TW201132367A - A process for producing inorganic interconnected 3D open-cell bone substitutes - Google Patents

Abstract

The present invention relates to a process of using a heat responsive mixture to produce inorganic interconnected 3D open-cell bone substitutes which can be applied in the orthopedic or dental field for treatment of bone damage. The invention provides a simple and easily-controlled process of preparing porous inorganic bone substitute materials.

Description

201132367 VI. Description of the Invention: [Technical Field] The present invention relates to a method for producing an inorganic three-dimensional space-connected porous bone substitute using a temperature-sensitive mixture, and is applied to the field of orthopedics and dentistry. Specifically, the temperature sensitive mixture used in the method of the present invention comprises a polyelectrolyte complex and a biomedical ceramic powder. [Prior Art] Bone defects are usually caused by tumor resection or bone trauma. In the United States, there are approximately 500,000 bone grafts per year. Bone grafting usually involves implanting bone substitutes to promote healing of bone tissue. Autografts can be divided into autologous bone grafts and allograft bones. Although graft bones usually have better efficacy, 'autologous, allogeneic, and xenograft bones have problems of insufficient source, disease infection, and immune rejection, respectively, thus limiting their related applications. In recent years, due to the development of bone tissue engineering, synthetic bone substitutes can be used as temporary cell growth scaffolds. With the degradation of materials and the regeneration of bone tissue, the implanted materials will gradually be replaced by new bone. Synthetic bone substitutes have the advantages of biocompatibility, bone conduction, and low risk of disease transmission, and are therefore preferred materials. Commercially available synthetic inorganic bone substitutes are mainly composed of base-filled limestone (Ha,

Ca丨. (P04)6(〇H)2), β_tricalcium phosphate (p_Tcp, p_Ca3(p〇4)2) and calcium sulfate (CS, CaS〇4) and by different forms (such as powder, granule, pellet) , mud or block) to facilitate the application of a variety of different bone defects. However, these materials do not have the interconnected porous structure required for cell and blood vessel ingrowth. The connected porous structure of biological scaffolds is important in tissue engineering for the growth of bone cells and neovascularization of 139374.doc 201132367. Appropriate pore configuration can promote the proliferation and differentiation of bone cells in addition to the growth of new bone tissue. . In general, the pore structure of the bioscaffold has a great influence on the nature of bone conduction. If the pore size is less than 100 μηι, bone tissue will only accumulate on the surface of the bone substitute. After transplantation, the porous structure can be classified into an open cell type or a closed type. The three-dimensional spatial communication hole is open-celled, and its structure is designed to simulate the three-dimensional spatial cavity environment of the extracellular matrix in the body to promote the ingrowth of bone cells and new blood vessels. After the bone graft is implanted, the bone graft will be It gradually degraded and was replaced by the recipient's autologous new bone. At present, a number of manufacturing methods for interconnecting porous bone substitutes have been developed and are summarized in Table 1. Most commercially available bone substitutes are in the form of particles and porous blocks. Some types of three-dimensional space-connected porous bone replace blocks that are synthesized and manufactured by lengthy and cumbersome procedures, or processed from animal bones or corals. The natural animal bone path is fixed in composition and usually consists of basal limestone or calcium carbonate. The degradation time of hydroxyapatite is too long, and the degradation of calcium carbonate is too short. Therefore, it is impossible to provide suitable degradation time by adjusting the components to meet the needs of different clinical applications. Table 1 Product Name _ (manufacturer, factory) Healos (Depuy Spine) ProOsteon (Interpore Int., USA) Collagraft (Zimmer Inc, USA) MBCP (Biomatlante) Triosite (Zimmer Europe Ltd, UK) Composition (content)

Shore coral (calcium carbonate) HA coated with 70% type I bovine collagen 60% HA > 40% TCP 60% HA ' 40% TCP BCP (Bioland)

60% HA ' 40% TCP

Ostilit (Stryker Howmedica Osteonics, UK) 20% HA, 80% TCP, no macropores BoneSave (Stryker Howmedica Osteonics, UK) 20% HA ' 80% TCP > 139374.doc 201132367

Cerasorb ORTHO(curasan) VitossTM Scaffold(curasan) ConduitTM TCP Particles (DePuy Spine)

CellpleXTMTCP Synthetic Loose Bone (Wright)

Ceros 82

Synthes (USA) chronOSTM (Synthes) Calciresorb (Ceraver Osteal, France) Synthograf (Milter, USA) Augmen (Milter, USA) SkeliteTM (Millenium Biologix) Norian Skeletal Repair System (SRS) Aperture: 400-600 μιη Pure β- TCP, micropore · < 80 μπι β-TCP, microporous: < 1-1000 μιη > 99% (p-TCP) Ca3(P04)2 · Hole size: 1-600 μιη β-TCP Porous padding, pore size: ι〇〇_ 400 μιη

