Artificial meniscus and preparation method therefor

Abstract

An artificial meniscus and a preparation method therefor. The preparation method comprises: uniformly punching an acellular matrix material, and filling the pores with collagen, wherein the acellular matrix material is the skin tissue of pigs, cattle, etc., has a high biomechanical performance, and is filled with collagen after punching, which is beneficial for cell adhesion, proliferation and migration, and can induce meniscus regeneration. In addition, the acellular matrix material and the filled collagen are subjected to a cross-linking treatment, thereby having a controllable biodegradability and being adaptable to the rate of meniscus tissue formation. The artificial meniscus prepared in the present invention has a good biocompatibility, biomechanical performance and biodegradability.

Description

Artificial meniscus and preparation method thereof Technical Field

[0001] The present invention belongs to the field of medical biomaterials and medical devices, and particularly relates to an artificial meniscus and a preparation method thereof. Background Art

[0002] Meniscus injury is one of the most common knee injuries. Clinically, appropriate treatment options are selected based on the location, severity, and type of meniscus injury, including Traditional Chinese Medicine (TCM) therapy, intra-articular injections, complete (or partial) meniscectomy, meniscus suture repair, and meniscus reconstruction with transplantation. These procedures can help alleviate pain, improve knee function, and mitigate cartilage degeneration. Among these, meniscus reconstruction primarily involves allograft meniscus transplantation and artificial replacement transplantation. While allograft meniscus transplantation can effectively alleviate pain and improve knee function, it is associated with numerous challenges, including limited graft availability, difficulty in tissue preservation, the risk of disease transmission, immune rejection, and high costs. Consequently, research on artificial menisci is gaining increasing attention, and the ability to effectively achieve in vivo meniscus regeneration or long-term meniscus replacement has become a hot research topic. Artificial menisci are primarily categorized as degradable meniscus scaffolds and replacement meniscus grafts. Once implanted, the degradable meniscus scaffold temporarily replaces the damaged meniscus. As the meniscus regenerates, it gradually degrades and metabolizes, ultimately achieving autologous meniscus repair. The replacement meniscus graft remains stable and long-term after implantation, without inducing tissue regeneration, replacing the meniscus and maintaining knee joint function.

[0003] Long-term follow-up of cases reveals that replacement meniscus implants, such as ultra-high molecular weight polyethylene meniscus prostheses, can experience severe wear and tear, leading to late loosening of the artificial joint and the need for revision surgery. Furthermore, wear debris can cause adverse reactions such as osteolysis in the surrounding body. Degradable meniscus scaffold materials, such as collagen, silk fibroin, chitosan, alginate, and absorbable polymers, offer excellent biocompatibility and biodegradability, but their biomechanical properties often fall short of maintaining normal joint function. Technical issues

[0004] In order to solve the problems in the prior art that ultra-high molecular weight polyethylene meniscus prostheses are easy to wear and the biomechanical properties of degradable meniscus scaffold materials cannot maintain normal joint function, the present invention provides an artificial meniscus and a preparation method thereof. Technical Solutions

[0005] The technical solution adopted by the present invention is: preparing acellular matrix material, uniformly punching holes in the material, filling the holes with collagen gel and then freeze-drying.

[0006] Furthermore, the preparation of the acellular matrix material and collagen gel includes the following steps:

[0007] (1) Pretreatment: Clean and disinfect the animal membrane and tissue, remove the villi, fibers, and large pieces of fat tissue, and cut the animal tissue into thin slices;

[0008] (2) Degreasing: Use organic solvent extraction to remove fat and fat-soluble impurities;

[0009] (3) Decellularization: Use 0.1-0.5% surfactant solution combined with 0.1-0.5% protease solution to ultrasonically treat animal membranes and tissues;

[0010] (4) Antigen removal: Use an active reagent to react with animal membrane materials and tissues at a pH of 7-8 for 4-48 hours, then use a strong hydrogen bond reagent to react with animal membrane materials and tissues at a pH of 7-8 for 4-48 hours to obtain de-antigenized acellular matrix materials and animal tissues;

[0011] (5) Collagen extraction: Soak the animal tissue after removing antigens in 0.05-0.5% acid solution, mash it, and then treat it with 0.1-1.0% protease solution under acidic conditions of pH 2-4, then salt out and wash with water to obtain collagen;

[0012] (6) Cross-linking: Use 0.5-5% non-aldehyde chemical cross-linking agent to react with the deantigenized acellular matrix material and extracted collagen at pH 7-10 under stirring conditions for 12 hours to 8 days;

[0013] (7) Punching: Punch holes uniformly in the direction perpendicular to the surface of the cross-linked acellular matrix;

[0014] (8) Filling and freeze-drying: Fill the cross-linked collagen gel into the pores of the punched acellular matrix material and vacuum freeze-dry;

[0015] (9) Cutting: Cut the freeze-dried decellularized matrix material into a meniscus shape to obtain an artificial meniscus.

[0016] Furthermore, the animal membrane material is pigskin, cowhide, etc. Furthermore, the animal tissue is Achilles tendon, skin, etc. of animals such as pigs and cows.

[0017] Furthermore, the non-aldehyde chemical cross-linking agent is at least one of epoxide, diamide, diisocyanate, polyethylene glycol and carbodiimide, with a concentration of 0.1-5.0%.

[0018] Furthermore, the diameter of the holes in the acellular matrix material is 0.3-2 mm, and the distance between the hole edges is 0.5-2 mm.

