Biomimetic material, super-tough biomimetic temporomandibular joint disc, and preparation method and application thereof

Abstract

The application discloses a biomimetic material, a super-strong and tough biomimetic temporomandibular joint disc and a preparation method and application thereof. The biomimetic material comprises: 1%-3% (w / v) of a first base material for photocuring, wherein the first base material for photocuring comprises a combination of one or more of alkyl acrylic anhydride modified hyaluronic acid, chitosan, chondroitin sulfate and silk fibroin; 7wt.%-9wt.% (w / v) of a second base material for photocuring; the second base material for photocuring comprises at least one of ortho-nitrobenzyl alcohol modified gelatin, ortho-nitrobenzyl alcohol modified polyethylene glycol and polyethylene glycol acrylate compounds; and the balance is deionized water or a PBS aqueous solution. The biomimetic material is prepared through a photocured double-network hydrogel system, realizes high tensile strength and compressive strength, and can effectively bear complex loads in daily activities such as chewing.

Description

Technical Field

[0001] This application belongs to the field of implantable bionic medical device technology, and in particular relates to a bionic material, an ultra-strong and tough bionic temporomandibular joint disc, its preparation method and application. Background Technology

[0002] like Figure 1 As shown, the temporomandibular disc 3 (TMD3) is a piece of fibrocartilage covering the condyle 21 of the mandible 2 in the human skull, playing an important role in cushioning and lubrication during daily activities such as chewing and eating. Due to unilateral chewing, bruxism, and other reasons, the TMD3 in some patients is prone to displacement, thinning, or even perforation, leading to temporomandibular joint disorders (TMD). Clinically, there are no good treatment methods for large and difficult-to-suture defects of the TMD3; the only option is TMD resection. Otherwise, the defect of the TMD3 will lead to further wear and tear of the exposed condyle 21 cartilage at the temporomandibular joint, or a patching procedure using autologous fascia tissue. For more severe TMD3 lesions, total temporomandibular joint replacement may be necessary. Even if it's just a temporomandibular joint disc lesion, it's necessary to replace the entire temporal bone's mandibular fossa 11, temporomandibular joint disc 3, and mandibular condyle 21.

[0003] Currently, there are no commercially available prostheses for implantation after temporomandibular joint disc resection (TMD3). TMD3 prostheses made with Proplast-Teflon were previously used clinically, but were banned by the U.S. Food and Drug Administration (FDA) due to serious complications. Current TMD3 replacements under development also suffer from insufficient mechanical properties, poor biocompatibility, low biomimicry, and lack of regenerative capacity, resulting in poor prospects for clinical application. Summary of the Invention

[0004] This application provides a biomimetic material, an ultra-tough biomimetic temporomandibular joint disc, its preparation method, and its application. The biomimetic material can achieve high tensile and compressive strength of the ultra-tough biomimetic temporomandibular joint disc through a photocurable dual-network hydrogel system. It has mechanical properties close to or even exceeding those of a natural temporomandibular joint disc and can effectively withstand complex loads in daily activities such as chewing.

[0005] In a first aspect, embodiments of this application provide a biomimetic material for preparing an ultra-strong and tough biomimetic temporomandibular joint disc, comprising: a first base material for photocuring, 1% to 3% (w / v), the first base material for photocuring comprising one or more combinations of alkyl acrylic anhydride modified hyaluronic acid, chitosan, chondroitin sulfate, and silk fibroin; and a second base material for photocuring, 7% to 9% (w / v), the second base material for photocuring comprising at least one of o-nitrobenzyl alcohol modified gelatin, o-nitrobenzyl alcohol modified polyethylene glycol, and polyethylene glycol acrylate compounds; the balance being deionized water or PBS aqueous solution.

[0006] According to an embodiment of the first aspect of this application, the PBS aqueous solution used is an aqueous solution with a PBS concentration of 0.005 mol / L to 0.02 mol / L and a pH of 7.2 to 7.4. Optionally, the PBS concentration is 0.01 mol / L to 0.015 mol / L.

[0007] According to an embodiment of the first aspect of this application, the first base material for photocuring includes one or more combinations of C1-C6 alkyl acrylic anhydride modified hyaluronic acid, chitosan, chondroitin sulfate, and silk fibroin.

[0008] According to the embodiments of the first aspect of this application, the C1-C6 alkyl acrylic anhydrides include one or more combinations of methacrylic anhydride, ethylacrylic anhydride, propylacrylic anhydride, isopropylacrylic anhydride, n-butylacrylic anhydride, isobutylacrylic anhydride, tert-butylacrylic anhydride, n-pentylacrylic anhydride, n-hexylacrylic anhydride, 3-methylpentylacrylic anhydride, and 2-ethylbutylacrylic anhydride.

[0009] According to an embodiment of the first aspect of this application, the mass ratio of o-nitrobenzyl alcohol modified gelatin to alkyl acrylic anhydride modified hyaluronic acid is (5-8):1.

[0010] According to an embodiment of the first aspect of this application, the mass ratio of o-nitrobenzyl alcohol-modified polyethylene glycol to alkyl acrylic anhydride-modified hyaluronic acid is (5-7):1.

[0011] According to the embodiments of the first aspect of this application, the polyethylene glycol acrylate compounds include one or more combinations of polyethylene glycol methacrylate, polyethylene glycol diacrylate, polyethylene glycol triacrylate, and multi-arm PEG-acrylate.

[0012] According to an embodiment of the first aspect of this application, the biomimetic material further includes a photoinitiator selected from LAP, wherein the LAP accounts for 0.01% to 0.1% (w / v) of the biomimetic material by mass volume.

[0013] According to an embodiment of the first aspect of this application, the biomimetic material used to prepare the ultra-strong and tough biomimetic temporomandibular joint disc further includes any one or a combination of several of the following: 0.5% to 4% (w / v) of umbilical cord Wharton glue, 2% to 4% (w / v) of umbilical cord ECM, 3% to 9% (w / v) of cartilage ECM, and 0.5% to 3% (w / v) of meniscus ECM.

[0014] According to an embodiment of the first aspect of this application, the biomimetic material used to prepare the ultra-strong and tough biomimetic temporomandibular joint disc includes a sustained-release factor, which includes at least one of CTGF and TGF-β.

[0015] According to an embodiment of the first aspect of this application, the sustained-release factor is loaded with polylactic acid-glycolic acid copolymer microspheres.

[0016] According to an embodiment of the first aspect of this application, the polylactic acid-glycolic acid copolymer microspheres include first copolymer microspheres with an average diameter of 200 nm to 1 μm and / or second copolymer microspheres with an average diameter of 20 μm to 50 μm.

[0017] According to the embodiments of the first aspect of this application, the polylactic acid-glycolic acid copolymer microspheres account for 0.1% (w / v) to 1% (w / v) of the solution by mass volume.

[0018] According to an embodiment of the first aspect of this application, in the polylactic acid-glycolic acid copolymer microspheres, the proportion of the first copolymer microspheres with an average diameter of 200 nm to 1 μm is 40% to 60%; and the proportion of the second copolymer microspheres with an average diameter of 20 μm to 50 μm is 40% to 60%.

[0019] According to an embodiment of the first aspect of this application, in the first copolymer microspheres with an average diameter of 200 nm to 1 μm, the molar percentage of polylactic acid is 45% to 55%, and the molar percentage of glycolic acid polylactic acid is 45% to 55%.

[0020] According to an embodiment of the first aspect of this application, CTGF cytokines are loaded using first copolymer microspheres with an average diameter of 200 nm to 1 μm.

[0021] According to an embodiment of the first aspect of this application, the loading concentration of CTGF cytokine is 20 ng / mg to 80 ng / mg.