β-TCP 'Adjusts the absorption rate by regulating the change of porosity, the absorption time is between 6-12 months β-TCP pore control: 100-500 μηη Porous β-TCP Small size dense TCP Large size dense TCP multiphase Porous Phosphate-Fed Calcium Phosphate Cement In the prior art, current methods of making porous bone substitutes can be divided into several categories: 1. Washing method WO 2006/099332 A2 discloses a method of making a man-made porous bone graft. The method comprises the use of a salt particle as a pore former, which is mixed with a linonic acid material, which is plasticized by extrusion and then sintered, and finally the salt particles are eluted to form a porous structure. However, this method has the disadvantage of being complicated in steps and the holes formed are lacking in connectivity. Therefore, the dissolution step after sintering does not effectively wash out the salt particles remaining therein. 2. Gasification WO 04/098457 A1 provides a process comprising the use of organic particles as a pore-forming agent. The method comprises mixing a pore former with a ceramic powder, forming a mixture by extrusion, and vaporizing the organic compound by a sintering step, and forming a porous structure by utilizing a space in which the organic substance is vaporized. Although 139374.doc 201132367 can effectively form a connected porous structure, the mechanical strength of the resulting product is insufficient. 3. Using a porous polyamine phthalate (PU) sponge as a model US 20060198939 provides a method for producing a connected porous ceramic composite body coated with a biodegradable polymer. This reference uses a highly porous polyurethane (PU) sponge as a template. The pu sponge was immersed in phosphoric acid several times in the bow slurry to ensure that the porous structure of the PU was attached to the disc sour slurry. After drying, the PU sponge was vaporized by a sintering procedure to obtain a strontium sulphate block having interconnected pores. However, the resulting porous block has insufficient mechanical strength. Therefore, the substrate needs to be immersed in a polycaprolactone (PCL) solution and dried at room temperature to enhance its mechanical properties by coating with PCL. 4. Foaming US 200702 1 8098 is a method for making a porous linseed mother. Its porous structure mainly produces foaming by heating C?2 produced by ammonium carbonate. Further, US 20080069852 provides a foaming method using a supercritical fluid. However, the foaming process is generally unstable and it is difficult to control its pore size. 5. Computer-Aided Design and Manufacturing Method US 69055 1 6 refers to the use of a computer-aided design of a specific mold method to make a porous bone substitute. The calcium phosphate bone cement slurry is injected into the mold. After the material is solidified to form hydroxyapatite, it is demolded to form a connected porous structure. However, molds and associated design equipment are often very expensive, and this method is time consuming and cumbersome. Although many researchers have made various improvements to the method of producing porous bone substitutes, the preparation of three-dimensionally interconnected porous bone substitutes is still quite complicated. Therefore, we need to produce a faster, simpler, cheaper, and more reliable method of producing bone substitute blocks with interconnected porous structures and good mechanical properties. SUMMARY OF THE INVENTION The present invention provides a method of forming an inorganic three-dimensional spatially connected pore bone substitute using a temperature sensitive mixture, wherein the temperature sensitive mixture comprises one or more polyelectrolyte complexes and one or more biomedical ceramic powders, The method comprises heating the mixture at a temperature ranging from 25 ° C to 100 t to expand the sample, further heating the resulting mixture to remove the polyelectrolyte complex and moisture contained therein to obtain a connected porous structure and simultaneously performing biomedical ceramics Sintering, followed by cooling the mixture provides a bone substitute for the inorganic interconnected pores. [Embodiment] The present invention utilizes a polyelectrolyte complex and a biomedical ceramic material to form a temperature sensitive mixture which, after heating the mixture, produces a three dimensional spatially connected pore bone substitute. The three-dimensional spatially connected porous bone substitute can be applied to the bone defect site in the orthopedic or dental field. The temperature sensitive mixture of the present invention will form an inorganic three-dimensional spatially connected porous structure after a special heating process, wherein the three-dimensional spatially connected porous structure is mainly produced by volume