Artificial ligament and method for its production

Abstract

The application provides an artificial ligament and a preparation method thereof, and relates to the technical field of artificial ligaments. In the application, a plurality of groups of silk bundles are provided first, each group of the silk bundles comprising a plurality of silk threads, then the plurality of silk threads are twisted into a bundle to form a silk bundle, a plurality of the silk bundles are woven to form a first woven rope with a first preset structure, then the first woven rope is subjected to a degumming treatment to obtain a second woven rope, and finally, the plurality of second woven ropes are woven according to a second preset structure by using a three-dimensional four-step method, so that the artificial ligament is prepared. The artificial ligament prepared by the above technical scheme has excellent mechanical properties and biocompatibility, and shows important clinical application value.

Description

Technical Field

[0001] This invention relates to the field of artificial ligament technology, and in particular to an artificial ligament and its preparation method. Background Technology

[0002] Repairing and reconstructing the anterior cruciate ligament (ACL) of the knee is a challenging problem jointly faced by joint surgery and regenerative medicine. Due to the ligament's limited self-healing ability, most injuries require surgical intervention. Currently, the most commonly used treatment options in clinical practice include autologous transplantation, allogeneic transplantation, and artificial ligaments. While autologous transplantation offers good tissue compatibility, it suffers from limitations such as limited donor sites, prolonged surgery time, and the risk of complications at the donor site. Allogeneic transplantation, on the other hand, has limitations in terms of sterilization and preservation, which can reduce the graft's mechanical properties and may trigger inflammatory reactions, spread disease, and incur high costs. Given these issues with autologous and allogeneic transplantation, artificial ligaments have become the preferred reconstruction method in most clinical surgeries.

[0003] However, existing artificial ligaments still have some technical problems. On the one hand, the biocompatibility of existing artificial ligaments is poor, and long-term implantation may cause rejection reactions or hinder the integration and healing of surrounding tissues. On the other hand, the mechanical properties of existing artificial ligaments are poor, making it difficult to match the biomechanical characteristics of the natural anterior cruciate ligament. Summary of the Invention

[0004] One objective of the first aspect of this invention is to provide a method for preparing an artificial ligament, thereby solving the technical problems of poor biocompatibility and poor mechanical properties of artificial ligaments in the prior art.

[0005] Another objective of this invention is to improve the stability of artificial ligaments.

[0006] A second aspect of the present invention aims to provide an artificial ligament.

[0007] According to a first aspect of the present invention, the present invention provides a method for preparing an artificial ligament, comprising the following steps: Multiple sets of silk bundles are provided, each set of silk bundles comprising multiple silk threads; Multiple silk threads in each group of silk bundles are twisted together to form a bundle of silk, and multiple bundles of silk are woven to form a first woven rope with a first preset structure; The first braided rope is degummed to obtain the second braided rope; An artificial ligament is prepared by weaving multiple second braided ropes according to a second preset structure using a three-dimensional four-step method.

[0008] Optionally, the step of twisting multiple silk threads together to form a bundle of silk, and weaving multiple bundles of silk to form a first braided rope having a first predetermined structure, further includes the following steps: The first braided rope is formed by weaving multiple bundles of filaments using a first braiding machine, wherein the rotation speed of the track turntable of the first braiding machine is any value between 10 r / min and 30 r / min.

[0009] Optionally, the braiding angle of the first braided rope is any value between 10° and 40°, the braiding diameter is any value between 0.6mm and 0.8mm, and the braiding pitch is any value between 2mm and 7mm.

[0010] Optionally, the three-dimensional four-step method is implemented by the yarn array of the second weaving machine, which consists of a first yarn carrier arranged radially and axially. The number of the first yarn carriers arranged along the axial direction and the number of the first yarn carriers arranged along the radial direction are calculated based on the total number N of the second braided ropes in the artificial ligament and the following formula: N = n × (m + 1); Wherein, n is the number of the first yarn carriers arranged along the axial direction in the yarn array, and is an even number; m+1 is the number of the first yarn carriers arranged along the radial direction in the yarn array; and m determines the number of layers of the preset structure of the artificial ligament arranged radially.

