CN111450316B

CN111450316B - Integrated bracket for simulating bone-tendon-bone mineralization-non-mineralization gradient structure

Info

Publication number: CN111450316B
Application number: CN202010144383.5A
Authority: CN
Inventors: 莫秀梅; 谢宪瑞
Original assignee: Shenzhen Meijie Life Science Co ltd
Current assignee: Shenzhen Meijie Life Science Co.,Ltd.
Priority date: 2020-03-04
Filing date: 2020-03-04
Publication date: 2021-07-20
Anticipated expiration: 2040-03-04
Also published as: CN111450316A

Abstract

The invention relates to an integrated bracket for simulating a bone-tendon-bone mineralization to non-mineralization gradient structure, wherein warps are nanofiber yarns with zero mineral content, wefts in the middle of the bracket are nanofiber yarns with zero mineral content, and the wefts in the middle to two ends are sequentially nanofiber yarns with gradually increased mineral content. The integrated bracket for simulating the bone-tendon-bone mineralization-non-mineralization gradient structure constructed by the invention provides an ideal microenvironment for repairing tendon defects, realizes the guidance of tendon tissue regeneration after implantation, promotes tendon-bone healing, and is very suitable for tendon-bone injury repair.

Description

Integrated bracket for simulating bone-tendon-bone mineralization-non-mineralization gradient structure

Technical Field

The invention belongs to the field of tendon/ligament scaffolds, and particularly relates to an integrated scaffold for simulating a gradient structure from bone-tendon-bone mineralization to non-mineralization.

Background

Tendon/ligament (T/L) refers to a tough bundle of regularly arranged connective tissue connecting muscles and bones, transferring force from the muscles to the bones, allowing stabilization and movement of joint function. Since the tendon has a rigidity between that of the muscle and the bone, it acts as an elastic buffer between the muscle and the bone. T/L injury, mainly caused by severe impact or frequent stretching at low intensity, including fracture, laceration, degeneration or inflammation, has become one of the most common injuries in humans. Currently, standard treatment methods for tendon repair require surgical intervention, most commonly autografts, allografts, artificial prostheses and sutures, and subsequent rehabilitation therapy. However, surgically repaired tendons fail to restore the functional, structural and biomechanical properties of healthy tendons. Tendon-bone junctions are natural 4-layer structures including tendon tissue, non-calcified fibrocartilage tissue, calcified fibrocartilage tissue and bone tissue. Clinically, tendon-bone healing is a slow process related to multiple factors, and the healing effect is poor, mainly because the interface between soft and hard tissues is easy to form scar, and the fibrous cartilage area is lack of blood supply and bone mass loss due to injury, so that the reconstruction of the 4-layer structure is more difficult. The tendon-bone interface tissue has a complex gradient structure and shows the characteristics of multi-tissue transition, composition, biological characteristics, cell phenotype and the like of multiple layers, so that the tendon-bone interface tissue is extremely challenging in engineering. At the same time, a challenging problem is presented: repair requires strength and flexibility to accommodate the forces of daily living activities and to avoid extension or breakage of the repair site. However, rivets and sutures used for repair cause stress concentration, limiting the adhesion strength. Therefore, the search for a novel method for promoting the physiological regeneration and repair of the tendon has extremely important clinical significance.

At present, the reconstructed bone tract segment and joint cavity segment between the artificial ligament and the host bone have certain difference in histological characteristics. Ligament prosthesis lacks osteoconductivity, autologous bone can not grow into the artificial ligament effectively, resulting in graft-bone healing disorder and serious bone canal enlargement; at the same time, the adhesion and proliferation of cells are influenced, so that the cells are difficult to self-assemble in joint cavities. The existing artificial ligament can not realize the complicated layered transition structure from mineralized to non-mineralized tissues at the tendon-bone interface, and is difficult to achieve the ideal repairing effect of the tendon-bone interface, thereby influencing the tendon/ligament function.

