1200 or nanoHAP-Lys, is 30% by weight of the total weight of the binary porous and solid composite.

Also of essence according to the invention is a method for producing elastomeric porous and solid biocomposites with osteoinductive properties based on chemically cross-linked poly(glycerol sebacate) using one of three chemical agents: LDI or IDI or ICPTES, and calcium phosphate particles of one type selected from four: HAP_B or HAP_B- Lys or nanoHAP1200 or nanoHAP-Lys, for use in bone defect filling and bone tissue regeneration.

Elastomeric composites according to the invention in the form of elastic porous and solid materials are obtained in two stages. The first step in the formation of the porous and solid biocomposite according to the invention is the preparation of the elastomeric matrix by preliminary chemical cross-linking of poly(glycerol sebacate) (obtained previously by a known method) with one of the agents selected from three: LDI or IDI or ICPTES, in a solvent. In a second step, pre-prepared calcium phosphate particles (previously prepared by a known method) of one of the four types: HAP_B or HAP_B-Lys or nanoHAP1200 or nanoHAP-Lys are added and further chemical cross-linking of the product is carried out at elevated temperature without solvent.

The chemical crosslinking reactions of the poly(glycerol sebacate) prepolymer with L- lysine diisocyanate ethyl ester, isophorone diisocyanate and 3-(triethoxysilyl)propyl isocyanate are shown schematically in formulas 1 - 4 and are further described.

Formula 1 Chemical reaction of the modification of poly(glycerol sebacate) prepolymer pPGS with 3-(triethoxysilyl)propyl isocyanate.

Formula 2 Chemical cross-linking reaction of poly(glycerol sebacate) pPGS modified with 3-(triethoxysilyl)propyl isocyanate. Formula 3 Chemical reaction of crosslinking of poly(glycerol sebacate) prepolymer with L- lysine diisocyanate ethyl ester

Formula 4 Chemical reaction of crosslinking of poly(glycerol sebacate) prepolymer with isophorone diisocyanate

The method of preparing an elastomeric composite in the form of a porous and solid material intended for bone defects filling and regenerating bone tissue consists in the first step of chemical modification of poly(glycerol sebacate) prepolymer pPGS (previously obtained by a known method) with LDI or IDI or ICPTES in anhydrous 1,4- dioxane, using a crosslinking agent in an amount of 2 mmol per 1 g of pPGS in the case of crosslinking with isophorone diisocyanate or with the ethyl ester of L-lysine diisocyanate, and 4 mmol per 1 g of pPGS in the case of crosslinking with 3-(triethoxysilyl)propyl isocyanate, at a temperature of 25°C to 70°C for 3 to 24 h, preferably at 50°C for 4 hours. For the porous composite, a solution of 15 to 25 %(m/v), preferably 20 %(m/v) of pPGS chemically modified with isophorone diisocyanate (pPGS-IDI) or L-lysine diisocyanate ethyl ester (pPGS-LDI) or propyl 3-(triethoxysilyl)isocyanate (pPGS-ICPTES) in an amount of 70 wt. % (m/v) to 85% wt. relative to the total dry weight of the composite is then prepared at room temperature. In the solution, the calcium phosphate particles previously obtained by the known method are then dispersed in an amount of from 15 wt.% to 30 wt.%, the dispersion components are thoroughly mixed and placed in the mold together with the porogen.

The solvent is then removed by lyophilisation. After solvent removal, specific crosslinking of the elastomeric matrix of the composite described above is carried out at 50°C to 90°C until the isocyanate band originating from LDI or IDI or ICPTES disappears. Porogen is then removed from the produced cross-linked samples by leaching with water. After porogen leaching, drying of the biocomposite samples is carried out in an atmospheric dryer at a temperature of 25 to 60°C, preferably 40°C, for at least 24 hours. The porous composites are obtained by preparing at room temperature a mixture of pPGS chemically modified with isophorone diisocyanate (pPGS-IDI) or L-lysine diisocyanate ethyl ester (pPGS-LDI) in amounts ranging from 70 wt.% to 80 wt.% in relation to the total weight of the composite and calcium phosphate particles previously obtained by a known method, in an amount of from 20 wt.% to 30 wt.%. The components of the mixture are mixed thoroughly and placed in a mold together with the porogen. The mixture is then poured into the molds and the elastomeric matrix of the above-described composite is crosslinked properly at 30°C to 90°C until the isocyanate band originating from isophorone diisocyanate or ethyl ester of L-lysine diisocyanate disappears.

The preparation of the porous and solid biocomposite according to the invention consists of the operations described below carried out in the order (steps):

1. Synthesis of poly(glycerol sebacate) prepolymer, by a known route, by the polycondensation reaction of sebacic acid and glycerol.

2. Synthesis of calcium phosphate particles. 2a. Synthesis of nanometric hydroxyapatite "HAP_B" by the known wet precipitation method from disodium hydrogen phosphate salt and hydrated calcium acetate at elevated temperature or the two-stage synthesis of calcium phosphate particles "nanoHAP1200" by the known method, in which wet precipitation from calcium hydroxide and orthophosphoric acid in an alkaline medium in a first step produces hydroxyapatite in the form of nanometric particles with a high surface development ("nanoHAP"), followed in a second step by calcination according to a known method at 1200°C.

2b. For "HAP_B-Lys" or "nanoHAP-Lys" : covalent functionalization of the surface of "HAP_B" or "nanoHAP" calcium phosphate particles obtained in step 2a by the known method of modification of the surface of hydroxyapatite particles with 3- aminopropyltriethoxysilane APTS and in a subsequent step the coupling reaction of the amino groups of APTS with the carboxyl groups of L-lysine using the coupling agent l-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and sulfo-N-hydroxysulfosuccinimide (sulfo-NHS) to increase the reaction yield leading to "HAP_B-Lys" in the case of "HAP_B" functionalization of "nanoHAP-Lys" in the case of "nanoHAP" functionalization. Chemical modification of poly(glycerol sebacate) prepolymer with isophorone diisocyanate or ethyl ester of L-lysine diisocyanate or 3-(triethoxysilyl)propyl isocyanate as shown in Formulas 1, 2, 3 consisting of the reaction of poly(glycerol sebacate) prepolymer with isophorone diisocyanate or 3-(triethoxysilyl)propyl isocyanate or ethyl ester of L-lysine diisocyanate carried out in anhydrous 1,4- dioxane or tetrahydrofuran at a temperature of 25°C to 70°C for a period of 3 to 24 hours and release of the reaction product by evaporation of the solvent.

For the porous composite, the next steps are: Preparation of a solution of 15 to 25 %(m/v), preferably 20 %(m/v) of poly(glycerol sebacate) chemically modified with isophorone diisocyanate or 3- (triethoxysi lyl)propyl isocyanate or ethyl ester of L-lysine diisocyanate, obtained in step 3, in anhydrous 1,4-dioxane. To the solution of poly(glycerol sebacate) chemically modified with 3-(triethoxysilyl)propyl isocyanate, water is also added in an amount of 12 mmol per 1 g of poly(glycerol sebacate) chemically modified with 3-(triethoxysilyl)propyl isocyanate and ethanol at 4 mmol per 1 g of poly(glycerol sebacate) chemically modified with 3-(triethoxysilyl)propyl isocyanate and one drop of concentrated hydrochloric acid is added.

5. Addition to the solution prepared in step 4 of one type of calcium phosphate particles in an amount of 15wt.% to 30wt.% relative to the total dry weight of the binary composite and thoroughly mixing the system using a laboratory magnetic stirrer.

6. Weighing into the mold porogen in the form of sodium chloride with a grain size of 200 to 800 pm, preferably 400 to 500 pm, in an amount of 0.6 to 1.3 g per 1 cm³ of mold volume, preferably 1 g per 1 cm³ of mold volume, and introducing into the mold the dispersion prepared in step 5, in an amount of 0.3 cm³ to 0.7 cm³ of dispersion per 1 cm³ of mold volume, preferably 0.5 cm³ per 1 cm³ of mold volume.

7. Removal of solvent from the mold prepared according to the procedure described in step 6 for poly(glycerol sebacate) chemically modified with 3- (triethoxysi lyl)propyl isocyanate, isophorone diisocyanate or ethyl ester of L-lysine diisocyanate was realized by leaving the plate at 25°C for 24 hours. Samples obtained from poly(glycerol sebacate) chemically modified with isophorone diisocyanate or L-lysine diisocyanate ethyl ester were then frozen at -20°C for 5 to 24 h, preferably 12 h, and lyophilised.

8. Specific cross-linking of biocomposite samples carried out at 50°C for 24 hours and then at 90°C for 96 hours.

9. Removal of porogen from the produced cross-linked samples by washing them at least four times with deionised water at a rate of at least 1 dm³ per 1 g of biocomposite, while controlling the conductivity of the water used for washing.

10. Drying of the biocomposite samples in an atmospheric dryer at 25 to 60 °C, preferably 40 °C, for at least 24 hours.

In the case of the solid composite, the next steps after step 3 are:

11) Preparation of a mixture of pPGS pre-cross-linked chemically with one agent selected from two: L-lysine diisocyanate ethyl ester or isophorone diisocyanate, in an amount of 70 wt%. to 80% w/w. relative to the total weight of the two- component composite, and calcium phosphate particles obtained in one of the previous steps : nanometric hydroxyapatite HAP_B or nanometric hydroxyapatite covalently functionalized with L-lysine HAP_B-Lys or biphasic calcium phosphate particles nanoHAP1200 or nanometric hydroxyapatite covalently functionalized with L-lysine nanoHAP-Lys, in an amount ranging from 20 wt.% to 30 wt.%. relative to the total weight of the composite - two-component mixture. The mixture of composite components is prepared at room temperature by thoroughly mixing the components and degassing the mixture in a vacuum dryer before the actual crosslinking step at elevated temperature.

12) Cross-linking of the mixture poured into the molds carried out at temperatures from 30° C to 90° C e.g. in an air-circulating dryer over a period of 5 days until the isocyanate band originating from the ethyl ester of L-lysine diisocyanate or isophorone diisocyanate disappears.

The object of the invention is illustrated in examples 1 to 32, and in the reaction diagrams for the chemical cross-linking of poly(glycerol sebacate) with L-lysine diisocyanate ethyl ester, isophorone diisocyanate and propyl 3-(triethoxysilyl)isocyanate in examples 1,2,3 and 4.

Figure 1 summarizes the biological characteristics of the HAP_B product in terms of cytocompatibility (a) against L929 fibroblasts and hFOB 1.19 osteoblasts, proliferation (b) of hFOB 1.9 cells under osteoinductive conditions, osteoconductive properties of the product against hFOB 1.19 cells under osteoinductive conditions: assessment of ALP production (c) and osteocalcin assay (d).

Figure 2 summarizes the biological characteristics of the nanoHAP product in terms of cytocompatibility (a) against L929 fibroblasts and hFOB 1.19 osteoblasts, proliferation (b) of hFOB 1.9 cells under osteoinductive conditions, osteoconductive properties of the product against hFOB 1.19 cells under osteoinductive conditions: assessment of ALP production (c) and osteocalcin assay (d).

Figure 3 summarizes the biological characteristics of the nanoHApl200 product in terms of cytocompatibility (a) against L929 fibroblasts and hFOB 1.19 osteoblasts, proliferation (b) of hFOB 1.9 cells under osteoinductive conditions, osteoconductive properties of the product against hFOB 1.19 cells under osteoinductive conditions: assessment of ALP production (c) and osteocalcin assay (d).