[0011] Optionally, the second preset structure of the artificial ligament is a multi-layer structure arranged radially, and the number of layers in each second preset structure is any one of 2, 3 or 5 layers.

[0012] Optionally, when the artificial ligament has two layers, the number of first yarn carriers arranged radially in the braided yarn array is three, and the number of first yarn carriers arranged circumferentially in the braided yarn array is sixteen. When the artificial ligament has 3 layers, the number of first yarn carriers arranged radially in the braided yarn array is 4, and the number of first yarn carriers arranged circumferentially in the braided yarn array is 12. When the artificial ligament has 5 layers, the number of first yarn carriers arranged radially in the woven yarn array is 6, and the number of first yarn carriers arranged circumferentially in the woven yarn array is 8.

[0013] Optionally, the step of degumming the second braided rope to obtain the third braided rope further includes the following steps: The first buffer solution containing papain is heated to any value between 50°C and 60°C, and the first braided rope is added to the heated first buffer solution to perform degumming treatment. The first braided rope, after being degummed, is subjected to high-temperature boiling, washing, and drying processes in sequence to prepare the second braided rope.

[0014] Optionally, the concentration of papain in the first buffer solution is any value between 2 g / L and 6 g / L, and the degumming time for the first braided rope is any value between 55 min and 65 min.

[0015] According to a second aspect of the present invention, the present invention provides an artificial ligament prepared using the above-described preparation method.

[0016] Optionally, the porosity of the artificial ligament is any value between 25% and 55%.

[0017] In this invention, multiple silk threads are twisted together to form a bundle, and then multiple bundles are woven to form a first woven rope with a first preset structure. The first woven rope is then degummed to obtain a second woven rope. Finally, a three-dimensional four-step method is used to weave multiple second woven ropes according to the second preset structure, thereby obtaining an artificial ligament. In the above technical solution, the first woven rope is degummed to remove sericin from the surface of the silk, resulting in an artificial ligament with good biocompatibility and controllable degradation characteristics. Furthermore, the multiple bundles of silk threads are woven into a first woven rope with the first preset structure, and then multiple second woven ropes are further woven according to the second preset structure using a three-dimensional four-step method. This multi-step weaving design significantly improves the mechanical strength, toughness, and fatigue durability of the artificial ligament, enabling it to better simulate the mechanical behavior of natural ligaments, promoting tissue integration and functional recovery after implantation, and demonstrating significant clinical application value.

[0018] Furthermore, in this invention, the rotation speed of the track turntable of the first braiding machine is set to 10r / min-30r / min, so as to weave multiple bundles of filaments into a first braided rope with structural stability, thereby improving the stability of the artificial ligament.

[0019] The above and other objects, advantages and features of the present invention will become more apparent to those skilled in the art from the following detailed description of specific embodiments of the invention in conjunction with the accompanying drawings. Attached Figure Description

[0020] The following sections will describe some specific embodiments of the invention in detail by way of example and not limitation, with reference to the accompanying drawings. The same reference numerals in the drawings denote the same or similar parts or portions. Those skilled in the art should understand that these drawings are not necessarily drawn to scale. In the drawings: Figure 1 This is a schematic flowchart of a method for preparing an artificial ligament according to an embodiment of the present invention; Figure 2 This is a schematic diagram of a first preset structure of a first braided rope with a rhomboid structure according to an embodiment of the present invention; Figure 3 This is a schematic diagram of a first preset structure with a regular structure of a first braided rope according to an embodiment of the present invention; Figure 4 This is a schematic diagram of a first braided rope with different braiding angles according to an embodiment of the present invention; Figure 5 These are morphological images of the artificial ligaments according to Embodiments 1 to 4 of the present invention; Figure 6 This is a statistical chart of cell proliferation on day 1 of culture for artificial ligaments according to Examples 1 to 4 and Comparative Example 2 of the present invention; Figure 7 This is a statistical chart of cell proliferation on the third day of culture of the artificial ligaments of Examples 1 to 4 and Comparative Example 2 according to the present invention; Figure 8 This is a statistical diagram of collagen fiber deposition in rats on day 58 after artificial ligament implantation according to Examples 1, 2 and Comparative Example 2 of the present invention; Figure 9 This is a stained tissue section of a rat on day 58 after artificial ligament implantation according to Example 1 of the present invention; Figure 10 This is a stained tissue section of a rat on day 58 after artificial ligament implantation according to Example 2 of the present invention. Detailed Implementation

[0021] Unless otherwise specified, all terms (including technical and scientific terms) used in the description of this embodiment have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0022] Figure 1 This is a schematic flowchart of a method for preparing an artificial ligament according to an embodiment of the present invention.