[0022] According to an embodiment of the first aspect of this application, in the second copolymer microspheres with an average diameter of 20 μm to 50 μm, the molar percentage of polylactic acid is 65% to 80%, and the molar percentage of glycolic acid is 20% to 35%.

[0023] According to an embodiment of the first aspect of this application, a second copolymer microsphere with an average diameter of 20 μm to 50 μm is used to load the TGF-β factor.

[0024] According to an embodiment of the first aspect of this application, the loading concentration of TGF-β factor is 5 ng / mg to 30 ng / mg.

[0025] Secondly, embodiments of this application provide a method for preparing an ultra-tough bionic temporomandibular joint disc, comprising: obtaining the three-dimensional shape of the temporomandibular joint disc; providing the bionic material provided in the embodiments of the first aspect for preparing the ultra-tough bionic temporomandibular joint disc; and forming the ultra-tough bionic temporomandibular joint disc using the bionic material provided in the embodiments of the first aspect according to the three-dimensional shape of the temporomandibular joint disc.

[0026] According to an embodiment of the second aspect of this application, based on the three-dimensional shape of the temporomandibular joint disc, the process includes: fabricating a central portion; and fabricating a peripheral portion outside the central portion, wherein both the central portion and the peripheral portion are made using the biomimetic material provided in the embodiment of the first aspect for preparing an ultra-strong and tough biomimetic temporomandibular joint disc.

[0027] Thirdly, embodiments of this application provide an ultra-strong and tough biomimetic temporomandibular joint disc, comprising: a central portion including a first fibroblast-like tissue arranged in an anterior-posterior direction; and a peripheral portion including a second fibroblast-like tissue arranged in a ring direction and surrounding the central portion; both the central portion and the peripheral portion are made of the biomimetic material provided in the first aspect embodiment for preparing the ultra-strong and tough biomimetic temporomandibular joint disc.

[0028] Fourthly, embodiments of this application provide the application of the biomimetic material provided in the first aspect of the embodiment for preparing ultra-tough biomimetic temporomandibular joint discs in the preparation of biomimetic cartilage tissue, such as for temporomandibular joint disc defect repair, replacement or regeneration induction.

[0029] The biomimetic material used in this application embodiment for preparing an ultra-tough biomimetic temporomandibular joint disc achieves high tensile and compressive strength of the ultra-tough biomimetic temporomandibular joint disc through a photocurable dual-network hydrogel system formed by the first and second base materials. The mechanical properties of the prepared ultra-tough biomimetic temporomandibular joint disc are close to or even exceed those of the natural temporomandibular joint disc, and it can effectively withstand the complex loads of the temporomandibular joint disc in daily activities such as chewing and biting. Attached Figure Description

[0030] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments of this application will be briefly introduced below. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0031] Figure 1 This is a schematic diagram showing the application location of the temporomandibular joint disc.

[0032] Figure 2 This is a three-dimensional scan of the ultra-strong and tough bionic temporomandibular joint disc provided in the embodiments of this application.

[0033] Figure 3 This is a three-dimensional scan of the ultra-strong and tough bionic temporomandibular joint disc provided in the embodiments of this application.

[0034] Figure 4 This is a schematic diagram of the structure of the anterior and posterior running tissues in the central part and the annular running tissues in the peripheral part of the ultra-strong and tough biomimetic temporomandibular joint disc provided in the embodiments of this application.

[0035] Figure 5 This is a schematic diagram of the structural fabrication sequence used in the preparation method of the ultra-strong and tough bionic temporomandibular joint disc provided in the embodiments of this application.

[0036] Figure 6 This is an MRI image of the temporomandibular joint disc obtained in this application.

[0037] Figure 7 This is a Sirius red staining image of a normal temporomandibular joint disc in a human.

[0038] Figure 8 yes Figure 7 A 40x magnified view of local region a and its quantitative analysis diagram, where the right side is the quantitative analysis diagram, the horizontal axis is the angle (°) and the vertical axis is the percentage (%).

[0039] Figure 9 yes Figure 7 A 40x magnified view of local region b and its quantitative analysis diagram, where the right side is the quantitative analysis diagram, the horizontal axis is the angle (°) and the vertical axis is the percentage (%).

[0040] Figure 10 This is a physical image of the ultra-strong and tough bionic temporomandibular joint disc provided in Embodiment 3 of this application in actual application.

[0041] Figure 11 This is a stress-strain diagram of the tensile strength test performed on the ultra-tough biomimetic temporomandibular joint disc prepared in Example 3 of this application.

[0042] Figure 12 This is a stress-strain diagram of the compressive strength test performed on the ultra-tough biomimetic temporomandibular joint disc prepared in Example 3 of this application.

[0043] Explanation of reference numerals in the attached figures: 1. Temporal bone; 11. Mandibular fossa; 2. Mandible; 21. Condyle; 3. Temporomandibular joint disc; 31. First concave portion; 32. Second concave portion; 4. Central portion; 40. First fibroblast-like tissue; 5. Peripheral portion; 50. Second fibroblast-like tissue. Figure 4 In the diagram, A represents the front, P represents the back, M represents the inside, and L represents the outside. Detailed Implementation

[0044] The features and exemplary embodiments of various aspects of this application will be described in detail below. To make the objectives, technical solutions, and advantages of this application clearer, the application will be further described in detail below with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are only intended to explain this application and not to limit it. For those skilled in the art, this application can be implemented without some of these specific details. The following description of the embodiments is merely to provide a better understanding of this application by illustrating examples.

[0045] It should be noted that, in this document, relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising..." does not exclude the presence of additional identical elements in the process, method, article, or apparatus that includes said element.

[0046] To address the problems of existing technologies, this application provides a biomimetic material for preparing an ultra-strong and tough biomimetic temporomandibular joint disc, the ultra-strong and tough biomimetic temporomandibular joint disc, its preparation method, and its application. The ultra-strong and tough biomimetic temporomandibular joint disc, its preparation method, and its application provided in this application are described below.

[0047] In a first aspect, embodiments of this application provide a biomimetic material for preparing an ultra-strong and tough biomimetic temporomandibular joint disc, comprising: a first base material for photocuring, 1% to 3% (w / v), the first base material for photocuring comprising one or more combinations of alkyl acrylic anhydride modified hyaluronic acid, chitosan, chondroitin sulfate, and silk fibroin; and a second base material for photocuring, 7% to 9% (w / v) based on the mass of the first base material; the second base material for photocuring comprises at least one of o-nitrobenzyl alcohol modified gelatin, o-nitrobenzyl alcohol modified polyethylene glycol, and polyethylene glycol acrylate compounds; the balance being deionized water or PBS aqueous solution.

[0048] The biomimetic material provided in this application for preparing an ultra-tough biomimetic temporomandibular joint disc is a photocurable hydrogel system. Through the photocurable dual-network hydrogel system formed by the first and second base materials, the ultra-tough biomimetic temporomandibular joint disc achieves high tensile and compressive strength. The mechanical properties of the prepared ultra-tough biomimetic temporomandibular joint disc are close to or even exceed those of the natural temporomandibular joint disc, and it can effectively withstand the complex loads in daily activities such as chewing.

[0049] The biomimetic material provided in this application for preparing an ultra-tough biomimetic temporomandibular joint disc uses one or more of the following naturally derived materials, such as gelatin, hyaluronic acid, chondroitin sulfate, and silk fibroin, as well as FDA-approved synthetic materials such as PLGA and PEG, to ensure that the material is safe and biodegradable and has good biocompatibility and biodegradability.

[0050] The biomimetic material provided in this application for preparing ultra-strong and tough biomimetic temporomandibular joint discs can be used to prepare superior temporomandibular joint discs with tensile strength and compressive strength ranging from 3.5 MPa to 30 MPa, meeting user needs.