Figure 4 summarizes the biological characteristics of the PGS-IDI product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 5 summarizes the biological characteristics of the PGS-IDI/15HAP_B product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 6 (a) summarizes the biological characteristics of the PGS-IDI/30HAP_B product in terms of cytoplasmic compatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay; (b) a representative photograph showing the histological analysis of the PGS-IDI product 7 days after implantation in vivo in a rat model in an assay of local tissue response to implantation.

Figure 7 summarizes the biological characteristics of the PGS-IDI/15HAP_B-Lys product in terms of cytoskeletal compatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 8 (a) summarizes the biological characteristics of the PGS-IDI/30HAP_B-Lys product in terms of cytoplasmic compatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay; (b) a representative photograph showing the histological analysis of the PGS-IDI product 7 days after implantation in vivo in a rat model in an assay of local tissue response to implantation.

Figure 9 summarizes the biological characteristics of the PGS-ICEPTES product in terms of cytoskeletal compatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 10 summarizes the biological characteristics of the PGS-ICEPTES/15HAP_B product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 11 summarizes the biological characteristics of the PGS-ICEPTES/30HAP_B product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. Figure 12 summarizes the biological characteristics of the PGS-ICEPTES/15HAP_B- Lys product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 13 summarizes the biological characteristics of the PGS-ICEPTES/30HAP_B- Lys product in terms of cytoskeletal compatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 14 summarizes the biological characteristics of the PGS-LDI product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 15 summarizes the biological characteristics of the PGS- LDI/30nanoHAP1200 product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 16 summarizes the biological characteristics of the PGS-LDI/30nanoHAP-Lys product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 17 summarizes the biological characteristics of the PGS-IDI product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 18 summarizes the biological characteristics of the PGS-IDI/30HAP_B product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 19 summarizes the biological characteristics of the PGS-IDI/30HAP_B-Lys product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 20 summarizes the biological characteristics of the PGS-LDI product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay.

Figure 21 summarizes the biological characteristics of the PGS- LDI_20%nanoHAP1200 product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The following abbreviations were used in the description of the¹ H NMR spectra: SA - sebacic acid residue; IT - terminal glyceride unit substituted in position 1; 2T - terminal glyceride unit substituted in position 2; 12L - double-substituted glyceride unit in positions 1 and 2; 13L - double-substituted glyceride unit in positions 1,1; D - triplesubstituted glyceride unit; sz.s - broad signal; m - multiplet.

The following abbreviations are used in the description of FTIR spectra: v - tensile vibrations, v_as - asymmetric tensile vibrations, v_s - symmetric tensile vibrations, p - swaying vibrations, 6 - deformation vibrations, 6_S - scissor vibrations, OJ - fan vibrations, T - torsional vibrations.

Glass transition temperatures of the polymers (T_g ) were determined from curves from DSC measurements, the thermal stability temperatures corresponding to the 10% mass loss temperature (T.i₀% ) were determined from curves from TGA measurements. DSC and TGA measurements were carried out at a nitrogen flow rate of 60 ml/min. and a heating and cooling rate of 10 °C/min.

Detailed description of the drawings:

Figure 1. The biological characteristics of the HAP_B product in terms of cytocompatibility (a) against L929 fibroblasts and hFOB 1.19 osteoblasts, proliferation (b) of hFOB 1.9 cells under osteoinductive conditions, osteoconductive properties of the product against hFOB 1.19 cells under osteoinductive conditions: assessment of ALP production (c) and osteocalcin assay (d). The results in graph (a) are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n

> 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 2. The biological characteristics of the nanoHAP product in terms of cytocompatibility (a) against L929 fibroblasts and hFOB 1.19 osteoblasts, proliferation (b) of hFOB 1.9 cells under osteoinductive conditions, osteoconductive properties of the product against hFOB 1.19 cells under osteoinductive conditions: assessment of ALP production (c) and osteocalcin assay (d). The results in graph (a) are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n

> 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material. Figure 3. The biological characteristics of the nanoHApl200 product in terms of cytocompatibility (a) against L929 fibroblasts and hFOB 1.19 osteoblasts, proliferation (b) of hFOB 1.9 cells under osteoinductive conditions, osteoconductive properties of the product against hFOB 1.19 cells under osteoinductive conditions: assessment of ALP production (c) and osteocalcin assay (d). The results in graph (a) are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 4. a) The biological characteristics of the PGS-IDI product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results in graph (a) are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material, b) Representative photo showing the histological analysis of PGS-IDI product after 7 days post-implantation in vivo in a rat model in the study of local tissue response to implantation.

Figure 5. The biological characteristics of the PGS-IDI/15HAP_B product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 6. (a) The biological characteristics of the PGS-IDI/30HAP_B product in terms of cytoplasmic compatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results in graph (a) are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material, (b) Representative photograph showing the histological analysis of the PGS-IDI/30HAP_B product 7 days after implantation in vivo in a rat model in an assay of local tissue response to implantation. Figure 7. The biological characteristics of the PGS-IDI/15HAP_B-Lys product in terms of cytoskeletal compatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 8. (a) The biological characteristics of the PGS-IDI/30HAP_B-Lys product in terms of cytoplasmic compatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results in graph (a) are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material, (b) Representative photograph showing the histological analysis of the PGS-IDI /30HAP_B-Lys product 7 days after implantation in vivo in a rat model in an assay of local tissue response to implantation.

Figure 9. The biological characteristics of the PGS-ICEPTES product in terms of cytoskeletal compatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 10. The biological characteristics of the PGS-ICEPTES/15HAP_B product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 11. The biological characteristics of the PGS-ICEPTES/30HAP_B product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 12. The biological characteristics of the PGS-ICEPTES/15HAP_B-Lys product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 13. The biological characteristics of the PGS-ICEPTES/30HAP_B-Lys product in terms of cytoskeletal compatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 14. The biological characteristics of the PGS-LDI product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 15. The biological characteristics of the PGS-LDI/30nanoHAP1200 product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 16. The biological characteristics of the PGS-LDI/30nanoHAP-Lys product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 17. The biological characteristics of the PGS-IDI product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 18. The biological characteristics of the PGS-IDI/30HAP_B product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 19. The biological characteristics of the PGS-IDI/30HAP_B-Lys product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 20. The biological characteristics of the PGS-LDI product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Figure 21. The biological characteristics of the PGS-LDI_20%nanoHAP1200 product in terms of cytocompatibility against L929 fibroblasts and hFOB 1.19 osteoblasts in the MTT reduction assay. The results are presented relative to the control assumed to be 100% viability. Data presented as mean ± standard deviation, n > 3. TC - control treated with hydrogen peroxide; NTC - untreated control of cells incubated in medium without product; R - biocompatible reference material.

Example 1. pPGS prepolymer synthesis

An equal volume of sebacic acid (65.85 g, 0.3258 mole) and glycerine (30 g, 0.3258 mole) was weighed into the reaction vessel. The reaction mixture was heated to 135°C until the sebacic acid melted while stirring. The temperature was then lowered to 130°C and the reaction was continued for 48 hours at atmospheric pressure together with continuous stirring. The reaction mixture was cooled and quantitatively transferred to a sterile container. The synthesis product is stored at 8°C.

The structure of the product was confirmed by Fourier transform infrared spectroscopy (FTIR) and proton magnetic resonance H NMR). The thermal properties of the product were characterized by differential scanning calorimetry (DSC) and thermal stability was determined by thermogravimetric analysis (TGA).

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3436 - v O-H, 2927 - v_as C-H in CH₂ , 2854 - v_s C-H in CH₂ , 1733 - v C=O, 1455 - 6_S C-H in CH₂, 1417 and 1210 - v C-0 in COOH, 1165 - v C-O, 1097 - 6 in 2° O-H, 1046 - 6 in 1° O-H, 945 - 6 O-H in COOH, 724 - p C-H in CH₂

^TH NMR ((CD )₃₂ CO), 400 MHz): 6 1.29 (sh.s, 8H, SA C4,C5,C6,C7); 1.57 (sh.s, 4H, sebacic acid C3 and C8); 2.30 (m, 4H, SA C2 and C9); 3.52 (m, 0.44H, -CH₂ OH IT); 3.65 (m, 0.44H, - CH₂ OH 2T and -CH₂ OH 12L); 3.80 (m, 0.22H, -CHOH IT); 4.07 (m, 2.28H, -CH₂ O- IT and 13L); 4.30 (m, 1.10H, CH₂ O- in D and CH₂ O- in 12L); 4.84 (m, 0.025H, CH in 2T); 5.04 (m, 0.17H, CH in 12L); 5.24 (m, 0.19H, CH in D).

DSC and TGA: Tg (_2nc| heating) ⁼ -16.6 C, T_io% ⁼ 382 C.

Example 2. pPGS-IDI synthesis

In a round-bottom flask, 18.00 g of pPGS from Example 1 was placed and dissolved in 90 ml of anhydrous 1,4-dioxane. Then, 7.60 ml of isophorone diisocyanate was added to the solution in small portions. The contents of the flask were heated at 50°C under a reflux condenser for 4 hours with continuous stirring on a magnetic stirrer. After this time, the solvent was evaporated on a rotary evaporator to give an oily liquid in 75% yield. ^XH NMR ((CD )₃₂ CO, 400 MHz): 0.99 (s), 1.01 (s) and 1.08 (s) (6H, CH₃ at 1,3,3- trimethylcyclohexane ring in C3 position); 1.05 (s) and 1.12 (s) (3H, CH₃ at 1,3,3- trimethylcyclohexane ring in Cl position); 1.34 (br, 16H, SA at positions C4,C5,C6,C7); 1.62 (br.s., 8H, SA in positions C3 and C8); 1.19-1.31 (m), 1.50 (m) and 1.53 (m) (4H, CH₂ in the 1,3,3-trimethylcyclohexane ring in positions C4 and C6); 1.83 (m), 1.96 (m) (2H, CH₂ at 1,3,3-trimethylcyclohexane ring at C2 position); 2.34 (m, 8H, SA at C2 and C9 positions); 3.20 (s) and 3.47 (m) (2H, at 1,3,3-trimethylcyclohexane ring at Cl position); 3.53 (m, 0.88H, -CH₂ OH in IT); 3.65 (m, 0.88H, -CH₂ OH in 2T and -CH₂ OH in 12L); 3.85 (m, 0.44H, -CHOH in IT); 3.89 (m) (1H, CH at 1,3,3-trimethylcyclohexane ring in C5 position); 4.07 (m, 4.56H, -CH₂ O- in IT and protons in 13L); 4.34 (m, 2.20H, CH₂ O- in D and CH₂ O- in 12L); 4.88 (m, 0.05H, CH in 2T); 5.08 (m, 0.34H, CH in 12L); 5.28 (m, 0.38H, CH in D)

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3338 - v_as in -OH and - NH-, 2957 - v_as CH₂ , 2852 - v_as CH in CH₂ , 2266 - v in -NCO, 1735 - v C=O, 1658 and 1533 - 6 in - NH-, 1454 - 6_S CH in CH₂ , 1417 and 1254 - v C-0 in COOH, 1168 - v in -C-O, 1082 - 6 in 2° OH, 1048 - 6 in 1° OH, 775 - p CH in CH₂.

Example 3. pPGS-ICPTES synthesis

Into a round-bottomed flask, 9 g of pPGS from Example 1 was weighed, 45 ml of anhydrous 1,4-dioxane was added and the whole was stirred until the prepolymer was completely dissolved. The content of the flask were then purged with nitrogen and 36 mmol of 3-(triethoxysilyl)propyl isocyanate was added. The whole was heated under a reflux condenser with continuous stirring at 50° C for 24 hours. After this time, the solvent was evaporated on a rotary evaporator. The product as an oily liquid was obtained in 90% yield.