[0023] like Figure 1 As shown, in one specific embodiment, a method for preparing an artificial ligament includes the following steps: Step S100: Provide multiple sets of silk bundles, each set of silk bundles including multiple silk threads; Step S200: Twist multiple silk threads from each group of silk bundles together to form a bundle of silk, and weave multiple bundles of silk to form a first braided rope with a first preset structure; Step S300: Degumming is performed on the first braided rope to obtain the second braided rope; In step S400, multiple second braided ropes are braided according to the second preset structure using a three-dimensional four-step method to prepare an artificial ligament.

[0024] In this embodiment, the first braided rope is degummed to remove sericin from the surface of the silk, thereby obtaining an artificial ligament with good biocompatibility and controllable degradation characteristics. Furthermore, multiple bundles of silk are braided into a first braided rope with a first preset structure, and then multiple second braided ropes are further braided according to a second preset structure using a three-dimensional four-step method. This multi-step braiding design significantly improves the mechanical strength, toughness, and fatigue durability of the artificial ligament, enabling it to better mimic the mechanical behavior of natural ligaments, promote post-implantation tissue integration and functional recovery, and demonstrates significant clinical application value.

[0025] Figure 2 This is a schematic diagram of a first predetermined structure of a first braided rope with a rhomboid shape according to an embodiment of the present invention. Figure 3 This is a schematic diagram of a first predetermined structure of a first braided rope with a regular structure according to an embodiment of the present invention. In step S200, 8-10 silk threads are spirally twisted at 10°-25° to obtain a bundle of silk threads. The number of silk threads in each bundle can be, for example, 8, 9, or 10 threads, and the twisting angle can be, for example, 10°, 15°, 20°, or 25°, or any other value within 10°-25°. The first predetermined structure can be a rhomboid structure or a regular structure. The rhomboid structure is formed by... Figure 2 It can be seen that the rule structure is composed of Figure 3 It can be seen that the rhomboid structure is formed by alternating single strands of filament, interlaced in a 1:1 ratio, passing successively above or below adjacent strands to form a typical rhomboid interlacing pattern. A regular structure refers to strands of filament arranged at orthogonal or oblique angles in the warp and weft directions, interlacing according to a specific weaving pattern to form a stable weaving shape. The weaving angle of the first weaving rope is any value between 10° and 40°, for example, 10°, 15°, 30°, or 40°, or any other value between 10° and 40°. The weaving pitch of the first weaving rope is any value between 2mm and 7mm, for example, 2mm, 2.74mm, 3.953mm, 5.987mm, or 7mm, or any other value between 2mm and 7mm.

[0026] Step S200 also includes the following steps: Step S210: Use the first braiding machine to weave multiple bundles of filaments into a first braided rope. The rotation speed of the track turntable of the first braiding machine is any value between 10 r / min and 30 r / min.

[0027] In step S210, the first braiding machine is a multi-spindle braiding machine, which includes multiple track turntables. Each track turntable is equipped with two second yarn carriers, and each second yarn carrier introduces a bundle of yarn. After the multi-spindle braiding machine is started, the multiple bundles of yarn move in a relatively staggered manner along the circular track formed by the multiple track turntables. Through the regular interweaving between the bundles of yarn, a first braided rope with a uniform structure is formed. The use of a multi-spindle braiding machine for weaving is well known to those skilled in the art and will not be described in detail here.

