RU2762511C1 - Composition of bioresorbable 3d matrix for restoring bone tissue defects - Google Patents

Abstract

FIELD: medicine.SUBSTANCE: invention relates to the field of medicine. A composition of a bioresorbable 3D matrix for the restoration of bone tissue defects is proposed. The composition includes the following components, parts by weight: casein 5% aqueous solution - 20; hyaluronic acid 2.5% aqueous solution - 40; chondroitin sulfate 0.2% aqueous solution - 10; licorice 3% water infusion - 20; polyethylene glycol 1500 - 3; agar-agar - 6.EFFECT: improving the structure and characteristics of the 3D matrix, increasing the safety of its use, increasing the ability to implant into tissue, as well as simplifying and accelerating the process of its manufacture.1 cl, 3 ex, 2 tbl, 9 dwg

Description

Technology area

The invention relates to medicine and is intended for planning and treatment of bone and osteochondral defects. This hybrid bioresorbable 3D bone matrix of a given shape based on hyaluronic acid is installed at the site of the defect and subsequently replaced by the body's bone tissue.

State of the art

Guided bone regeneration is one of the most widespread and demanded osteoplastic surgeries [1].

Known composite scaffold for the restoration of bone defects (RF patent for invention No. 2624854, class IPC A61L 27/44, A61L 27/12, A61L 27/14, A61P 19/00, publ. 07.07.2017), which contains a polymer solution with a concentration of 9 wt. % from granular powder of polycaprolactone and chloroform, powder of silicon-containing hydroxyapatite Ca10 (PO4) 6-x (SiO4) x (OH) 2-x with substitution x = 0.5 with a concentration of 5-15 wt. % in solution.

Known composite material for the restoration of bone tissue defects (RF patent for invention No. 2476236, class IPC A61K 38/39, A61K 33/42, A61P 9/00, publ. 10.06.2007), containing not reconstructed collagen in the form of a gel and a suspension nanohydroxyapatite in an aqueous solution of glucose with a weight ratio of collagen to nanohydroxyapatite from 5: 1 to 1: 5.

Known composite implant (RF patent for invention No. 2218895, class IPC A61F 2/28, A61F 2/30, A61L 27/00, A61L 27/02, A61L 27/10, A61L 27/40, A61L 27/56, publ . 20.12.2003), which contains a porous matrix and a dense composition, and the porous matrix is made of porous ceramic material, and the dense composition contains bone cement, or bone glue, or alloplant, or dense ceramic material, or other dense biocompatible material filling or covering the pores of one or more surface layers of the porous matrix.

The existing analogues are xenomaterials and allomaterials of Russian and foreign manufacturers. The main disadvantages of the above-described manufacturers are: standardized size range, lack of individual development of implants.

Allomaterials are made from human tissue. The disadvantage of this technology is the complexity of a high technological level of purification from contamination of bacterial agents, fat, protein. Although the probability of transmission of the AIDS virus, hepatitis, tuberculosis by these materials is low, it cannot be completely ruled out.

The raw materials for the production of xenomaterials are animal tissues such as cattle and pigs. This technology also requires a high degree of purification. The literature describes that materials of xenogenic origin can be sources of infection with viruses that carry neuroregenerative disorders [2].

The disadvantages of the known technical solutions are the insufficient degree of cleaning from contamination, the impossibility of individual selection of the required size, as well as the relative complexity, laboriousness and duration of the technology for the manufacture of materials for the restoration of bone tissue defects.

Disclosure of invention

The objective of the invention is to develop a formulation of a 3D bone matrix for the restoration of bone tissue defects.

The technical result of the invention is to improve the structure and characteristics of the 3D matrix: viscoelasticity and porosity, increase the safety of its use, increase the ability to implant into tissue, as well as simplify and accelerate the process of its manufacture.

The specified technical result is achieved by the fact that the composition of the bioresorbable 3D-matrix for the restoration of bone tissue defects includes the following components, in wt. parts:

Casein 5% aqueous solution - 20

Hyaluronic acid 2.5% aqueous solution - 40

Chondroitin sulfate 0.2% aqueous solution - 10

Licorice 3% water infusion - 20

Polyethylene glycol 1500 - 3

Agar-agar - 4.

The developed 3D matrix for the restoration of bone defects makes it possible to obtain implants of a given shape and structure, realizing the principle of personalized medicine. Modeling is carried out on the basis of the results of digital methods of patient examination (computed tomography, magnetic resonance imaging, ultrasound) provided by the customer. Three-dimensional bioresorbable matrices do not require special storage conditions, as they are made individually, immediately before use, hermetically packed and sterilized. The implant is produced under aseptic conditions and delivered to the customer in a sterile package.

Brief Description of Drawings

FIG. 1-3 are photos of the obtained hydrogel, sample No. 1-3, respectively.

