WO2023143337A1 - 一种肩袖补片及其制备方法 - Google Patents

一种肩袖补片及其制备方法 Download PDF

Info

Publication number
WO2023143337A1
WO2023143337A1 PCT/CN2023/073024 CN2023073024W WO2023143337A1 WO 2023143337 A1 WO2023143337 A1 WO 2023143337A1 CN 2023073024 W CN2023073024 W CN 2023073024W WO 2023143337 A1 WO2023143337 A1 WO 2023143337A1
Authority
WO
WIPO (PCT)
Prior art keywords
rotator cuff
solution
fibrinogen
cuff patch
patch
Prior art date
Application number
PCT/CN2023/073024
Other languages
English (en)
French (fr)
Inventor
何红兵
尹荣鑫
周星宇
杨莉
闫侃
Original Assignee
上海松力生物技术有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 上海松力生物技术有限公司 filed Critical 上海松力生物技术有限公司
Publication of WO2023143337A1 publication Critical patent/WO2023143337A1/zh

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
    • A61L27/225—Fibrin; Fibrinogen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/56—Porous materials, e.g. foams or sponges
    • D—TEXTILES; PAPER
    • D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
    • D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
    • D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
    • D04H1/40—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties
    • D04H1/42—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties characterised by the use of certain kinds of fibres insofar as this use has no preponderant influence on the consolidation of the fleece
    • D—TEXTILES; PAPER
    • D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
    • D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
    • D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
    • D04H1/70—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres
    • D04H1/72—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged
    • D04H1/728—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged by electro-spinning
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/41—Anti-inflammatory agents, e.g. NSAIDs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/10—Materials or treatment for tissue regeneration for reconstruction of tendons or ligaments

Definitions

  • the invention belongs to the technical field of medical devices, and in particular relates to a rotator cuff patch and a preparation method thereof.
  • the high recurrence rate after rotator cuff repair is a great challenge for surgeons.
  • the recurrence rate is about 26%; for large (3-5cm) and extra-large (>5cm) tears, the recurrence rate can be as high as 94%.
  • Approaches to the treatment of rotator cuff tears include various suturing techniques (eg, single- and double-row anchors with suture), tissue grafting (allograft, xenograft, autograft, acellular matrix), and tissue regeneration.
  • the normal tendon healing process is divided into three overlapping phases, including 1) the inflammatory phase, with collagen deposition and migration of inflammatory cells to the recovery site within 4-7 days postoperatively; 2) proliferation and repair of fibroblasts and tenocytes, forming a temporary 3) 6 to 8 weeks after operation, the tendon tissue composed of collagen arranged in order is formed.
  • the repair of rotator cuff tear mainly uses threaded anchors to fix the tendon to the bone end, and its limitations can be summarized into three categories: (1) The normal healing process of the tendon cannot be reproduced, and the tissue formed after repair is mainly composed of fibrovascular scar tissue composition. (2) At the interface where the tendon attaches to the bone tissue (origin and end points) Four areas where tendon-bone healing is ultimately not reproducible. The origin-point is a complex composite biological junction that allows the transfer of mechanical stress between different materials. The mechanical properties of the fibrovascular scar tissue formed at the repaired interface differ from normal tendon structures, making the repair site prone to failure. (3) Sufficient mechanical strength cannot be obtained after repair.
  • Tissue regeneration techniques mainly include the use of cells, growth factors, and biomaterial matrices, alone or in combination with biomimetic extracellular matrix fiber structures, to eventually restore the mechanical and physiological properties of the rotator cuff tendon.
  • tissue regeneration techniques mainly include the use of cells, growth factors, and biomaterial matrices, alone or in combination with biomimetic extracellular matrix fiber structures, to eventually restore the mechanical and physiological properties of the rotator cuff tendon.
  • the clinical outcome of large or giant rotator cuff defects is often inferior to that of small or moderate tears.
  • Related research focuses on alternative reconstruction techniques for large or very large tears. A variety of technical strategies have been proposed, and patch repair is one of the most popular research topics.
  • Sano et al used autologous fascia transplantation to repair supraspinatus defect in rabbits. Cell density gradually increased during survival, and the patch was found histologically similar to normal tendon structure at 8 weeks postoperatively. The problem with this study is that the use of autografts causes secondary damage to the donor site.
  • Synthetic grafts have achieved some success in the repair of rotator cuff tears, and their greatest advantage is that they can avoid secondary damage to the donor site.
  • Polytetrafluoroethylene (PTFE) is used for reconstruction of irreparable rotator cuff tears with mixed clinical results. Some studies have reported that it has a good clinical outcome of pain relief, and it has also been found that bone resorption occurs at the tissue limba-graft bone attachment. Kimura et al. used PTFE as a graft to repair an irreparable infraspinatus tear in a canine model and found that the pull-off test failed within PTFE immediately after implantation. After 12 weeks, all animals failed at the PTFE-bone interface.
  • Absorbable biological patches are also used in clinical practice.
  • Porcine small intestinal submucosa (SIS) is a biomaterial that serves as a scaffold for tendon, ligament, and fascia reconstruction with minimal host response.
  • Dejardin et al. used SIS to induce repair of a rotator cuff tear in a canine model.
  • the mesh-tendon was significantly thinner, similar to the contralateral tendon, and the bone-mesh-tendon tissue had continuity.
  • Fibrous hyperplasia and collagen deposition were found in sham-operated and SIS specimens at 3 months postoperatively. within 6 months can be observed close to Regenerated structure of normal tendon.
  • sham and SIS patches had much lower maximal force at all time points (16.57%-49.73%).
  • the purpose of the present invention is to provide a rotator cuff patch and a preparation method thereof.
  • the rotator cuff patch has a nano-scale three-dimensional structure similar to extracellular matrix and good hydrophilicity. It can form its own tendon in the shoulder joint cavity after implantation for about 6 weeks, and form tendon-bone healing with shoulder joint bone tissue ; About 24 weeks after implantation, the maximum force of the regenerated rotator cuff tissue returned to 95-100% of the normal side; the rotator cuff patch can be used for bridging or strengthening treatment of large and super large rotator cuff tissue tears.
  • the present invention provides the application of a hydrophilic electrospun biocomposite scaffold material in the preparation of an implant for treating rotator cuff tissue tear
  • the composite scaffold material is composed of fibrinogen, L-arginine
  • the aqueous solution of its hydrochloride and P(LLA-CL) solution are blended and prepared by electrospinning technology; wherein, the mass ratio of the fibrinogen to L-arginine or its hydrochloride is 1.2 :1 ⁇ 12.5:1;
  • the aqueous solution of the fibrinogen, L-arginine or its hydrochloride wherein the solvent is selected from one or more of pure water, water for injection, saline solution, buffer; the saline solution is selected from chlorine Sodium chloride solution, potassium chloride solution;
  • the buffer is selected from phosphate buffer, Tris-HCl buffer, glycine buffer, D-Hank's solution.