In view of the above problems, there is a need for an improved structure of the existing artificial tendon/ligament, which has fine adjustment of its components. In recent years, researches prove that the electrospun nanofiber can simulate a filamentous structure of an extracellular matrix of a tissue, has a larger specific surface area, is beneficial to adsorbing more proteins, can provide more adhesion sites for receptors on a cell membrane, promotes adhesion, proliferation and differentiation of cells, regulates a signal path for controlling transcriptional activity and gene expression in the cells, guides directional arrangement of cytoskeletal proteins, and is widely applied to tissue repair scaffolds, particularly tendon/ligament repair. The nano yarn is similar to collagen fiber formed by a plurality of triple-helical structures, and well simulates a nano filamentous structure of tendon extracellular matrix. The three-dimensional scaffold obtained by combining the electrospinning nano yarn technology with the traditional weaving technology can simulate a multilayer structure of a natural tendon/ligament tissue from a nanometer level to a macroscopic level, better simulate the shape of the natural tendon/ligament tissue and promote the regeneration of the tendon tissue.

CN 208481528U discloses a mixed artificial ligament, but the ligament material is only composed of biodegradable and non-degradable parts, and can not simulate the mineralization of a natural tendon-bone combination part to a non-mineralized gradient structure.

Disclosure of Invention

The invention aims to solve the technical problem of providing an integrated bracket for simulating a bone-tendon-bone mineralization to non-mineralization gradient structure, which fills the blank that an artificial tendon/ligament bracket cannot simulate the integrated bracket for simulating the bone-tendon-bone mineralization to the non-mineralization gradient structure.

The integrated bracket is characterized in that the warps are nanofiber yarns (J) with zero mineral content, the wefts in the middle of the bracket are nanofiber yarns (W1) with zero mineral content, and the wefts in the middle to two ends are sequentially nanofiber yarns (W2, W3 and W4) with gradually increased mineral content.

The integrated stent appearance includes, but is not limited to, an elongated shape, a flat shape, a cylinder.

The nanofiber yarn is of a core-spun structure, wherein a core layer is a polymer yarn, an outer layer is an electrospun nanofiber, the nanofiber wraps the polymer yarn to form the nanofiber yarn, and in the nanofiber yarn with zero mineral content, the electrospun nanofiber on the outer layer contains a polymer and an electrospun nanofiber of a natural polymer, but does not contain minerals; in the mineral-loaded nanofiber yarn, the outer layer electrical discharge nanofiber contains minerals, polymers and natural macromolecules.

The polymer yarn is a single yarn with the diameter of 0.03-0.1 mm formed by merging and drafting a plurality of polymer single fibers with the diameter of 20-50 microns.

The polymer thread is one or more of polyethylene terephthalate (PET) thread, polylactic acid (PLA) thread, Polycaprolactone (PCL) thread, lactic acid-glycolic acid copolymer (PLGA) thread and Silk Fibroin (SF) thread; the polymer is one or more of lactic acid-caprolactone copolymer PLCL, polycaprolactone PCL and lactic acid-glycolic acid copolymer PLGA; the natural polymer is one or more of silk fibroin SF, Collagen (COL) and gelatin.

The mineral is one or more of hydroxyapatite HA, bioactive glass, silicon dioxide, calcium carbonate and calcium phosphate mineral.

Further, the nano-fiber with zero mineral content comprises polymer and natural polymer nano-fiber; wherein, preferably, the polymer comprises one or more of lactic acid-caprolactone copolymer, polycaprolactone and polylactic acid-glycolic acid copolymer; the natural polymer comprises one or more of silk fibroin, collagen and gelatin.

Further, the mineral nanofiber comprises a polymer, a natural polymer and an added mineral blending nanofiber; wherein, preferably, the polymer comprises one or more of lactic acid-caprolactone copolymer, polycaprolactone and polylactic acid-glycolic acid copolymer; the natural polymer comprises one or more of silk fibroin, collagen and gelatin; the mineral is hydroxyapatite, bioactive glass, silicon dioxide, calcium carbonate, or calcium phosphate.

The integrated bracket comprises a mineralized part and a non-mineralized part, wherein the non-mineralized part is positioned in the middle of the bracket, and the mineralized part is positioned at two ends of the bracket.

