CN110141398B

CN110141398B - Method for preparing artificial ligament and product thereof

Info

Publication number: CN110141398B
Application number: CN201910443812.6A
Authority: CN
Inventors: 王富友; 张颖; 陈光兴; 杨柳
Original assignee: First Affiliated Hospital of PLA Military Medical University
Current assignee: First Affiliated Hospital of PLA Military Medical University
Priority date: 2019-05-27
Filing date: 2019-05-27
Publication date: 2021-06-22
Anticipated expiration: 2039-05-27
Also published as: CN110141398A

Abstract

The invention relates to a preparation method of an artificial ligament, which is characterized in that a filiform artificial material is braided and twisted into a rope-shaped object and then twisted into the artificial ligament. The twist plait is formed by twisting 15-25 filiform artificial materials into one strand by using 1 strand and 3-4 strands; the rope-shaped object is formed by twisting 15-25 filiform artificial materials into 1 bundle, wherein 3 bundles are twisted into one strand, and 3 strands are twisted into 1 rope. The artificial ligament obtained by the artificial ligament weaving method has good mechanical property, the mechanical property of the artificial ligament is closer to the autologous ACL, and the stretching length of the ligament is better than the autologous ACL. The artificial ligament braided structure can ensure that ligaments respectively meet the requirements of maintaining the front and back and complex rotation stabilizing effects of a knee joint.

Description

Method for preparing artificial ligament and product thereof

Technical Field

The invention belongs to the technical field of medical materials, and relates to a preparation method of an artificial ligament and a product thereof.

Background

With the increasing national force and the improvement of living standard, the incidence of ligament injury is increased year by year. Taking the most common Anterior Cruciate Ligament (ACL) injury as an example, the number of injured ligaments per year in China is conservatively estimated to be over 1 million. The anterior cruciate ligament is an inner ligament of the knee joint, connects the tibia with the femur, controls the rotation of the knee joint and prevents the tibial plateau from excessively moving forwards or the femur from moving backwards, has the functional importance of maintaining and coordinating the stability of the knee joint to maintain the performance stability of the knee joint, helps the human body to complete various actions with great difficulty, and has important physiological functions. Long periods of intense lower limb movement or a variety of other factors can lead to rupture of the anterior cruciate ligament. The rupture of the anterior cruciate ligament can lead to knee joint instability, and if not treated in time, can cause articular cartilage, meniscus damage and osteoarthritis due to repeated sprains. The conservative treatment effect of ligament injury is poor, and the self-healing capability of the totally fractured ACL is low, so that most of the ACL cannot be recovered. Therefore, clinically, reconstructed ACL is mostly used as a treatment direction, and grafts used for reconstructing ACL mainly include autologous tendons, allogeneic tendons and artificial ligaments.

The benefit of autologous tissue transplantation is that there is no rejection, but often complications are caused at the donor site, and secondary operations are required; the materials are limited, the wound healing period of the material-taking part is longer, and the like; allogenic tissue transplantation can avoid self-trauma, but the source is very limited, the risk of infectious diseases such as hepatitis B, AIDS and the like exists, the problem of immunological rejection also exists, and the allogenic tissue transplantation is gradually replaced by artificial ligaments. The most clinically used artificial ligament at present is the LARS ligament (ligament) system, which is made of polyethylene terephthalate (PET) fiber. However, the structure of the PET molecule is highly symmetrical, the cell is difficult to attach and enter into the PET fiber due to low surface energy, the osteoinductivity is poor, the problem of poor healing of the graft and the host bone exists, and the bioactivity of the PET molecule is still unsatisfactory. The poor mechanical property of the material causes the ligament to be woven too densely and the fiber gap is small, which is also not beneficial to the growth of the surrounding tissues. This can cause a series of problems such as bone expansion, graft abrasion, and loosening of internal fixation, resulting in surgical failure, and thus it is critical to enhance the tissue-like inducibility of the artificial ligament and promote graft-host self-like healing.