expansion, water evaporation and gasification of polyelectrolyte complex, and the pore size thereof It can also be controlled by heating rate. The inorganic interconnected three-dimensionally interconnected porous bone substitute can be readily prepared by heating the temperature sensitive mixture of the present invention, which is simple and easy to control. The present invention provides a method for forming an inorganic three-dimensional space using a temperature-sensitive mixture 139374.doc 201132367 to connect a porous bone substitute in which the temperature-sensitive mixture comprises one or more polyelectrolyte mismatched or sinister biomedical ceramics a powder comprising heating the mixture at a temperature of from 25 C to 1 〇0 〇C, further heating the resulting mixture to remove water and polyelectrolyte complexes contained in the sulphide, and then cooling the mixture, I7 producing an inorganic three The space is connected to the porous bone substitute. The polyelectrolyte mismatch system of the present invention is produced by ionic crosslinking of one or more cationic polyelectrolytes or cation anionic polyelectrolytes, the oppositely charged polymers being attracted to each other and irreversibly bonded together. According to the invention, "polyelectrolyte" means a polymer with an electrically functional group. The cationic polyelectrolyte is mainly a positively charged polyelectrolyte knives and comprises a polyelectrolyte having a net positive charge; the anionic polyelectrolyte is mainly a negatively charged polyelectrolyte polymer and comprises a polyelectrolyte having a net negative charge. According to a preferred embodiment of the invention, the cationic polyelectrolyte can be selected from the group consisting of chitosan, polyarginine, polyornosine, agarose gel (DEAE), polybrene ( Polybrene), polylysine, amine «polyesterimine resin and mixtures thereof. According to a preferred embodiment of the present invention, the anionic polyelectrolyte may be selected from the group consisting of acetaminophen, γ-poly glutamic acid (y_PGa), hydroxypropyl fluorenyl cellulose (HPMC), carboxymethyl fiber. (CMC), sodium polyphosphate, pectin, hyaluronic acid, alginate and mixtures thereof. According to the present invention, the biomedical ceramic powder can be a calcium phosphate-based ceramic powder 139374.doc 201132367, a sulfuric acid-based ceramic powder, an oxide-based, nitride-based ceramics. Lushan exempts the end of the Wenshu Shuo, the sulphate-based ceramic powder, the knife chemistry has the oxidation of Ilu oxidized for W" emulsified bismuth or alumina dispersed with titanium oxide. The ceramic material is preferably phosphate sulphuric acid Calcium or oxidatively dominated material or: i. In one embodiment, the ceramsite E powder, which is mainly squaric acid, may be selected from the group consisting of: base apatite (HA), β- Phosphate Tris (Bu TCP) 'Amorphous Calcium Phosphate (ACp) and mixtures thereof. In another embodiment, 'Tao sulfur powder based on sulfuric acid feed can be selected from the following group: Dihydrate sulfur gn semi-water Sulfur (4) and anhydrous sulfuric acid dance. In another example, the oxide-based ceramic powder may be selected from the group consisting of oxidized zirconia, zirconia and titania. In other examples, nitride-based The ceramic powder is selected from the group consisting of nitride rock, titanium nitride and aluminum nitride. In another embodiment, The carbide-based ceramic powder is tantalum carbide. According to the present invention, the weight percentage of the polyelectrolyte complex and the biomedical ceramic material of the system (the polyelectrolyte or the dry weight/temperature-sensitive mixture of the biomedical ceramic material) The dry weight is in the range of 2% to 40% and 10% to 75%, respectively, and the balance is water. The weight percentage (dry weight) of the polyelectrolyte complex is preferably 2% to 40%, 2% to 30%, And 2% to 20% /. The weight percentage (dry weight) of biomedical ceramic materials is 10% to 75%, 20% to 75%, 30% to 75%, 40% to 75%, 50% to 75%, And 60 〇 / 〇 to 75%. More preferably, the weight percentage (dry weight) of the polyelectrolyte complex is 2% to 20% and the percentage of the weight of the raw material (dry weight) is 15 ° / to 50 The remaining composition is water and related biologically active substances. 139374.doc 201132367 According to the invention, the porous structure is affected by the concentration of the cationic and anionic polyelectrolytes. According to the invention, the temperature of the temperature sensitive mixture is at 25 ° C to 100. Within the range of 〇. Those skilled in the art can choose the appropriate temperature depending on the type of polyelectrolyte complex. The heating temperature is preferably in 38〇c to l〇〇〇c, to 55.〇