  • the fibrinogen is mammalian-derived fibrinogen.
  • the mammal is human, pig, cow, sheep or horse.
  • the mass ratio of polylactic acid and polycaprolactone in the P(LLA-CL) is 20:80 ⁇ 95:5.
  • the solvent in the P(LLA-CL) solution is selected from one or more of hexafluoroisopropanol, chloroform, dimethylformamide, tetrahydrofuran, chloroform or acetone kind.
  • L-arginine or its hydrochloride is blended with the P(LLA-CL) solution, wherein the fibrinogen: P(LLA-CL)
  • the mass ratio is 0.2:1 to 2.1:1.
  • the equilibrium contact angle of the hydrophilic electrospun biocomposite scaffold material is less than 55°.
  • the total volume shrinkage rate is not greater than 20%; the porosity is not lower than 30%.
  • the aqueous solution of fibrinogen, L-arginine or its hydrochloride is further loaded with antibacterial substances, and the antibacterial substances are selected from penicillins, cephalosporins, Carbapenems, aminoglycosides, tetracyclines, macrolides, glycosides, sulfonamides, quinolones, nimidazoles, lincoamines, fosfomycin, chloramphenicol, myxomyces One or more of vitamin B and bacitracin.
  • antibacterial substances are selected from penicillins, cephalosporins, Carbapenems, aminoglycosides, tetracyclines, macrolides, glycosides, sulfonamides, quinolones, nimidazoles, lincoamines, fosfomycin, chloramphenicol, myxomyces One or more of vitamin B and bacitracin.
  • the penicillins are selected from penicillins, ampicillin, carbenicillin; the cephalosporins are selected from cephalexin, cefuroxime sodium, ceftriaxone, cefpirome; Penenzymes are thiamycins; the aminoglycosides are selected from gentamicin, streptomycin, kanamycin; the tetracyclines are selected from tetracyclines, aureomycin; the macrolides Selected from erythromycin and azithromycin; the glycosides are vancomycin; the sulfonamides are selected from sulfadiazine and trimethoprim; the quinolones are selected from pipemidic acid and ciprofloxacin; The imidazoles are selected from metronidazole and tinidazole; the lincoamines are selected from lincomycin and clindamycin.
  • the release amount of the antibacterial substance is not less than 30% of the total loaded amount.
  • the invention provides a rotator cuff patch.
  • the rotator cuff patch has a three-dimensional nano-network structure and is made of raw materials containing fibrinogen complex and polylactic acid-caprolactone.
  • the fibrinogen complex includes the following components in parts by weight:
  • 0.1-20 parts of fibrinogen such as 0.1 part, 0.5 part, 1 part, 1.5 part, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 12 parts, 13 parts, 15 parts, 18 parts or 20 parts, etc.
  • 0.1-10 parts of arginine hydrochloride such as 0.1 parts, 0.5 parts, 0.8 parts, 1 part, 1.2 parts, 1.5 parts, 1.8 parts parts, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts or 10 parts, etc.
  • sodium chloride 0.01-10 parts such as 0.01 parts, 0.1 parts, 0.2 parts , 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 parts, etc.
  • 1-10 parts of sodium citrate such as 1 part, 1.2 parts, 1.5 parts, 1.8 parts, 2 parts, 2.2 parts, 2.5 parts, 2.8 parts, 3 parts, 4 parts, 5 parts, 6 parts parts, etc
  • the fibrinogen complex includes the following components in parts by weight:
  • the source of the fibrinogen there is no special requirement for the source of the fibrinogen.
  • it can be derived from mammals, such as humans, pigs, cattle, sheep, horses, etc. Unless otherwise specified, it is generally preferably derived from pig blood
  • the main components of fibrinogen and fibrinogen complex are coagulable protein, various excipients or protective agents, etc.
  • the Mn of the polylactic acid-caprolactone is 30000-80000 (for example, it can be 30000, 40000, 50000, 60000, 70000 or 80000, etc.), and the Mw is 80000-160000 (for example, it can be 80000 .
  • the polylactic acid segment in the polylactic acid-caprolactone and The mass ratio of polycaprolactone segment is 20:80 ⁇ 95:5; for example, it can be 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90 :10 or 95:5 etc.
  • the thickness of the rotator cuff patch is 1.6-10mm, for example, it can be 1.6mm, 2mm, 2.5mm, 3mm, 3.5mm, 4mm, 5mm, 6mm, 7mm, 8mm, 9mm or 10mm etc.; preferably 2 to 3 mm.
  • the protein content in the rotator cuff patch is 100-220mg/g, such as 100mg/g, 120mg/g, 130mg/g, 150mg/g, 160mg/g, 180mg/g. g, 200mg/g or 220mg/g, etc.; residual protein content ⁇ 12mg/g, such as 11mg/g, 10mg/g, 9mg/g, 8mg/g, 7mg/g, 6mg/g, 5mg/g, 3mg/g, 2mg/g or 1mg/g etc.
  • the pH of the water extract of the rotator cuff patch is 6-8.
  • the porosity of the rotator cuff patch is 41% to 100%, such as 41%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%, etc.; preferably 41 to 80%.
  • the water absorption rate of the rotator cuff patch is 35% to 200%, such as 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 90%, 100%, 120%, 130%, 150%, 160%, 180%, or 200%, etc.; preferably 55 to 80%.
  • the tensile strength of the rotator cuff patch is 2-5 MPa, such as 2 MPa, 2.5 MPa, 3 MPa, 3.5 MPa, 4 MPa, 4.5 MPa or 5 MPa.
  • the elongation at break of the rotator cuff patch is 100% to 200%; %, 180%, 190% or 200%, etc.
  • the rotator cuff patch also contains drugs with antibacterial and/or anti-inflammatory effects.
  • the present invention provides a method for preparing the rotator cuff patch as described in the second aspect, the preparation method comprising the following steps:
  • fibrinogen will be denatured after electrospinning, and the denatured fibrinogen is a water insoluble protein, which only retains the primary structure (amino acid sequence) of fibrinogen. and loss of tertiary and quaternary structure.
  • the protein contained in the rotator cuff patch in the present invention is mainly the denatured fibrinogen, and there may be a small amount of undenatured water-soluble fibrinogen.
  • the residual protein in the present invention is the water-soluble fibrinogen.