Disclosure of Invention

In view of the above, the present invention aims to provide a method for preparing an artificial ligament and a product thereof.

In order to achieve the purpose, the invention provides the following technical scheme:

1. a preparation method of an artificial ligament is characterized in that a filiform artificial material is braided and twisted into a rope-shaped object and then twisted into the artificial ligament.

Furthermore, the preparation method comprises the step of twisting the filamentous artificial material into a rope-shaped object by 1 twist braid and 2 twists, and then twisting the rope-shaped object to form the artificial ligament.

Further, the twist braid is four-ply braided.

Furthermore, the twist braid is a circular twist braid braided with four strands.

Furthermore, the twist plait is a filiform artificial material, 15-25 strands are used as 1 strand, 3 strands are twisted into one strand, and then 3-4 strands are woven into the twist plait.

Furthermore, the twist plait is a filiform artificial material, 15-20 strands are used as 1 strand, 3 strands are twisted into one strand, and then 3-4 strands are woven into the twist plait.

Furthermore, the twist plait is a filiform artificial material, 20 strands are used as 1 strand, 3 strands are twisted into one strand, and then 3-4 strands are woven into the twist plait.

Furthermore, the twisted rope-shaped object is formed by twisting 15-25 filiform artificial materials into 1 bundle, 3 bundles are twisted into one strand, and 3 strands are twisted into 1 rope.

Furthermore, the twisted rope-shaped object is formed by twisting 15-20 filiform artificial materials into 1 bundle, 3 bundles are twisted into one strand, and 3 strands are twisted into 1 rope.

Furthermore, the twisted rope-shaped object is formed by twisting 15 artificial materials in a thread shape into 1 bundle, 3 bundles are twisted into one strand, and 3 strands are twisted into 1 rope.

Furthermore, the filamentous artificial material is prepared from raw materials of type I collagen, silk fibroin, polyvinyl alcohol and chondroitin sulfate.

Further, the raw material of the filamentous artificial material contains, in terms of mass ratio, type I collagen: silk fibroin: polyvinyl alcohol: the chondroitin sulfate is 15-20: 20-25: 40-50: 0.02-0.03.

Further, the ratio of type I collagen: the ratio of the silk fibroin is 1: 1-1.5.

Further, the ratio of type I collagen: the ratio of silk fibroin is 1: 1.

Furthermore, the preparation method of the silk fibroin comprises the following steps: degumming silk for 3 times at 100 ℃ in a Na2CO3 solution with a material-to-liquid ratio of 1: 20-30, removing sericin protein for 30-40min each time, obtaining degummed fibroin protein, cleaning the obtained fibroin protein with deionized water, and drying at 50-60 ℃ to constant weight for later use.

Further, the preparation method of the filamentous artificial material comprises the following steps: dissolving silk fibroin in calcium chloride-ethanol-water solution, adding polyvinyl alcohol and chondroitin sulfate, uniformly stirring until the mixture is completely dissolved, adding I type collagen, slowly stirring until the mixture is dissolved to form spinning solution, and preparing the regenerated fiber by adopting a wet spinning technology.

Further, in the calcium chloride-ethanol-aqueous solution, the ratio of calcium chloride: ethanol: the ratio of water is 1:2: 8.

Furthermore, the wet spinning technology is as follows: extruding the spinning solution into a spinning nozzle by a pressure pump at the room temperature of 20-25 ℃; the spinning solution from the spinning nozzle enters a coagulating bath which is 10% alcohol, and the length of the coagulating bath is 1 m; the spinning pressure is 0.1 MPa; the spinning speed is 5-7 mL/h; the winding speed is 8-10 r/min.

2. An artificial ligament obtained by the method of any one of the above methods of making an artificial ligament.

Still further, the artificial ligament is an ACL ligament.