[0051] It should be noted that the mass-volume content of both the first and second substrates used for photocuring are the contents after dissolution in deionized water or PBS aqueous solution. Taking the first substrate for photocuring as an example, each 100 mL of water or PBS aqueous solution contains 1 to 3 grams of the first substrate as a solute, expressed in g / 100 mL. Similarly, each 100 mL of water or PBS aqueous solution contains 7 to 9 grams of the second substrate as a solute. It should be noted that the mass-volume content (m / v) of both the first and second substrates is calculated using the same solvent system, not by preparing separate aqueous solutions from different solvent systems and then mixing them. The concentration of PBS in the PBS aqueous solution is 0.005 mol / L to 0.02 mol / L, and the pH is 7.2 to 7.4. Optionally, the concentration of PBS in the PBS aqueous solution is 0.01 mol / L to 0.015 mol / L.

[0052] In some embodiments, the first base material for photocuring includes one or more combinations of C1-C6 alkyl acrylic anhydride modified hyaluronic acid, chitosan, chondroitin sulfate, and silk fibroin.

[0053] In some embodiments, C1-C6 alkyl acrylic anhydrides include one or more combinations of methacrylic anhydride, ethylacrylic anhydride, propylacrylic anhydride, isopropylacrylic anhydride, n-butylacrylic anhydride, isobutylacrylic anhydride, tert-butylacrylic anhydride, n-pentylacrylic anhydride, n-hexylacrylic anhydride, 3-methylpentylacrylic anhydride, and 2-ethylbutylacrylic anhydride.

[0054] In some embodiments, modified products such as C1-C6 alkyl acrylic anhydride-modified hyaluronic acid, chitosan, chondroitin sulfate, and silk fibroin can all be purchased from EFL.

[0055] In some embodiments, the biomimetic material further includes a photoinitiator selected from LAP, wherein the LAP content in the biomimetic material is 0.01% to 0.1% (w / v). The proportion of photoinitiator LAP also refers to the corresponding weight of material as solute per 100 mL of solution, expressed in g / 100 mL.

[0056] In some embodiments, o-nitrobenzyl alcohol-modified gelatin, abbreviated as GelNB, is used. Here, o-nitrobenzyl is the photosensitive group in "o-nitrobenzyl alcohol". In the modification reaction, a derivative of this alcohol (such as o-nitrobenzyl bromide) typically reacts with an amino group on the gelatin (such as the ε-amino group of lysine), attaching the o-nitrobenzyl group to the gelatin backbone. This gives it photocurable properties, meaning it cross-links into a network under ultraviolet light. The photocrosslinking speed of NB-modified materials is much faster than that of traditional MA-modified materials, and they are less prone to collapse after molding, thus offering better printability.

[0057] In some embodiments, gelatin is a natural biopolymer obtained from the hydrolysis of collagen, exhibiting good biocompatibility and cell adhesion. The gelatin may be derived from porcine or human collagen gelatin. Optionally, the human collagen is selected from recombinant human type I collagen gelatin.

[0058] In some embodiments, the mass ratio of o-nitrobenzyl alcohol-modified gelatin to alkyl acrylic anhydride-modified hyaluronic acid in the biomimetic material is (5-8):1. Exemplarily, the mass ratio of o-nitrobenzyl alcohol-modified gelatin to alkyl acrylic anhydride-modified hyaluronic acid can be 5.2:1, 5.4:1, 5.5:1, 5.6:1, 5.75:1, 5.8:1, 6:1, 6.3:1, 6.5:1, 6.7:1, 6.9:1, 7:1, 7.2:1, 7.5:1, 7.6:1, 7.8:1, or 8:1.

[0059] In some embodiments, the mass ratio of o-nitrobenzyl alcohol-modified polyethylene glycol to alkyl acrylic anhydride-modified hyaluronic acid in the biomimetic material is (5-7):1. Exemplarily, the mass ratio of o-nitrobenzyl alcohol-modified polyethylene glycol to alkyl acrylic anhydride-modified hyaluronic acid can be 5.2:1, 5.5:1, 5.6:1, 5.8:1, 6:1, 6.3:1, 6.5:1, 6.7:1, or 6.9:1.

[0060] The biomimetic material provided in this application for preparing an ultra-tough biomimetic temporomandibular joint disc includes o-nitrobenzyl alcohol-modified gelatin. The material is in a mixed solution state and can form a very strong and tough double-network hydrogel under ultraviolet light irradiation. This results in the ultra-tough biomimetic temporomandibular joint disc 3 having a tensile strength exceeding 8 MPa, close to the strength of a human temporomandibular joint disc 3. Furthermore, the ultra-tough biomimetic temporomandibular joint disc 3 has a compressive strength exceeding 25 MPa, far exceeding the compressive strength of a human temporomandibular joint disc 3.

[0061] The inventors of this application discovered during their research on materials for temporomandibular joint discs that a mixed solution containing o-nitrobenzyl alcohol-modified gelatin and alkyl acrylic anhydride-modified hyaluronic acid exhibits relatively poor rheological properties and lacks printability. Therefore, it is necessary to inject this biomimetic material into a mold for fabricating the temporomandibular joint disc 3 and then cure it under ultraviolet light to construct the first biomimetic temporomandibular joint disc 3. This biomimetic material is based entirely on natural materials, is slowly degradable, possesses excellent mechanical properties, and exhibits good biocompatibility. However, the inventors of this application found that the injection molding method results in the biomimetic temporomandibular joint disc 3 only replicating the external shape of the disc, but not the fiber orientation, resulting in insufficient biomimicry and precision.

[0062] Based on this, this application also provides another biomimetic material for preparing an ultra-strong and tough biomimetic temporomandibular joint disc 3. Specifically, it is a biomimetic material containing polyethylene glycol modified with o-nitrobenzyl alcohol as a second base material for photocuring. This utilizes the fact that polyethylene glycol (PEG) is a synthetic polymer compound that is non-toxic, non-irritating, and possesses good lubricity and water solubility.

[0063] In some embodiments, o-nitrobenzyl alcohol-modified polyethylene glycol, or Poly(ethylene glycol) o-Nitrobenzyl ether, abbreviated as PEGNB, is used as a photosensitive crosslinking agent. Hydrogels prepared with PEGNB no longer undergo random hydrolysis during degradation; instead, their degradation can be programmed through light-induced patterns. For example, a tissue scaffold can be printed first, and then microvascular channels can be sculpted at specific times and locations using light to guide cell growth. Drugs or growth factors can be optically linked to the hydrogel network via PEGNB. Precise, timed, and quantitative release can be achieved by irradiating the desired location and site, mimicking complex signal transduction processes in vivo.

[0064] The inventors of this application discovered that a mixed solution containing polyethylene glycol modified with o-nitrobenzyl alcohol and hyaluronic acid modified with alkyl acrylic anhydride produces an ultra-tough biomimetic temporomandibular joint disc 3 that is also quite strong, with a tensile strength exceeding 3.5 MPa and a compressive strength exceeding 12 MPa, surpassing the strength of a human temporomandibular joint disc 3. Furthermore, this mixed solution has excellent fluidity and can be used as an ink for 3D printing. It can be used for layer-by-layer photopolymerization 3D printing to fabricate the temporomandibular joint disc 3. During printing, the nozzle of the ink containing the mixed solution of polyethylene glycol modified with o-nitrobenzyl alcohol and hyaluronic acid modified with alkyl acrylic anhydride moves along the fiber orientation of the natural temporomandibular joint disc 3, constructing a highly precise biomimetic temporomandibular joint disc 3 with a high degree of biomimicry. This biomimetic material used to form the ultra-tough biomimetic temporomandibular joint disc 3 is also slowly degradable and has good biocompatibility.