The integrated bracket can simulate the histological characteristics of the bone tract section and the joint cavity section, and the mineralized part (W2, W3 and W4) has osteoconductivity and accelerates the effective growth of autogenous bones of the bone tract section; the non-mineralised (W1) portion is capable of promoting its automization within the joint cavity. (the mineralized part of the scaffold is W2, W3 and W4; the non-mineralized part is W1; the warp is J containing no mineral substance; J and W1 are the same material and are completely the same, but J and W1 are used for distinguishing the warp from the weft respectively, and can be represented by the same letter)

The length of the integrated bracket is 5-50 mm, the width of the integrated bracket is 5-20 mm, and the length of the non-mineralized part is 5-30 mm; the unilateral mineralization length is 10-20 mm.

The mineralized part is woven by nano fiber yarns with different mineral contents, and the non-mineralized part is woven by nano fiber yarns with zero mineral content.

The length of the support is 5-50 mm, and the width of the support is 5-20 mm.

The support is as follows: the nanofiber yarn with zero mineral content is used as warp yarns, the distance between the two warp yarns is 0.5-2 mm, and 10 warp yarns are adopted for each support; the middle of the bracket uses nanofiber yarns with zero mineral content as wefts, the nanofiber yarns with gradually increased mineral content are sequentially used as wefts from the middle to two sides, the weaving length of each weft is 2-10 mm, and the nanofiber yarns in the warp direction and the weft direction are interwoven to form the bracket.

Further, the nanofiber yarn with zero mineral content is: the PLCL/SF nanofibers are twisted onto the core PLA string.

Further, nanofiber yarns with increasing mineral content gradient: PLA-PLCL/SF/HA nanofiber yarn, the content of loaded HA is 5%, 10% and 15%.

The invention discloses a preparation method of an integrated bracket, which comprises the following steps:

(1) respectively preparing polymer-containing and natural polymer blending spinning solutions; the blended spinning solution contains polymers, natural polymers and minerals with different contents;

(2) passing a core layer line through a hollow rotating funnel, then connecting the blended spinning solution obtained in the step (1) to two opposite-spraying spinning heads, respectively applying positive and negative electric fields to the spraying spinning heads, spraying the nanofibers on the middle rotating funnel and the core layer line, twisting the nanofibers to the core layer line by the rotating funnel, and receiving by a receiving roller to obtain nanofiber yarns with zero mineral content and nanofiber yarns loaded with minerals with different content;

(3) the mineral content of the nanofiber yarns is zero, the mineral content of the nanofiber yarns is different, the nanofiber yarns in the warp direction and the weft direction are interwoven by a weaving method, the mineral content of the nanofiber yarns is zero in the weft of the middle of the bracket, the mineral content of the nanofiber yarns gradually increasing from the middle to the two ends of the bracket is sequentially adopted as the weft, and the integrated bracket with the mineralized to non-mineralized gradient structure is obtained.

The preferred mode of the above preparation method is as follows:

the mass fraction of the spinning solution in the step (1) is between 5 and 20 percent. Further, 6%, 8%, 9%, 10.5%, 15%, etc., more preferably 8%.

The mass ratio of the polymer to the natural polymer in the step (1) is 90: 10-10: 90; preferably, the mass ratio of the polymer to the natural macromolecule is 90:10, 80:20, 70:30, 75:25, 60:40, 50:50, 40:60, 25:75, 30:70, 20:80 or 10: 90.

The mass fraction of the added minerals in the blended spinning solution is 0-20%, preferably 5%, 10% and 15%.

The solvent for preparing the spinning solution in the step (1) is one or more of hexafluoroisopropanol, trifluoroethanol, dichloromethane and trifluoroacetic acid.

The specific processes of electrostatic spinning in the step (2) are as follows: adding the spinning solution into an injector, respectively connecting spinning nozzles at the left end and the right end of a rotating funnel, respectively applying positive pressure and negative pressure of 8KV, respectively, wherein the speed of a propulsion pump is 1.2mL/h, the receiving distance is 12cm, the rotating speed of the rotating funnel is 400 rpm, and the receiving roller is 8 rpm.

The invention provides an integrated bracket prepared by the method.