L〇〇°C, 55. (: to 85 ° C, 55 ° C to 8 (TC, or 55. (: to 75. (: within the range. Heating temperature is better at 55. (: to 10 (TC, 55 ° C to 85 ° C , in the range of 55 ° C to 8 (TC, or 55 C to 75 ° C. After heating the temperature sensitive mixture of the invention, the mixture will expand and form a three dimensional spatially connected pore structure. In one embodiment, After raising the temperature, the method further comprises a temperature maintaining phase of 0.25 to 1 hour; the preferred temperature maintaining phase is 1 to 8 hours, 1 to 6 hours, 2 to 8 hours, 2 to 6 hours, 3 to 8 hours, And 3 to 6 hours; 0.25 to 4 hours; 0.5 to 3 hours; 0.5 to 2 or 0.5 to 1 hour. The preferred temperature retention period is 1 hour. The heating rate is 〇.1 to 2〇 °C/min, 0.3 To 15 ° C / min, 0.3 to l 〇 C / min, 〇 .3 to 5 t: / min, 〇 3 to rc / min or 〇 3 to 2 〇 c / min. The better heating rate is 1.67 ° C / niiii 〇.63t: /min and 0.42 ° C / min. Further heating 'the expanded mixture for drying to evaporate the water contained therein and form a three-dimensional space-connected porous bone substitute. According to one of the present invention For example, the polyelectrolyte malogen can be gasified by high temperature heating and the biomedical ceramics can be sintered. In another embodiment, the further heating process can be completed in one or more stages. The temperature used in the further heating step is from 85t to 150%. Depending on the selected heating stage, the temperature is preferably selected from one or more 139374.doc -10- 201132367 or less temperature range: 85 ° C To 30 (TC, 100 ° C to 250 ° (:, 100 ° C to 200 ° C or 100 ° C to 150 ° C, 300 ° C to 1400 ° C, 300. (: to 1300. (:, 300 ° C to 1200 ° C, 30 (TC to 1150 ° C, 300 ° C to 1100 ° C, 30 (TC to 1000 ° C, 500 ° C to 1400 ° C, 500 ° C to 1300 ° C, and 500 ° C To 1200 ° C. More preferably in the range of 1 〇〇 〇 to 250 ° C, l 〇〇 t: to 200 ° C, or 100 ° C to 150 ° C, 300. (: to 13000 ° C, or 300 ° C to 1150 ° C. In another embodiment, the heating rate is 0.1 to 2 ° C / min, 0.3 to 15 ° C / min, 0.3 to 10 ° C / min, 0.3 to 5 ° C / min, 0.3 to 3 ° C / min or 0 · 3 to 2 ° C / min. Better heating rate is ι.ππ / π ή η, 0.6 3 ° C / min and 0.42 ° C / min. In another embodiment, the method further comprises a temperature holding phase of 0.25 to 10 hours after raising the temperature; a preferred temperature maintaining phase is 丨 to 8 hours, 1 to 6 hours, 2 to 8 hours, 2 to 6 Hours, 3 to 8 hours, and 3 to 6 hours; 0.25 to 4 hours; 0.5 to 3 hours; 0.5 to 2 or 0.5 to 1 hour. The preferred temperature maintenance period is 1 hour. In another embodiment, the sintering temperature ranges from 300 ° C to 1,5 〇 (rc. The preferred sintering temperature ranges from 300 ° C to 1400 ° C, 300 ° C to 1300. (:, 300 ° C to 1200 °C, 3 ton to 1150 ° C, 300. (: to 1100 ° C, 300 ° C to 1000 ° C, 500 ° C to 1400 ° C, 500 ° C to 1300 ° C, and 500. (: to 1200 ° C. More preferably in the range of 300 ° C to 1150 ° C and 300 ° C to 1300 ° C. The heating rate of sintering ranges from 〇 1 to 20 ° C / min. The preferred sintering heating rate is 0.5 to 15 ° C/min, 0.5 to l〇t/min, 0.5 to 5 °C/min, 1 to 15 °C/min, 1 to l〇°C/min, 1 to 5 °C/min, 3 to 15 °C /min, 3 to 10 ° C / min, and 3 to 5 ° C / min. More preferably, the range is 3.5 ° C / min. Sintering step 139374.doc 11 201132367 The step contains 1 to 1 hour, Tian / Jtt Degree of maintenance; preferred temperature maintenance period is 1 to 8 hours, 1 to 6 hours, 2 $ 8 I 0 is 2 to 8 hours, 2 to ό hours, 3 to 8 hours, and 3 to 6 hours. 5 hours. The pore size of the space-connected porous bone substitute in this month can be controlled by the rate of .... The pore becomes smaller as the heating rate increases. According to the present invention, the bone-replacement material formed by the present invention has 〇·05 to 5 mm, 〇.05 to 3 mm, 〇.〇5 to 2 mm, 0.05 L mm 'mm 0.1 to 3 mm or 〇 a pore size range of 3 to 〇5 mm and having a pore size range of G.1 to 30 μm, G1 to MM, q丨 to job or 〇(1) micron. The porosity of the bone substitute material is 50% to 95%. According to the present invention, the method of the present invention further comprises the step of attaching a polymer or a biologically active substance to the monthly replacement material of the present invention by coating or addition. According to the present invention The substance is selected from the group consisting of demineralized bone matrix, growth factor +, bone morphogenetic protein, antibiotic agent, vitamin, collagen, mesenchymal stem cell, antitumor agent, cell attachment agent, immunosuppressant , coagulation activators, platelet-rich plasma, platelet-rich fibrin glue, and silk proteins. The method of the present invention can be exemplified by the following flow chart. 139374.doc •12- 201132367 g-sound xinqin electrolyte I.::, . .···: V: :· · Powder .:::,::Thirsty thirsty waterfalls········.