[0065] In some embodiments, a mixed solution of polyethylene glycol modified with o-nitrobenzyl alcohol and hyaluronic acid modified with alkyl acrylic anhydride is used to make an ultra-tough biomimetic temporomandibular joint disc 3 with a tensile strength of more than 5 MPa and a compressive strength of more than 15 MPa.

[0066] In some embodiments, when polyethylene glycol acrylate compounds (abbreviated as PEGDA) are used as the second base material, PEGDA+HAMA needs to be first irradiated to form a stable PEGDA network, and then immersed in HAMA solution for full penetration and a second irradiation, so as to obtain a biomimetic temporomandibular joint disc 3 that has structural strength and mechanical strength, good biocompatibility and biodegradability, and also has a high degree of fine fiber tissue orientation.

[0067] In some embodiments, the polyethylene glycol acrylate compounds include one or a combination of several of polyethylene glycol methacrylate, polyethylene glycol diacrylate, polyethylene glycol triacrylate, and multi-arm PEG-acrylate.

[0068] It is understood that, in some embodiments, o-nitrobenzyl alcohol-modified gelatin, o-nitrobenzyl alcohol-modified polyethylene glycol, and polyethylene glycol acrylate compounds can be mixed in pairs or in all three proportions as biomimetic materials to prepare the ultra-strong and tough biomimetic temporomandibular joint disc 3. This allows for adjustment of the required tensile and compressive strength of the ultra-strong and tough biomimetic temporomandibular joint disc 3, providing users with an ultra-strong and tough biomimetic temporomandibular joint disc 3 with suitable mechanical strength.

[0069] In some embodiments, the biomimetic material used to prepare the ultra-strong and tough biomimetic temporomandibular joint disc 3 further includes any one or more combinations of the following: 0.5%–4% (w / v) of umbilical cord Wharton's glue, 2%–4% (w / v) of umbilical cord ECM, 3%–9% (w / v) of cartilage ECM, and 0.5%–3% (w / v) of meniscus ECM. It should be noted that the mass-volume content of the above materials refers to the corresponding weight of material as solute per 100 mL of solution, expressed in g / 100 mL.

[0070] In some embodiments, the biomimetic material used to prepare the super-strong biomimetic temporomandibular joint disc 3 further includes any one of the following: 0.5% to 4% (w / v) of umbilical cord Wharton glue, 2% to 4% (w / v) of umbilical cord ECM, 3% to 9% (w / v) of cartilage ECM, and 0.5% to 3% (w / v) of meniscus ECM, accounting for 0.5% to 3% (w / v) of the biomimetic material by weight and volume.

[0071] Wharton's Jelly is a gel-like connective tissue that surrounds the umbilical cord vessels (two umbilical arteries and one umbilical vein), named after the English anatomist Thomas Wharton, who first described it in 1656. It should be noted that different users may experience varying degrees of rejection. Therefore, we preferentially use the user's own umbilical cord Wharton's Jelly from the ultra-strong and resilient biomimetic temporomandibular joint disc 3 for fabrication.

[0072] In some embodiments, the biomimetic material used to prepare the super-strong biomimetic temporomandibular joint disc 3 also includes umbilical Wharton glue, which accounts for 0.5% to 4% (w / v) of the biomimetic material by weight and volume. For example, the Wharton's umbilical cord adhesive can account for 0.50%, 0.54%, 0.58%, 0.6%, 0.7%, 0.72%, 0.75%, 0.8%, 0.9%, 1.0%, 1.2%, 1.5%, 1.75%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.28%, 2.3%, 2.38%, 2.4%, 2.45%, 2.48%, 25%, 2.55%, 2.56%, 2.6%, 2.63%, 2.65%, 2.68%, 2.7%, 2.72%, 2.75%, 2.8%, 2.84%, 2.88%, 2.9%, 3.0%, 3.2%, 3.5%, 3.8%, and 4.0% (w / v) of the biomimetic material.

[0073] In some embodiments, the biomimetic material used to prepare the ultra-strong biomimetic temporomandibular joint disc 3 further includes umbilical cord ECM accounting for 2% to 4% (w / v) of the biomimetic material's mass-volume content. Exemplarily, the mass-volume content of umbilical cord ECM in the biomimetic material can be 2.1%, 2.3%, 2.4%, 2.5%, 2.75%, 2.8%, 3.0%, 3.2%, 3.3%, 3.45%, 3.5%, 3.6%, 3.75%, 3.8%, or 3.9% (w / v). Umbilical cord ECM, or Umbilical Cord Extracellular Matrix, is abbreviated as umbilical cord ECM.

[0074] In some embodiments, the biomimetic material used to prepare the ultra-strong biomimetic temporomandibular joint disc 3 further includes cartilage ECM accounting for 3% to 9% (w / v) of the biomimetic material's mass-volume content. Exemplarily, the cartilage ECM content of the biomimetic material can be 3.0%, 3.2%, 3.3%, 3.45%, 3.5%, 3.6%, 3.75%, 3.8%, 3.9%, 4.1%, 4.3%, 4.5%, 4.8%, 5.0%, 5.3%, 5.5%, 5.8%, 6.0%, 6.3%, 6.5%, 6.8%, 7.0%, 7.2%, 7.5%, 7.8%, 8.0%, 8.3%, 8.5%, or 8.8% (w / v). Cartilage ECM, or cartilage extracellular matrix, is abbreviated as cartilage ECM. Cartilage ECM specifically refers to the extracellular matrix of articular cartilage. It is composed primarily of type II collagen and abundant proteoglycans. This unique composition makes it the gold standard biomimetic material for repairing joint surface defects and regenerating hyaline cartilage (rather than fibrous cartilage). Cartilage ECM scaffolds can maximally mimic the microenvironment of natural cartilage cells.

[0075] In some embodiments, the biomimetic material used to prepare the super-strong biomimetic temporomandibular joint disc 3 further includes meniscus ECM accounting for 0.5% to 3% (w / v) of the biomimetic material's mass-volume content. Exemplarily, the mass-volume content of meniscus ECM in the biomimetic material can be 0.52%, 0.53%, 0.55%, 0.6%, 0.63%, 0.65%, 0.68%, 0.7%, 0.72%, 0.75%, 1.0%, 1.2%, 1.5%, 1.6%, 1.65%, 1.7%, 1.8%, 2.0%, 2.15%, 2.2%, 2.3%, 2.5%, 2.8%, or 3.0% (w / v). Meniscus ECM, or meniscus extracellular matrix, is abbreviated as meniscus ECM. The meniscus extracellular matrix (ECM) is a unique fibrocartilage-derived ECM, which can be selected from the extracellular matrix of the medial meniscus, the extracellular matrix of the lateral meniscus, or a combination of both. The collagen in the meniscus ECM is predominantly type I collagen, giving it excellent tensile strength and enabling it to withstand the complex stresses of the knee joint, unlike articular cartilage which is predominantly type II collagen. The meniscus ECM can be processed into porous scaffolds or hydrogels to serve as templates for guiding tissue regeneration and to provide a biomimetic microenvironment for stem cells.

[0076] In some embodiments, the biomimetic material used to prepare the ultra-strong and tough biomimetic temporomandibular joint disc 3 includes a sustained-release factor, which includes at least one of CTGF and TGF-β.

[0077] The biomimetic material used in this application embodiment for preparing the super-strong biomimetic temporomandibular joint disc 3 supports tissue regeneration through the addition of sustained-release factors, that is, it has the ability to assist endogenous regeneration.

[0078] In some embodiments, the sustained-release factor is loaded with polylactic-co-glycolic acid copolymer (PLGA) microspheres.