The invention provides an application of the integrated stent as a tendon and ligament stent.

The invention firstly prepares a core-spun nanofiber yarn, a spraying nozzle is respectively added with a positive electric field and a negative electric field, a core layer wire passes through a hollow rotating funnel to be used as a receiving device, after two nozzles spray nanofibers to the rotating funnel in the middle and the core layer wire, the rotating funnel twists the nanofibers to the core layer wire, and a receiving roller receives the nanofibers to obtain the continuous nanofiber yarn. Then, the integrated bracket is prepared by interweaving the nanofiber yarns along the warp direction and the weft direction by a weaving method. The integrated bracket is prepared by adopting nanofiber yarns with zero mineral content as warps, adopting nanofiber yarns with zero mineral content as wefts in the middle part, and sequentially adopting nanofiber yarns with gradually increased mineral content as wefts from the middle part to two ends.

Advantageous effects

(1) The invention provides an integrated bracket for simulating a bone-tendon-bone mineralization to non-mineralization gradient structure, which consists of a mineralization part and a non-mineralization part; the mineralized part has osteoconductivity, promotes the autologous bone of the bone tract section to effectively grow in, and the non-mineralized part guides and promotes the endogenous repair of the tendon, so that the joint cavity is self-formed (as shown in an alizarin red test data figure 8 in the example).

(2) The integrated scaffold for simulating the bone-tendon-bone mineralization-non-mineralization gradient structure provided by the invention has the advantages that the pore size of the scaffold is suitable (the pore size of the scaffold is 40-70 mu m), the problems that the pore size of a nanofiber scaffold is small and cells are difficult to grow in are solved, the three-dimensional cell growing-in capacity is improved, the scaffold is reorganized and fibrillated along with the proliferation and growing of tendon/ligament self tissues, and a community is formed by the scaffold and the human body self tissues, so that a better motion function and motion feeling are provided for a patient.

(3) The polymer and the natural polymer used in the invention are biodegradable materials, can be gradually degraded in a certain period, and are beneficial to the formation of regenerated tendon/ligament tissues. The degradation product can be discharged along with the normal metabolism of the human body, has no side effect on the human body, and achieves the ideal tendon/ligament reconstruction and repair (the used material is degradable material, and has no side effect after in vivo degradation).

(4) The composite material nanofiber yarn is used as the minimum weaving unit, and compared with the single material yarn, the composite material nanofiber can exert the advantages of various repair materials. Meanwhile, the woven structure is finer, the strength is higher, and higher mechanical strength can be provided. (As shown in the SEM pictures, it can be seen that the structure is dense, which results in high mechanical strength.)

(5) The bone-tendon-bone mineralization simulation to non-mineralization gradient structure integrated scaffold provided by the invention can be added with anti-inflammatory and tissue adhesion prevention medicines or growth factors in the preparation process according to clinical requirements, so that the inflammatory reaction is relieved, the tissue adhesion is avoided, and the tendon repair and normal function recovery are promoted.

Drawings

FIG. 1 is a schematic diagram of a process for preparing an electrospun nanofiber yarn;

FIG. 2 is a photograph of an integrated scaffold simulating mineralization of bone-tendon-bone to non-mineralization gradient structure;

fig. 3 is a mic-CT scan picture of a scaffold integrating a gradient structure simulating bone-tendon-bone mineralization to non-mineralization, wherein (a) is a front view, (b) is an oblique view, and (c) is a side view;

FIG. 4 is a scanning electron microscope (a) and a transmission electron microscope (b) of a single nanofiber yarn surface loaded with hydroxyapatite of different contents;

FIG. 5 is a scanning electron microscope image of the non-mineralized portion (PPS-0% HA) and the gradient mineralized portion (PPS-5% HA, PPS-10% HA, PPS-15% HA) of the integrated scaffold;

FIG. 6 is a graph showing water contact angles of portions of the integrated scaffold simulating mineralization of bone-tendon-bone to non-mineralization gradient structure;

FIG. 7 shows the activity of each part of the integrated scaffold simulating the mineralization of bone-tendon-bone to non-mineralization gradient structure on rat bone marrow mesenchymal stem cells (BMSC) at various time points (1 day, 4 days, 7 days);

FIG. 8 shows calcium content expression of each part of the alizarin red quantitative determination integrated scaffold acting on BMSC cells, respectively;

FIG. 9 is a schematic view of an integrated bracket structure of the present invention; wherein (J) is the nano fiber yarn with zero mineral content as warp yarn, (W1, W2, W3 and W4) are the nano fiber yarn with 0%, 5%, 10% and 15% mineral content as weft yarn.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.