^TH NMR (CD₃)₂ CO, 400 MHz): 0.67 (m, 2H, CH₂ in the 3-triethoxysilylpropyl group at position C3), 1.19 (t, 9H, CH₃ in the ethoxyl group), 1.33 (br.s., 8H, SA at positions C4, C5, C6, C7), 1.61 (br.s., 4H, SA in positions C3 and C8); 1.72 (m, 2H, CH₂ in the 3- triethoxysilylpropyl group in position C2); 2.33 (m, 4H, SA in positions C2 and C9); 3.36 (t, 2H, CH₂ in 3-triethoxysilylpropyl group at Cl position); 3.59 (m, 0.44H, -CH₂ OH at IT); 3.68 (m, 0.44H, -CH₂ OH at 2T and -CH₂ OH at 12L); 3.81 (m, 6H, CH₂ in the ethoxyl group); 3.85 (m, 0.22H, -CHOH in IT); 4.11 (m, 2.28H, -CH₂ O- in IT and protons in 13L); 4.34 (m, 1.10H, CH₂ O- in D and CH₂ O- in 12L); 4.87 (m, 0.025H, CH in 2T); 5.07 (m, 0.17H, CH in 12L); 5.28 (m, 0.19H, CH in D); 8.01 (br.s., 1H, NH in urethane group)

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3368 - v in -OH , 2929 - v_as CH in CH₂ , 2855 - v_s CH in CH₂ , 1984 - v_s in SiO₂ , 1732 - v C=O, 1456 - 6_S CH in CH₂ , 1416 and 1240 - v C-0 in COOH, 1193 - v in-C-O, 1097 - 6 in 2° OH and v_as in SiO₂ , 774 - p CH in CH₂

Example 4. Synthesis of pPGS-LDI

5.00g of pPGS from Example 1 was dissolved in 50ml of anhydrous tetrahydrofuran. 2.02ml of L-lysine diisocyanate ethyl ester was dropped into the solution. The reaction solution was stirred on a magnetic stirrer (250 rpm) and heated at 50°C under a reflux condenser in a nitrogen atmosphere for 5h. The tetrahydrofuran was then evaporated on a rotary evaporator. The oily liquid product was obtained in 100% yield.

FTIR: 3365cm"¹ and 1417cm"¹ - v and 6 -OH; 2950cm"¹ , 2869cm"¹ and 1456cm"¹ - v and 6 in C-H located in the PGS main chain; 1723cm"¹ - band is associated with the presence of a carbonyl group and is shifted towards lower wavenumbers relative to the pPGS spectrum (Example 1); 2252cm"¹ - v of unreacted isocyanate groups of the ethyl ester of L-lysine diisocyanate; 1531cm’¹ - II amide band -CO-NH, confirming the formation of an amide bond in the reaction of the isocyanate group of the ethyl ester of L-lysine diisocyanate with the hydroxyl group of pPGS; 1021cm’¹ - v of C-O-C ether bonds from the ethyl ester of L-lysine diisocyanate.

TGA: T_₁₀% = 338°C.

Example 5. Synthesis of nanometric hydroxyapatite HAP_B

HAP_B was obtained by a wet precipitation method conducted at the boiling point of the ingredients. Distilled water and Na₂HPO₄ (0.32 mol/L) were poured into a 500 mL triplenecked flask and placed on a heating bowl with magnetic stirring. The pH of the medium was brought to a pH of 11 with the addition of 25 % ammonia water. After bringing the system to a boil, (CH₃ COO)₂ Ca (0.128 mol/L) was dropped in at a rate of 1 drop/sec. After completion of the dropping, the suspension was stirred for one hour, then cooled and allowed to mature the precipitate for 24 h. After this time, the precipitate was washed thoroughly with distilled water, brought to neutral pH and lyophilised. XRD: X-ray diffraction analysis was used to image the structure of HAP_B. The resulting material is phase pure, with hydroxyapatite being the only phase identified in X-rays. The result is consistent with the phases listed in the ICDD database (catalogue card number 01-080-7085), and the XRD reflections were assigned to a hexagonal structure (space group P63/m) of hydroxyapatite with lattice parameters a = 9.4172 A and c = 6.8799 A.

FTIR: Spectroscopic analysis of HAP_B revealed the presence of phosphate groups, as evidenced by distinct bands in the range 550-1016 cm"¹ . The intense peaks characteristic of hydroxyapatite associated with asymmetric P-0 stretching vibrations are attributed to values of 960 cm"¹ and 1026 cm"¹ . The broad peaks for values of 2900-3640 cm"¹ indicate the presence of valence vibrations of the OH" group; these broad absorption bands are attributed to the presence of H₂O bound to HAp molecules. The band in the wavelength range from 558 cm"¹ to 605 cm"¹ was associated with a triple degenerate O-P-O bending mode in the PO₄ ³" groups , which occupy two sites in the crystal lattice. The band observed at 885 cm"¹ corresponds to the presence of carbonate ions.

Molar ratio Ca/P: Based on standards PN-80/C-87015 and on PN-97/R-64803, the calcium and phosphorus contents of HAP_B powder were determined. The calcium content is 37.51 ± 0.65 (wt. %) and the phosphorus content is 17.26 ± 0.19 (wt. %). The molar ratio of Ca/P powder was determined to be 1.6795, indicating that the resulting material is a stoichiometric hydroxyapatite.

Study of specific surface area and porosity: Nitrogen adsorption and desorption isotherms (77 K) for HAP_B were determined experimentally. The investigated powder reveals a mesoporous and microporous structure. The pore volume distribution profile shows a single narrow peak with a maximum at 8-12 nm. The physisorption parameters characterizing the resulting HAP_B ceramic powder are shown in Table 1.

Table 1. N-physisorption parameters characterizing the resulting HAP_B powder. DLS and zeta potential: the particle size of HAP_B was determined using the DLS technique. Prior to the DLS measurement, an aqueous suspension of HAP_B was subjected to ultrasound (T=25°C, 15minutes, 160W). The suspension is characterized by a bimodal distribution. Subjecting the HAP_B solution to ultrasound at 160 W showed a size distribution covering the range of 43 nm - 531 nm. The solution contains predominantly particles with an average size of approximately 86 nm (80%) and a smaller proportion of particles with an average size of 280 nm (20%).

A zeta potential of -21.7 ± 1.33 was determined. The zeta potential is closely related to the mechanism of formation of new apatite layers during incubation in SBF-type fluids. A negative result was obtained and a negative zeta potential was described in favor of osteointegration, apatite nucleation and bone regeneration. Negative zeta potentials are thought to favour the adsorption of Ca ions²⁺ , which are involved in the deposition of the extracellular matrix necessary for cell adhesion.

The cytocompatibility of the HAP_B product was tested at the level of mitochondrial activity of mouse fibroblast L929 cells and human osteoblast hFOB.1.19 cells according to ISO 10993-5:2009 standards using the MTT reduction assay after treatment with HAP_B particles at a concentration of 1 mg/mL for 24 hours. Proliferative potential of hFOB 1.19 cells cultured under osteoinductive conditions (39 °C; differentiation medium: DMEM F- 12 without phenol red with 1% bovine serum supplement, containing genetin G148 and the stimulators p-glycerophosphate, ascorbic acid, dexamethasone, and the addition of the test product in HEPES-buffered collagen at a concentration of 62.5 mg/mL) with the product was tested after 21 days with a CyQuant® assay based on fluorescence cell count determination. The osteoconductive properties of the test product were assessed after incubation with hFOB 1.19 cells under osteoinductive conditions (as above) for up to 21 days. Osteoconductive properties were assessed by alkaline phosphatase (ALP) activity, an indicator of osteogenic cell differentiation, bone formation and matrix mineralisation, and by testing the concentration, by ELISA, of soluble mediators, i.e. osteocalcin, a specific marker responsible for bone mineralisation.

Biological characterisation of HAP_B demonstrated the following properties of the product. HAP_B at the concentration tested is not cytotoxic to mouse fibroblasts of the L929 lineage and human osteoblasts of the hFOB 1.19 lineage, as supported by the results of cell viability after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993:5 2006 standard (Figure la). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 hours incubation with 1 mg/mL product was 118.3 % ± 12.8 % and 84.4 % ± 5.7 %, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests. In addition, the product at a concentration of 62.5 mg/mL showed no proliferation-promoting effect on hFOB 1.19 cells under osteoinductive conditions (Fig. lb).

At 62.5 mg/mL under osteoinductive conditions, the product caused: increase in alkaline phosphatase (ALP) activity to 0.05 ± 0.04 iU/mL after 21 days (Fig. lc); increase in osteocalcin (OC) production by hFOB 1.19 cells over time to a level of 432.6 ± 0.4 pg/mL after 21 days of incubation (Fig. Id).

Example 6: Synthesis of nanoHAP particles

In a 250 mL spherical flask, 0.25 moles of Ca(OH)₂ was prepared. 200 mL of deionised water was added and placed on a magnetic stirrer for 20 min at 50°C. Using a 100 mL spherical flask, 0.15 mol of H₃PO₄ was measured and diluted in 50 mL of deionised water. The solution prepared in this way was infused for 45 min into a Ca(OH)₂ solution placed on a magnetic stirrer operated with continuous stirring at 600 rpm. During the addition of H3PO4, pH=ll was maintained by adding ammonia water in 25% solution if necessary. All reaction steps were carried out at 50°C. The solution was stirred for a further 2 hours. After synthesis, the solution was centrifuged and the product was washed with deionised water until neutral. The reaction product was transferred to a ceramic vessel and dried for 24 hours at 90°C.

SEM/EDS: Microstructure, grain size and elemental composition analysis were determined by SEM imaging with EDS analysis. The reaction yielded a product with a homogeneous structure and spherical morphology, with grain sizes ranging from 40.9 to 48.8 nm.

XRD: The phase composition of the obtained product was determined by X-ray diffraction (XRD). The obtained product is phase-pure hydroxyapatite. FTIR: The chemical structure was confirmed by Fourier transform infrared spectroscopy (FTIR). The spectrum shows characteristic bands from the stretching vibrations of the hydroxyl groups (OH) (3568 cm"¹ ), bands from the asymmetric stretching vibrations of the P-0 bonds in the phosphate groups (PO₄ ^3- ) (1092 cm"¹ , 1033 cm"¹ , 962 cm"¹ ) and bands originating from bending vibrations of O-P-O bonds of phosphate groupings (PO₄ ³" ) (603 cm"¹ , 565 cm"¹ , 477 cm"¹ ). A broad band at 3422 cm"¹ derived from water bound to hydroxyapatite particles and bands derived from bending vibrations of H-O-H in water are visible (1634 cm )^-1

BET: The degree of surface development of the particles obtained was assessed by determining the size of the specific surface area using the BET method. The size of the specific surface area of the grains is: 80.7m /g.²

The cytocompatibility, proliferation and osteoconductive properties of the product were investigated as in example 5.

NanoHAP at the concentrations tested (1 and 5 mg/mL) is not cytotoxic to mouse fibroblasts of the L929 lineage and human osteoblasts of the hFOB 1.19 lineage, as supported by the cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed in accordance with the relevant ISO 10993-5:2009 standard (Fig. 2a). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation with 1 mg/mL product was 117.5% ± 1.2% and 89.5% ± 5.8%, respectively, and for 5 mg/mL, 88.0% ± 1.5% and 86.0% ± 8.0%, indicating the cytocompatibility of the products. In addition, the product at 5 mg/mL showed no proliferation-promoting effect on hFOB 1.19 cells under osteoinductive conditions (Fig. 2b). At 5 mg/mL under osteoinductive conditions, the product caused: increase in alkaline phosphatase (ALP) activity to 0.8 ± 0.01 iU/mL after 21 days (Fig. 2c); increase in osteocalcin (OC) production by hFOB 1.19 cells over time to a level of 346.3 ± 48.8 pg/mL after 21 days of incubation (Fig. 2d).