[0079] The biomimetic material used in this application embodiment for preparing the ultra-strong biomimetic temporomandibular joint disc 3 is prepared by loading CTGF and TGF-β factors onto polylactic acid-glycolic acid copolymer microspheres, and then printing and photocuring them together with the biomimetic material for preparing the ultra-strong biomimetic temporomandibular joint disc 3 as needed. This achieves the sequential release of sustained-release factors, effectively recruits endogenous stem cells and induces them to differentiate into fibrochondrocytes, and is expected to achieve long-term functional regeneration of the temporomandibular joint disc 3.

[0080] In some embodiments, the polylactic acid-glycolic acid copolymer microspheres include first copolymer microspheres with an average diameter of 200 nm to 1 μm and / or second copolymer microspheres with an average diameter of 20 μm to 50 μm. For example, the polylactic acid-glycolic acid copolymer microspheres include first copolymer microspheres with average diameters of 220, 250, 300, 330, 350, 360, 380, 400, 500, 540, 560, 580, 590, 600, 610, 635, 655, 688, 700, 720, 750, 770, 800, 840, 860, 880, 900, 940, 960, 980, and 1000 nm, and second copolymer microspheres with average diameters of 22, 24, 26, 28, 30, 32, 33, 35, 38, 40, 41, 43, 44, 45, 46, 48, and 50 μm.

[0081] In some embodiments, in the polylactic acid-glycolic acid copolymer microspheres, the proportion of the first copolymer microspheres with an average diameter of 200 nm to 1 μm is 40% to 60%, and the proportion of the second copolymer microspheres with an average diameter of 20 μm to 50 μm is 40% to 60%.

[0082] In some embodiments, the polylactic acid-glycolic acid copolymer microspheres constitute 0.1% (w / v) to 1% (w / v) of the solution by mass volume. It should be noted that the solution here refers to a mixed solution of the biomimetic material comprising at least a first photocurable binder, a second photocurable binder, and a photocuring agent. In other embodiments, the solution further includes one or more of the above-mentioned umbilical cord Wharton's adhesive, umbilical cord ECM, cartilage ECM, and meniscus ECM.

[0083] In some embodiments, in the first copolymer microspheres with an average diameter of 200 nm to 1 μm, the molar percentage of polylactic acid is 45% to 55%, and the molar percentage of glycolic acid-polylactic acid is 45% to 55%. Polylactic acid-glycolic acid copolymers that meet these molar percentages degrade more rapidly.

[0084] In some embodiments, CTGF cytokines are loaded using first copolymer microspheres with an average diameter of 200 nm to 1 μm.

[0085] In some embodiments, the loading concentration of CTGF cytokine is 20 ng / mg to 80 ng / mg.

[0086] In some embodiments, in the second copolymer microspheres with an average diameter of 20 μm to 50 μm, the molar percentage of polylactic acid is 65% to 80%, and the molar percentage of glycolic acid is 20% to 35%.

[0087] In some embodiments, a second copolymer microsphere with an average diameter of 20 μm to 50 μm is used to load the TGF-β factor.

[0088] In some embodiments, the loading concentration of TGF-β factor is 5 ng / mg to 30 ng / mg.

[0089] Figure 2 and Figure 3 A physical image of the ultra-strong and tough bionic temporomandibular joint disc 3 provided in the embodiments of this application is shown.

[0090] Secondly, such as Figure 4 and Figure 5 As shown, an embodiment of this application provides an ultra-strong and tough biomimetic temporomandibular joint disc 3, comprising: a central portion 4, including a first fibroblast-like tissue 40, the first fibroblast-like tissue being arranged in an anterior-posterior direction; and a peripheral portion 5, including a second fibroblast-like tissue 50, the second fibroblast-like tissue being arranged in a ring direction and surrounding the central portion 4.

[0091] The ultra-strong and tough biomimetic temporomandibular joint disc 3 provided in this application embodiment can precisely control the fiber direction through 3D printing technology, restoring the dual-gradient structure of the temporomandibular joint disc 3. This involves a second fibroblast-like tissue arranged in a ring-like pattern on the periphery and a first fibroblast-like cartilage tissue arranged anteroposteriorly in the center, significantly improving the biomimetic degree and structural integrity of the ultra-strong and tough biomimetic temporomandibular joint disc 3. Based on obtaining a good external shape of the temporomandibular joint disc 3, its corresponding tissue structure is also obtained, making the structure of the ultra-strong and tough biomimetic temporomandibular joint disc 3 more suitable for human application conditions, thus ensuring that the deformation and mechanical strength of the ultra-strong and tough biomimetic temporomandibular joint disc 3 meet the usage requirements.

[0092] In some embodiments, such as Figure 6As shown, the ultra-strong and tough bionic temporomandibular joint disc 3 includes a first concave part 31 and a second concave part 32 connected to each other. Its whole body is in the shape of a double concave shallow bowl, that is, it has a double concave bionic shape, so as to connect well with the condyle 21 of the mandible 2, thereby playing a role in stabilizing the joint operation order.

[0093] Thirdly, the method for preparing the ultra-tough bionic temporomandibular joint disc 3 provided in the embodiments of this application includes: obtaining the three-dimensional shape of the temporomandibular joint disc 3; providing the bionic material for preparing the ultra-tough bionic temporomandibular joint disc 3 provided in the embodiments of the first aspect; and forming the ultra-tough bionic temporomandibular joint disc 3 using the bionic material for preparing the ultra-tough bionic temporomandibular joint disc 3 provided in the embodiments of the first aspect according to the three-dimensional shape of the temporomandibular joint disc 3.

[0094] The method for preparing the ultra-strong and tough bionic temporomandibular joint disc 3 provided in this application embodiment offers a complete preparation process for the ultra-strong and tough bionic temporomandibular joint disc 3, providing two preparation paths: injection molding or 3D printing. The appropriate process can be selected according to the material properties and clinical needs, or the two preparation paths can be combined to obtain an ultra-strong and tough bionic temporomandibular joint disc 3 that balances mechanical properties and structural accuracy.

[0095] The method for preparing the ultra-strong and tough bionic temporomandibular joint disc 3 provided in this application first obtains the three-dimensional shape of the temporomandibular joint disc 3. This can be obtained by performing a three-dimensional scan of the shape of the patient's existing temporomandibular joint disc 3, or by modeling the patient's temporomandibular joint position. This application does not explicitly limit this method.

[0096] In some embodiments, a biomimetic material for preparing an ultra-strong and tough biomimetic temporomandibular joint disc 3 is provided, comprising: dissolving a first base material and a second base material for photocuring using a solvent to obtain a hydrogel solution. As mentioned above, deionized water or PBS solution can be used as a solvent to dissolve the first base material or the second base material for photocuring, or to dissolve a mixture of the first base material and the second base material, to prepare the hydrogel solution.

[0097] In some embodiments, based on the three-dimensional shape of the temporomandibular joint disc 3, the ultra-tough biomimetic temporomandibular joint disc 3 is formed using the biomimetic material provided in the first aspect embodiment for preparing the ultra-tough biomimetic temporomandibular joint disc 3, including: forming a central portion 4; and forming a peripheral portion 5 outside the central portion 4.

[0098] In the preparation method of the ultra-tough bionic temporomandibular joint disc 3 provided in this application embodiment, exemplarily, o-nitrobenzyl alcohol modified gelatin is used as the second base material and mixed with the first base material in the preparation of the ultra-tough bionic temporomandibular joint disc 3 by injection molding. When o-nitrobenzyl alcohol modified polyethylene glycol or polyethylene glycol acrylate compounds are used as the second base material and mixed with the first base material to prepare the ultra-tough bionic temporomandibular joint disc 3, a 3D printing method is used. It can be understood that in the preparation of the ultra-tough bionic temporomandibular joint disc 3, o-nitrobenzyl alcohol modified polyethylene glycol or polyethylene glycol acrylate compounds can first be used as the second base material and mixed with the first base material to prepare the central part 4 of the ultra-tough bionic temporomandibular joint disc 3 by 3D printing. Then, the outer peripheral part 5 of the ultra-tough bionic temporomandibular joint disc 3 is prepared by injection molding using o-nitrobenzyl alcohol modified gelatin as the second base material and the first base material. Alternatively, the manufacturing sequence can be reversed. First, use o-nitrobenzyl alcohol modified gelatin as the second base material and mix it with the first base material. Then, use o-nitrobenzyl alcohol modified polyethylene glycol or polyethylene glycol acrylate compounds as the second base materials and mix them with the first base material. Finally, use 3D printing to prepare the outer peripheral part 5.