The invention has the beneficial effects that: the regenerated fiber prepared by the invention consists of collagen and silk fibroin, greatly improves the histocompatibility of the fiber, has no toxicity to cells, and is beneficial to the adhesion and growth of the cells; after the prepared artificial ligament is transplanted, the growth of autologous ACL ligament-like neogenetic tissue is facilitated in vivo, and the neogenetic tissue is fused with the ligament more closely. The prepared artificial ligament has good histocompatibility, the outer synovial membrane tissue does not thicken after 12 weeks, and a new blood vessel grows in the middle of the synovial membrane, which fully shows that the artificial ligament prepared from the regenerated cellosilk provided by the invention can well form an autologous ligament. Further deeply researching the weaving process of the regenerated fiber yarns, and screening out the optimal weaving process by combining the mechanical test result; the mechanical property of the artificial ligament prepared by the weaving method provided by the invention is closer to the autologous ACL, the stretching length of each ligament is superior to the autologous ACL, the stretching length is in a certain relation with the weaving method of the twisted braid, and the rigidity of the artificial ligament is well limited by directly twisting the other 2 strands into a rope shape. Experiments prove that the artificial ligament formed by weaving the regenerated fiber yarns after the regenerated fiber yarns are braided by a twist braid and then are directly twisted into a rope has a certain internal space structure, the internal pore diameter of the artificial ligament reaches 40-60nm, and cell migration and matrix formation are facilitated; and the maximum load is improved by the four-strand braided circular twisted braid, the effect of dispersing the tensile force is achieved by the cooperation of the four-strand braided circular twisted braid and the four-strand braided circular twisted braid, the average diameter of the artificial ligament is reduced, and the mechanical requirement of anterior cross reconstruction can be met. The artificial ligament prepared by the regenerated fiber has good mechanical property and slow degradability, and does not cause the reduction of the mechanical property of the tissue engineering ligament during degradation. The artificial ligament braided structure can ensure that ligaments respectively meet the requirements of maintaining the front and back and complex rotation stabilizing effects of a knee joint. Compared with tissue engineering ACL and LARS (tissue Advanced repair System), the artificial ligament provided by the invention has obvious advantages, can realize ACL regeneration repair without inoculating cells and implanting simple artificial materials, and is simple and convenient.

Drawings

In order to make the object, technical scheme and beneficial effect of the invention more clear, the invention provides the following drawings for explanation:

FIG. 1 is an illustration of a circular twist braid braiding procedure 1;

FIG. 2 is an exemplary circular twist braid knitting procedure 2;

FIG. 3 is an exemplary circular twist braid knitting procedure 3;

fig. 4 shows an exemplary circular twist braiding process 4.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. The experimental procedures, in which specific conditions are not specified in the examples, are generally carried out under conventional conditions or under conditions recommended by the manufacturers. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

In the quantitative tests in the following examples, three replicates were set, and the data are the mean or the mean ± standard deviation of the three replicates.

Example 1

Preparation of silk fibroin solution:

degumming 200g silk in 4-6L Na2CO3 solution with mass concentration of 1g/L at 100 deg.C for 3 times, each for 30-40min, removing sericin to obtain degummed fibroin, washing the obtained fibroin with deionized water, and drying at 50-60 deg.C to constant weight.

Preparing a calcium chloride-ethanol-water (CaCl2-EtoH-H2O) solution according to the mass ratio of 1:2:8, dissolving silk fibroin in the calcium chloride-ethanol-water (CaCl2-EtoH-H2O) solution, dissolving degummed silk fibroin in a constant-temperature water bath kettle at 60 ℃ for 4-5H, cooling the silk fibroin solution to room temperature, centrifuging for 5-10min at 10000r/min by using a centrifuge 8000, and obtaining the silk fibroin solution.