[0019] Furthermore, the collagen gel is compounded with glycosaminoglycan. Beneficial effects

[0020] The artificial meniscus involved in the present invention is a highly biomechanically compatible acellular matrix material with good biocompatibility, degradability, suitable mechanical strength, good sutureability, and easy processing and molding. Best Mode for Carrying Out the Invention

[0021] To more clearly illustrate the overall concept of this application, the following is a detailed description using examples. It should be noted that many specific details are set forth in the following description to facilitate a full understanding of this application. However, this application may also be implemented in other ways different from those described herein. Therefore, the scope of protection of this application is not limited to the specific embodiments disclosed below.

[0022] An embodiment of a method for preparing an artificial meniscus according to the present invention comprises the following steps:

[0023] (1) Pretreatment: Clean and disinfect the pig skin and Achilles tendon, remove the hair, fiber, and large pieces of fat tissue, and cut the Achilles tendon into thin slices;

[0024] (2) Degreasing: Use organic solvent extraction to remove fat and fat-soluble impurities;

[0025] (3) Decellularization: Pig skin and Achilles tendon were ultrasonically treated using 0.2% surfactant solution combined with 0.1% protease solution;

[0026] (4) Antigen removal: using an active reagent to react with pig skin and pig Achilles tendon at pH 7 for 24 hours, and then using a strong hydrogen bond reagent to react with pig skin and pig Achilles tendon at pH 7 for 24 hours to obtain a deantigenized acellular matrix material and pig Achilles tendon slices;

[0027] (5) Collagen extraction: The antigen-removed pig Achilles tendon slices were soaked in 0.5% acid solution, crushed, and then treated with 0.5% pepsin solution under acidic conditions of pH 3, followed by salting out and washing with water to obtain collagen;

[0028] (6) Cross-linking: Use 2% non-aldehyde chemical cross-linking agent to react with the deantigenized acellular matrix material and extracted collagen at pH 9 with stirring for 1 day;

[0029] (7) Punching: Punch holes uniformly in the direction perpendicular to the surface of the cross-linked acellular matrix;

[0030] (8) Filling and freeze-drying: Fill the cross-linked collagen gel into the pores of the punched acellular matrix material and vacuum freeze-dry;

[0031] (9) Cutting: The freeze-dried acellular matrix material is cut into a meniscus shape to obtain an artificial meniscus. The resulting artificial meniscus has a perforation diameter of 200 μm and a spacing of 1.5 mm. The collagen porosity is 70-90% and the pore size is 10-120 μm. The perforated acellular matrix material is filled with collagen, which is conducive to cell adhesion, proliferation and migration, promoting meniscus tissue regeneration; at the same time, it has strong biomechanical properties to maintain normal joint function requirements. Industrial Applicability

[0032] The artificial meniscus prepared by the present invention has good biocompatibility, biomechanical properties and biodegradability, and can be widely used in the repair and treatment of knee joint injuries.

Claims

1. A method for preparing an artificial meniscus, characterized in that: The following steps are involved: (1) Pretreatment: Clean and disinfect the animal membrane and tissue, remove the villi, fibers, large pieces of fat tissue, etc., and cut the animal tissue into thin slices; (2) Degreasing: Use organic solvents to extract and remove fat and fat-soluble impurities; (3) Decellularization: Use 0.1-0.5% surfactant solution combined with 0.1-0.5% protease solution to ultrasonically treat animal membranes and tissues; (4) Antigen removal: Use an active reagent to react with animal membrane materials and tissues at a pH of 7-8 for 4-48 hours, and then use a strong hydrogen bonding reagent to react with animal membrane materials and tissues at a pH of 7-8 for 4-48 hours to obtain de-antigenized acellular matrix materials and animal tissues; (5) Collagen extraction: Soak the antigen-removed animal tissue in 0.05-0.5% acid solution, mash it, and then treat it with 0.1-1.0% protease solution under acidic conditions of pH 2-4, then salt out and wash with water to obtain collagen; (6) Cross-linking: Use 0.5-5% non-aldehyde chemical cross-linking agent to react with the deantigenized acellular matrix material and extracted collagen at pH 7-10 with stirring for 12 hours to 8 days; (7) Punching: Punching holes uniformly in the direction perpendicular to the surface of the cross-linked decellularized matrix material; (8) Filling and freeze-drying: filling the cross-linked collagen gel into the pores of the punched acellular matrix material and vacuum freeze-drying; (9) Cutting: Cut the freeze-dried decellularized matrix material into a meniscus shape to obtain an artificial meniscus.

2. The method for preparing an artificial meniscus according to claim 1, characterized in that: The animal membrane material is pig skin, cow skin, etc.; the animal tissue is Achilles tendon, skin, etc. of pigs, cows, etc.

3. The method for preparing an artificial meniscus according to claim 1, characterized in that: The non-aldehyde chemical cross-linking agent is at least one of epoxide, diamide, diisocyanate, polyethylene glycol and carbodiimide, with a concentration of 0.1-5.0%.

4. The method for preparing an artificial meniscus according to claim 1, characterized in that: The diameter of the holes in the decellularized matrix material is 0.3-2 mm, and the distance between the holes is 0.5-2 mm.

5. The method for preparing an artificial meniscus according to claim 1, characterized in that: The collagen filler has a porosity of 70-90% and a pore size of 10-120 microns.

6. The method for preparing an artificial meniscus according to claim 1, characterized in that: The collagen gel is complexed with glycosaminoglycan.

7. An artificial meniscus, characterized in that: The artificial meniscus is manufactured using the artificial meniscus preparation method described in any one of claims 1 to 6 above.