·- -,·.. f ..... . . . expansion/heating: : .... . . . . . . . . . t | $: 呑 耷 耷 离 ▲ ▲ ▲ ▲ 架 架 架 架 架 架 架 架 架 架 架 本 本 本 本 本 本 本 本 本 本 本The method only requires mixing the biomedical ceramics into the polyelectrolyte complex to form a temperature sensitive mixture. The mixture is preferably in a colloidal form. The three-dimensional (3D) network structure of the polyelectrolyte complex and the bioceramic is formed by a physical crosslinking mechanism which is formed by the ionic crosslinking of the two phases & charge "polyelectrolyte" in the hydrazine solution. During the heating, the expansion ratio of the mixture, the evaporation rate of the water, and the vaporization rate of the polymer and the body can be controlled, thereby forming a porous structure with different pore sizes and pores. The inorganic materials and sintering conditions can be selected in the process. Preparation of bone substitutes with different compositions, physical properties or crystallinity. This procedure can be based on specific clinical applications and occasional partial requirements for the same absorption time of the bone; F can be used as a H polyelectrolyte complex in the sintering day Q '' Significantly enhance the mechanical properties of the ceramic samples after sintering. This issue (4) is for the _ species to promote the regeneration of the ^ (four) connected more than 1 139374.doc • 13 · 201132367 bone to replace the block. EXAMPLES Example 1 Expansion of the Thermally Reactive Mixture of the Invention A cationic polyelectrolyte (10% chitosan) solution was mixed with an anionic polyelectrolyte (2% HPMC) solution to form a polyelectrolyte complex (PEC), and then Medical ceramic powder (HA: P-TCP = 1:9) was added to the mixed body to form a mixture by using a mixer, and the weight percentages of PEC, biomedical ceramic powder and water in the system were 6% and 25%, respectively. 69%. The volume of the mixture expands from a temperature of about 55 °C. After heating the resulting mixture to 75 ° C, the volume of the mixture was expanded to 3.3 times the volume of the mixture at 25 t (Fig. 1, (C) and (D)). Comparative Example 1 Expansion of polyelectrolyte complex A cationic polyelectrolyte (1% chit〇san) solution was mixed with an anionic polyelectrolyte (2% HPMC) solution to form a polyelectrolyte complex. After heating the miscellaneous body to 75 C, its volume is 25. (The next difference in volume of the combined body did not change significantly (Figure 1, (A) and (B)). Example 2 produced a three-dimensional open cell unit substitute with a 1:1 (W/W, dry weight) ratio Mixing a cationic polyelectrolyte (chitosan) with an anionic polyelectrolyte (HPMC), followed by mixing the raw f Taobao powder. The weight percentages of PEC, biomedical ceramic powder and water in the system are 6%, 25% and 69%, respectively. The material is placed in a simmering pot and placed in a high temperature furnace. The heating process can be divided into three stages. In the first stage, the temperature is ramped from chamber/dish to 100 C and then held for J hours. The heating rate is 67 C/min from 1 〇〇. 〇 is increased to 3 〇〇. 〇, under 3 〇〇 it is maintained at 139374.doc -14 - 201132367 for 1 hour, and then for the third stage with S fc The rate of /min is from 3 〇〇. The enthalpy is raised to 1150 ° C and maintained at 115 〇t for 5 hours. After sintering, the product is naturally cooled in the furnace. Figure 2 shows chitosan / HPMC polyelectrolyte complex (comparative example) Volume and temperature of 1} and chit〇san/HPMC polyelectrolyte complex and thermophilic mixture of biomedical ceramic powder The relationship between the obtained products was observed by SEM as shown in Fig. 3. ImageJ j 37c image processing software (National Institutes 〇f Health (NIH), Bethesda, MD, USA; from http://rsb The free software of .inf0.nih.gov/ij) is used to calculate the pore size. The calculated sample has a pore diameter of 9 ± 7 μηη and a pore size of ± 220 μχη. It is known by Archimedes measurement. The obtained product has a porosity of 91.9%. Comparative data between Examples 2 to 12 and Examples 2 to 12 and comparative examples Other examples are obtained by using steps similar to those of Example 2. The parameters of the preferred examples and comparative examples are listed in Table 2 to Table 6. According to Table 2', all the examples have a porous structure which is superior to the example. In the case of fixing other parameters, it is indeed possible to show that the (tetra) ionic polyelectrolyte and the anionic polyelectrolyte can be arid. ^ #J 夕 hole structure of inorganic bone filler. When using only 嗒拙:7 β in Comparative Examples 2 to 4, using % ion polyelectrolyte or using only anionic polyelectrolyte, it is impossible to obtain 1 right 玄 q q 八 有 porous A sample of the structure. In addition, the aperture 〇 can be changed by using different ionic polyelectrolytes in Examples 2 to 5. 15 374 374 374 374 374 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 〇 Electrolyte chitosan chitosan chitosan chitosan chitosan (% by weight) 10 l〇l〇l〇l〇"" (% by weight) 2 2 2 2 " 2 " β-β-TCP: HA TCP: HA β· TCP: HA p-TCP: HA (9:1) β-β-TCP: HA TCP: HA P-TCP: HA (9:1) (9:1) (9:1) (9:1) (9:1) (9:1) 6%, 25% '69% 6% ' 25% 6% ' 25% 6% ' 25% , 69% , 69% ' 69% 3% , 25% , 72 % 3% ' 25 % ' 72% No No No No NaCl (50°/〇) 1.67 1.67 1.67 1.67 1.67 1.67 1.67 1 1 1 1 1 1 1 1.67 1.67 1.67 1.67 1.67 1.67 1.67 1 1 1 1 1 1 1 3.5 3.5 3.5 3.5 3.5 3.5 3.5 5 5 5 5 5 5 5 431 ± 220 1,538 ± 634 1,151 661 552 ± 146 NDNDND 9 士 7 15 ± 10 9 ± 8 30 ± 20 NDNDND