[0028] In this embodiment, the rotational speed of the turntable of the first braiding machine is, for example, 10 r / min, 15 r / min, 20 r / min, or 30 r / min, or any other value within the range of 10 r / min to 30 r / min. Setting the rotational speed of the turntable of the first braiding machine to 10 r / min to 30 r / min ensures the efficiency of the first braided rope preparation while enabling multiple strands of yarn to interweave stably and orderly along a circular track, forming a first braided rope with a first preset structure. When the rotational speed of the turntable of the first braiding machine is lower than 10 r / min, the interlacing points of the first braided rope become too sparse and the structure becomes loose, making it difficult to form the first preset structure. When the rotational speed of the turntable of the first braiding machine is higher than 30 r / min, the turntable drives the strands of yarn too fast, resulting in structural disorder and decreased uniformity of the first braided rope, and even strand entanglement or breakage.

[0029] Figure 4 This is a schematic diagram of a first braided rope with different braiding angles according to an embodiment of the present invention. The braiding angle of the first braided rope is any value between 10° and 40°, for example, it can be 10°, 15°, 30° or 40°, or any other value between 10° and 40°. The braiding angle refers to the angle between the axis of the first braided rope and the bundle of filaments. Schematic diagrams showing braiding angles of 15°, 25° and 35° for the first braided rope are provided. Figure 4 It can be seen that the weaving diameter of the first braided rope is 0.6mm-0.8mm, for example, it can be 0.6mm, 0.726mm or 0.8mm, or any other value among 0.6mm-0.8mm. The weaving pitch of the first braided rope is any value among 2mm-7mm, for example, it can be 2mm, 2.74mm, 3.953mm, 5.987mm or 7mm, or any other value among 2mm-7mm. The weaving pitch is the distance the bundle of yarn moves along the axial direction of the first braided rope when the second yarn carrier completes one full rotation.

[0030] In step S300, the silk comprises fibroin fibers and sericin coating the surface of the fibroin fibers. While fibroin itself possesses good mechanical properties, biocompatibility, and controllable degradation characteristics, sericin, as a foreign protein, has high immunogenicity, which can trigger adverse inflammatory and foreign body reactions at the implantation site. Furthermore, premature and uncontrollable degradation of sericin leads to a decrease in mechanical properties and the release of inflammatory irritants. The first braided rope is degummed to remove sericin from the surface of the silk. International standards stipulate that any material in contact with blood must have a hemolysis rate below 5% to ensure its safety. The hemolysis rate of fibroin is less than 2%, indicating that fibroin has superior blood compatibility, thus resulting in a second braided rope with better biocompatibility and controllable degradation characteristics.

[0031] Step S300 also includes the following steps: Step S310: Heat the first buffer solution containing papain to any value between 50°C and 60°C, and add the first braided rope to the heated first buffer solution for degumming. Step S320: The first braided rope after degumming is subjected to high-temperature boiling, rinsing and drying treatment in sequence to prepare the second braided rope.

[0032] In step S310, a predetermined weight of the first braided rope is weighed. An appropriate amount of the second buffer solution is taken, with a volume ratio of the first braided rope to the second buffer solution of 1:45-1:55. The second buffer solution is a papain-free buffer solution prepared by mixing 0.1 mol / L-0.3 mol / L anhydrous disodium hydrogen phosphate and 0.08 mol / L-0.15 mol / L citric acid (pH 6-7) in a volume ratio of 14.55:5.45. Papain is added to the second buffer solution to obtain a papain-containing first buffer solution. The first buffer solution is heated to 50℃-60℃, for example, 50℃, 55℃, or 60℃, or any other value within the 50℃-60℃ range. Then, the weighed first braided rope is added to the heated first buffer solution for degumming, until all the first braided ropes are degummed. The volume ratio of the first braided rope to the second buffer solution can be, for example, 1:45, 1:50, or 1:55, or any other value between 1:45 and 1:55. The pH of the second buffer solution can be, for example, 6, 6.6, or 7, or any other value between 6 and 7. The concentration of anhydrous disodium hydrogen phosphate can be, for example, 0.05 mol / L, 0.2 mol / L, or 0.3 mol / L, or any other value between 0.1 mol / L and 0.3 mol / L. The concentration of citric acid can be, for example, 0.08 mol / L, 0.1 mol / L, or 0.15 mol / L, or any other value between 0.08 mol / L and 0.15 mol / L.