Example 7: Synthesis of nanoHAP1200 particles

In a 250 mL spherical flask, 0.25 moles of Ca(OH)₂ was placed, 200 mL of deionised water was added and the contents were stirred for 20 min on a magnetic stirrer. In a 100 mL spherical flask, 0.15 moles of H₃ PO₄ was placed and 50 mL of deionised water was added, and the prepared solution was then dropped into a Ca(OH)₂ solution placed on a magnetic stirrer (800 rpm) for 45 min. During the addition of H₃PO₄, pH=ll was maintained by adding 25% ammonia water solution. All reaction steps were carried out at 48°C. The solution was stirred for a further 3 hours, then centrifuged and the product was washed with deionised water to a neutral reaction. The reaction product was dried for 24 hours at 90°C. After drying, the resulting product was calcined at 1200°C for 1 h according to the following calcination conditions: heating 10°C /min to 1200°C, holding 1200°C for 1 h, cooling 10°C /min to 20°C, air cooling at room temperature. After calcination, the product was ground to completely crumble and obtain a homogeneous powder.

SEM/EDS: The reaction yielded a product with a spherical morphology and a grain size ranging from 692.4 nm to 958.0 nm.

XRD: The phase product obtained consisted of approximately 94 % hydroxyapatite and approximately 6 % -TCP.

FTIR: 3571 cm"¹ - v -OH, 1091 cm"¹ , 1048 cm"¹ , 961 cm"¹ - v_as P-0 in phosphate groupings (PO₄ ³" ), 601 cm"¹ , 570 cm"¹ , 438 cm"¹ )- 6 -0-P-0 phosphate groupings (PO₄ ³" ), 3434 cm"¹ - derived from bound water.

BET: The size of the specific surface area of the grains is: 0.33 m /g.²

The cytocompatibility, proliferation and osteoconductive properties of the product were investigated as in example 5. nanoHAP1200 at the concentrations tested (1, 5, and 10 mg/mL) is not cytotoxic to mouse fibroblast lineage L929 and human osteoblast lineage hFOB 1.19, as supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed in accordance with the relevant ISO 10993:5 2006 standard) (Fig. 3a). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 hours incubation with 1 mg/mL product was 120.4% ± 7.4% and 101.9% ± 2.9%, respectively, and for 5 mg/mL, 109.0% ± 3.4% and 93.3% ± 9.8%, respectively, and for 10 mg/mL, 103.5% ± 9.4% and 104.8% ± 6.1%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests. In addition, the product at 62.5 mg/mL showed no proliferation-promoting effect on hFOB 1.19 cells under osteoinductive conditions (Fig. 3b). At 62.5 mg/mL under osteoinductive conditions, the product caused: increase in alkaline phosphatase (ALP) activity to 1.1 ± 0.2 iU/mL after 21 days (Fig. 3c); increase in osteocalcin (OC) production by hFOB 1.19 cells over time to a level of 1299.0 ± 41.0 pg/mL after 21 days of incubation (Fig. 3d).

Example 8. Production of nano-sized hydroxyapatite covalently functionalised with L- lysine (HAP_B-Lys)

Prior to modification, HAP_B hydroxyapatite obtained by the method described in Example 5 was annealed at 600° C in a muffle furnace for 3h. In the first modification step, 1.5g of HAP_B hydroxyapatite was dispersed in 20ml of anhydrous toluene. The dispersion was sonicated for 5 min (0.5 cycle; 50% amplitude). Then 2.03 mg of 3- aminopropyltriethoxysilane (APTES) was injected into the dispersion and purged with argon. After APTES was injected, the reaction system was stirred on a magnetic stirrer at 130° C for 24 h under an argon atmosphere. The reaction product was then separated on a ceramic funnel and washed with anhydrous toluene (3 times 10ml each). The precipitate was dried for 24 h in a vacuum dryer at 100° C. 1.45g of product was obtained (97% yield).

FTIR of the reaction product of HAP_B with APTES (ATR, diamond crystal): 3571 cm"¹ and 630 cm"¹ - v and 6 of O-H bonds in hydroxyl groups of hydroxyapatite crystal phase; 1088 cm"¹ and 1026 cm"¹ - v₃ P-0 bonds; 963 cm"¹ - v i P-0 bonds; 2929 cm"¹ and 2868 cm"¹ - v_as and v_s C-H bonds in the -CH₂ - groups of the aminopropyl chain of APTES; 1571 cm"¹ from the -NH₃ ⁺ (HCO )₃~ formed by the amino groups of APTES.

TGA: The amount of APTES silane precursor introduced on the surface of the hydroxyapatite particles, based on thermogravimetric measurements, determined from the difference in total mass loss at 900° C between the unmodified HAP_B particles and the product of the first modification step, is 1.50 wt.%. In the second step, the APTES-modified HAP_B particles are reacted with L-lysine using the coupling agent l-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and the coupling reaction enhancing agent N-hydroxysulfo-succinimide (Sulfo - NHS). For this purpose, 14.8mg of L-lysine, 17.3mg of EDC, 31.5mg of Sulfo - NHS are dissolved in 10ml of MES buffer (pH 6) in a round-bottomed flask. The solution is stirred on a magnetic stirrer 30min to activate the carboxyl groups of L-lysine. After this time, APTES-modified HAP_B particles (1.45g) are dispersed in 20ml of MES buffer (pH=6), sonicated for 5min (0.5 cycle; 50% amplitude), then transferred to a solution of L-lysine, EDC and sulfo - NHS. The whole is mixed using a magnetic stirrer for 24hr at room temperature. After this time, the suspension is centrifuged to separate the modified particles, washed 3 times with MES buffer (pH 6) and dried under vacuum at 60°C for 24 hours. A 1.27g product HAPB_Lys was obtained as a white powder (87% yield).

FTIR of the reaction product with L-lysine: (ATR, diamond crystal): The presence of L- lysine covalently attached by an amide bond is confirmed by the disappearance of the band at 1571cm"¹ from the amine groups, and the presence on the FTIR-ATR spectrum of a characteristic I amide band at 1646cm"¹ from C=O bond vibrations and the II amide band at 1457cm’¹ from N-H bond deformation vibrations.

TGA: The content of L-lysine covalently bound to the surface of HAP_B particles, calculated from the difference between the total mass loss in the range 25°C-900°C for APTES-modified and L-lysine-modified HAP_B particles and for APTES-modified hydroxyapatite particles, is 5.65 wt%.

Example 9. Production of nanoHAP particles covalently functionalized with L-lysine (nanoHAP-Lys)

In the first modification step, 2.50g of nanoHAP hydroxyapatite obtained by the method described in Example 6 was dispersed in 20ml of anhydrous toluene. The dispersion was sonicated for 5min. (0.5 cycle; 50% amplitude). Then 3.84mg of 3- aminopropyltriethoxysilane (APTES) was injected into the dispersion and purged with argon. After APTES was injected, the reaction system was stirred on a magnetic stirrer at 130° C for 24 h under an argon atmosphere. The reaction product was then separated on a ceramic funnel and washed with anhydrous toluene (3 times 10ml each). The precipitate was dried for 24 h in a vacuum dryer at 100° C. 2.42g of product nanoHAP-Lys was obtained (97% yield).

FTIR of the reaction product of nanoHAP with APTES: (ATR, diamond crystal): 3567 cm"¹ and 632 cm"¹ - v and 6 O-H bonds in hydroxyl groups of hydroxyapatite crystalline phase; 3252 cm"¹ and 602 cm"¹ - from water adsorbed on nanoHAP surface; 1090 cm"¹ and 1026 cm"¹ - v₃ P-0 bonds; 602 cm"¹ - v₄ P-0 bonds; 963 cm"¹ - Vi P-0 bonds; 1384 cm"¹ and 1457 cm"¹ corresponding to residual NO₃ ²" and CO₃ ²" groups ; 29296 cm"¹ and 2871 cm"¹ v_as and v_s C-H bonds in the -CH₂ - groups of the aminopropyl chain of APTES; 1573 cm"¹ from the - NH₃ ⁺ (HCO )₃~ formed by the amino groups of APTES.

TGA: The amount of APTES silane precursor introduced onto the surface of the hydroxyapatite particles, based on thermogravimetric measurements, determined from the difference in total mass loss at 900° C between the unmodified nanoHAP particles and the stage I modification product, is 1.45wt.%.

In the second step, the APTES-modified nanoHAP particles are reacted with L-lysine using the coupling agent l-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and the coupling reaction enhancing agent N-hydroxysulfo-succinimide (Sulfo - NHS). For this purpose, 12.7mg of L-lysine, 14.8mg of EDC, 27.0mg of Sulfo - NHS are dissolved in 20ml of MES buffer (pH 6) in a round-bottomed flask. The solution is stirred on a magnetic stirrer 30min to activate the carboxyl groups of L-lysine. After this time, the APTES- modified nanoHAP particles (2.42g) are dispersed in 20ml of MES buffer (pH=6), sonicated for 5min. (0.5 cycle; 50% amplitude), then transferred to a solution of L-lysine, EDC and sulfo - NHS. The whole is mixed using a magnetic stirrer for 24hr at room temperature. After this time, the suspension is centrifuged to separate the modified particles, washed 3 times with MES buffer (pH 6) and dried under vacuum at 60°C for 24 hours. A 2.14g product was obtained as a white powder (88% yield).

FTIR of the reaction product with L-lysine: (ATR, diamond crystal): The presence of L- lysine covalently attached by an amide bond is confirmed by the disappearance of the band at 1573 cm"¹ from the amino groups and the presence on the FTIR-ATR spectrum of the characteristic I amide band at 1644 cm"¹ from the vibrations of the C=O bonds.

TGA: The content of L-lysine covalently bound to the surface of nanoHAP particles via APTES, calculated from the difference between the total mass loss in the range 25°C- 900°C for nanoHAP particles modified with APTES and L-lysine and for nanoHAP particles modified with APTES, is 1.60 wt%.

Example 10. Production of porous material PGS-IDI

3.00 g of the product described in Example 2 was dissolved in 15 ml of 1,4-dioxane. Porous rock samples were then prepared by thermally induced phase separation combined with porogen leaching (TIPS-SL) consisting of the following steps:

1. Weighing 1.50 g of NaCI with a grain size of 400-500 pm as porogen into each well of a silicon 37-well plate.

2. Adding 0.75 ml of the previously prepared solution (dispersion) to each of the wells and leaving the plate in the air for 24 hours.

3. Freezing and freeze-drying for 24 hours.

4. Cross-linking: 50°C for 24 hours, 90°C for 96 hours.

5. Porogen washing - washing the foams with a large quantity of deionised water while controlling the conductivity of the washing medium.

6. Drying at 50°C to constant weight.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3369 - v -OH and - NH-, 2927 - v_as CH in CH₂ , 2855 - v_s CH in CH₂ , 1732 - v C=O, 1658 and 1533 - 6 -NH-, 1460 - 6_S CH in CH₂ , 1417 and 1233 - v C-0 in COOH, 1160 - v -C-0, 1097 - 6 in 2° OH, 1043 - 6 in 1° OH, 724 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) ⁼ 15.4°C, T_io% = 302.3°C

The cytocompatibility of the PGS-IDI product was tested at the level of mitochondrial activity of mouse fibroblast L929 cells and human osteoblast hFOB.1.19 cells according to ISO 10993-5:2009 standards using the MTT reduction assay after 24 hours incubation with biomaterial l/10th of the bottom well of a 96-well plate.