[0099] like Figure 1 As shown, the temporomandibular joint disc 3 is located at the junction of the mandibular fossa 11 of the temporal bone 1 and the condyle 21 of the mandible 2, i.e., the movable fibrocartilage at the temporomandibular joint. The ultra-strong and tough biomimetic temporomandibular joint disc 3 prepared in this application embodiment is used to replace the defective temporomandibular joint disc 3 to connect the mandibular fossa 11 of the temporal bone 1 and the condyle 21 of the mandible 2, i.e., the temporomandibular joint. Figure 2 , Figure 3 and Figure 6 As shown, the ultra-strong and tough bionic temporomandibular joint disc 3 is shaped like an inverted bowl, which acts as a buffer pad.

[0100] Fourthly, embodiments of this application provide the application of the biomimetic material provided in the first aspect of the embodiment for preparing ultra-tough biomimetic temporomandibular joint disc 3 in the preparation of biomimetic cartilage tissue, which can at least be used for temporomandibular joint disc 3 defect repair, replacement or regeneration induction.

[0101] The technical solution and beneficial effects of this application will be further explained below through specific embodiments and comparative examples. The following are the specifications, requirements, and available sources of some raw materials and organisms; any parts not mentioned can be obtained through commercial channels and will not be elaborated upon here.

[0102] o-Nitrobenzyl alcohol modified gelatin and o-nitrobenzyl alcohol modified polyethylene glycol were both purchased from Shanghai Lingjiu Medical Technology Co., Ltd.; HAMA was purchased from EFL, whose website link can be found at: https: / / www.efl-tech.com / product / cpmlc79 / p1.html.

[0103] Example Preliminary preparations 1. MRI and microCT scans were performed on the human temporomandibular joint disc 3 to obtain its three-dimensional shape, and then 3D reconstruction was performed. MRI and microCT scans were also performed on the temporomandibular joint disc 3 of New Zealand white rabbits and beagles to obtain their three-dimensional shapes, and then 3D reconstruction was performed. It was determined that the temporomandibular joint disc 3 in humans, New Zealand white rabbits, and beagles is a biconcave structure. Among them, such as... Figure 6 As shown, it is a magnetic resonance imaging image of the human mandibular disc, which has a first concave portion 31 and a second concave portion 32.

[0104] 2. Histological staining of the human temporomandibular joint disc 3, including Sirius red staining, was performed. Images were taken using a polarized light microscope, followed by quantitative analysis. (e.g.) Figure 7 The image shown is a Sirius red staining image of the normal temporomandibular joint disc 3. Based on existing technical literature and staining results, the regional divisions, collagen distribution, and fiber orientation of the temporomandibular joint disc 3 were clarified. The results indicate that the natural temporomandibular joint disc 3 has abundant and densely arranged collagen fibers, with certain regional orientation differences. Specifically, the peripheral portion 5 exhibits a circumferential orientation, with the most prominent fibers being... Figure 7 The red and yellow mixed areas in the middle show that it is mainly composed of type I collagen; the central part 4 has an approximately front-to-back orientation, and has both... Figure 7 The yellow area shows type I collagen, and the green area shows type II collagen, thus providing a basis for the design of the bionic articular disc in this application. A detailed analysis follows: Figure 8 yes Figure 7 A 40x magnified view of local region a and its quantitative analysis diagram. Figure 8 The left side shows a 40x magnified view of the peripheral region, and the right side shows a quantitative analysis of the tissue. Figure 8 The red fibers indicate that 70% of the periphery of the temporomandibular joint disc 3 contains only type I collagen. The direction of the collagen fibers is concentrated near the horizontal direction of 0°, which indicates that the collagen fibers are circularly arranged fibrous tissue, that is, the second fibroblast-like tissue 50 arranged in a circular direction.

[0105] Figure 9 yes Figure 7 A 40x magnified view of a local area in the central region b and its quantitative analysis diagram. Figure 9In the image, the left side shows a 40x magnified view of a portion of the central region b, and the right side shows a quantitative analysis of the tissue. Figure 9 The green fibers located in the central region indicate that the central 30% contains both type I and type II collagen. The collagen fibers are concentrated around 90° or -90°, running anteroposteriorly, forming the first fibroblast-like tissue 40 arranged anteroposteriorly. This means the temporomandibular joint disc 3 is a fibrocartilaginous tissue combining anteroposterior and circumferential running patterns, exhibiting a two-gradient structure.

[0106] Example 1 A super-strong and resilient biomimetic temporomandibular joint disc, such as... Figure 4 and Figure 5 As shown, it includes: a central portion 4, including a first fibroblast-like tissue 40, which is arranged in a front-to-back direction; and an outer peripheral portion 5, including a second fibroblast-like tissue 50, which is arranged in a ring direction and surrounds the central portion 4.

[0107] Both the anterior and posterior running structures and the annular running structure are made of biomimetic materials used to prepare the ultra-strong and tough biomimetic temporomandibular joint disc 3. The biomimetic materials used to prepare the ultra-strong and tough biomimetic temporomandibular joint disc 3 include: a first base material for photocuring, 1.5% (w / v), which includes methacrylic anhydride-modified hyaluronic acid, i.e., HAMA; the content of the first base material is its mass content in an aqueous solution. HA refers to hyaluronic acid, a polysaccharide found in the extracellular matrix of chondrocytes. HAMA is HA (hyaluronic acid) modified with MA (methacrylic anhydride) groups, possessing photocuring properties.

[0108] The second binder for photocuring, based on the mass of the first binder, is 10% (w / v); the second binder for photocuring is o-nitrobenzyl alcohol modified gelatin, i.e., GelNB. The content of photoinitiator LAP is 0.05% (w / v).

[0109] Example 2 Example 2 provides a method for preparing a human ultra-tough bionic temporomandibular joint disc 3, comprising: obtaining the three-dimensional shape of the temporomandibular joint disc 3; providing the bionic material provided in Example 1 for preparing the ultra-tough bionic temporomandibular joint disc 3; according to the three-dimensional shape of the temporomandibular joint disc 3, using the bionic material provided in the first aspect of the example for preparing the ultra-tough bionic temporomandibular joint disc 3, comprising: fabricating a mold for forming the temporomandibular joint disc 3 according to the three-dimensional shape of the temporomandibular joint disc 3; pouring a mixed solution of the bionic material provided in Example 1 for preparing the ultra-tough bionic temporomandibular joint disc 3 into the mold; and forming the ultra-tough bionic temporomandibular joint disc 3 by photocuring. The light intensity is 60 mW / cm², the layer exposure time is 10 s, and the layer thickness is 50 μm.

[0110] Example 3 A super-strong and tough biomimetic temporomandibular joint disc 3 includes: a central part 4, including a first fibroblast-like tissue 40, which is arranged in an anterior-posterior direction; and a peripheral part 5, including a second fibroblast-like tissue 50, which is arranged in a ring direction and surrounds the central part 4.