FIG. 4 shows a general view of the scaffold in three projections - sample 1.

FIG. 5 shows 3D visualization of pores in sample 1 (volume ≈200 mm3).

FIG. 6 shows a general view of the scaffold in three projections - sample 2.

FIG. 7 shows 3D visualization of pores in sample 2 (volume ≈200 mm3).

FIG. 8 shows a general view of the scaffold in three projections - sample No. 3.

FIG. 9 shows 3D visualization of pores in sample 3 (volume ≈200 mm3).

Implementation of the invention

The starting materials for the manufacture of the new product are: hyaluronic acid, chondroitin, casein and licorice, from which a three-dimensional implant is printed. The composition of a bioresorbable 3D matrix for the restoration of bone tissue defects includes the following components, wt. parts: 5% aqueous solution of casein - 20; 2.5% aqueous solution of hyaluronic acid –40; 0.2% aqueous solution of chondroitin sulfate - 10; 3% aqueous liquorice infusion - 20; polyethylene glycol (PEG) 1500 - 3; agar-agar - 4.

This implant is installed at the site of the defect and subsequently replaced by the bone tissue of the body. The advantage of the new product is a personalized approach to the production of 3D bone matrices, which will improve postoperative regeneration and reduce the number of complications by 30-50%. This technology does not require a high technological level of purification and eliminates the risk of transmission of viruses AIDS, hepatitis, tuberculosis.

For the manufacture of the hydrogel, casein was chosen as a structurant, which provides good suspension, gelation in an aqueous medium, and the ability to withstand high temperatures.

Hyaluronic acid makes it possible to obtain a hydrogel with the following characteristics:

- Good ability to push it through needles and, due to this, it can be used for the manufacture of 3D bone matrices;

- The absence of the transition of one or more constitutive phases of the hydrogel into particles during cross-linking (but the product is characterized by a low extrusion force);

- The absence of co-crosslinked particles in the product, which leads to a significant improvement in the safety profile (fewer side effects and complications in the short and / or medium / long term), increased ability to integrate / implant the hydrogel into the tissue (more even distribution of the product in the tissue) , and also provides a much shorter and less complex manufacturing method (since there are no laborious grinding steps in the manufacturing process);

- Exceptional viscoelasticity properties derived from the special structure of the obtained hydrogel;

- Ability to implant tissue in a simple way in the area to be treated;

- The complex structure of the matrix of the obtained hydrogel, characterized as "multi-structured", due to which it is possible to optimize the controlled release of active and / or biological substances.

To improve the healing process, chondroitin sulfate and licorice infusion were introduced into the hydrogel.

To achieve porosity, an infusion of licorice root and powder of licorice root were used. When extracting active substances from licorice root by the method of infusion and use as a filler in the structure of the hydrogel, the best results were obtained; therefore, an infusion of licorice root was included in the hydrogel.

Experimenting

Preparation of liquorice infusion 3%.

The crushed raw material of licorice root was sifted out of dust, infused with water in an infundir apparatus (boiling water bath) and cooled at room temperature, the extraction was filtered with squeezing of the raw material, and the extract was brought to the described volume with water.

Casein - 0.5 g was placed in a beaker, the calculated amount of purified water (10 ml) was added, stirred with a glass rod until uniformity was formed, and chondroitin sulfate - 0.01 g dissolved in 5 ml of water was added.

Hyaluronic acid was dispersed in a beaker in a calculated amount of purified water (20 ml) - 0.5 g until a homogeneous dispersion was obtained.

In a water bath, a base was prepared, consisting of a 3% infusion of licorice root - 10 ml, agar-agar - 3.0 g, PEG 1500 - 1.5 g (for samples No. 1-3).

Solutions of casein, chondroitin sulfate, and hyaluric acid were introduced into the finished bases with continuous stirring. The mixture was thoroughly emulsified with a homogenizer until a homogeneous mass was formed. Assessed visually - appearance.

When developing the optimal composition of the hydrogel, 3 compositions were investigated, composed by varying the following pharmaceutical factors: the nature and amount of the base, the homogeneity of the mass, the technological operations and equipment used in the manufacture of hydrogels, as well as taking into account their in vitro bioavailability:

Sample No. 1

Casein - 0.5 g (10 ml H2O)

Hyaluronic acid - 0.5 g (20 ml H2O)

Chondroitin sulfate - 0.01 g (5 ml H2O)

Infusion of licorice - 0.3 g (10 ml H2O) = 3%

PEG 1500 - 1.5 g

Agar-agar - 2.0 g

Sample No. 2

Casein - 0.5 g (10 ml H2O)

Hyaluronic acid - 0.5 g (20 ml H2O)

Chondroitin sulfate - 0.01 g (5 ml H2O)

Infusion of licorice - 0.3 g (10 ml H2O) = 3%

PEG 1500 - 1.5 g

Agar-agar 3.0 g

Sample No. 3

Casein - 0.5 g (10 ml H2O)

Hyaluronic acid - 0.5 g (20 ml H2O)

Chondroitin sulfate - 0.01 g (5 ml H2O)

Infusion of licorice - 0.3 g (10 ml H2O) = 3%

PEG 1500 - 1.5 g

Agar-agar - 3.0 g

The organoleptic characteristics of the gels are presented in table 1.