  • the concentration of the polylactic acid-caprolactone solution is 5-8g/100mL; for example, it can be 5g/100mL, 5.5g/100mL, 6g/100mL, 6.5g/100mL, 7g/100mL , 7.5g/100mL or 8g/100mL, etc.
  • the solvent of the polylactic acid-caprolactone solution is selected from one or a combination of at least two of hexafluoroisopropanol, chloroform, dimethylformamide, tetrahydrofuran and acetone .
  • the solvent of the fibrinogen complex solution is water or an aqueous solution.
  • the concentration of the fibrinogen complex solution is 8-29g/100mL; 100mL, 20g/100mL, 22g/100mL, 25g/100mL, 27g/100mL or 29g/100mL, etc.
  • the mass ratio of the fibrinogen complex in the fibrinogen complex solution to the polylactic acid-caprolactone solution and the polylactic acid-caprolactone is 0.48-1.1:1; for example, it can be 0.48:1, 0.5:1, 0.6:1, 0.7:1, 0.8:1, 0.9:1, 1:1 or 1.1:1, etc.
  • the condition of the electrospinning is: the voltage difference is 15-140kV (such as 15kV, 20kV, 30kV, 40kV, 50kV, 60kV, 70kV, 80kV, 90kV, 100kV, 110kV, 120kV , 130kV or 140kV, etc.), the solution flow rate is 3-399mL/h or 401-960mL/h (for example, it can be 3mL/h, 10mL/h, 20mL/h, 50mL/h, 80mL/h, 100mL/h, 200mL /h, 300mL/h, 399mL/h, 401mL/h, 500mL/h, 600mL/h, 700mL/h, 800mL/h, 900mL/h and 960mL/h, etc.), the distance between the needle and the mandrel is 10-50cm (for example, it can be 10cm, 20cm, 30
  • the preparation method further includes: coating a drug with antibacterial and/or anti-inflammatory effects on the surface of the rotator cuff patch.
  • the present invention has the following beneficial effects:
  • the rotator cuff patch provided by the present invention has a three-dimensional nano-reticular structure, its protein content is 100-220mg/g, residual protein content ⁇ 12mg/g, porosity is 41-100%, water absorption rate is 35-200 %, the tensile strength is 2-5MPa, the elongation at break is 100-200%, the pH of the water extract is 6-8, and has good hydrophilicity and tensile strength.
  • the rotator cuff patch can regenerate tendon tissue in the joint cavity at about 6 weeks after implantation, and form tendon-bone healing at the junction of bone and tendon, thereby replacing the injured tendon to play a role.
  • the rotator cuff patch provided by the present invention can be completely degraded in an in vivo environment, and at the same time induce rotator cuff tissue regeneration in situ.
  • the rotator cuff patch provided by the present invention is prepared from synthetic absorbable polymers and natural macromolecular biomaterials, without any chemical or biological cross-linking agent for cross-linking and fixing,
  • the absence of bioactive factors and living cells eliminates any tissue culture or tissue preparation processes, reducing cost and logistical complexity.
  • the use of electrospinning technology to manufacture implant materials can effectively design and control the quality parameters of the product by controlling raw materials (such as composition, ratio), equipment parameters (such as voltage, concentration, distance, etc.), including adjusting The characteristics of the implant (such as mechanical strength, degradation rate), induced functionality, remodeling regeneration rate, etc., are very conducive to mass production and the promotion and application of innovative products.
  • Fig. 1 is the shape schematic diagram of the rotator cuff patch used in the animal experiment of the present invention
  • Fig. 2 is the schematic diagram of the suturing mode of the rotator cuff patch in the animal experiment of the present invention
  • a and B are the opposite sides of the rotator cuff patch.
  • Fig. 3 is a diagram of the sampling site in the animal experiment of the present invention.
  • Fig. 4 is a cross-sectional view of the humerus of the surgical side sample taken at each predetermined postoperative observation time.
  • Fig. 5 is a comparison chart of the maximum breaking force of samples taken on the operation side and the normal side at each predetermined observation time after the operation.
  • Fig. 6 is a comparison chart of the tensile strength of samples taken on the operation side and the normal side at each predetermined observation time after operation.
  • Fig. 7 is the optical micrographs of HE staining and Masson staining of the tendon tissue regenerated in the joint cavity at each observation time after operation.
  • Fig. 8 is the HE staining optical micrograph of the tendon-bone healing structure regenerated at each observation time after the predetermined operation
  • the part raw material source that adopts is as follows:
  • Polylactic acid-caprolactone (PLCL): Mn is 50000, Mw is 100000, Mw/Mn 2.0, the mass ratio of polylactic acid segment and polycaprolactone segment is 70:30, purchased from Netherlands Purac company;
  • Fibrinogen common porcine fibrinogen.
  • the present embodiment provides a kind of rotator cuff patch, and its preparation method is as follows:
  • F-Fg fibrinogen complex
  • component concentrations are as follows: fibrinogen (Fg) 12.61g/100mL, arginine hydrochloride 3.29g/ 100mL, sodium chloride 3.27g/100mL and sodium citrate 9.83g/100mL;
  • Examples 2-5 each provide a rotator cuff patch, the difference between the preparation method and Example 1 is that the concentration of F-Fg in the mixed electrospinning solution and the mass ratio of F-Fg to PLCL are different, as shown in the following table 1.
  • the physical parameter of the rotator cuff patch that above-mentioned embodiment obtains carries out characterization test, and test method is as follows:
  • Tests were performed using a high temperature mechanical property tester (Instron-5567) with a gauge length of 20 mm and a crosshead speed of 20 mm/min. According to the method of above-mentioned embodiment and comparative example, prepare the sample that size is 1.5cm * 7cm, thickness is 0.2-0.4mm, test is carried out in the room temperature that relative humidity is 60%, 5 readings of each group of samples, take the average value.
  • a high temperature mechanical property tester Instron-5567
  • Porosity was tested by dipping method. Soak the sample in absolute ethanol for 12 hours, measure the mass of the sample before and after soaking, and calculate the porosity according to the following formula:
  • W is the mass of the sample after immersion
  • W 0 is the mass of the sample before immersion
  • ⁇ is the density of absolute ethanol
  • V is the apparent volume of the sample.
  • Sample preparation cut 3 samples with a mass of about 0.5 g/piece for measurement.
  • Determination of samples 1) Put the sample into a weighing bottle, place it in an oven at 50.0°C ⁇ 2.0°C, dry it for at least 24 hours, then cool it to room temperature in a desiccator, weigh each sample, accurate to 0.1mg, repeat In this step, the mass change of the sample is within ⁇ 1 mg.