Example 2

At room temperature, 100ml of 20% (mass volume fraction) silk fibroin solution is added with 50g of polyvinyl alcohol and 30mg of chondroitin sulfate and evenly stirred until the mixture is completely dissolved, then 20g I type collagen is added and slowly stirred until the mixture is dissolved to form spinning solution, and the preparation of regenerated fibers is carried out by adopting a wet spinning technology. The spinning process comprises the following steps: extruding the spinning solution into a spinning nozzle by a pressure pump at the room temperature of 20-25 ℃; the spinning solution from the spinning nozzle enters a coagulating bath which is 10% alcohol, and the length of the coagulating bath is 1 m; the spinning pressure is 0.1 MPa; the spinning speed is 5 mL/h; the winding rate was 8 r/min. By comparing the spinning process parameters, the coagulation bath has the largest influence factor, and the spinning speed and the pressure have the smallest influence factor. Through the experimental test of water and alcohol coagulation baths with different concentrations, the forming effect and the performance of the regenerated fiber yarn are best when the coagulation bath is 10 percent alcohol. The solidification bath with alcohol content of more than 50% has poor molding effect or cannot mold. The regenerated fiber filament prepared in this example was designated NF 1.

Example 3

At room temperature, 100ml of 25% (mass volume fraction) silk fibroin solution is added with 50g of polyvinyl alcohol and 30mg of chondroitin sulfate and evenly stirred until the mixture is completely dissolved, then 15g I type collagen is added and slowly stirred until the mixture is dissolved to form spinning solution, and the preparation of regenerated fibers is carried out by adopting a wet spinning technology. The spinning process comprises the following steps: extruding the spinning solution into a spinning nozzle by a pressure pump at the room temperature of 20-25 ℃; the spinning solution from the spinning nozzle enters a coagulating bath which is 10% alcohol, and the length of the coagulating bath is 1 m; the spinning pressure is 0.1 MPa; the spinning speed is 5 mL/h; the winding rate was 8 r/min. The regenerated fiber filament prepared in this example was designated NF 2.

Example 4

At room temperature, 100ml of 25% (mass volume fraction) silk fibroin solution is added with 40g of polyvinyl alcohol and 20mg of chondroitin sulfate and evenly stirred until the chondroitin sulfate is completely dissolved, then 15g I type collagen is added and slowly stirred until the chondroitin sulfate is dissolved to form spinning solution, and the preparation of regenerated fibers is carried out by adopting a wet spinning technology. The spinning process comprises the following steps: extruding the spinning solution into a spinning nozzle by a pressure pump at the room temperature of 20-25 ℃; the spinning solution from the spinning nozzle enters a coagulating bath which is 10% alcohol, and the length of the coagulating bath is 1 m; the spinning pressure is 0.1 MPa; the spinning speed is 5 mL/h; the winding rate was 8 r/min. The regenerated fiber filament prepared in this example was designated NF 3.

Example 5

Placing the regenerated cellosilk of the artificial ligament material prepared in the examples 2-4 into a 24-well plate after sterilization, and arranging a PET fiber group (the material is added with PET cellosilk with the same weight) and a blank control group (without any material), wherein each group is provided with 3 repeated plates; l-929 mouse fibroblast cells in the exponential growth phase are digested by 0.25% trypsin, blown, resuspended and diluted 4 times, inoculated into each sample well, and put into a 5% CO2 incubator at 37 ℃ for CO-culture, wherein the culture medium is DMEM with 10% FBS. And (3) taking out the 24-well plate after 2, 4 and 7 days of culture, adding 100ul of CCK-8 reagent into each well, continuously culturing for 4 hours, transferring the solution in each well into a 96-well enzyme label plate, adding 100ul of reagent into each well, and measuring the absorbance value at the wavelength of 450nm by using an enzyme label instrument, wherein the test results are shown in table 1. And proliferation of L-929 cells in the cell culture plate and on each material was observed using an inverted microscope.