Example 1

(1) Extracting silk fibroin: weighing 60g of silkworm cocoon, cutting into a plurality of layers, putting the silkworm cocoon into 0.5% sodium bicarbonate water solution, boiling for 30 minutes, repeating twice to remove surface sericin, fully cleaning with deionized water, and airing to obtain the silk fibroin fiber. Placing silk fibroin fiber into CaCl₂/H₂Dissolving in ternary solvent of O/anhydrous ethanol (molar ratio of 1:8:2) at 70 deg.C for 40 min, dialyzing in dialysis bag with molecular weight cutoff of 14000Da at room temperature for 7 days, filtering to obtain pure silk fibroin aqueous solution, and freeze drying to obtain spongy silk fibroin.

(2) Preparing a spinning solution: 0.6g of P (LLA-CL) (75:25) (available from the bio-technology company, Inc. of the handle of the Tibei, Jinan, Ltd.) having a molecular weight of about twenty thousand and 0.2g of silk fibroin were dissolved in 10mL of hexafluoroisopropanol (available from the fine chemical company, Ltd. of the Shanghai, Dai, Ltd.) to prepare a PLCL/SF spinning solution having a mass concentration of 8%.

(3) Preparing a loaded mineral spinning solution: firstly, preparing PLCL/SF spinning solution with the mass concentration of 8% according to the step (2), then respectively adding hydroxyapatite (purchased from Shanghai Aladdin Biotechnology GmbH, purity is more than or equal to 97%, and particle size is less than 100nm) with the mass percentage of 5%, 10% and 15%, and ultrasonically stirring and uniformly dispersing.

(4) Preparation of PLA-PLCL/SF (PPS-0% HA) nanofiber yarn: a PLA thread (a single thread with the diameter of about 180 microns formed by merging and drafting a plurality of PLA single fibers with the diameter of about 30 microns; from China center laboratory of fiber material modification of Donghua university) passes through a rotating funnel of electrospinning nanofiber yarn preparation equipment to serve as a core layer, the PLCL/SF spinning solution in the step (2) is sprayed oppositely, the PLCL/SF nanofibers are twisted onto the PLA thread of the core layer by the rotating funnel, and a receiving roller collects continuous PPS-0% HA nanofiber yarn.

(5) Preparing PLA-PLCL/SF/HA nanofiber yarns: and (3) allowing a PLA line to pass through a rotating funnel of electrospinning nanofiber yarn preparation equipment to serve as a core layer, oppositely spraying PLCL/SF/HA spinning solutions in the step (3), twisting PLCL/SF/HA nanofibers onto the PLA line of the core layer by using the rotating funnel, collecting continuous PLA-PLCL/SF/HA nanofiber yarns by using a receiving roller, wherein the loaded HA content is 5%, 10% and 15% of the yarns are respectively marked as PPS-5% HA, PPS-10% HA and PPS-15% HA.

(6) The electrostatic spinning process of the nano yarn in the steps (4) to (5) comprises the following steps: adding the spinning solution into an injector, then respectively connecting spinning nozzles at the left end and the right end of the yarn, respectively applying positive pressure and negative pressure of 8KV, respectively, wherein the speed of a propulsion pump is 1.2mL/h, the receiving distance is 12cm, the rotating speed of a rotating funnel is 400 r/min, and the receiving roller is 8 r/min.