  • m 2 the quality of the sample after soaking, in mg
  • m 1 the mass of the sample after drying before soaking, in mg.
  • the protein molecule contains more than two peptide bonds, and will form a purple-red complex with Cu 2+ in an alkaline solution.
  • the color depth is proportional to the protein concentration within a certain range.
  • the standard curve is made with the protein reference solution , using a colorimetric method to measure the protein content (mg/mL) in the test sample, and convert it into the protein content (mg/g) in the sample.
  • Determination of the sample Cut the sample into pieces, accurately weigh 100mg, put it into the sample tube attached to the biological sample homogenizer (with 16 3mm and 2 5mm magnetic beads built-in), and add 3mL of biuret test solution. Set the homogenization speed: 6m/sec, time: 20sec x 5 times, control the temperature below 37°C, and prepare the sample as a suspension. Centrifuge at 4000rpm for 15min until the precipitate is removed, take the supernatant as the test solution, and use it within 120min. Parallel samples were prepared in the same way.
  • C protein content (mg/g) in the sample
  • W the weight (mg) of the sample in the sample tube attached to each biological sample homogenizer
  • V The volume (mL) of the biuret test solution added to the sample tube attached to each biological sample homogenizer.
  • the protein reduces Cu 2+ to Cu + , and Cu + forms a blue-purple complex with BCA reagent. Measure its absorbance at 562nm and compare it with the standard curve to calculate the protein concentration.
  • Determination of the sample Cut the sample into pieces, weigh 20mg of the sample fragments and place them in a homogeneous sample tube (built-in with 8 3mm magnetic beads), add 1mL of lysate containing PMSF with a final concentration of 1mM, set the homogenization speed: 6m /sec, time: 30sec ⁇ 2 times, the temperature is controlled below 37°C, and the sample is prepared as a suspension. Crack at 25°C ⁇ 5°C for 3h, shake the sample every 30min to mix it evenly. Centrifuge at 4000 rpm for 10 min after the lysis is complete, and take the supernatant as the test solution. 3 parallel samples with the legal system.
  • C is the content (mg/g) of residual protein in the sample
  • C ts is the final concentration of residual protein in the test solution (mg/mL);
  • V is the volume of the lysate added in the homogeneous sample tube (being 1mL in the formula);
  • M is the mass (g) of the sample.
  • Beagle dogs were placed in the lateral position, intravenously anesthetized with Shutai, and the left or right shoulder joint was randomly prepared for skin preparation.
  • the non-skin-prepared side was the normal control side without any treatment; the skin-prepared side was the experimental side.
  • the surgical procedure is as follows: A 4-5 cm incision is made in the skin above the shoulder and then dissected to the deltoid muscle, dividing this muscle parallel to the muscle fibers to expose the infraspinatus tendon. The tendon and greater tuberosity were sharply dissected, and a transverse cut was made at the junction of the tendon.
  • the infraspinatus tendon was completely removed from the tendon junction of the greater tuberosity of the humerus, resulting in a defect of about 20 mm in length.
  • the resected portion of the tendon is removed with the capsule, fully exposing the glenoid joint.
  • Two 5 ⁇ 5mm bone tunnels were made next to the greater tuberosity with a distance of 5mm.
  • Get the rotator cuff patch (long: 50mm, wide: 30mm, thickness: 1.6mm), cut it into the shape shown in Figure 1, where A and B are the opposite sides of the rotator cuff patch.
  • Side A of the rotator cuff patch was fixed to the humeral head through a bone tunnel, and side B was sutured to the muscle tissue.
  • the suture method is shown in Figure 2.
  • the wound was sutured intermittently layer by layer with Johnson & Johnson 4-0vicryl absorbable suture, and the skin was sutured continuously with nonabsorbable suture.
  • each Beagle dog was injected intramuscularly with ceftiofur sodium 8 mg/kg for 5 consecutive days.
  • the cross-sectional area of the tendon was calculated by treating the cross-section as a rectangle and measuring the width and thickness of the tendon in the middle region, and the results are described in Table 2 below.
  • Fig. 4 is the cross-sectional view of the humerus on the operated side at 6 weeks (A), 12 weeks (B) and 24 weeks (C) after operation. It can be seen that at each time point, the tendon forms a tendon-bone interface at the entrance of the bone tunnel and the surrounding cortex (see the place marked with an asterisk in the figure). At weeks 6 and 12, the rotator cuff patch material was clearly visible due to the low mechanical tension in the bone tunnel (arrows).
  • the momentary tensile force when the sample is broken on the tensile machine is the maximum breaking force (unit: N). 6-24 weeks after the implantation of the rotator cuff patch, the results of the maximum force measurement of the normal side and the surgical side samples are shown in Figure 5.
  • the maximum force of the sample on the operation side was 140.54 ⁇ 23.06N, which was 40.15% of the normal control side (354.96 ⁇ 79.30N); at 12 weeks after operation, the maximum force of the sample on the operation side was 188.04 ⁇ 51.31N, which was normal 42.76% of the normal side (443.89 ⁇ 62.22N); and 24 weeks after surgery, the maximum force of the surgical side was 506.72 ⁇ 129.36N, which was not statistically different from the normal side (485.54 ⁇ 148.66N) (P>0.05). normal side 95%-100%.
  • the tensile strength is the maximum force per unit area of the sample (in MPa). 6-24 weeks after the implantation of the rotator cuff patch, the results of the tensile strength measurement of the samples of the normal side and the operated side are shown in Fig. 6 . It can be seen from Figure 6 that since the cross-section of the tendon on the operated side was thicker than that on the normal control side, the corresponding tensile strength was lower than that on the normal control side. Among them, at 6 weeks, 12 weeks and 24 weeks after operation, the tensile strength of the samples on the operation side were 24.25%, 15.28% and 49.98% of the normal control side, respectively. It can be seen that the tensile strength of the surgical side samples is gradually increasing.
  • Tendons and regenerated tendons were harvested from the proximal end of the tendon junction (including the greater tuberosity) on the humerus; contralateral normal control specimens were also harvested to examine the histological appearance of the intrinsic infraspinatus tendon.
  • the shoulder joint specimen was cut longitudinally to fully expose the repaired tissue, muscle and tendon-bone interface. Fix with 10% neutral formalin and decalcify with 5% nitric acid solution. All histological sections were stained with HE and trichrome, and observed under a light microscope.
  • Figure 7 is the optical micrographs of HE staining and Masson staining of the tendon tissue regenerated in the joint cavity at various observation times after operation. From Figure 7, it can be seen that from the 6th postoperative week, all the tendon tissues on the operated side had been formed. An infiltration of inflammatory cells was seen near the surgical sutures on the adjacent muscle side. The rotator cuff patch material has almost completely degraded.