[0111] Both the anterior and posterior running structures and the annular running structure are made of biomimetic materials used to prepare the ultra-strong and tough biomimetic temporomandibular joint disc 3. The biomimetic materials used to prepare the ultra-strong and tough biomimetic temporomandibular joint disc 3 include: a first base material for photocuring, 1.5% (w / v), which includes methacrylic anhydride-modified hyaluronic acid, i.e., HAMA; the content of the first base material is its mass content in an aqueous solution. HA refers to hyaluronic acid, a polysaccharide found in the extracellular matrix of chondrocytes, and HAMA is hyaluronic acid modified with MA (methacrylic anhydride) groups, possessing photocuring properties.

[0112] The second binder for photocuring, based on the mass of the first binder, is 8.5% (w / v); the second binder for photocuring is o-nitrobenzyl alcohol-modified polyethylene glycol, i.e., PEGNB. The photoinitiator LAP content is 0.05% (w / v).

[0113] like Figure 10 As shown, this is a physical image of the final printed human ultra-strong bionic temporomandibular joint disc 3 of Example 3. The final product's dimensions are: major axis 20mm × minor axis 14mm × edge thickness 4mm × central thickness 2mm. It should be noted that the annular pattern of the ultra-strong bionic temporomandibular joint disc 3 is not specifically designed for this application, but rather a characteristic of DLP printing. Because DLP printing involves layer-by-layer photopolymerization, there is a difference in height between layers, naturally forming a concentric circle appearance, which is morphologically similar to the fibrous tissue orientation of a natural human temporomandibular joint disc 3 (i.e., TMJ disc). Figure 11 As shown, this is a stress-strain diagram of the ultra-strong and tough bionic temporomandibular joint disc 3 prepared in Example 3 during tensile strength testing. The test results show that the ultimate tensile strength of the ultra-strong and tough bionic temporomandibular joint disc 3 reaches 5 MPa. Figure 12 As shown, this is a stress-strain diagram of the compressive strength test performed on the ultra-tough bionic temporomandibular joint disc 3 prepared in Example 3. The test results show that the ultimate tensile strength of the ultra-tough bionic temporomandibular joint disc 3 reaches 15 MPa. It should be noted that, due to limitations of the testing equipment, the ultimate tensile strength test of the ultra-tough bionic temporomandibular joint disc 3 only tested up to the compressive strength limit value at 60% deformation.

[0114] Example 4 Example 4 provides a method for fabricating a human ultra-strong and tough bionic temporomandibular joint disc 3, comprising: obtaining the three-dimensional shape of the temporomandibular joint disc 3; providing the bionic material provided in Example 3 for fabricating the ultra-strong and tough bionic temporomandibular joint disc 3; and, based on the three-dimensional shape of the temporomandibular joint disc 3, using the bionic material provided in the first aspect of the example for fabricating the ultra-strong and tough bionic temporomandibular joint disc 3, performing 3D printing to form the ultra-strong and tough bionic temporomandibular joint disc 3. Wherein, as... Figure 4 and Figure 5 As shown, during 3D printing, the front and rear running structures 40 are pre-printed to form the central part 4, and then the annular running structures 50 are printed around the central part 4 to form the outer peripheral part 5. DLP printing parameters: light intensity 60mW / cm², layer exposure time 10s, layer thickness 50μm.

[0115] Example 5 Example 5 provides a super-tough bionic temporomandibular joint disc 3, which differs from Example 1 in that the bionic material used to prepare the super-tough bionic temporomandibular joint disc 3 also includes 2% (w / v) of umbilical cord Wharton's adhesive. The 2% (w / v) umbilical cord Wharton's adhesive improves the biocompatibility of the bionic material and has little impact on the photocuring properties of the bionic material mixture.

[0116] Example 6 Example 6 provides a super-strong and tough biomimetic temporomandibular joint disc 3, which differs from Example 3 in that: the biomimetic material used to prepare the super-strong and tough biomimetic temporomandibular joint disc 3 further includes 0.5% (w / v) polylactic acid-glycolic acid copolymer microspheres, which are used to load sustained-release factors. The average particle sizes are 30 μm and 200 nm, respectively. Specifically, the first copolymer microspheres with an average particle size of 200 nm account for 0.25% (w / v) and are used to load the sustained-release factor CTGF at a concentration of 40 ng / mg; the second copolymer microspheres with an average particle size of 30 μm account for 0.25% (w / v) and are used to load the sustained-release factor TGF-β at a concentration of 10 ng / mg. In the polylactic acid-glycolic acid copolymer microspheres with an average diameter of 200 nm, the molar percentage of polylactic acid is 50%, and the molar percentage of glycolic acid-polylactic acid is 50%. In the polylactic acid-glycolic acid copolymer microspheres with an average diameter of 30 μm, the molar percentage of polylactic acid is 75% and the molar percentage of glycolic acid is 25%. The content of photoinitiator LAP is 0.05% (w / v). DLP printing parameters: light intensity 60 mW / cm², layer exposure time 10 s, layer thickness 50 μm.

[0117] Example 7 Example 7 provides a human ultra-tough bionic temporomandibular joint disc 3, which differs from Example 3 in that: the ultra-tough bionic temporomandibular joint disc 3 uses the material of Example 3 to 3D print the anterior and posterior running tissue 40 to form the central part 4. Then, the central part 4 is fixed with a mold, and the material of the human ultra-tough bionic temporomandibular joint disc 3 of Example 1 is used for infusion to form a ring-shaped running tissue 50 around the central part 4 to form the peripheral part 5, thus obtaining the ultra-tough bionic temporomandibular joint disc 3. DLP printing parameters: light intensity 60mW / cm², layer exposure time 10s, layer thickness 50μm.

[0118] It should be noted that the temporomandibular joint discs 3 in Examples 1, 3, 5-7 and Comparative Examples 1-3 were all prepared with deionized water to form corresponding mixed solutions for photocuring.

[0119] Comparative Example 1 Comparative Example 1 provides an existing human bionic temporomandibular joint disc 3, which differs from Example 7 in that it is made of only Proplast-Teflon material to create the temporomandibular joint disc 3 prosthesis. In Comparative Example 1, a portion of the surgical suture is inserted into the hydrogel solution and embedded after the hydrogel solution is cross-linked and light-cured. The two ends of a surgical suture are connected to the human super-strong bionic temporomandibular joint disc 3 formed by the hydrogel, which can be used for knotting during suturing.

[0120] Comparative Example 2 Comparative Example 2 provides an existing human biomimetic temporomandibular joint disc 3, which is a high-performance, non-degradable temporomandibular joint disc 3 prosthesis made of polycaprolactone (PCL) material.

[0121] Comparative Example 3 Comparative Example 3 provides an existing human biomimetic temporomandibular joint disc 3, which is made of polycaprolactone (PCL) and polyvinyl alcohol (PVA) materials to create the temporomandibular joint disc 3 prosthesis. PCL powder is dissolved in CH... Cl PCL implants were fabricated using a layer-by-layer deposition technique with a 3D Bioprinter V2.0 (Hangzhou Genofe Biotechnology Co., Ltd.). The printed PCL structure was then injected into a 15wt% PVA hydrogel solution, and the PVA hydrogel was subjected to three freeze-thaw cycles (-20°C for 12 hours, thawed at room temperature for 2 hours) to achieve network cross-linking. Finally, the PCL implants filled with PVA hydrogel were immersed in a 33wt% NaCl solution for 12 hours. PCL samples (without PVA) for Comparative Example 2 were prepared in a similar manner.

[0122] The temporomandibular joint discs 3 of Examples 1, 3, 5-7 and Comparative Examples 1-3 were subjected to performance tests. The test items are as follows and recorded in Table 1 below.

[0123] (1) Tensile strength and compressive strength test Tensile strength was tested according to ASTM F2451-05(2020), and compressive strength was tested according to ASTM F2118-14 / GB / T 1041.