Bio-compatibility at the in vivo level was performed using a rat model (Wandering rat - Rattus norvegicus, breed Wistar, age - adult specimens minimum 10 weeks of age, weight - minimum 220g) in a study of local tissue response and generalized response to implantation of the product according to PN-EN ISO 10993-6:2017 (consent of the bioethics committee for the experiments number 42/£B 192/2021). After subcutaneous implantation of the test product, observation of the animals' behavior, the implantation site, and histopathological analysis of the implanted product were carried out.

Biological characterisation of PGS-IDI showed that the test material is not cytotoxic to mouse fibroblast line L929 and human osteoblast line hFOB 1.19, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993-5:2009 standard (Figure 4a). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 110.2% ± 13.9% and 105.6% ± 6.0%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Seven days after subcutaneous implantation of PGS-IDI material in vivo, no development of inflammation was observed and the histological image of the implanted material (Figure 4b) confirmed the absence of signs of inflammatory reaction (normal histological appearance). Macrophage cells present in the field of view invade the site of product implantation to clear accumulated postoperative fluid. The results indicate the biocompatibility of the PGS-IDI material in an in vivo system.

Example 11. Production of porous composite PGS-IDI/15HAP_B

3.00 g of the product described in example 2 was dissolved in 15 ml of 1,4-dioxane. To the solution, 0.53 g of the hydroxyapatite HAP_B described in example 5 was added (15 wt.% to the polymer) and stirred vigorously until a homogeneous system was obtained. Crosslinked porous scaffolds were then obtained from the resulting dispersion using the TIPS- SL method by the method described in example 10 in steps 1-6.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3367 - v in -OH and - NH-, 2928 - v_as in CH₂ , 2855 - v_as CH in CH₂ , 1732 - v in C=O, 1646 and 1537 - 6 in - NH-, 1461 - 6_S in CH₂ , 1417 and 1234 - v C-0 in COOH, 1163 - v -C-O, 1089 - 6 in 2° OH, 1030 - 6 in 1° OH, 725 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) ⁼ 11.6°C, T.io% = 325.8°C

The cytocompatibility of the product was tested as in example 10.

The biological characterisation of PGS-IDI/15HAP_B showed that the test material is not cytotoxic to mouse fibroblasts of the L929 lineage and human osteoblasts of the hFOB 1.19 lineage, which was supported by the results of cell viability after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993:5 2006 standard) (Fig. 5). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 hours incubation was 104.2% ± 9.1% and 96.6% ± 3.7%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 12. Production of porous composite PGS-IDI/30HAP_B

3.00 g of the product described in example 2 was dissolved in 15 ml of 1,4-dioxane. To the solution, 1.29 g of the hydroxyapatite HAP_B described in example 5 (30% by weight of the polymer) was added and stirred vigorously until a homogeneous system was obtained. Cross-linked porous scaffolds were then obtained from the resulting dispersion using the TIPS-SL method by the method described in example 10 in steps 1-6.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3368 - v in -OH and - NH-, 2928 - v CH in CH_{as 2} , 2855 - v_s CH in CH₂ , 1732 - v C=O, 1651 and 1538 - 6 - NH-, 1461 - 6_S CH in CH₂ , 1417 and 1235 - v in C-0 in COOH, 1164 - v -C-0, 1089 - 6 in 2° OH, 1030 - 6 in 1° OH, 724 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) = 23.1 °C, T_io% = 313.6 °C

The in vitro cytocompatibility and in vivo biocompatibility of the product were tested analogously to Example 10.

Biological characterisation of PGS-IDI/30HAP_B showed that the test material is not cytotoxic to mouse fibroblasts of the L929 lineage and human osteoblasts of the hFOB 1.19 lineage, which was supported by the results of cell viability after 24-hour exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993-5:2009 standard (Fig. 6a). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 94.8% ± 5.3% and 86.1% ± 17.4%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Seven days after subcutaneous implantation of PGS-IDI/30HAP_B material in vivo, no development of inflammation was observed and the histological image of the implanted material (Fig. 6b) was normal. Single macrophage cells present in the field of view infiltrated the implantation site to clear the accumulated postoperative fluid. The results indicate the biocompatibility of the PGS-IDI/30HAP_B material in an in vivo system. Example 13. Production of porous composite PGS-IDI/15HAP_B-Lys

4.50 g of the product described in Example 2 was dissolved in 22.50 ml of 1,4-dioxane. To the solution, 0.79 g (15 wt.% relative to the polymer) of hydroxyapatite HAP_B covalently functionalized with L-lysine, described in example 8, was added and stirred vigorously until a homogeneous system was obtained. Cross-linked porous scaffolds were then prepared from the resulting dispersion using the TIPS-SL method by the method described in Example 10 in steps 1-6.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3368 - v -OH and - NH-, 2926 - v_as CH in CH₂ , 2854 - v_s CH in CH₂ , 1732 - v C=O, 1651 and 1537 - 6 - NH-, 1457 - 6_S CH in CH₂ , 1417 and 1235 - v C-0 in COOH, 1163 - v -C-O, 1088 - 6 in 2° OH, 1030 - 6 in 1° OH, 724 - p CH in CH₂ .

DSC and TGA: Tg _{(lst heating)} = 23.4 °C, T__10% = 295.9 °C

The cytocompatibility of the PGS-IDI/15HAP_B-Lys product was tested analogously to Example 10.

Biological characterisation of PGS-IDI/15HAP_B-Lys showed that the test material is not cytotoxic to mouse fibroblasts of the L929 lineage and human osteoblasts of the hFOB 1.19 lineage, which was supported by the results of cell viability after 24 h exposure to the test product, in an MTT reduction assay performed according to the releva nt ISO 10993:5 2006 standard) (Fig. 7). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 hours incubation was 88.3% ± 3.7% and 77.8% ± 3.8%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 14. Production of porous composite PGS-IDI/30HAP_B-Lys

4.50 g of the product described in Example 2 was dissolved in 22.50 ml of 1,4-dioxane. To the solution, 1.87 g (30% by weight relative to the polymer) of HAP_B hydroxyapatite covalently functionalized with L-lysine, described in Example 8, was added and stirred vigorously until a homogeneous system was obtained. Cross-linked porous scaffolds were then prepared from the resulting dispersion using the TIPS-SL method by the method described in Example 10 in steps 1-6. FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3368 - v -OH and - NH-, 2925 - v_as CH in CH₂ , 2854 - v_s CH in CH₂ , 1732 - v C=O, 1646 and 1563 - 6 in -NH-, 1457 - 6_S CH in CH₂ , 1417 and 1235 - v C-0 in COOH, 1163 - v -C-O, 1088 - 6 in 2° OH, 1027 - 6 in 1° OH, 722 - p CH in CH₂

DSC and TGA: Tg (i_st heating) ⁼ 27.6 °C, T_io% =310.6 °C

The in vitro cytocompatibility and in vivo biocompatibility of the product were tested as in example 10.

Biological characterisation of PGS-IDI/30HAP_B-Lys showed that the test material is not cytotoxic to mouse fibroblasts of the L929 lineage and human osteoblasts of the hFOB 1.19 lineage, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993- 5:2009 standard (Fig. 8a). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 103.0% ± 5.8% and 98.9% ± 4.6%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Seven days after subcutaneous implantation of PGS-IDI/30HAP_B-Lys material in vivo, no development of inflammation was observed and the histological image of the implanted material (Fig. 8b) was normal. The few macrophage cells present in the field of view invaded the implantation site to clear the accumulated postoperative fluid. The results indicate the biocompatibility of the PGS-IDI/30HAP_B-Lys material in an in vivo system.

Example 15. Production of porous material PGS-ICPTES

3.00 g of the product described in Example 3 was dissolved in 15 ml of 1,4-dioxane, 36 mmol of deionised water, 12 mmol of ethanol and 1 drop of concentrated HCI were added. The resulting dispersion was used to form porous scaffolds using the porogen leach solution casting (SCPL) method described in steps 1-6:

1. Weighing 1.50 g of NaCI with a grain size of 400-500 pm into each of the wells of a hexagonal silicon 37-well plate (single hole diameter: 15 mm, hole depth: 10 mm),

2. Introduction of a 1.5 ml solution (dispersion) of the pPGS derivative into each well,

3. Leaving the plate for 24 hours at room temperature,

4. Cross-linking: 50°C for 24 hours, 90°C for 96 hours, 5. Sodium chloride washing - washing the foams with a large quantity of deionised water while controlling the conductivity of the wash medium,

6. drying at 50 °C.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3339 - v -OH , 2930 - v_as CH in CH₂ , 2856 - v_s CH in CH₂ , 1980 - v_s SiO₂ , 1733 - v C=O, 1457 - 6_S CH in CH₂ , 1416 and 1241 - v C-0 in COOH, 1193 - v -C-O, 1093 - 6 in 2° OH and v_as SiO₂ , 1012 - 6 in 1° OH and v_as SiO₂ , 774 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) =-7.6 °C, T_io% =314.6 °C

The cytocompatibility of the PGS-ICEPTES product was tested as in example 10.

Biological characterisation of PGS-ICEPTES showed that the test material is not cytotoxic to mouse fibroblast line L929 and human osteoblast line hFOB 1.19, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993-5:2009 standard (Fig. 9). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 92.1% ± 4.7% and 98.1% ± 6.8%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 16. Production of porous composite PGS-ICEPTES/15HAP_B

3.00 g of the product described in Example 3 was dissolved in 15 ml of 1,4-dioxane, 0.53 g of HAP_B hydroxyapatite described in Example 5 was added (15 wt.% to the polymer) and stirred vigorously until a homogeneous system was obtained. Then 36 mmol (0.65 g) of deionised water, 12 mmol (0.55 g) of ethanol and 1 drop of concentrated HCI were introduced successively into the mixture. The dispersion was used to produce porous scaffolds by solution-casting with porogen leaching (SCPL) by the method described in Example 15 in steps 1-6.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3341 - v -OH , 2930 - v_as CH in CH₂ , 2856 - v_s CH in CH₂ , 1980 - v_s SiO₂ , 1733 - v C=O, 1456 - 6_S CH in CH₂ , 1416 and 1240 - v C-0 in COOH, 1192 - v -C-O, 1087 - 6 in 2° OH and v_as SiO₂ , 1027 - 6 in 1° OH and v_as SiO₂ , 774 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) =-8.8 °C, T_io% =346.6 °C The cytocompatibility of the PGS-ICEPTES/15HAP_B product was tested similarly to Example 10.

Biological characterisation of PGS-ICEPTES/15HAP_B showed that the test material is not cytotoxic to mouse fibroblasts of the L929 lineage and human osteoblasts of the hFOB 1.19 lineage, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993- 5:2009 standard (Fig. 10). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 111.9% ± 7.3% and 102.9% ± 4.7%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 17. Production of porous composite PGS-ICEPTES/30HAP_B

3.00 g of the product described in Example 3 was dissolved in 15 ml of 1,4-dioxane, 0.53 g of HAP_B hydroxyapatite described in Example 5 was added (15 wt.% to the polymer) and stirred vigorously until a homogeneous system was obtained. Then 36 mmol (0.65 g) of deionised water, 12 mmol (0.55 g) of ethanol and 1 drop of concentrated HCI were introduced successively into the mixture and mixed thoroughly. The dispersion was used to fabricate porous scaffolds by solution-casting with porogen leaching (SCPL) by the method described in Example 15 in steps 1-6.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3339 - v -OH , 2930 - v_as CH in CH₂ , 2856 - v_s CH in CH₂ , 1980 - v_s SiO₂ , 1733 - v C=O, 1456 - 6_S CH in CH₂ , 1416 and 1239 - v C-0 in COOH, 1192 - v -C-O, 1089 - 6 in 2° OH and v_as SiO₂ , 1027 - 6 in 1° OH and v_as SiO₂ , 773 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) ⁼~7 -2 °C, T_io% =376.4 °C

The cytocompatibility of the PGS-ICEPTES/30HAP_B product was tested analogously to Example 10.