  • Figure 8 is the HE-stained optical micrographs of the regenerated tendon-bone healing structure at each observation time after operation, among which, 6 weeks after operation: A ( ⁇ 20) and D ( ⁇ 40); 12 weeks after operation: B ( ⁇ 20) and E( ⁇ 40); 24 weeks after operation: C( ⁇ 20) and F( ⁇ 40).
  • Typical tendon-bone healing consists of four layers of tissue: tendon tissue and Sharpy's fibers, non-calcified fibrous structures, calcified fibrous structures, and bone tissue. From the 6th week, it can be clearly distinguished under the microscope. 6-12 weeks after operation, residual rotator cuff patch and chronic inflammatory mediators can be seen in the bone marrow; The tract is composed of disordered collagen-like structures.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Dermatology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Transplantation (AREA)
  • Engineering & Computer Science (AREA)
  • Textile Engineering (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Dispersion Chemistry (AREA)
  • Materials For Medical Uses (AREA)

Abstract

本发明提供了一种肩袖补片及其制备方法。所述肩袖补片具有三维纳米网状结构，由包含纤维蛋白原复合物和聚乳酸-己内酯的原料制成。所述肩袖补片是通过将纤维蛋白原复合物溶液和聚乳酸-己内酯溶液加入静电纺丝机的同一容量管中，或者分别加入到静电纺丝机的不同容量管中，进行静电纺丝，然后干燥的方法制备得到。本发明提供的肩袖补片具有类似细胞外基质的纳米级三维结构和良好的亲水性；其在植入6周左右即可在肩关节腔内形成自身肌腱，并与肩关节骨组织形成腱骨愈合；植入后24周左右，再生形成的肩袖组织的最大力恢复到正常侧的95-100％。该肩袖补片可用于大型及超大型肩袖组织撕裂的桥接或加强性治疗。

Description

一种肩袖补片及其制备方法 技术领域
本发明属于医疗器械技术领域，具体涉及一种肩袖补片及其制备方法。
背景技术
肩关节的稳定性和功能很大程度上依赖肩袖肌肉群的完整性。急性肩袖撕裂常见于运动员，慢性退行性肌腱改变和随后的撕裂常见于老年人和久坐的患者。随着年龄的增长，肩袖撕裂的频率和大小随之增加。肩袖损伤不易自行愈合，这与肩关节的复杂解剖结构、活动范围大、肌腱的相对弱化和低血管化有关。未经治疗的部分肩袖撕裂，无论是创伤性的还是退行性的，会发展成大型及超大型撕裂。表现为局部疼痛和肌肉动作反射抑制，运动能力下降，甚至危及日常活动。随着病情的发展，肩关节的稳定性和力学性能会进一步受损，最终导致不可逆的完全肩袖撕裂关节病综合征。
目前，肩袖修复术后的高复发率是外科医生面临的极大挑战。小(<1cm)到中型(1-3cm)撕裂，复发率为26％左右；大(3-5cm)和超大(﹥5cm)型撕裂，复发率可高达94％。
治疗肩袖撕裂的方法包括各种缝合技术(如单排和双排带线锚钉)、组织移植(同种异体移植、异种移植、自体移植、脱细胞基质)和组织再生。正常的肌腱愈合过程分为三个相互重叠的阶段，包括1)炎症阶段，术后4-7天内，胶原沉积和炎症细胞迁移到恢复部位；2)成纤维细胞和腱细胞增殖修复，形成临时性胶原蛋白组成的细胞外基质；3)术后6到8周，形成有序排列的胶原蛋白组成的肌腱组织。
目前肩袖撕裂修复主要是使用带线锚钉将肌腱固定于骨端，其局限性可归纳为三类：(1)无法再现肌腱正常愈合过程，修复后形成的组织主要由纤维血管瘢痕组织组成。(2)在肌腱附着于骨组织的界面 (起止点)最终无法再现腱骨愈合的四个区域。起止点是一个复杂的复合生物连接，允许机械压力在不同材料之间转移。修复后的界面形成的纤维血管瘢痕组织的力学性质与正常肌腱结构不同，从而使修复部位易发生故障。(3)修复后无法获得足够的机械强度。
在临床上，常用的缝合技术和组织移植都不能令人满意地治疗大和超大型肩袖撕裂，因而催生了组织再生技术的发展。组织再生技术主要包括采用细胞、生长因子和生物材料基质，单独或联合仿生细胞外基质的纤维结构，以最终恢复肩袖肌腱的力学和生理特性。在骨科实践中，对大或巨大肩袖缺损的临床治疗效果往往劣于小或中度撕裂。相关的研究重点在大或超大撕裂的替代重建技术。已经提出多种技术策略，补片修补是其中热门的研究课题之一。
Sano等采用自体筋膜移植修复兔冈上肌缺损。存活期间细胞密度逐渐增加，术后8周在组织学上发现补片与正常肌腱结构相似。该研究的问题在于，使用自体移植物会造成供体部位的二次损伤。
合成移植物用于肩袖撕裂修复取得一定的成功，其最大优点是可以避免供体部位的二次损伤。聚四氟乙烯(PTFE)用于重建不可修复的肩袖撕裂，临床结果报道不一。一些研究报道其具有缓解疼痛的良好临床结果，也有发现在固有组织-移植物骨附着处有骨吸收的发生。Kimura等人使用PTFE作为修复犬模型中不可修复的冈下肌撕裂的移植物，植入后立即就发现拉断实验在PTFE内失败。12周后，所有动物均在PTFE-骨界面失败。