[0124] (2) Biocompatibility and biodegradability The biodegradability of the material was evaluated according to ASTM F2150-19, "Standard Guidelines for Characterization and Testing of Biomaterial Scaffolds for Regenerative Medicine and Tissue Engineering Medical Products". The biocompatibility of the ultra-tough biomimetic temporomandibular joint disc 3 was evaluated for in-cartilage implantation according to YY / T 0606.10-2008, "Tissue Engineering Medical Products Part 10: Guidelines for In-Vitro Evaluation of Implants for Repairing or Regenerating Articular Cartilage".

[0125] The following are partial test results of the temporomandibular joint disc 3 in Examples 1, 3 and 5-7 of this application and Comparative Examples 1-3.

[0126] Table 1

[0127] As can be seen from Table 1, the ultra-tough bionic temporomandibular joint disc 3 prepared in Examples 1, 3 and 5-7 of this application has tensile strength and compressive strength that meet the requirements of human bionic temporomandibular joint disc 3. It also has good biocompatibility and good degradability. With the addition of sustained-release factors, it has excellent tissue regeneration ability and can further be compatible with the tissues around the temporomandibular joint disc 3.

[0128] The ultra-strong and tough biomimetic temporomandibular joint disc 3 prepared in Examples 1, 3 and 5-7 of this application, as an implantable medical device, achieves high biocompatibility and high degree of biomimicry while conforming to the joint disc morphology of Chinese people and meeting mechanical performance requirements, and provides certain assistance for the endogenous regeneration of the temporomandibular joint.

[0129] The above description is merely a specific implementation of this application. Those skilled in the art will clearly understand that, for the sake of convenience and brevity, the specific working process described above can be referred to the corresponding process in the foregoing method embodiments, and will not be repeated here. It should be understood that the protection scope of this application is not limited thereto. Any person skilled in the art can easily conceive of various equivalent modifications or substitutions within the technical scope disclosed in this application, and these modifications or substitutions should all be covered within the protection scope of this application.

Claims

1. A biomimetic material for preparing ultra-strong and tough biomimetic temporomandibular joint discs, characterized in that, include: The first base material for photocuring is 1% to 3% (w / v); said first base material for photocuring includes one or more combinations of alkyl acrylic anhydride-modified hyaluronic acid, chitosan, chondroitin sulfate, and silk fibroin; and... The second base material for photocuring is 7% to 9% (w / v); the second base material for photocuring includes at least one of o-nitrobenzyl alcohol modified gelatin, o-nitrobenzyl alcohol modified polyethylene glycol, and polyethylene glycol acrylate compounds. The remainder is deionized water or PBS aqueous solution.

2. The biomimetic material for preparing an ultra-strong and tough biomimetic temporomandibular joint disc according to claim 1, characterized in that, The first base material for photocuring includes one or more combinations of C1-C6 alkyl acrylic anhydride modified hyaluronic acid, chitosan, chondroitin sulfate, and silk fibroin. Optionally, the mass ratio of the o-nitrobenzyl alcohol-modified gelatin to the alkyl acrylic anhydride-modified hyaluronic acid is (5-8):1; Optionally, the mass ratio of the o-nitrobenzyl alcohol-modified polyethylene glycol to the alkyl acrylic anhydride-modified hyaluronic acid is (5-7):1; Optionally, the biomimetic material further includes a photoinitiator selected from LAP, wherein the LAP accounts for 0.01% to 0.1% (w / v) of the biomimetic material by mass.

3. The biomimetic material for preparing an ultra-strong and tough biomimetic temporomandibular joint disc according to claim 2, characterized in that, The C1-C6 alkyl acrylic anhydrides include one or more combinations of methacrylic anhydride, ethylacrylic anhydride, propylacrylic anhydride, isopropylacrylic anhydride, n-butylacrylic anhydride, isobutylacrylic anhydride, tert-butylacrylic anhydride, n-pentylacrylic anhydride, n-hexylacrylic anhydride, 3-methylpentylacrylic anhydride, and 2-ethylbutylacrylic anhydride.

4. The biomimetic material for preparing an ultra-strong and tough biomimetic temporomandibular joint disc according to claim 1, characterized in that, The polyethylene glycol acrylate compounds include one or more combinations of polyethylene glycol methacrylate, polyethylene glycol diacrylate, polyethylene glycol triacrylate, and multi-arm PEG-acrylate.

5. The biomimetic material for preparing an ultra-strong and tough biomimetic temporomandibular joint disc according to claim 1, characterized in that, It also includes any one or a combination of several of the following: 0.5% to 4% (w / v) of umbilical cord Wharton glue, 2% to 4% (w / v) of umbilical cord ECM, 3% to 9% (w / v) of cartilage ECM, and 0.5% to 3% (w / v) of meniscus ECM.

6. The biomimetic material for preparing an ultra-strong and tough biomimetic temporomandibular joint disc according to claim 1, characterized in that, It also includes a sustained-release factor, which includes at least one of CTGF and TGF-β, and the sustained-release factor is loaded onto polylactic acid-glycolic acid copolymer microspheres.

7. The biomimetic material for preparing an ultra-strong and tough biomimetic temporomandibular joint disc according to claim 6, characterized in that, The polylactic acid-glycolic acid copolymer microspheres include first copolymer microspheres with an average diameter of 200 nm to 1 μm and / or second copolymer microspheres with an average diameter of 20 μm to 50 μm. Optionally, the polylactic acid-glycolic acid copolymer microspheres account for 0.1% (w / v) to 1% (w / v) of the solution by mass volume. Optionally, in the polylactic acid-glycolic acid copolymer microspheres, the proportion of the first copolymer microspheres with an average diameter of 200 nm to 1 μm is 40% to 60%, and the proportion of the second copolymer microspheres with an average diameter of 20 μm to 50 μm is 40% to 60%. Optionally, in the first copolymer microspheres with an average diameter of 200 nm to 1 μm, the molar percentage of polylactic acid is 45% to 55%, and the molar percentage of glycolic acid polylactic acid is 45% to 55%. Optionally, CTGF cytokines are loaded onto first copolymer microspheres with an average diameter of 200 nm to 1 μm. Optionally, the loading concentration of CTGF cytokine is 20 ng / mg to 80 ng / mg; Optionally, in the second copolymer microspheres with an average diameter of 20 μm to 50 μm, the molar percentage of polylactic acid is 65% to 80%, and the molar percentage of glycolic acid is 20% to 35%. Optionally, a second copolymer microsphere with an average diameter of 20 μm to 50 μm is used to load the TGF-β factor; Optionally, the loading concentration of TGF-β factor is 5 ng / mg to 30 ng / mg.

8. A super-strong and tough biomimetic temporomandibular joint disc, characterized in that, include: The central portion includes a first fibroblast-like tissue, which is arranged in a front-to-back direction. The peripheral portion includes a second fibroblast-like tissue arranged in a ring-shaped pattern, surrounding the central portion; both the central portion and the peripheral portion are made of the biomimetic material for preparing an ultra-strong and tough biomimetic temporomandibular joint disc as described in any one of claims 1-7.

9. A method for preparing an ultra-strong and tough biomimetic temporomandibular joint disc, characterized in that, include: Obtain the three-dimensional shape of the temporomandibular joint disc; Provide biomimetic materials for fabricating ultra-strong and tough biomimetic temporomandibular joint discs; Based on the three-dimensional shape of the temporomandibular joint disc, an ultra-tough biomimetic temporomandibular joint disc is formed using the biomimetic material described in any one of claims 1-7 for preparing an ultra-tough biomimetic temporomandibular joint disc.

10. The application of the biomimetic material for preparing an ultra-tough biomimetic temporomandibular joint disc as described in any one of claims 1-7 in the preparation of biomimetic cartilage tissue.

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