阴 though t electrolyte HPMC γ-PGA CMC alginate HPMC medicinal ceramic composition (weight ratio) mixture composition (PEC, biomedical ceramics and water) (% by weight) ligating agent from 25 ° C to 100 ° C. Heating rate plus (°C/min) Heat step retention time (hours) Segment from 100°c to 300°C. Heating rate plus (°C/min) G at 300°C Time (hours) From 300 ° C to two l, 15 ° C heating rate ( ° C / min) g at 1,150 ° C period of holding time (hours) knot giant? Micropores (μηι) According to Table 3, it is shown that the porous structure can be controlled by the heating rate of the heating process. With other parameters fixed, the pore size increases as the heating rate decreases. SEM photographs of Examples 6 and 7 are shown in Figures 4 and 5, respectively. 139374.doc -16- 201132367 Table 3 Example 2 Example 6 Example 7 chitosan chitosan chitosan 10 10 10 HPMC HPMC HPMC 2 2 2 P-TCP: HA p-TCP: HA p-TCP: HA (9:1) (9: 1) (9:1) 6% ' 25% ' 69% 6% ' 25% ' 69% 6% ' 25% ' 69% (A) Cationic polyelectrolyte (% by weight) (B) Anionic polyelectrolyte (% by weight ) Material (C) Composition of biomedical ceramics (weight ratio) Composition of the mixture (PEC, biomedical ceramics and water X% by weight) Pore-forming agent without or without heating rate from 25 ° C to 100 t in the first heating stage (° C/min) Hold time (hours) 1.67 from 100 °C to 300 °C 1 1.67 0.63 1 1.67 0.42 1 1.67 Second heating rate (°C/min) Stage 1 at 300 °C (hours) 1 1 1 3.5 3.5 from 300 ° C to 1,150 ° C 3.5 Third heating heating rate ( ° C / min) Stage at 1,150 ° C to maintain r C period (hours) JJJ structure giant孔ίμπι) 431 ± 220 1,033±278 2,320 ± 778 micropores (μηι) 9±7 9±7 14± 13 Table 4 shows that the porous structure is affected by the concentration of cations and anions. In Example 8, with 15% chitosan and 3% HPMC, the final product had a macropore pore size of 514 178 μm and a pore diameter of 7 ± 4 μm. In Example 9, the final product of 200/〇chitosan and 4% HPMC' had a pore diameter of 367 117 μηη and a pore diameter of 6 ± 4 μηι. Alternatively, the pore size can be varied in Examples 2, 8 and 9 by using different anionic polyelectrolytes. An SEM photograph of Example 9 is shown in FIG. 139374.doc -17- 201132367 Table 4 Example 2 Example 8 Example 9 Material First Heating Stage Second Heating Stage Third Heating Stage Structure (VIII) Cationic Polyelectrolyte (% by Weight) (B) Anionic Polyelectrolyte (% by Weight) (C) Composition of biomedical ceramics (weight ratio) Composition of the mixture (PEC, biomedical ceramics and water) (% by weight) Heating rate of pore former from 25 ° C to l ° ° C (°C / min) retention time ( Hour) Heating rate from 100°C to 300°C (°C/min) Holding period at 300°C (hours) Heating rate from 300°C to 1,150°C (°C/min) Hold time at 1,150 °C (hours) Giant 孑L〇im) Microporous 〇m) chitosan 10 chitosan 15 chitosan 20 HPMC 2 HPMC 3 HPMC 4 p-TCP:HA (9:1) p-TCP:HA (9:1) p-TCP:HA (9:1) 6% ' 25% ' 69% 9% ' 25% > 66% 12% ' 25% ' 63% No No No 1.67 1.67 1.67 1 1 1 1.67 1.67 1.67 1 1 1 3.5 3.5 3.5 5 5 5 431 ±220 9±7 514± 178 7±4 367±117 6±4 Table 5 shows that the porous structure is affected by the ratio of the calcium phosphate-based material. In Example 10, the content of the biomedical ceramic material was increased to increase the ratio to 1:1:0.93, and as a result, the product had a pore diameter of 390 mound 314 μηη and a pore diameter of 10 ± 9 μηι. 139374.doc 18- 201132367 Table 5 Example 2 Example 10 (eight) chitosan chitosan cationic polyelectrolyte (% by weight) 10 10 (Β) HPMC HPMC material anionic polyelectrolyte (% by weight) 2 2 (C) P-TCP: HA p-TCP :HA Biomedical ceramic composition (weight ratio) (9:1) (9:1) Composition of the mixture (PEC, biomedical 6% > 25% '69% 6%, 32%, 62% ceramics and water) ( Weight %) First heating step 15 - Heating rate from 25 ° C to 100 ° C (°C / min) 1.67 1.67 Holding time (hours) 1 1 Heating rate from 100 ° C to 300 ° C 1.67 1.67 Second Heating rate (.C/min) The holding time of the section at 300 °C (small 1 1) 1 1 Heating rate from 300 °C to 1,150 °C 3.5 3.5 Third heating rate (.C/min) The retention time of the segment at 1150 ° C (small C) J structure giant 孑ΐΧ μιη) 431 ± 220 390 ± 314 micropores (μηι) 9 ± 7 10 ± 9 Table 6 shows the preparation of samples with various biomedical ceramic compositions. In Example 11, a mixture of β-TCP and CaS04 was used as the biomedical ceramic, and the product had a pore diameter of 656 ± 407 μηη and a pore diameter of 13 ± 12 μηη. Further, ZrO 2 was used as a ceramic material in Example 12 and its product had a macropore diameter of 285 ± 259 μη. 139374.doc 19- 201132367 Table 6 Example 9 Example 11 Example 12 (A) Cationic polyelectrolyte (weight chitosan 20 chitosan 20 chitosan 20% by weight) Material (B) Anionic polyelectrolyte (weight ° / 〇) (C) Biomedical ceramics Composition (weight ratio) HPMC 4 HPMC 4 HPMC 4 p-TCP: HA (9:1) p-TCP: CaS04 (1:1) Composition of Zr02 mixture (PEC, biomedical ceramics and water) (12%, 25 %, 63% 12%, 25%, 63% 12%, 25%, 63% by weight) The first heating stage is from 25 ° C to 1 Torr. (: heating rate (°C/min) 1.67 1.67 1.67 second heating step holding time (hours) from loot: to 30 (TC 1 1 1 heating rate fC/min) 1.67 1.67 1.67 segment at 30 (Tc Hold time (hours) 1 1 1 Third heating step heating rate fC/min) 3.5 3.5 3.5 segments (300-1,150°C) (300-1,150°C) (300-1,400°C) Hold period (°c ) 1,150 (5 hours) 1,150 (5 hours) 1,400 (5 hours) Structure macropores (μηι) 367±117 656 ± 407 285 ± 259 — micropores (μιη) 6±4 13± 12 ND The present invention does not The above-described embodiments are limited by the examples, which are presented by way of example only, but may be modified in various ways within the scope of protection defined by the scope of the appended patent application. [Simplified illustration of the drawings] Figure 1 shows the chit〇san/HPMC Comparison of volume change between polyelectrolyte complex and ehUGsan/HPMc polyelectrolyte complex and thermophilic mixture of biomedical ceramic powder (HA: P-TCP = 1:9). (A) Polyelectrolyte at 251 Mismatched; polyelectrolyte complex at 75 °c; (C) temperature-sensitive mixture at 25 ° C; and (D) at 75 Figure 129 shows the composition of the Chitosan/HPMC polyelectrolyte complex (Comparative Example 1) and the chitosan/HPMC polyelectrolyte complex and the biomedical ceramic powder (HA$- TCP = 1:9) (Example 1) · Relationship between volume and temperature. Figure 3 shows the porous structure of the bone-substituted material of the present invention observed by SEM (Example 2); wherein (A) refers to the test (B) refers to the SEM photograph (20X) of the sample; and (C) refers to the SEM photograph (1,000X) of the sample. Figure 4 shows the bone replacement of the present invention observed by SEM The porous structure of the material (Example 6); wherein (A) refers to the optical photograph of the sample; (B) refers to the SEM photograph (20X) of the sample, and (C) refers to the SEM photograph of the sample (1,000X) Figure 5 shows the porous structure C of the bone-substituting material of the present invention observed by SEM. Example 7) _; · where (_A>-- refers to the optical photograph of the sample; (B) refers to the SEM of the sample Photographs (20X), and (C) refer to SEM photographs (1,000X) of the sample. Figure 6 shows the porous structure of the bone-substituting material of the present invention observed by SEM (Example 9); ([Alpha]) refers to an optical photograph of the sample; (Beta) mean SEM photograph of a sample (20Χ), and (C) means a SEM photograph of a sample (1,000Χ). 139374.doc -21 -