可吸收生物补片(如同种异体移植、异种移植)也用于临床实践中。猪小肠黏膜下层(SIS)是一种生物材料，可作为肌腱、韧带和筋膜重建的支架，具有最小的宿主反应。Dejardin等人使用SIS在犬模型诱导修复肩袖撕裂。术后3个月和6个月，补片-肌腱明显变薄，与对侧肌腱相似，骨-补片-肌腱组织具有连续性。术后3个月，在假手术和SIS标本中发现纤维增生和胶原沉积。6个月内可以观察到接近 正常肌腱的再生结构。与正常的冈下肌相比，假手术和SIS补片在所有时间点的最大力均小得多(16.57％-49.73％)。
Julie E报道的使用人皮脱细胞基质的狗动物试验结果表明，尽管术后3个月(538.6N)和6个月(552.4N)的最大力比手术后即时(65.5N)的高出8-9倍，但是，仅仅恢复到正常对照(918.7N)的58-60％。
目前的补片对于肩袖撕裂的修复效果均不理想，因此仍然有待研究具有更好的肩袖撕裂治疗效果的植入物。
发明内容
针对现有技术存在的不足，本发明的目的在于提供一种肩袖补片及其制备方法。该肩袖补片具有类似细胞外基质的纳米级三维结构和良好的亲水性，其在植入6周左右即可在肩关节腔内形成自身肌腱，并与肩关节骨组织形成腱骨愈合；植入后24周左右，再生形成的肩袖组织的最大力恢复到正常侧的95-100％；该肩袖补片可用于大型及超大型肩袖组织撕裂的桥接或加强性治疗。
为达此目的，本发明采用以下技术方案：
第一方面，本发明提供一种亲水性静电纺生物复合支架材料在制备治疗肩袖组织撕裂的植入物中的应用，所述复合支架材料是由纤维蛋白原、L-精氨酸或其盐酸盐的水溶液与P(LLA-CL)溶液共混，采用静电纺技术制备而得到的；其中，所述纤维蛋白原与L-精氨酸或其盐酸盐的质量比为1.2:1～12.5:1；
所述纤维蛋白原、L-精氨酸或其盐酸盐的水溶液，其中的溶剂选自纯水、注射用水、盐溶液、缓冲液中的一种或多种；所述盐溶液选自氯化钠溶液、氯化钾溶液；所述缓冲液选自磷酸盐缓冲液、Tris-HCl缓冲液、甘氨酸缓冲液、D-Hank’s液。
在本发明一些实施方式中，所述纤维蛋白原是来源于哺乳动物的纤维蛋白原。
在本发明一些实施方式中，所述哺乳动物为人、猪、牛、羊或马。
在本发明一些实施方式中，所述P(LLA-CL)中聚乳酸和聚己内酯的质量比为20:80～95:5。
在本发明一些实施方式中，所述的P(LLA-CL)溶液中的溶剂选自六氟异丙醇、三氯甲烷、二甲基甲酰胺、四氢呋喃、氯仿或丙酮中的一种或者多种。
在本发明一些实施方式中，所述纤维蛋白原、L-精氨酸或其盐酸盐的水溶液与P(LLA-CL)溶液共混后，其中纤维蛋白原:P(LLA-CL)的质量比为0.2:1～2.1:1。
在本发明一些实施方式中，所述亲水性静电纺生物复合支架材料的平衡接触角小于55°。
在本发明一些实施方式中，所述亲水性静电纺生物复合支架材料与水溶液接触后，总体积皱缩率不大于20％；孔隙率不低于30％。
在本发明一些实施方式中，，所述纤维蛋白原、L-精氨酸或其盐酸盐的水溶液中，还进一步加载有抗菌物质，所述抗菌物质选自青霉素类、头孢菌素类、碳青酶烯类、氨基糖甙类、四环素类、大环内脂类、糖甙类、磺胺类、喹诺酮类、硝咪唑类、林克胺类、磷霉素、氯霉素、对粘菌素B、杆菌肽中的一种或多种。
在本发明一些实施方式中，所述青霉素类选自青霉素、氨苄西林、羧苄西林；所述头孢菌素类选自头孢氨苄、头孢呋辛钠、头孢曲松、头孢匹罗；所述碳青酶烯类为硫霉素；所述氨基糖甙类选自庆大霉素、链霉素、卡那霉素；所述四环素类选自四环素、金霉素；所述大环内脂类选自红霉素、阿奇霉素；所述糖甙类为万古霉素；所述磺胺类选自磺胺嘧啶、甲氧苄啶；所述喹诺酮类选自吡哌酸、环丙沙星；所述硝咪唑类选自甲硝唑、替硝唑；所述林克胺类选自林可霉素、克林霉素。
在本发明一些实施方式中，在该支架材料植入体内后15分钟内，所述抗菌物质的释放量不低于总加载量的30％。
本发明提供一种肩袖补片，所述肩袖补片具有三维纳米网状结构，由包含纤维蛋白原复合物和聚乳酸-己内酯的原料制成。
在本发明一些实施方式中，所述纤维蛋白原复合物包括如下重量份数的组分：
纤维蛋白原0.1-20份(例如可以是0.1份、0.5份、1份、1.5份、2份、3份、4份、5份、6份、7份、8份、9份、10份、12份、13份、15份、18份或20份等)、精氨酸盐酸盐0.1-10份(例如可以是0.1份、0.5份、0.8份、1份、1.2份、1.5份、1.8份、2份、3份、4份、5份、6份、7份、8份、9份或10份等)、氯化钠0.01-10份(例如可以是0.01份、0.1份、0.2份、0.3份、0.4份、0.5份、0.6份、0.7份、0.8份、0.9份、1份、2份、3份、4份、5份、6份、7份、8份、9份或10份等)和枸橼酸钠1-10份(例如可以是1份、1.2份、1.5份、1.8份、2份、2.2份、2.5份、2.8份、3份、4份、5份、6份、7份、8份、9份或10份等)。
优选的，所述纤维蛋白原复合物包括如下重量份数的组分：
纤维蛋白原3-15份、精氨酸盐酸盐0.5-5份、氯化钠0.3-5份和枸橼酸钠1-10份。
本发明中，对于所述纤维蛋白原的来源没有特殊要求，示例性地，可以是来源于哺乳动物，例如人、猪、牛、羊、马等，如无特殊说明，一般优选为猪血来源的纤维蛋白原，纤维蛋白原复合物主要组成成分为可凝固蛋白，各种赋形剂或保护剂等。
在本发明一些实施方式中，所述聚乳酸-己内酯的Mn为30000-80000(例如可以是30000、40000、50000、60000、70000或80000等)，Mw为80000-160000(例如可以是80000、90000、100000、110000、120000、130000、140000、150000或160000等)，Mw/Mn<3.0(例如可以是2.9、2.6、2.5、2.2、2.0、1.8或1.5等)。
在本发明一些实施方式中，所述聚乳酸-己内酯中聚乳酸链段与 聚己内酯链段的质量比为20:80～95:5；例如可以是20:80、30:70、40:60、50:50、60:40、70:30、80:20、90:10或95:5等。