Example 2

(1) Preparing an integrated bracket with a mineralized to non-mineralized gradient structure: PPS-0% HA nanofiber yarns are used as warp yarns, PLA-PLCL/SF/HA nanofiber yarns loaded with different mineral contents are used as weft yarns, the PLA-PLCL/SF yarns with zero mineral content are used as weft yarns in the middle of the bracket, the PPS-5% HA, the PPS-10% HA and the PPS-15% HA nanofiber yarns are sequentially used as weft yarns from the middle to two ends, and the integrated bracket with the mineralized to non-mineralized gradient structure is prepared by interweaving the nanofiber yarns in the warp and weft directions by a weaving method.

(2) The weaving process in the step (1) comprises the following steps: on a semi-automatic weaving machine, PPS-0% HA nanofiber yarns are used as warps, the distance between the two warps is 0.5-2 mm, and each support adopts 10 warps; the bracket is characterized in that PPS-0% HA is used as weft yarns in the middle of the bracket, PPS-5% HA, PPS-10% HA and PPS-15% HA are used as weft yarns from the middle to two sides in sequence, the weaving length of each weft yarn is 2-10 mm, and the weft yarns are interwoven by nanofiber yarns in the warp direction and the weft direction.

(3) After weaving is finished, the prepared ribbon-shaped integrated scaffold is subjected to ultrasonic cleaning and freeze drying, and then ethylene oxide sterilization treatment is carried out for 24 hours, so that the integrated scaffold simulating mineralization of bone-tendon-bone to a non-mineralized gradient structure is finally obtained, and the integrated scaffold can be used for artificial tendon/ligament.

The integrated scaffold object with the bone-tendon-bone mineralization-non-mineralization gradient structure obtained in the embodiment 2 is shown in fig. 2, the scaffold is formed by interweaving nanofiber yarns in the warp direction and the weft direction, the processing is relatively simple, and the length, the width, and the size of the mineralized part and the non-mineralized part of the scaffold can be adjusted according to actual requirements.

The surface scanning electron microscope (a) and the transmission electron microscope (b) loaded with the hydroxyapatite nanofibers with different contents obtained in the above example 2 are shown in fig. 4, the diameter of the nanofiber yarn is 200-300 μm, the diameter of a single fiber is 500-800nm, and the pore diameter of the scaffold is 40-70 μm. The surface of the yarn without HA was free of HA deposition, and TEM showed no HA inside the single nanofiber; with the increase of HA in the spinning solution, the deposition of HA on the surface of the yarn is increased, and the HA inside the nanofiber gradually becomes more, which shows that HA is not only deposited on the surface of the fiber, but also successfully electrospun into the inside of the nanofiber.

Scanning electron microscopy of the non-mineralized portion (PPS-0% HA) and the gradient mineralized portion (PPS-5% HA, PPS-10% HA, PPS-15% HA) of the integrated scaffold obtained in example 2 is shown in FIG. 5, and the scaffold HAs a three-dimensional woven structure, which facilitates cell growth into the interior of the scaffold and can simulate the fibrous structure of extracellular matrix of tendon tissue.

The integrated holder obtained in example 2 was measured for the water contact angle of the surface of each part using a contact angle measuring instrument (DSA 100). Respectively and horizontally placing part of water on the support on an objective table of a contact angle meter, adjusting the size (the diameter is 5 mu L) of the distilled water drop, dropping the distilled water drop on the flat part of the surface of the support, measuring the size of the contact angle within 5s after dropping the distilled water and recording data. The surface water contact angles of all parts of the integrated bracket are shown in figure 6, the contact angle of a non-mineralized part (PPS-0% HA) is 111.8 degrees, the contact angles of mineralized parts (PPS-5% HA, PPS-10% HA and PPS-15% HA) are gradually reduced to 93.5 degrees, 69.3 degrees and 51.2 degrees along with the increase of the HA content, and the contact angles are favorable for cell adhesion growth.