[0033] In step S310, the concentration of papain in the first buffer solution is any value between 2 g / L and 6 g / L, for example, it can be 2 g / L, 5 g / L, or 6 g / L, or any other value between 2 g / L and 6 g / L. The degumming treatment time for the first braided rope is any value between 55 min and 65 min, for example, it can be 55 min, 60 min, or 65 min, or any other value between 55 min and 65 min.

[0034] In step S320, the degummed first braided rope is first heated in boiling deionized water for 12-20 minutes to inactivate the papain remaining on the surface of the silk in the first braided rope. Then, the first braided rope is washed multiple times with warm and room temperature deionized water to remove residual sericin and papain. Step S310 is then repeated once to completely remove the sericin from the surface of the silk in the first braided rope. Finally, the rope is washed and dried to obtain the second braided rope. The drying temperature is 55℃-60℃, for example, 55℃, 58℃, or 60℃, or any other value within the 55℃-60℃ range.

[0035] In step S400, the three-dimensional four-step method is implemented by the yarn array of a second braiding machine. The second braiding machine is preferably a circular braiding device. The yarn array of the circular braiding device has multiple first yarn carriers arranged radially and axially, with each first yarn carrier introducing a second braiding rope. The three-dimensional four-step method, through the alternating and reverse movements of the first yarn carriers in the radial and axial directions, causes multiple second braiding ropes to continuously interweave and intertwine in space, gradually forming a uniformly structured and tightly interwoven artificial ligament. This three-dimensional four-step method is a mature three-dimensional braiding technology in the prior art, and its braiding process will not be described in detail.

[0036] In step S400, the number of first yarn carriers arranged axially and radially in the woven yarn array can be calculated using the number N of the second braided ropes and the formula, thereby preparing artificial ligaments with 2, 3, and 5 layers. The artificial ligaments exhibit excellent tensile strength, elongation at break, tensile strength, elastic recovery, and cyclic tensile properties. Specifically, the tensile strength and tensile strength of the artificial ligaments are higher than those of the anterior cruciate ligament of the human knee joint, and the elongation at break is close to that of the human anterior cruciate ligament. Existing technologies typically improve the mechanical properties of artificial ligaments by immersing the degummed braided framework in a collagen solution, but this method has limitations such as uneven collagen distribution or easy peeling. This invention, through a braiding process and the introduction of a second pre-designed structure, enables the artificial ligaments to possess excellent mechanical properties and a long service life, meeting the needs of the human body. It demonstrates significant clinical application value. In another embodiment, other numbers of second braided cords can be selected adaptively according to the specific mechanical performance requirements of the artificial ligament to meet different clinical implantation and functional reconstruction needs.

[0037] In another embodiment, the difference from the above embodiment is that multiple bundles of filaments are directly combined together to obtain strands; the other steps are the same as in the above embodiment. The artificial ligament prepared can have, for example, 2, 3, or 5 layers arranged radially, all exhibiting good tensile strength, elongation at break, tensile strength, elastic recovery, and cyclic tensile properties. Furthermore, its tensile strength and tensile strength are higher than those of the anterior cruciate ligament of the human knee joint. The artificial ligament of this embodiment can also be designed with other numbers of layers as needed.

[0038] In some embodiments, the present invention also provides an artificial ligament prepared using the above-described preparation method. The porosity of the artificial ligament is any value between 25% and 55%, for example, it can be 25%, 45%, 50%, or 55%, or any other value between 25% and 55%.

[0039] The following detailed description uses specific embodiments and comparative examples.