在本发明一些实施方式中，所述肩袖补片的厚度为1.6～10mm，例如可以是1.6mm、2mm、2.5mm、3mm、3.5mm、4mm、5mm、6mm、7mm、8mm、9mm或10mm等；优选为2～3mm。
在本发明一些实施方式中，所述肩袖补片中的蛋白质含量为100-220mg/g，例如可以是100mg/g、120mg/g、130mg/g、150mg/g、160mg/g、180mg/g、200mg/g或220mg/g等；残留蛋白含量<12mg/g，例如可以是11mg/g、10mg/g、9mg/g、8mg/g、7mg/g、6mg/g、5mg/g、3mg/g、2mg/g或1mg/g等。
在本发明一些实施方式中，所述肩袖补片的水浸提液的pH为6-8。
在本发明一些实施方式中，所述肩袖补片的孔隙率为41～100％，例如可以是41％、45％、50％、55％、60％、65％、70％、75％、80％、85％、90％、95％或100％等；优选为41～80％。
在本发明一些实施方式中，所述肩袖补片的吸水率为35～200％，例如可以是35％、40％、45％、50％、55％、60％、65％、70％、75％、80％、90％、100％、120％、130％、150％、160％、180％或200％等；优选为55～80％。
在本发明一些实施方式中，所述肩袖补片的拉伸强度为2～5MPa，例如可以是2MPa、2.5MPa、3MPa、3.5MPa、4MPa、4.5MPa或5MPa等。
在本发明一些实施方式中，所述肩袖补片的断裂伸长率为100～200％；例如可以是100％、110％、120％、130％、140％、150％、160％、170％、180％、190％或200％等。
在本发明一些实施方式中，所述肩袖补片还含有具有抗菌和/或消炎作用的药物。
第三方面，本发明提供一种如第二方面所述的肩袖补片的制备方法，所述制备方法包括如下步骤：
将纤维蛋白原复合物溶液和聚乳酸-己内酯溶液混合后加入静电纺丝机的同一容量管中，进行静电纺丝；或者，将纤维蛋白原复合物溶液和聚乳酸-己内酯溶液分别加入到静电纺丝机的不同容量管中，同时进行静电纺丝；干燥后得到所述肩袖补片。
需要说明的是，本发明中，纤维蛋白原经过静电纺丝后会发生变性，变性的纤维蛋白原为水难溶性蛋白，其仅保留有纤维蛋白原的一级结构(氨基酸序列)，二级和三级和四级结构丧失。本发明中所述肩袖补片中所含的蛋白主要为该变性的纤维蛋白原，可能还有少量未变性的水溶性纤维蛋白原。本发明中残余蛋白即为该水溶性纤维蛋白原。
在本发明一些实施方式中，所述聚乳酸-己内酯溶液的浓度为5～8g/100mL；例如可以是5g/100mL、5.5g/100mL、6g/100mL、6.5g/100mL、7g/100mL、7.5g/100mL或8g/100mL等。
在本发明一些实施方式中，所述聚乳酸-己内酯溶液的溶剂选自六氟异丙醇、三氯甲烷、二甲基甲酰胺、四氢呋喃和丙酮中的一种或至少两种的组合。本发明中，优选控制上述有机溶剂在肩袖补片中的残留量<0.55mg/g。
在本发明一些实施方式中，所述纤维蛋白原复合物溶液的溶剂为水或水溶液。
在本发明一些实施方式中，所述纤维蛋白原复合物溶液的浓度为8-29g/100mL；例如可以是8g/100mL、9g/100mL、10g/100mL、12g/100mL、15g/100mL、18g/100mL、20g/100mL、22g/100mL、25g/100mL、27g/100mL或29g/100mL等。
在本发明一些实施方式中，所述纤维蛋白原复合物溶液与聚乳酸-己内酯溶液中的纤维蛋白原复合物与聚乳酸-己内酯的质量比为 0.48～1.1:1；例如可以是0.48:1、0.5:1、0.6:1、0.7:1、0.8:1、0.9:1、1:1或1.1:1等。
在本发明一些实施方式中，所述静电纺丝的条件为：电压差为15-140kV(例如可以是15kV、20kV、30kV、40kV、50kV、60kV、70kV、80kV、90kV、100kV、110kV、120kV、130kV或140kV等)，溶液流速为3-399mL/h或401-960mL/h(例如可以是3mL/h、10mL/h、20mL/h、50mL/h、80mL/h、100mL/h、200mL/h、300mL/h、399mL/h、401mL/h、500mL/h、600mL/h、700mL/h、800mL/h、900mL/h和960mL/h等)，针头与芯轴之间的距离为10-50cm(例如可以是10cm、20cm、30cm、40cm或50cm等)。
在本发明一些实施方式中，所述制备方法还包括：在所述肩袖补片表面涂覆具有抗菌和/或消炎作用的药物。
与现有技术相比，本发明具有以下有益效果：
(1)本发明提供的肩袖补片具有三维纳米网状结构，其蛋白质含量为100-220mg/g，残留蛋白含量<12mg/g，孔隙率为41～100％，吸水率为35～200％，拉伸强度为2～5MPa，断裂伸长率为100～200％，水浸提液的pH为6-8，具有良好的亲水性和拉伸强度。该肩袖补片能够在植入6周左右，在关节腔内再生出肌腱组织，并在骨与肌腱的结合部位形成腱骨愈合，从而代替损伤的肌腱发挥作用。
(2)本发明提供的肩袖补片在体内环境中能够完全降解，同时原位诱导肩袖组织再生。通过动物实验结果，我们惊奇地发现，随着肩袖补片植入体内时间的延长，再生肌腱的拉断最大力和拉伸强度逐渐增大，术后24周，再生肌腱的拉断最大力达到正常对照组水平(>95％)以上，远远大于Dejardin报道的SIS材料(16.57％-49.73％)和Julie E报道的使用人皮脱细胞基质(58-60％)。
(3)本发明提供的肩袖补片由合成可吸收聚合物和天然高分子生物材料制备而成，无需采用任何化学或生物交联剂进行交联固定， 不含有生物活性因子和活细胞，省去了任何组织培养或组织制备的过程，从而降低了成本和物流复杂性。此外，采用静电纺丝技术制造植入物材料，可以有效地通过控制原料(如组成、比例)，设备参数(如电压，浓度，距离等)等对产品的质量参数进行设计和控制，包括调节植入物的特性(例如机械强度、降解速率)、诱导的功能性、重塑再生速率等，从而十分有利于大规模量产，及创新产品的推广应用。
附图说明
图1为本发明动物实验中使用的肩袖补片的形状示意图；