Biological characterisation of PGS-ICEPTES/30HAP_B showed that the test material is not cytotoxic to mouse fibroblasts of the L929 lineage and human osteoblasts of the hFOB 1.19 lineage, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993- 5:2009 standard (Fig. 11). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 98.3% ± 3.4% and 106.9% ± 4.5%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 18. Production of porous composite PGS-ICPTES/15HAP_B-Lys

3.00 g of the product described in Example 3 was dissolved in 15 ml of 1,4-dioxane, 0.53 g of the hydroxyapatite HAP_B covalently functionalized with L-lysine described in Example 8 (15 wt.% relative to the polymer) was added and stirred vigorously until a homogeneous system was obtained. Then 36 mmol (0.65 g) of deionised water, 12 mmol (0.55 g) of ethanol and 1 drop of concentrated HCI were introduced successively into the mixture and mixed thoroughly. The dispersion was used to fabricate porous scaffolds by solutioncasting with porogen leaching (SCPL) by the method described in Example 15 in steps 1-6. FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3359 - v -OH , 2929 - v_as CH in CH₂ , 2856 - v_s CH in CH₂ , 1980 - v_s SiO₂ , 1732 - v C=O, 1456 - 6_S CH in CH₂ , 1416 and 1239 - v C-0 in COOH group, 1089 - 6 in 2° OH and v_as SiO_2; 1028 - 6 in 1° OH and v_as SiO₂ , 773 - p CH in CH₂

DSC and TGA: Tg (i_st heating) =10.7 °C, T_io% =322.6 °C

The cytocompatibility of the product PGS-ICEPTES/15HAP_B-Lys was tested similarly to example 10.

Biological characterisation of PGS-ICEPTES/15HAP_B-Lys showed that the test material is not cytotoxic to mouse fibroblasts of the L929 lineage and human osteoblasts of the hFOB 1.19 lineage, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993- 5:2009 standard (Fig. 12). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 80.1% ± 3.3% and 86.6% ± 5.1%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 19. Production of porous composite PGS-ICPTES/30HAP_B-Lys

3.00 g of the product described in Example 3 was dissolved in 15 mL of 1,4-dioxane, 1.29 g of HAP_B hydroxyapatite covalently functionalized with L-lysine described in Example 8 (30% by weight relative to the polymer) was added and stirred vigorously until a homogeneous system was obtained. Then 36 mmol (0.65 g) of deionised water, 12 mmol (0.55 g) of ethanol and 1 drop of concentrated HCI were introduced successively into the mixture. The entire mixture was mixed and the dispersion was then used to produce porous scaffolds by solution casting with porogen leaching (SCPL) by the method described in Example 15 in steps 1-6.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3359 - v -OH , 2929 - v_as CH in CH₂ , 2855 - v_s CH in CH₂ , 1733 - v C=O, 1456 - 6 CH in CH_s2 , 1417 and 1239 - v C-0 in COOH, 1088 - 6 2° OH and v_as SiO₂, 1026 - 6 1° OH and v_as SiO₂ , 773 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) ⁼6.8 °C, T_io% =348.4 °C

The cytocompatibility of the product PGS-ICEPTES/30HAP_B-Lys was tested analogously to Example 10.

Biological characterisation of PGS-ICEPTES/30HAP_B-Lys showed that the test material is not cytotoxic to mouse fibroblasts of the L929 lineage and human osteoblasts of the hFOB 1.19 lineage, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993- 5:2009 standard (Fig. 13). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 91.1% ± 7.0% and 98.4% ± 6.3%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 20. Production of porous PGS-LDI material

3.00 g of the product described in Example 4 was dissolved in 15 ml of 1,4-dioxane. Crosslinked, porous scaffolds were then obtained from the resulting solution using the TIPS-SL method by the method described in Example 10 in steps 1-6.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3360 - v -OH and - NH-, 2929 - v_as CH in CH₂ , 2855 - v_s CH in CH₂ , 1732 - v C=O, 1659 and 1533 - 6 - NH-, 1456 - 6_S CH in CH₂ , 1417 and 1236 - v C-0 in COOH, 1163 - v -C-O, 1095 - 6 in 2° OH, 1026 - 6 in 1° OH, 725 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) ⁼ 0-7 °C, T_io% ⁼ 304.8 °C

The cytocompatibility of the PGS-LDI product was tested as in example 10.

Biological characterisation of PGS-LDI showed that the test material is not cytotoxic towards mouse fibroblast line L929 and human osteoblast line hFOB 1.19, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993-5:2009 standard (Fig. 14). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 89.2% ± 13.0% and 91.1% ± 10.0%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 21. Production of porous composite PGS-LDI/30nanoHAP1200

3.00 g of the product described in Example 4 was dissolved in 15 ml of 1,4-dioxane. To the solution, 1.29 g of the Ca-P nanoHAP1200 particles described in Example 7 (30 wt% to the polymer) were added and stirred vigorously until a homogeneous dispersion of particles was obtained. The resulting dispersion was then used to obtain cross-linked porous scaffolds from the obtained dispersion using the TIPS-SL method as described in Example 10 in steps 1-6.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3365 - v -OH and - NH-, 2929 - v_as CH in CH₂ , 2855 - v_s CH in CH₂ , 1732 - v C=O, 1658 and 1537 - 6 - NH-, 1456 - 6_S CH in CH₂ , 1417 and 1259 - v C-0 in COOH, 1163 - v -C-O, 1088 - 6 in 2° OH, 1022 - 6 in 1° OH, 725 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) ⁼ 14.7 °C, T_io% ⁼ 335.7 °C

The cytocompatibility of the PGS-LDI/30nanoHAP1200 product was tested similarly to Example 10.

Biological characterisation of PGS-LDI/30nanoHAP1200 showed that the test material is not cytotoxic to mouse fibroblast line L929 and human osteoblast line hFOB 1.19, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993-5:2009 standard (Fig. 15). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 93.2% ± 7.8% and 78.2% ± 11.8%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 22. Production of porous composite PGS-LDI/30nanoHAP-Lys 3.00 g of the product described in Example 4 was dissolved in 15 ml of 1,4-dioxane. To the solution, 1.29 g of nanometric nanoHAP hydroxyapatite particles covalently functionalized with L-lysine nanoHAP-Lys described in Example 9 (30% by weight relative to the polymer) were added and stirred vigorously until a homogeneous dispersion of particles was obtained. The resulting dispersion was then used to prepare cross-linked, porous scaffolds using the TIPS-SL method as described in Example 10 at 1-6. The resulting materials were characterized using FTIR, DSC and TGA.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3363 - v -OH and - NH-, 2928 - v_as CH in CH₂ , 2855 - v_s CH in CH₂ , 1732 - v C=O, 1651 and 1533 - 6 - NH-, 1456 - 6_S CH in CH₂ , 1417 and 1257 - v C-0 in COOH, 1163 - v -C-O, 1093 - 6 in 2° OH, 1027 - 6 in 1° OH, 725 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) ⁼ 13.4 °C, T_io% =338.3 °C

The cytocompatibility of the PGS-LDI/30nanoHAP-Lys product was tested analogously to Example 10.

Biological characterisation of PGS-LDI/30nanoHAP-Lys showed that the test material is not cytotoxic to mouse fibroblasts of the L929 line and human osteoblasts of the hFOB 1.19 line, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993- 5:2009 standard (Fig. 16). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 87.7% ± 11.4% and 85.6% ± 13.5%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 23. Production of PGS-IDI solid material

From the product obtained as described in example 2, which is in the form of a viscous liquid, solid discs with a diameter of 2 mm and a height of 1 mm were produced by the following method: the solution (dispersion) was placed in a teflon tube with an internal diameter of 2 mm, the tube was placed in an atmospheric dryer and heated at 50°C for 24 hours and then at 90°C for a further 96 hours. The cross-linked material was then removed from the tubing and the resulting rod was cut into discs measuring 2 mm in diameter and 1 mm high. FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3368 - v -OH and - NH-, 2926 - v_as CH in CH₂ , 2854 - v_s CH in CH₂ , 1732 - v C=O, 1650 and 1532 - 6 -NH-, 1462 - 6_S CH in CH₂ , 1417 and 1233 - v C-0 in COOH, 1158 - v -C-0, 1097 - 6 in 2° OH, 1028 - 6 in 1° OH, 724 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) =29.6 °C, T_io% =321.3 °C

The cytocompatibility of the PGS-IDI product was tested as in example 10.

Biological characterisation of PGS-IDI showed that the test material is not cytotoxic to mouse fibroblast line L929 and human osteoblast line hFOB 1.19, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993-5:2009 standard (Fig. 17). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 99.9% ± 6.4% and 109.2% ± 6.2%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 24. Production of solid composite PGS-IDI/30HAP_B

The product obtained as described in example 2 (pPGS-IDI) in an amount of 3.00 g and 1.29 g of hydroxyapatite HAP_B obtained as described in example 5 (30wt.% in relation to the polymer) were mixed thoroughly to obtain a homogeneous dispersion. The resulting dispersion was used to produce discs of 2 mm diameter and 1 mm height by the method described in example 23.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3367 - v -OH and - NH-, 2924 - v_as CH in CH₂ , 2854 - v_s CH in CH₂ , 1723 - v C=O, 1650 and 1526 - 6 -NH-, 1462 - 6_S CH in CH₂ , 1417 and 1232 - v C-0 in COOH, 1156 - v -C-0, 1090 - 6 in 2° OH, 1029 - 6 in 1° OH, 723 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) ⁼ 49.8 °C, T_io% =333.6 °C

The cytocompatibility of the PGS-IDI/30HAP_B product was tested similarly to Example 10.

Biological characterisation of PGS-IDI/30HAP_B showed that the test material is not cytotoxic to mouse fibroblasts of the L929 lineage and human osteoblasts of the hFOB 1.19 lineage, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993- 5:2009 standard (Fig. 18). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 103.9% ± 8.8% and 100.3% ± 9.3%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 25. Production of a solid composite PGS-IDI/30HAP_B-Lys

The product obtained as described in Example 2 (pPGS-IDI) in an amount of 3.00g and 1.29 g of hydroxyapatite covalently functionalized with L-lysine obtained as described in Example 8 (HAP_B-Lys) (30% by weight in relation to the polymer) were placed in a round-bottomed flask and mixed thoroughly to obtain a homogeneous dispersion. From the resulting dispersion, discs with a diameter of 2 mm and a height of 1 mm were produced by the method described in example 23.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3366 - v -OH and - NH-, 2924 - v_as CH in CH₂ , 2854 - v_s CH in CH₂ , 1732 - v C=O, 1647 and 1526 - 6 -NH-, 1457 - 6_S CH in CH₂ , 1417 and 1233 - v C-0 in COOH, 1158 - v -C-0, 1095 - 6 in 2° OH, 1031 - 6 in 1° OH, 721 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) ⁼ 40.8 °C, T_io% ⁼ 347.9 °C

The cytotoxicity of the PGS-IDI/30HAP_B-Lys product was tested analogously to Example 10. The biological characterisation of PGS-IDI/30HAP_B-Lys showed that the test material is not cytotoxic to mouse fibroblasts of the L929 lineage and human osteoblasts of the hFOB 1.19 lineage, which was supported by the cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO standard 10993-5:2009 (Fig. 19). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 105.2% ± 4.9% and 100.3% ± 10.9%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 26. Production of solid material from PGS-LDI in the form of 6.5mm diameter, 1.0mm high discs and 10.0mm diameter, 0.6mm thick discs 5.0 g of the product described in Example 3 (pPGS-LDI) was poured into Teflon molds with 6.5mm diameter and 1.0mm high wells and cross-linked at 40 °C in an air-circulating dryer for 4 days until the absorption band from the isocyanate group completely disappeared. FTIR-ATR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3351 - v -OH and - NH-, 2928 - v_as CH in CH₂ , 2855 - v_s CH in CH₂ , 1731 - v C=O, 1650 and 1538 - 6 -NH-, 1460 - 6_S CH₂ , 1416 and 1257 - v C-0 in COOH, 1163 - v -C-O, 1096 - 6 2° OH, 1025 - 6 1° OH, 724 - p CH in CH₂

DSC and TGA: Tg (i_st heating) =-18.0 °C, T.io% =297.9 °C

DMA: The composite complex shear modulus (G*) of a 10.0mm diameter, 0.6mm thick disc material determined at 25° C, at a frequency of 10Hz and a force of 4N is 924kPa.