[0040] Example 1 provides a method for preparing an artificial ligament, the method comprising: Step S111: Provide 288 sets of silk bundles, each set of silk bundles including 10 silk threads; In step S121, 10 silk threads are twisted at 15° to form a single strand bundle, resulting in 288 bundles. Six bundles are then woven into a first braided rope with a diamond structure using a 12-spindle braiding machine, thus producing 48 first braided ropes. The braiding angle of the first braided rope is 15°, the braiding diameter is 0.673±0.027mm, and the braiding pitch is 5.987±0.030mm. Step S131: Weigh multiple sets of woven fabrics, each set including 4g of the first woven rope. Take an appropriate volume of the second buffer solution, prepared by mixing 0.2mol / L anhydrous disodium hydrogen phosphate (pH=6.6) and 0.1mol / L citric acid at a volume ratio of 14.55:5.45. Add papain to the second buffer solution to obtain a first buffer solution containing papain, achieving an enzyme concentration of 2g / L. Heat the second buffer solution to 55℃, then add the pre-weighed first woven rope for degumming treatment for 60 minutes. Step S132: Immerse the degummed first braided rope in boiling deionized water and heat for 15 minutes. Then, wash it three times each with warm and room temperature deionized water. Repeat step S131 once. Finally, after washing and drying at 60°C, the second braided rope is obtained. Step S141: The 48 second braiding ropes are braided into an artificial ligament with 2 layers using a three-dimensional four-step method. The braiding angle of the artificial ligament is 32°±3°. The three-dimensional four-step method is implemented by a braiding yarn array of a circular braiding device. The number of first yarn carriers arranged radially in the braiding yarn array is 3, and the number of first yarn carriers arranged circumferentially in the braiding yarn array is 16.

[0041] Example 2: The difference between Example 2 and Example 1 is that the 6 bundled filaments are not woven using a 12-spindle braiding machine, but are instead twisted together to form a twisted yarn. The other steps are the same as in Example 1 to form an artificial ligament with 2 layers.

[0042] Example 3: The difference between Example 3 and Example 1 is that the 48 second braiding ropes are braided into an artificial ligament with 3 layers using a three-dimensional four-step method. The number of first yarn carriers arranged radially in the braided yarn array is 4, and the number of first yarn carriers arranged circumferentially in the braided yarn array is 12.

[0043] Example 4: The difference between Example 4 and Example 3 is that the 6 bundled filaments are not woven using a 12-spindle braiding machine, but are instead twisted together to form a ply yarn, thereby forming an artificial ligament with 3 layers.

[0044] Comparative Example 1: Comparative Example 1 is the anterior cruciate ligament of the human knee joint.

[0045] Comparative Example 2: The difference between Comparative Example 2 and Example 1 is that the material used to weave the artificial ligament is polyethylene terephthalate (PET), thus obtaining an artificial ligament with the same structure as in Example 1.

[0046] Figure 5 These are morphological images of the artificial ligaments according to Embodiments 1 to 4 of the present invention. Figure 5 As shown, the artificial ligaments of Examples 1 to 4 exhibit a good thickness distribution and a uniform weave structure, which helps to distribute stress evenly within the artificial ligament, reduces local stress concentration, and thus enhances the strength and durability of the artificial ligament. Furthermore, the uniform structure makes the surface of the artificial ligament smoother, reduces foreign body reactions and inflammation, improves post-implantation biocompatibility, and promotes good integration with surrounding tissues.

[0047] Table 1 below lists a comparison of the mechanical properties of the artificial ligaments of Examples 1 to 4 of the present invention with those of the anterior cruciate ligament of the human knee joint in Comparative Example 1.

[0048] Table 1 As shown in Table 1, the tensile strength of the artificial ligaments prepared in Examples 1 to 4 is higher than that of the human anterior cruciate ligament of the knee joint in Comparative Example 1, and the tensile strength of Examples 1 to 4 is also higher than that of Comparative Example 1, indicating that the artificial ligaments prepared in Examples 1 to 4 have excellent performance in terms of tensile strength and tensile strength. Furthermore, the elongation at break of Examples 1 to 4 is close to the maximum elongation at break of 60N of the human anterior cruciate ligament of the knee joint, indicating that the artificial ligaments prepared in Examples 1 to 4 have good toughness and ductility, and can withstand a certain amount of tension without easily breaking.

[0049] Table 2 below lists the mechanical performance parameters of the artificial ligaments of Examples 1 to 4 of the present invention.