Table 1 - Organoleptic characteristics of gels of samples No. 1, 2, 3

N / a Compositions Description one Hydrogel test No. 1 Homogeneous hydrogel of soft, elastic consistency. Yellow color. Elastic can relatively easily change its volume upon absorption or release of the solvent. 2 Hydrogel test No. 2 Homogeneous hydrogel of dense consistency, yellow color, thick, viscous mass. 3 Hydrogel test No. 3 Homogeneous hydrogel of soft consistency, yellow color, the thickest, viscous, elastic mass. Pronounced porosity. It holds its shape better than samples 1 and 2.

After the mutual contact of the active substances, the most optimal for the formation of the scaffold structure is sample No. 3, the other two samples also showed good results. Fillers in the structure of the hydrogel during its introduction are not distributed evenly enough, therefore, the increase in mechanical properties throughout the volume is uneven.

As a study of the structuring of the hydrogel, a microstructural analysis of 3D bone matrices based on hyaluronic acid, chondroitin, casein and licorice was carried out using microcomputer tomography.

The objects of study were hydrogels with different carriers.

Scanning parameters in the Skyscan 1176 control program (10.0.0.0, Bruker-microCT, Belgium): X-ray tube voltage 45 kV, current 550 µA, Al filter 0.2 mm, pixel size 8.87 µm, sample rotation at each step by 0 , 3, frame averaging 4.

The scanned objects were reconstructed in the Nrecon program (1.7.4.2, Bruker-microCT, Belgium) with the following main reconstruction parameters: correction of ring artifacts (20), beam hardening (20) is included.

The orientation in space (x, y, z) and the selection of individual areas of the reconstructed materials were carried out using the DataViewer software (1.5.6.2, Bruker-microCT, Belgium).

Data visualization and analysis were performed using the CT-analyzer software (1.18.4.0, Bruker-microCT, Belgium).

3D visualization of the results obtained depending on the X-ray density was carried out using the CTvox software (3.3.0r1403, Bruker-microCT, Belgium).

Table 2 - General 3D analysis of samples

Parameters 1 sample 2 sample 3 sample The total volume of the investigated sample, mm3 200 200 200 Total porosity,% 2.81 6.6 3.31

Conclusions:

1. Microstructural analysis of 3D bone matrices of three compositions was carried out, showing that all three compositions have a positive result.

2. Based on the results of the experiment, the optimal composition of 3D bone matrices was proposed, in wt. parts:

Casein 5% aqueous solution - 20

Hyaluronic acid 2.5% aqueous solution - 40

Chondroitin sulfate 0.2% aqueous solution - 10

Licorice 3% water infusion - 20

Polyethylene glycol 1500 - 3

Agar-agar - 4

For the manufacture of 3D bone matrices, the authors have chosen the optimal composition in accordance with sample No. 2.

Comparative analysis of the claimed composition of a bioresorbable 3D matrix shows that the totality of its essential features is unknown from the prior art and, therefore, meets the “Novelty” patentability condition.

In the prior art, no features have been identified that coincide with the distinctive features of the claimed composition of a 3D matrix and affect the achievement of the claimed technical result, therefore, the claimed invention complies with the patentability condition "Inventive level".

The above information confirms the possibility of using the claimed technical solution for the restoration of bone tissue defects, and therefore complies with the condition of patentability "Industrial applicability".

List of used literature

1. Dolgalev A.A., Arakelyan N.G. Tissue engineering for the restoration of bone tissue defects. Scientific and practical journal Bulletin of a young scientist No. 4 (23) 2018. P. 19-24.

2. Zhdanov A.V., Aleshin D.S., Khasanov Sh.M., Gabrielyan N.G., Eldashev D.S.-A., Boyko E.M., Dolgalev A.A. Evaluation of the effectiveness of xenogenic material at the site of the osteoplastic procedure on the alveolar ridge of the jaw in case of exposure of the dPTFE membrane. Medical Alphabet No. 2, 2021. P. 47-53.

Claims (2)

Composition of a bioresorbable 3D matrix for the restoration of bone tissue defects, including the following components, parts by weight: Casein 5% aqueous solution twenty Hyaluronic acid 2.5% aqueous solution 40 Chondroitin sulfate 0.2% aqueous solution 10 Licorice 3% water infusion twenty Polyethylene glycol 1500 3 Agar agar 6

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