The cytocompatibility of the PGS-LDI product was tested as in Example 10.

Biological characterisation of PGS-LDI showed that the test material is not cytotoxic to mouse fibroblast line L929 and human osteoblast line hFOB 1.19, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993-5:2009 standard (Fig. 20). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 83.3% ± 8.7% and 88.7% ± 8.8%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 27. Production of a solid composite PGS-LDI_20%nanoHAP1200

4.0g of the product described in Example 4 (pPGS-LDI) and 1.0g of nanometric nanoHAP1200 biphasic calcium phosphate particles described in Example 7 were weighted into an agate mortar. The ingredients were mixed in the mortar until a homogeneous dispersion of the ceramic particles in the polymer matrix was achieved. The mixture was then degassed in a vacuum dryer and poured into Teflon molds with wells 6.5mm in diameter and 1.0mm high and cross-linked at 40°C in an air-circulating dryer for 4 days until the absorption band originating from the isocyanate group completely disappeared.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3365 - v -OH and -NH-, 2929 - v_as CH in CH₂ , 2856 - v_s CH in CH₂ , 1732 - v C=O, 1651 and 1538 - 6 -NH-, 1460 - 6_S CH in CH₂ , 1417 and 1236 - v C-0 in COOH, 1164 - v -C-O, 1090 - 6 in 2° OH, 1025 - 6 in 1° OH, 962 - Vi P-O, 725 - p CH in CH₂ , 602 - v₃ P-O.

DSC and TGA: Tg (i_st heating) ⁼ -8.0 °C, T_io% ⁼ 322.4 °C

DMA: The composite complex shear modulus (G*) of a 10.0mm diameter, 0.6mm thick disc material determined at 25° C, at a frequency of 10Hz and a force of 4N is 2.806MPa. The cytocompatibility of the product PGS-LDI_20%nanoHAP1200 was tested analogously to Example 10.

Biological characterisation of PGS-LDI_20%nanoHAP1200 showed that the test material is not cytotoxic to mouse fibroblasts of the L929 line and human osteoblasts of the hFOB 1.19 line, which was supported by cell viability results after 24 h exposure to the test product, in an MTT reduction assay performed according to the relevant ISO 10993- 5:2009 standard (Fig. 21). The viability of L929 fibroblasts and hFOB 1.19 osteoblasts after 24 h incubation was 95.3% ± 6.2% and 104.5% ± 7.6%, respectively, indicating the absence of deleterious effects of the developed material in in vitro tests.

Example 28. Production of a solid composite PGS-LDI_20%HAP_B

4.0g of the product described in Example 4 (pPGS-LDI) and 1.0g of nanometric hydroxyapatite HAP_B particles obtained as described in Example 5 were weighted into an agate mortar. The ingredients were mixed in the mortar until a homogeneous dispersion of the ceramic particles in the polymer matrix was achieved. The mixture was then degassed in a vacuum dryer and poured into Teflon molds with wells 6.5mm in diameter and 1.0mm high and cross-linked at 40°C in an air-circulating dryer for 4 days until complete disappearance of the absorption band at 2252 cm"¹ originating from the isocyanate group.

FTIR (ATR, diamond crystal, wavenumber [cm ¹ ]): 3370 - v -OH and -NH-, 2929 - v_as CH in CH₂ , 2855 - v_s CH in CH₂ , 1732 - v C=O, 1658 and 1544 - 6 -NH-, 1457 - 6_S CH in CH₂ , 1417 and 1234 - v C-0 in COOH, 1166 - v -C-O, 1089 - 6 in 2° OH, 1031 - 6 in 1° OH, 962 - Vi P-O, 724 - p CH in CH₂ , 602 - v₃ P-O.

DSC and TGA: Tg (i_st heating) =-25.0 °C, T_io% = 361.0 °C

DMA: The composite complex shear modulus (G*) of a 10.0mm diameter, 0.6mm thick disc material determined at 25° C, at a frequency of 10Hz and a force of 4N is 1.068MPa. Example 29. Production of a solid composite PGS-LDI_20%HAP_B-Lys

Into an agate mortar were weighted 4.0g of pPGS-LDI as described in Example 4 and 1.0g of hydroxyapatite particles covalently modified with L-lysine HAP_B-Lys, obtained as described in Example 8. The ingredients were mixed in the mortar until a homogeneous dispersion of the ceramic particles in the polymer matrix was obtained. The mixture was then degassed in a vacuum dryer and poured into Teflon molds with wells 6.5mm in diameter and 1.0mm high and cross-linked at 40°C in an air-circulating dryer for 4 days until complete disappearance of the absorption band at 2252cm"¹ originating from the isocyanate group.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3370 - v -OH and -NH-, 2928 - v_as CH in CH₂ , 2855 - v_s CH in CH₂ , 1732 - v C=O, 1651 and 1545 - 6 -NH-, 1457 - 6_S CH in CH₂ , 1417 and 1234 - v C-0 in COOH, 1165 - v -C-O, 1090 - 6 in 2° OH, 1032 - 6 in 1° OH, 963 - Vi P-O, 724 - p CH in CH₂ , 602 - v₃ P-O.

DSC and TGA: Tg (i_st heating) ⁼-21.7 °C, T_io% ⁼ 333.0 °C

DMA: The composite complex shear modulus (G*) of a 10.0mm diameter, 0.6mm thick disc material determined at 25° C, at a frequency of 10Hz and a force of 4N is 1.271MPa.

Example 30. Production of PGS-LDI solid material in the form of 2 mm diameter discs

The product obtained as described in Example 4 (pPGS-LDI), which is in the form of a viscous liquid, was transformed into solid discs of 2 mm diameter and 1 mm height by the method described in Example 23.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3359 - v -OH and -NH-, 2929 - v_as CH in CH₂ , 2856 - v_s CH in CH₂ , 1722 - v C=O, 1651 and 1526 -6 -NH-, 1456 - 6_S CH in CH₂ , 1417 and 1235 - v C-0 in COOH, 1163 - v -C-O, 1095 - 6 in 2° OH, 1026 - 6 in 1° OH, 725 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) = 9.1 °C, T_io% = 323.8 °C

Example 31. Production of a solid composite PGS-LDI_30nanoHAP1200

The product obtained as described in Example 4 (pPGS-LDI) in an amount of 3.00 g and 1.29 g of nanoHAP1200 hydroxyapatite as described in Example 7 (30% by weight relative to the polymer) was mixed thoroughly to obtain a homogeneous dispersion. The dispersion was used to produce discs of 2 mm diameter and 1 mm height by the method described in Example 23.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3369 - pv -OH and -NH-, 2925 - v_as CH in CH₂ , 2855 - v_s CH in CH₂ , 1731 - v C=O, 1647 and 1532 - 6 -NH-, 1456 - 6_S CH in CH₂ , 1417 and 1235 - v C-0 in COOH, 1163 - v -C-O, 1093 - 6 in 2° OH, 1025 - 6 in 1° OH, 725 - p CH in CH₂ .

DSC and TGA: Tg _(lst heating) = H.l °C, T_₁₀% = 354.2 °C

Example 32. Production of a solid composite PGS-LDI_30nanoHAP-Lys

The product obtained as described in Example 4 (pPGS-LDI) in an amount of 3.00g and 1.29g of hydroxyapatite covalently functionalized with L-lysine (nanoHAP-Lys) as described in Example 8 (30% by weight relative to the polymer) was thoroughly mixed to produce a homogeneous dispersion. The dispersion was used to produce discs of 2 mm diameter and 1 mm height by the method described in Example 23.

FTIR: (ATR, diamond crystal, wavenumber [cm ¹ ]): 3362 - v -OH and -NH-, 2927 - v_as CH in CH₂ , 2855 - v_s CH in CH₂ , 1732 - v C=O, 1650 and 1532 - 6 -NH-, 1456 - 6_S CH in CH₂ , 1417 and 1236 - v C-0 in COOH, 1165 - v -C-O, 1092 - 6 in 2° OH, 1027 - 6 in 1° OH, 725 - p CH in CH₂ .

DSC and TGA: Tg (i_st heating) ⁼ 10.9 °C, T_io% = 340.2 °C

Claims

1. Biocompatible solid elastomeric material intended for use as an implantable material for filling bone defects, bone tissue regeneration and as a coating material for biomedical bone applications, based on poly(glycerol sebacate), characterized in that it is poly(glycerol sebacate) cross-linked with a chemical agent in the form of L-lysine diisocyanate ethyl ester or isophorone diisocyanate.

2. Method of producing a solid elastomer for use as an implantable material for bone defects filling, bone tissue regeneration and as a coating material for biomedical applications, based on a biodegradable polymer, characterized in that in a first step a chemical pre-crosslinking of the poly(glycerol sebacate) pPGS with L-lysine diisocyanate ethyl ester (LDI) or isophorone diisocyanate (IDI) in tetra hydrofuran or 1,4-dioxane is carried out, using LDI or IDI in an amount of 1 to 2.5 mmol per 1 g pPGS, preferably 2.0 mmol per 1 g pPGS, at a temperature of 25°C to 60°C for a period of 5 to 24 hours., preferably from 40° C to 60° C for 5 hours to 24 hours, followed by appropriate thermal cross-linking of pPGS pre-crosslinked with L-lysine diisocyanate ethyl ester (pPGS-LDI) or isophorone diisocyanate (pPGS-IDI) at 30°C to 90°C until the isocyanate band originating from the chemical cross-linking agent disappears.

3. Method according to claim 2, wherein the initial chemical cross-linking of pPGS with L- lysine diisocyanate ethyl ester or isophorone diisocyanate is carried out in tetrahydrofuran or 1,4-dioxane as solvent, at 50° C for 4 h.

4. Method according to claim 2, wherein the specific cross-linking is carried out at an elevated temperature over a period of 3 to 14 days.

5. Solid composite with osteoinductive properties for bone defects filling and regenerating bone tissue based on calcium phosphate particles and biodegradable polymers, characterized in that the composite comprises poly(glycerol sebacate) chemically cross-linked using a chemical agent such as: L-lysine diisocyanate ethyl ester or isophorone diisocyanate, and further comprises, as filler, calcium phosphate particles such as hydroxyapatite in the form of nanometric particles "HAP_B" or nanometric hydroxyapatite particles covalently functionalized with L-lysine "HAP_B-Lys" or nanometric particles of biphasic calcium phosphate "nanoHAP1200" or hydroxyapatite in the form of nanometric particles with a high surface development covalently modified with L-lysine "nanoHAP-Lys", whereby the chemically cross-linked poly(glycerol sebacate) is 70 wt.%. to 80% by weight relative to the total weight of the two-component composite, apatite particles are between 20% by weight and 30% by weight relative to the total weight of the two-component composite.