The proliferation of mouse fibroblasts (L-929) was observed under a microscope at 1 st to 7 th days of cell culture. On the 4 th day of cell culture, the cells in the blank control group are in fusiform shape under a microscope, have good growth state and proliferate to more than about 80% of the area of the plate bottom; the cell forms of the NF1, NF2 and NF3 groups are fusiform, the cells are also proliferated in large quantity and have normal forms, and a small amount of fusiform cells are attached to and grow on each material; the PET fiber group cells are mostly fusiform in shape, and have a small amount of proliferation, and no cells are attached to and grow on the material.

Cell proliferation rate (RGR)% (OD average of experimental group/OD average of blank control group) × 100%

Grading cytotoxicity: grade 0, RGR is more than or equal to 100%; grade 1, 99% more than RGR more than or equal to 75%; grade 2, 74% > RGR is more than or equal to 50%; grade 3, 49% more than RGR more than or equal to 25%; grade 4, more than 24% and RGR more than or equal to 1%; grade 5, RGR equals 0%.

Table 1 groups 2d, 4d and 7d cell proliferation results and cytotoxicity ranking

As can be seen from table 1, the cells co-cultured with the PET material proliferated less and increased more significantly over time compared to the blank control. The cells co-cultured with each regenerated cellosilk group obviously proliferate more, the RGR is 101-113%, the cytotoxicity grades are 0, which indicates that the artificial ligament can be further prepared without cytotoxicity and can be used clinically.

Example 6

a. 20 prepared regenerated fiber yarns are twisted into one strand by 1 strand and 3 strands, and then 4 strands are taken to be woven into a circular twisted braid;

b. 20 prepared regenerated fibers are twisted into one strand by 1 strand of 20 fibers, and 3 strands are twisted into 1 rope;

c. and (c) twisting 1 round twisted braid prepared in the step (a) and 2 ropes prepared in the step (b) into 1 rope, and preparing the artificial ligament.

The weaving method of the circular twist braid is as shown in figures 1 to 4 of the accompanying drawings, wherein A, B, C, D four strands are sequentially arranged, as shown in figure 1, the circular twist braid is divided into a left group and a right group, the outer strand on the left side is firstly used, the inner strand on the right side is picked, namely, the strand A is used for picking the strand C, and the strand A presses the strand C from the upper side, as shown in figure 2; then, the outside lines on the right side are used for picking the inside lines on the left side, namely, the strand D is used for picking the strand A from the top to the left side, which is shown in figure 3; then, the outer line on the right side is used for picking the inner line on the left side, as shown in figure 4; and repeating the steps continuously until the length is proper.

This method is labeled as braiding method a.

Example 7

a. Twisting 15 prepared regenerated fiber yarns into one strand with 3 strands as 1 strand, and weaving the other 4 strands into a circular twisted braid;

b. 15 prepared regenerated fibers are twisted into one strand by 1 strand, 3 strands are twisted into 1 rope by 3 strands;

This method is labeled as weaving method B.

Example 8

a. Twisting 3 bundles of 25 prepared regenerated fiber yarns into one strand of 1 strand, and weaving 4 strands into a circular twisted braid;

b. 25 prepared regenerated fibers are twisted into one strand by 1 strand of 25 fibers, and the other strand of 3 strands of the regenerated fibers are twisted into 1 rope;

This method is labeled as knitting method C.

Example 9

This method is labeled as weaving method D.

Example 10

This method is labeled as braiding method E.

Example 11

20 prepared regenerated fibers are twisted into one strand by 1 strand by 3 strands, 1 rope is twisted into 3 strands, and 1 artificial ligament is twisted into 1 rope by 3 ropes.

This method is labeled as weaving method F.

The above weaving method is also applicable to other thread-like artificial materials, which are various materials that can be used for medical purposes. Such as CN 106048765B, an artificial ligament material obtained by the preparation method of the artificial ligament material. Also such as one mentioned in CN 108728931 a, can be used for artificial ligament.