[0050] Table 2 As shown in Table 2, the three-dimensional structures of the artificial ligaments prepared in Examples 1 to 4 exhibit high elastic recovery performance due to their relatively flexible network characteristics. In practical use, they can effectively restore their original shape and meet the mechanical requirements of the anterior cruciate ligament of the human knee joint. In particular, the artificial ligament of Example 2 has the highest elastic recovery rate of 46%. Cyclic tensile performance tests show that the artificial ligaments prepared in Examples 1 to 4 have relatively low fracture strength attenuation rates and can maintain stability for a relatively long time, making them suitable for long-term implantation in the human body. The porosities of Examples 1 to 4 are 44.16%, 49.16%, 27.5%, and 31.5%, respectively, indicating that the pore structure of the artificial ligaments is reasonable. After implantation in the human body, this helps promote cell growth and tissue fusion while ensuring the mechanical properties of the artificial ligaments.

[0051] Figure 6 This is a statistical chart showing the number of cells proliferating on day 1 of culture for the artificial ligaments of Examples 1 to 4 and Comparative Example 2 according to the present invention. Figure 7 This is a statistical chart showing the number of cells proliferating on the third day of culture of the artificial ligaments according to Examples 1 to 4 and Comparative Example 2. The artificial ligaments prepared in Examples 1 to 4 and Comparative Example 2 were made into segments with a diameter of 0.5 cm. Rat fibroblasts were inoculated onto the artificial ligaments after high-temperature and high-pressure sterilization, and placed in a culture medium for cell culture. Cell viability was tested on days 1 and 3. The fluorescence intensity of the rat fibroblasts on the artificial ligaments was detected by an enzyme-linked immunosorbent assay (ELISA) reader. Higher fluorescence intensity indicated a higher number of proliferating rat fibroblasts, and correspondingly better cell compatibility of the artificial ligament. Figure 6 and Figure 7 It can be seen that as the culture time is extended from day 1 to day 3, the number of cells proliferating in Examples 1 to 4 and Comparative Example 2 all show an increasing trend. However, the number of cells proliferating in Examples 1 to 4 is higher than that in Comparative Example 2, indicating that the artificial ligaments prepared in Examples 1 to 4 of the present invention have significant advantages in terms of cell compatibility. Figure 6 and Figure 7 In the middle, asterisk The number of asterisks indicates the statistical significance of the differences between groups; the more asterisks, the greater the differences between the groups.

[0052] Figure 8 This is a statistical diagram of collagen fiber deposition in rats on day 58 after implantation of artificial ligaments according to Examples 1, 2, and Comparative Example 2 of the present invention. Collagen fibers are new tissues secreted by host cells on the artificial ligament. Figure 8It can be seen that after 58 days of implantation of artificial ligaments into rats, the collagen fiber deposition rates of the artificial ligaments prepared in Example 1 and Example 2 were 53.38±5.18% and 60.60±1.99%, respectively, both higher than the collagen fiber deposition rate of the artificial ligament in Comparative Example 2. This indicates that the artificial ligaments in Example 1 and Example 2 can effectively promote the metabolic activity of fibroblasts and accelerate biological revascularization and tendon-bone healing processes. Figure 8 The asterisk in This indicates that Example 2 differs from Comparative Example 2, and the artificial ligament of Example 2 has advantages over Comparative Example 2 in terms of regeneration and repair.

[0053] Figure 9 This is a stained tissue section of a rat on day 58 after artificial ligament implantation according to Example 1 of the present invention. Figure 10 This is a stained tissue section of a rat on day 58 after artificial ligament implantation according to Example 2 of the present invention. Figure 9 (a) is a schematic diagram of a cross-section along the radial direction of the artificial ligament in Example 1. Figure 9 The image in the middle (bd) shows the staining of the artificial ligament in Example 1 at different magnifications. Figure 10 Image (e) is a schematic diagram of a cross-section along the radial direction of the artificial ligament in Example 2. Figure 10 (fh) shows the staining images of the artificial ligament in Example 2 at different magnifications. Figure 9 (a) and Figure 10 (e) It can be seen that the artificial ligaments prepared in Example 1 and Example 2 both have a tubular hollow structure, which provides a good infiltration space for cells and supports cell migration and growth. Figure 9 (d) and Figure 10 The arrow in (h) points to a location where new blood vessels have formed, and a large amount of blue-reacting collagen fiber deposits are observed inside the gaps, indicating that the artificial ligaments of Examples 1 and 2 have excellent tissue repair and regeneration capabilities.