6. Composite according to claim 5, wherein the poly(glycerol sebacate) is chemically cross-linked using a chemical agent such as ethyl ester of L-lysine diisocyanate or isophorone diisocyanate.

7. Composite according to claim 5, wherein the composite contains chemically crosslinked poly(glycerol sebacate) in an amount of 70%wt. to 80 wt. relative to the total weight of the two-component composite.

8. Composite according to claim 5, wherein the filler is calcium phosphate particles selected from: non-surface-modified "HAP_B" or "nanoHAP1200" particles or calcium phosphate particles covalently modified with L-lysine "HAP_B-Lys" and "nanoHAP-Lys" particles containing from 1 to 10 wt.% of L-lysine relative to the total weight of the filler.

9. Composite according to claim 5, wherein the filler particle content is between 20 wt.% and 30 wt.% in relation to the total weight of the two-component composite.

10. Method for producing a solid composite for bone defects filling and regenerating bone tissue based on calcium phosphate particles and biodegradable polymers, characterized in that chemical crosslinking of poly(glycerol sebacate) pPGS with ethyl ester of L-lysine diisocyanate (LDI) or isophorone diisocyanate (IDI) in tetra hydrofuran is carried out in a first step, using LDI or IDI in an amount of 2 mmoles per 1 g of pPGS, at a temperature of 25°C to 60°C for a period of 5 to 24 hours., preferably from 40° C to 60° C for 5 hours to 24 hours, followed by the preparation at room temperature of a mixture of the components of the composite consisting of pPGS pre-crosslinked with L-lysine diisocyanate ethyl ester (pPGS-LDI) or isophorone diisocyanate (pPGS-IDI) in an amount of 70%wt. to 80 wt. relative to the total weight of the binary composite, calcium phosphate particles of one type of the four: hydroxyapatite in the form of nanometric particles "HAP_B" or nanometric hydroxyapatite particles covalently functionalized with L-lysine "HAP_B-Lys" or biphasic Ca-P particles "nanoHAP1200" or and nanometric hydroxyapatite particles with high surface development covalently functionalized with L-lysine "nanoHAP-Lys" in an amount of from 20 wt. %. to 30 wt. %. relative to the total weight of binary composite, and the substrates are thoroughly mixed and degassed, followed by thermal crosslinking of the mixture at 30°C to 90°C until the isocyanate band from the chemical crosslinking agent disappears.

11. Method according to claim 10, wherein the initial chemical cross-linking of PGS with L- lysine diisocyanate ethyl ester or isophorone diisocyanate is carried out in tetrahydrofuran or 1,4-dioxane as solvent, at 50° C for 4h.

12. Method according to claim 10, wherein thermal crosslinking of the mixture is carried out until the disappearance of the isocyanate band originating from the ethyl ester of L- lysine diisocyanate or isophorone diisocyanate.

13. Method according to claim 10, wherein the thermal cross-linking of the mixture is carried out at an elevated temperature over a period of 5 days.

14. Porous elastomeric material with in vitro and in vivo biocompatible properties for bone defects filling and regenerating bone tissue for biomedical bone applications, based on biodegradable polymers poly(glycerol sebacate), characterized in that the material is composed of poly(glycerol sebacate) chemically cross-linked using a chemical agent of one of three choices: L-lysine diisocyanate ethyl ester or isophorone diisocyanate or 3- (triethoxysilyl)propyl isocyanate.

15. Method of producing a porous material for bone defects filling and regenerating bone tissue for biomedical bone applications, based on a biodegradable polymer, characterized in that the chemical cross-linking of poly(glycerol sebacate) pPGS with L-lysine diisocyanate ethyl ester (LDI) or isophorone diisocyanate (IDI) or 3-(triethoxysilyl)propyl isocyanate (ICPTES) in anhydrous 1,4-dioxane is carried out in a first step, using ethyl ester of L-lysine diisocyanate (LDI) or isophorone diisocyanate (IDI) in an amount of 1 to 2.5 mmol per 1 g pPGS, preferably 2 mmol per 1 g pPGS, and 3-(triethoxysilyl)propyl isocyanate in an amount of 2 to 5 mmol per 1 g pPGS, preferably 4 mmol per 1 g pPGS. The reaction is carried out at a temperature of 25°C to 60°C for a period of 5 to 24 hrs, preferably from 40° C to 60° C for 5 to 24 h. A solution of 20%(m/v) poly(glycerol sebacate) chemically modified with isophorone diisocyanate or 3-(triethoxysilyl)propyl isocyanate or ethyl ester of L-lysine diisocyanate in anhydrous 1,4-dioxane is then prepared at room temperature. To the solution of poly(glycerol sebacate) chemically modified with 3-(triethoxysilyl)propyl isocyanate, water is also added in an amount of 6 to 18 mmol per 1 g of pPGS, preferably 12 mmol per 1 g of poly(glycerol sebacate) chemically modified with 3-(triethoxysilyl)propyl isocyanate, and ethanol in an amount of 2 to 6 mmol per 1 g of pPGS, preferably 4 mmol per 1 g of poly(glycerol sebacate) chemically modified with 3-(triethoxysilyl)propyl isocyanate, and one drop of concentrated hydrochloric acid is added. The solution is then placed in the mold wells previously filled with porogen in the form of NaCI, the mold is left in air for 24 hours, then in the case of pPGS modified with IDI or LDI it is additionally subjected to a freeze-drying process. This is followed by a thermal crosslinking process at 50°C for 24 hours and then at 90°C for 96 hours. Porogen is then removed from the cross-linked porous materials by repeatedly washing with deionised water and dried at 40°C

16. Method according to claim 15, wherein the initial chemical cross-linking of pPGS with L-lysine diisocyanate ethyl ester or isophorone diisocyanate or 3-(triethoxysilyl)propyl isocyanate is carried out in anhydrous 1,4-dioxane as a solvent, at 50° C for 4 h.

17. Method according to claim 15, wherein a crosslinking process specific to the disappearance of the isocyanate band, originating from the ethyl ester of L-lysine diisocyanate or isophorone diisocyanate or 3-(triethoxysilyl)propyl isocyanate, is carried out.

18. The method according to claim 15, wherein the specific cross-linking is carried out at an elevated temperature over a period of 5 days.

19. Porous composite with osteoinductive properties for filling bone defects and regenerating bone tissue based on calcium phosphate particles and biodegradable polymers, characterized in that the composite comprises poly(glycerol sebacate) chemically cross-linked with a chemical agent such as ethyl ester of L-lysine diisocyanate or isophorone diisocyanate or 3-(triethoxysilyl)propyl isocyanate, and calcium phosphate particles as a filler, whereby the chemically cross-linked poly(glycerol sebacate) is between 70 wt. % and 85 wt. % relative to the filler, to 85 wt. % of the total weight of the two-component composite, while calcium phosphate particles are present in an amount of 15 wt. % to 30 wt. % of the total weight of the two-component composite.

20. Composite according to claim 19, wherein the filler is calcium phosphate particles selected from: non-surface-modified "HAP_B" or "nanoHAP1200" particles or calcium phosphate particles covalently modified with L-lysine "HAP_B-Lys" and "nanoHAP-Lys", containing from 1 to 10 wt.% of L-lysine in relation to the total weight of the filler.

21. Method of preparing a two-component porous composite for filling bone defects and bone tissue regeneration based on calcium phosphate particles and biodegradable polymers, wherein the first step a chemical modification of poly(glycerol sebacate) pPGS prepolymer is carried out and the product of the chemical modification reaction is isolated, then a solution of the chemically modified poly(glycerol sebacate) prepolymer in anhydrous 1,4-dioxane is prepared at room temperature, then calcium phosphate particles are added to the prepared solution and the whole is mixed thoroughly, then the obtained dispersion is placed in a mold filled with porogen, then the solvent is removed by freeze-drying, then the process of crosslinking the elastomeric matrix of the biocomposite is carried out at elevated temperature, then the porogen is removed from the crosslinked composite material by repeated washing with deionised water and the obtained composite material is dried.

22. Method according to claim 21, wherein the chemical modification of the poly(glycerol sebacate) prepolymer is carried out with ethyl ester of L-lysine diisocyanate (LDI) or isophorone diisocyanate (IDI) or 3-(triethoxysilyl)propyl isocyanate (ICPTES) in anhydrous 1,4-dioxane.

23. Method according to claim 21, wherein an ethyl ester of L-lysine diisocyanate (LDI) or isophorone diisocyanate (IDI) in an amount of 2 mmol per 1 g of pPGS is used for chemical modification of the poly(glycerol sebacate) prepolymer.

24. Method according to claim 21, wherein 3-(triethoxysilyl)propyl isocyanate (ICPTES) in an amount of 4 mmol per 1 g pPGS is used for chemical modification of the poly(glycerol sebacate) prepolymer.

25. Method according to claim 21, wherein the chemical modification of poly(glycerol sebacate) is carried out at a temperature between 25°C and 70°C for 3 to 24 hours, preferably at 50°C for 4 hours.

26. Method according to claim 21, wherein the reaction product of chemical modification of poly(glycerol sebacate) prepolymer with ethyl ester of L-lysine diisocyanate (LDI) or isophorone diisocyanate (I DI) or 3-(triethoxysilyl)propyl isocyanate (ICPTES) is dissolved in anhydrous 1,4-dioxane to obtain a solution with a concentration of 15%(m/v) to 25%(m/v), preferably 20%(m/v).

27. Method according to claim 21 in which to a solution of poly(glycerol sebacate) prepolymer chemically modified with 3-(triethoxysilyl)propyl isocyanate (ICPTES) in anhydrous 1,4-dioxane is added, immediately after dissolution, water in an amount of 12 mmol per 1 g of pPGS chemically modified with 3-(triethoxysilyl)propyl isocyanate and ethanol in an amount of 4 mmol per 1 g of pPGS chemically modified with 3- (triethoxysilyl)propyl isocyanate and ethanol in an amount of 4 mmol per 1 g of pPGS chemically modified with 3-(triethoxysilyl)propyl isocyanate and one drop of concentrated hydrochloric acid is finally added.

28. Method according to claim 21, wherein calcium phosphate particles are added to a solution of pPGS chemically modified with L-lysine diisocyanate ethyl ester (LDI) or isophorone diisocyanate (IDI) or 3-(triethoxysilyl)propyl isocyanate (ICPTES) in anhydrous 1,4-dioxane: hydroxyapatite in the form of nanometric particles "HAP_B" or nanometric hydroxyapatite particles covalently functionalized with L-lysine "HAP_B-Lys" or biphasic Ca-P particles "nanoHAP1200" or nanometric hydroxyapatite particles with a large surface development covalently functionalized with L-lysine "nanoHAP-Lys" in an amount ranging from 15 wt.%. to 30 wt. %. relative to the total dry weight of the binary composite, preferably in an amount of 30wt.%. relative to the total dry weight of the two-component composite.

29. Method according to claim 21, wherein sodium chloride with a grain size of 200 to 800 pm, preferably 400 to 500 pm, is used as porogen.

30. Method according to claim 21, wherein the specific crosslinking of the mixture is carried out at 50°Cfor 24 hours and then at 90 °Cfor 96 hours.