Example 12

The regenerated filaments NF1, NF2, and NF3 prepared in examples 2 to 4 were prepared into artificial ligaments for biocompatibility experiments according to the knitting methods a to F of examples 6 to 11, respectively, and were xenografted using 15 rabbits.

The weight of 32 healthy adult New Zealand rabbits is controlled to be (3 +/-0.50) kg, and 16 females and males are randomly distributed, but each artificial ligament is guaranteed to be female and male. Anaesthetizing 3% pentobarbital at a dose of 1.5ml/Kg, fixing on an operating table, shaving the back in a small range, operating under aseptic condition, disinfecting the shaved part with iodophor, cutting for 3-4cm, and blunt-separating subcutaneous tissues to two sides with forceps and vascular forceps to form a capsular bag-like structure; implanting each artificial ligament, sewing with absorbable suture, marking each rabbit, sterilizing, sewing skin, and wrapping dressing; penicillin is injected into muscles for 5 consecutive days to prevent incision infection, the injection frequency is 1 time/d, and the local part of the incision is closely observed to have inflammatory reaction. Rabbits were sacrificed for observation 12 weeks post-surgery. HE staining clearly showed no thickening of the outer synovial tissue, fibroblasts and neovascularization in the middle of the synovium on each implanted ligament. Some newly generated tissues grow like autologous ACL ligament-like tissues, and the newly generated tissues are fused with the ligament tightly. No obvious inflammatory reaction and tissue necrosis are seen in the whole experimental test process, and no other adverse reactions are caused in each experimental animal. The biocompatibility of the artificial ligament of the invention is proved to be good.

Example 13

The regenerated fibers NF1, NF2 and NF3 prepared in examples 2 to 4 were prepared into artificial ligaments for animal experiments according to the weaving methods of examples 6 to 11, weaving methods A to F, respectively. And autologous ACL was used as a positive control. And (4) carrying out tensile mechanical test on the woven artificial ligament by using a universal mechanical testing machine.

54 healthy adult New Zealand rabbits (3 +/-0.50) kg in weight and 30 male and female rabbits are bred in cages. Rabbits were randomly divided into groups of 3 animals each, ensuring at least one male and female for each experimental group. When in grouping, the anterior knee joint drawer test and the Lachman test of each rabbit are negative so as to ensure the functional normality of the knee joint before the operation of the experimental rabbit. The reconstruction of bilateral ACL is carried out on each experimental rabbit, 3% pentobarbital is anesthetized according to the dose of 1.5ml/Kg before the operation, and the bilateral joint is preserved and disinfected. The knee joint incision of patella inner edge under strict aseptic condition sequentially incises skin and joint cavity, excises partial patella lower fat pad, exposes anterior crossing primary band, completely cuts off autologous anterior crossing ligament in the middle of ligament, respectively drills femur and tibia bone tunnels by using an orthopedic electric drill provided with a 2.0mm Kirschner wire drill bit, the drilling angles of the femur and tibia tunnels are both 45 degrees, and the prepared sterilized artificial ligament is pulled into the bone tunnels by using steel wires through a traction wire. The tail end of the implant is fixed through a spongy bone screw peg, the artificial ligament is tensioned at the buckling position, and screws at two ends are screwed down. After operation, the rabbits are separately fed in the independent rabbit cages and are allowed to move freely. The knee joint incision is changed every other day and the healing condition of the incision is observed, penicillin is continuously given for intramuscular injection for 5 days to prevent the infection of the incision, and the injection frequency is 1 time/d. The mental state, the activity of the four limbs, the diet of the rabbit, the dislocation of the patella and other conditions are carefully observed every day, and the problems are timely treated.