Claims (1)

201132367 VII. Patent Application Range: 1. A method for forming an inorganic three-dimensional space-connected porous bone substitute using a temperature-sensitive mixture, wherein the temperature-sensitive mixture comprises one or more polyelectrolyte complexes and one or more biomedical ceramics a powder comprising heating the mixture at a temperature ranging from 25 C to 100 ° C, further heating the resulting mixture to remove water and polyelectrolyte complexes therein and sintering the biomedical ceramic, and then cooling The mixture produces an inorganic second degree space interconnected porous bone substitute. 2. The method of claim 1, wherein the cationic polyelectrolyte is selected from the group consisting of chitosan, polyarginine, polyornosine, agarose gel (DEAE), polybrene (polybrene) ), polyaminic acid, amino cellulose to extend ethyleneimine resin and mixtures thereof. 3. The method according to the item, wherein the anionic polyelectrolyte is selected from the group consisting of: acetaminophen, γ-polyglutamic acid (γ_ρ(5Α), hydroxypropyl fluorenyl cellulose (HPMC), carboxymethyl cellulose (CMC), sodium polyphosphate, hyaluronic acid, pectin, sodium alginate and mixtures thereof. 4. The method of claim 1, wherein the raw ceramic powder is selected Free group consisting of: Weiguan powder based on Wei office, ceramics knife with sulfuric acid as the main ingredient, ceramic powder based on oxide, ceramic powder based on nitride, carbonized The main material is the Tao powder, the oxidized cerium oxide, the titanium oxide dispersed in the titanium oxide, and the mixture thereof. 5 · ga&quot; jj? 1g λ method 'In which the acid-based pottery Man powder can be selected freely -I- Λ J·, lower, and into groups: hydroxyapatite (HA), β-tricalcium phosphate (β_ 139374.doc 201132367, amorphous π 畈 calcium (ACP) and The mixture. 6. If the request item 4 &gt; 士, j_ , ^ which is mainly based on calcium sulphate ff, choose the following έ 日 _ ^ between ,, into the group: calcium sulfate dihydrate, calcium sulfate half-jaws and anhydrous calcium Wen 8. 9. For the method of claim 4, freely consist of the following group: If the method of claim 4 is selected, the group consisting of the following components, such as the method of claim 4, carbon carbide. Among them, oxide-based ceramic powders are available in aluminum oxide, cerium oxide and titanium oxide. Among them, the nitride-based ceramic powders are tantalum nitride, titanium nitride, and aluminum nitride. Among them, the carbide-based ceramic powder is a method of 10.::request: i, wherein the polyelectrolyte complex and the percentage of the raw material of the biomedical pottery material (w/w, dry weight) are respectively 2% to 40%. /❶ and to the range of 75 ° / 0, the rest of the composition is water. 11. The method of claim 1, wherein the polyelectrolyte complex has a weight ratio of 2% to 4〇〇/0, 2〇/〇 to 3〇〇/0, and 2% by dry weight. Up to 20%. 12. The method of claim 1, wherein the weight percentage of the biomedical ceramic material is from 10% to 75 on a dry weight basis. /. 20% to 75. /. , 30°/. Up to 75%, 40% to 75%, 5〇〇/❶ to 75%, and 6〇% to 75〇/〇. 13. The method of claim </ RTI> wherein the temperature of the thermally reactive mixture is between 38 C and 100 ° C, 55 ° C to 1 Torr. (:, 55 ° C to 85 ° C, 55. (: to 8 ° ° C, or 55 ° C to 75 ° C. 14. The method of claim ,, where the further heating step is used The temperature is 85. (: to 150 CTC and the heating can be accomplished in more than one stage. 15. The method of claim 1, wherein the 139374.doc 201132367 temperature used in the further heating step is selected from one or more of the following Temperature: 85 ° C to 300 ° C, 100 ° C to 250 ° C, 100 ° C to 200 ° C, or l ° ° C to 150 ° C, 300 ° C to 1400 ° C, 300 ° C to 1300 °C, 300°C to 12〇〇°C, 300°C to 1150°C, 300°C to 1100°C, 300°C to 1000°C, 500°C to 1400°C, 500°C to 1300 And C. The method of claim 1, wherein the temperature used in the further heating step is selected from one or more of the following temperatures: 100. (: to 250 ° C, 100 ° C to 200 ° C, or 100 ° C to 150 ° C, 300 ° C to 1150 ° C, and 300 ° C to 1300 ° C. 17. The method of claim 1, wherein the heating and the further use One or more stages are completed. 18. If request item 1 The method wherein the heating rate of each section can be 〇丨 to 2〇C/min, 0.3 to 15 ° C/min, 0.3 to 1 (Tc/min, 〇.3 to 5 C/min, 0.3 to 3 ° C/min Or 0_3 to 2 ° C / min. 19. The method of claim 1, further comprising a temperature maintaining phase of 〇25 to 1 hour after the heating and the step of heating. The method of claim 18, wherein the heating and the further heating step are performed in the temperature maintaining phase of 1 to 8 hours, 1 to 6 hours, to (1), hour, 2 to 6 hours, 3 to 8 hours, and 3 to 6 respectively. Hours; 〇25 to 4 hours; to 3 hours; 0.5 to 2 or (^ to 丨 hours. 21. The method of claim 1, wherein the bone-replacement material has 0.05 mm to 5 mm, 〇.〇5 to 3 mm 〇.〇5 to 2 亳m, 〇〇5u mm, 〇" to 5 mm, 0.1 to 3 mm or 〇3 to 〇5 mm of the macropore pore size range, and have 〇1 to 30 μm, H2 〇 micron 〇 1 to 1 〇 micron or 〇 (1) 139374.doc 201132367 micron pore size range 22. The method of claim 1, wherein the bone replacement material has a porosity of 50% to 95%. The method of the requested item 1, which further comprises a polymer into 〆 or biologically active substance is attached to the bone of a porous material such substitution step. 24. The method of claim 22, wherein the agent is selected from the group consisting of demineralized bone matrix, growth factors, bone morphogenetic proteins, antibiotic agents, vitamins, collagen, mesenchymal stem cells, antineoplastic agents, Cell adhesion agents, immunosuppressants, coagulation activators and platelet-rich plasma (PRP), platelet-rich fibrin glue (PRF), and silk proteins. 139374.doc