[0054] Therefore, those skilled in the art should recognize that although numerous exemplary embodiments of the present invention have been shown and described in detail herein, many other variations or modifications conforming to the principles of the present invention can be directly determined or derived from the disclosure of the present invention without departing from the spirit and scope of the invention. Thus, the scope of the present invention should be understood and construed as covering all such other variations or modifications.

Claims

1. A method for preparing an artificial ligament, characterized in that, Includes the following steps: Multiple sets of silk bundles are provided, each set of silk bundles comprising multiple silk threads; Multiple silk threads in each group of silk bundles are twisted together to form a bundle of silk, and multiple bundles of silk are woven to form a first woven rope with a first preset structure; The first braided rope is degummed to obtain the second braided rope; An artificial ligament is prepared by weaving multiple second braided ropes according to a second preset structure using a three-dimensional four-step method.

2. The preparation method according to claim 1, characterized in that, The step of twisting multiple silk threads together to form a bundle of silk, and weaving multiple bundles of silk to form a first braided rope with a first predetermined structure, further includes the following step: The first braided rope is formed by weaving multiple bundles of filaments using a first braiding machine, wherein the rotation speed of the track turntable of the first braiding machine is any value between 10 r / min and 30 r / min.

3. The preparation method according to claim 2, characterized in that, The first braided rope has a braiding angle of any value between 10° and 40°, a braiding diameter of any value between 0.6mm and 0.8mm, and a braiding pitch of any value between 2mm and 7mm.

4. The preparation method according to claim 3, characterized in that, The three-dimensional four-step method is implemented by the yarn array of the second weaving machine, which consists of a first yarn carrier arranged along its radial and axial directions; The number of the first yarn carriers arranged along the axial direction and the number of the first yarn carriers arranged along the radial direction are calculated based on the total number N of the second braided ropes in the artificial ligament and the following formula: N = n × (m + 1); Wherein, n is the number of the first yarn carriers arranged along the axial direction in the yarn array, and is an even number; m+1 is the number of the first yarn carriers arranged along the radial direction in the yarn array; and m determines the number of layers of the preset structure of the artificial ligament arranged radially.

5. The preparation method according to claim 4, characterized in that, The second preset structure of the artificial ligament is a multi-layer structure arranged radially, and the number of layers in each second preset structure is any one of 2, 3 or 5 layers.

6. The preparation method according to claim 5, characterized in that, When the artificial ligament has 2 layers, the number of first yarn carriers arranged radially in the braided yarn array is 3, and the number of first yarn carriers arranged circumferentially in the braided yarn array is 16. When the artificial ligament has 3 layers, the number of first yarn carriers arranged radially in the braided yarn array is 4, and the number of first yarn carriers arranged circumferentially in the braided yarn array is 12. When the artificial ligament has 5 layers, the number of first yarn carriers arranged radially in the woven yarn array is 6, and the number of first yarn carriers arranged circumferentially in the woven yarn array is 8.

7. The preparation method according to claim 6, characterized in that, The step of degumming the second braided rope to obtain the third braided rope also includes the following steps: The first buffer solution containing papain is heated to any value between 50°C and 60°C, and the first braided rope is added to the heated first buffer solution to perform degumming treatment. The first braided rope, after being degummed, is subjected to high-temperature boiling, washing, and drying processes in sequence to prepare the second braided rope.

8. The preparation method according to claim 7, characterized in that, The concentration of papain in the first buffer solution is any value between 2 g / L and 6 g / L, and the degumming time for the first braided rope is any value between 55 min and 65 min.

9. An artificial ligament, characterized in that, It is prepared using the preparation method described in any one of claims 1-8.

10. The artificial ligament according to claim 9, characterized in that, The porosity of the artificial ligament is any value between 25% and 55%.

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