And (3) biomechanical testing: 3 rabbits of each group were sacrificed 20 weeks after surgery by air embolism, the activity of the knees of the experimental rabbits was carefully observed before sacrifice, and the anterior drawer test and Lachman test of the knees were carefully examined. Cutting off the tibia end and the femur end at a position 4cm away from the knee joint, removing all soft tissues such as muscles and fascia around the knee joint, only reserving the artificial ligament, respectively embedding the femur end and the tibia end by using bone cement to prepare a femur-artificial ligament-tibia complex specimen, respectively placing the femur end and the tibia end on a clamp of an electronic universal mechanical testing machine for fixation, adjusting a bone tunnel to be consistent with the direction of tension, and carrying out ultimate tensile mechanical testing. Firstly, applying 0-5N pulling force at the speed of 2mm/min, and circulating for 5 times, so as to completely pull out the tendon fibers of the artificial ligament and prevent the tortuosity of the tendon fibers from influencing the result of the biomechanical test. And applying a tensile force at a tensile speed of 10mm/min until the specimen is damaged, and determining the maximum load, the rigidity and the tensile length of the artificial ligament according to a load-displacement curve drawn by an electronic universal mechanical testing machine. The test results are shown in table 2.

TABLE 2 ligament mechanics test results

As can be seen from table 2, the mechanical properties of the artificial ligament using the braiding method E are relatively close to those of the ACL of the subject, and the tensile length of each ligament is superior to that of the ACL of the subject, which is related to the braiding method of the twisted braid, while the rigidity of the artificial ligament is well limited by the fact that the other 2 strands are directly twisted into a rope. Experiments prove that the artificial ligament formed by weaving the regenerated fiber yarns after the regenerated fiber yarns are braided by a twist braid and then are directly twisted into a rope has a certain internal space structure, the internal pore diameter of the artificial ligament reaches 40-60nm, and cell migration and matrix formation are facilitated; and the maximum load is improved by the four-strand braided circular twisted braid, the effect of dispersing the tensile force is achieved by the cooperation of the four-strand braided circular twisted braid and the four-strand braided circular twisted braid, the average diameter of the artificial ligament is reduced, and the mechanical requirement of anterior cross reconstruction can be met. The artificial ligament prepared by the regenerated fiber has good mechanical property and slow degradability, and does not cause the reduction of the mechanical property of the tissue engineering ligament during degradation. The artificial ligament braided structure can ensure that ligaments respectively meet the requirements of maintaining the front and back and complex rotation stabilizing effects of a knee joint.

General observation

After the ACL of each experimental group is reconstructed, no obvious effusion in the joint cavity and synovium tissue hyperplasia are seen, the screws for fixing are firm in place, the new ligament tissue is formed at the original ACL part, and the shape and the color are basically the same as the normal ACL. The prepared artificial ligament contains silk, and the silk fibroin fiber structure is obtained by removing matrix collagen, so that the biocompatibility of the artificial ligament is further increased, and the adhesion, migration and proliferation of cells are facilitated.

Finally, it is noted that the above-mentioned preferred embodiments illustrate rather than limit the invention, and that, although the invention has been described in detail with reference to the above-mentioned preferred embodiments, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the scope of the invention as defined by the appended claims.

Claims

1. The preparation method of the artificial ligament is characterized in that 1 twist braid and 2 rope-shaped objects are twisted into the artificial ligament from filiform artificial materials, the twist braid is a circular twist braid woven by 4 strands, the rope-shaped objects are 15-25 strands of the filiform artificial materials which are twisted into 1 strand, 3 strands of the rope-shaped objects are twisted into 1 rope.

2. The method of claim 1, wherein the filamentous artificial material is prepared from type I collagen, silk fibroin, polyvinyl alcohol, and chondroitin sulfate.

3. The method for producing an artificial ligament according to claim 2, wherein the ratio of type I collagen: silk fibroin: polyvinyl alcohol: the chondroitin sulfate is 15-20: 20-25: 40-50: 0.02-0.03.

4. An artificial ligament obtainable by a method of manufacture of an artificial ligament according to any one of claims 1 to 3.