The proliferation activity of the integrated scaffold on BMSC cells is tested by adopting a CCK-8 method, each part of the scaffold is cut into a round piece with the diameter of 14mm, and the round piece is placed in a hole of a 24-hole cell culture plate after being fumigated by alcohol and subjected to ultraviolet sterilization. 2.5 million BMSC cells are planted in each well and put into a carbon dioxide incubator for culture, and the absorbance value of each well is measured at 450nm by using a CCK-8 reagent on days 1, 4 and 7 respectively. As shown in FIG. 7, the OD values of the groups were significantly increased from day 1 to day 7, and the OD values of the PPS-0% HA group at day 7 were significantly higher than those of PPS-5% HA, PPS-10% HA and PPS-15% HA. The scaffold has no obvious cytotoxicity, good cell proliferation condition and good cell compatibility; meanwhile, the addition of hydroxyapatite has certain influence on the cell proliferation rate.

And (3) detecting the bone differentiation capacity of each part of the integrated scaffold for inducing BMSC cells by using an alizarin red quantitative detection kit, cutting each part of the scaffold into round pieces with the diameter of 14mm, fumigating and ultraviolet sterilizing by using alcohol, and placing the round pieces into holes of 24-hole cell culture plates. 2.5 ten thousand BMSC cells are planted in each hole and put into a carbon dioxide incubator for culture, and the absorbance value of each hole is measured at 562nm by using an alizarin red quantitative detection kit on the 21 st day. As shown in FIG. 8, the OD values of the PPS-5% HA, PPS-10% HA, and PPS-15% HA groups were significantly higher than those of the PPS-0% HA group at 21 days, and the OD values gradually increased as the HA content increased. The integrated bracket mineralized part is shown to have the function of promoting the differentiation of stem cells and bones, and the bracket has the potential of guiding the regeneration of tendon tissues and promoting the healing of tendon and bones after being implanted.

Claims

1. The integrated support comprises warps and wefts and is characterized in that the warps are nanofiber yarns with zero mineral content, the wefts in the middle of the support are nanofiber yarns with zero mineral content, and the wefts in the middle of the support are sequentially nanofiber yarns with gradually increased mineral content.

2. The stent of claim 1, wherein the mineral-content zero nanofiber yarn and the mineral-loaded nanofiber yarn are of a core-spun structure, wherein the core layer is a polymer thread and the outer layer is an electrospun nanofiber.

3. The stent according to claim 2, wherein the polymer thread is one or more of polyethylene terephthalate (PET) thread, polylactic acid (PLA) thread, Polycaprolactone (PCL) thread, lactic acid-glycolic acid copolymer (PLGA) thread and Silk Fibroin (SF) thread; the outer layer of the nanofiber yarn with zero mineral content is electrospun with nanofiber: electrospun nanofibers comprising polymer and natural macromolecules, but no minerals; outer layer of mineral-loaded nanofiber yarn electrospun nanofibers: contains mineral, polymer and natural polymer.

4. The stent of claim 3, wherein the polymer in the outer electrospun nanofiber is one or more of poly (lactic-co-caprolactone) PLCL, Polycaprolactone (PCL) and poly (lactic-co-glycolic acid) (PLGA); the natural polymer is one or more of silk fibroin SF, Collagen (COL) and gelatin; the mineral is one or more of hydroxyapatite HA, bioactive glass, silicon dioxide, calcium carbonate, and calcium phosphate.

5. The stent of claim 1 wherein the stent has an appearance of one of an elongated shape, a flat shape, and a cylindrical shape; the length of the support is 5-50 mm, and the width is 5-20 mm.

6. The stent of claim 1, wherein the stent is: the nanofiber yarn with zero mineral content is used as warp yarns, the distance between the two warp yarns is 0.5-2 mm, and 10 warp yarns are adopted for each support; the middle of the bracket uses nanofiber yarns with zero mineral content as wefts, the nanofiber yarns with gradually increased mineral content are sequentially used as wefts from the middle to two sides, the weaving length of each weft is 2-10 mm, and the nanofiber yarns in the warp direction and the weft direction are interwoven to form the bracket.

7. A method of making an integrated stent, comprising:

8. The preparation method according to claim 7, wherein the mass ratio of the polymer to the natural polymer in the step (1) is 90: 10-10: 90; the mass fraction of the added minerals in the blended spinning solution is 0-20%.

9. An integrated stent made by the method of claim 7.

10. Use of the integrated scaffold of claim 1 in the preparation of a tendon or ligament scaffold.