图2为本发明动物实验中肩袖补片的缝合方式示意图；
其中，A、B为肩袖补片相对的两侧。
图3为本发明动物实验中的取样部位图。
图4为预定术后各观察时间所取手术侧样品肱骨的剖面图。
图5为预定术后各观察时间所取手术侧和正常侧样品的拉断最大力对比图。
图6为预定术后各观察时间所取手术侧和正常侧样品的抗拉强度对比图。
图7为预定术后各观察时间关节腔内再生的肌腱组织的HE染色和Masson染色光学显微照片。
图8为预定术后各观察时间再生的腱骨愈合结构的HE染色光学显微照片；
其中，术后6周：A(×20)和D(×40)；术后12周：B(×20)和E(×40)；术后24周：C(×20)和F(×40)。
具体实施方式
下面结合附图并通过具体实施方式来进一步说明本发明的技术方案。本领域技术人员应该明了，所述具体实施方式仅仅是帮助理解本发明，不应视为对本发明的具体限制。
本发明实施例中，采用的部分原料来源如下：
聚乳酸-己内酯(PLCL)：Mn为50000，Mw为100000，Mw/Mn＝2.0，聚乳酸链段与聚己内酯链段的质量比为70:30，购自荷兰Purac公司；
纤维蛋白原：普通猪纤维蛋白原。
实施例1
本实施例提供一种肩袖补片，其制备方法如下：
(1)将PLCL溶于六氟异丙醇中，形成浓度为8g/100mL的PLCL溶液；
将纤维蛋白原复合物(以下简称F-Fg)溶于蒸馏水中，形成F-Fg溶液，其组分浓度如下：纤维蛋白原(Fg)12.61g/100mL、精氨酸盐酸盐3.29g/100mL、氯化钠3.27g/100mL和枸橼酸钠9.83g/100mL；
(2)将所述PLCL溶液和F-Fg溶液按0.77:0.23的体积比搅拌混合5h，形成混合静电纺溶液，其中，F-Fg与PLCL的质量比为1.08:1；
(3)将所述混合溶液转移到量产静电纺机(型号：NS1WS 500Elmarco捷克)的载液装置中进行静电纺丝，工艺条件为：电压差100kV，静电纺液流速为60mL/h，纺丝电极与基材距离14cm；制备的静电纺丝膜在室温下真空干燥48h，得到具有三维网状结构的肩袖补片，厚度为1.6mm。
实施例2-5
实施例2-5各提供一种肩袖补片，其制备方法与实施例1的区别仅在于，混合静电纺溶液中F-Fg的浓度、F-Fg与PLCL的质量比不同，具体如下表1所示。
表1

物理表征
对上述实施例得到的肩袖补片的物理参数进行表征测试，测试方法如下：
1、拉伸强度和断裂伸长率
使用20毫米轨距长度和20毫米/分钟十字头速度的高温力学性能测试仪(Instron-5567)进行测试。按照上述实施例和对比例的方法，制备尺寸为1.5cm×7cm，厚度为0.2-0.4mm的样品，测试在相对湿度为60％的室温中进行，每组样品5个读数，取平均值。
2、孔隙率
孔隙率：通过浸渍法测试孔隙率(P)。将样品在无水乙醇中浸泡12h，测定浸泡前后样品的质量，根据如下公式计算孔隙率：
其中，W为浸泡后样品质量，W0为浸泡前样品质量，ρ为无水乙醇的密度，V为样品的表观体积。
3、吸水率
原理：将样品浸入23.0±1.0℃蒸馏水中，放置一定时间，测定样品开始试验时与吸水后的质量差异，用质量差异相对于初始质量的百分比表示。
试样的准备：剪取3个质量为约0.5g/片的样品进行测定。
样品的测定：1)将样品放入称量瓶中，置于50.0℃±2.0℃烘箱内，至少干燥24h，然后在干燥器内冷却至室温，称量每个样品，精确至0.1mg，重复本步骤直至试样的质量变化在±1mg内。
2)将样品放入盛有蒸馏水的容器中，水温控制在23.0℃±1.0℃。
3)浸泡1±0.1h后，取出样品，用清洁干布或滤纸轻抚迅速擦去样品表面的水，再次称量每个样品，精确至0.1mg。样品从水中取出后，应该在1min内完成称量。
用下式计算样品的吸水率：
c(％)＝(m2-m1)/m1×100％；
式中：c:样品的吸水质量分数，数值以％表示；
m2:浸泡后样品的质量，单位为mg；
m1:浸泡前干燥后样品的质量，单位为mg。
4、蛋白含量
原理：蛋白质分子中含有两个以上肽键，在碱性溶液中会与Cu2+形成紫红色络合物，在一定范围内其颜色深浅与蛋白质浓度呈正比，以蛋白质对照品溶液做标准曲线，采用比色法测定供试品中蛋白质的含量(mg/mL)，并换算成样品中的蛋白质含量(mg/g)。
样品的测定：将样品剪碎，精确称取100mg，放入生物样品均质器附带的样品管内(内置16粒3mm和2粒5mm磁珠)，加入双缩脲试液3mL。设定均质速度：6m/sec，时间：20sec×5次，温度控制在37℃以下，将样品制备为混悬液。4000rpm离心15min直至沉淀去除，取上清为供试液，在120min内使用。同法制备平行样。
分别精密移取供试液1mL置于3支试管中(做平行样)，加入4mL双缩脲试液，混匀，室温放置30分钟，参照《中国药典》(2015年版)四部0401紫外-可见分光光度计法，在540nm的波长处测定吸光度；同时以生理盐水作为空白样。
结果计算：
根据吸光度和标准曲线计算供试液的蛋白浓度，按照下式换算样品中的蛋白含量(mg/g)：
其中，C:样品中蛋白质含量(mg/g)；
Cts:供试液中蛋白质终浓度(mg/mL)；
W：每管生物样品均质器附带的样品管中样品的重量(mg)；
V：每管生物样品均质器附带的样品管中加入的双缩脲试液的体积(mL)。
5、残留蛋白含量
原理：在碱性条件下，蛋白将Cu2+还原为Cu+，Cu+与BCA试剂形成蓝紫色的络合物，测定其在562nm处的吸光度，并与标准曲线对比，即可计算待测蛋白的浓度。
样品的测定：将样品剪碎，称取20mg样品碎片置于匀质样品管(内置8颗3mm的磁珠)中，加入1mL含PMSF最终浓度为1mM的裂解液，设定均质速度：6m/sec，时间：30sec×2次，温度控制在37℃以下，将样品制备为混悬液。25℃±5℃裂解3h，每隔30min，摇动样品使其混匀。裂解完毕后在4000rpm转速下离心10min，取上清液作为供试液。同法制3个平行样。
量取供试液0.4mL置于3支洁净西林瓶中，用移液器吸取BCA工作液4mL，用橡胶塞盖紧，用漩涡振荡器混匀后，置于37℃水浴槽中放置22min后，立即用紫外分光光度计测量562nm处吸光度。
结果计算：
根据吸光度和标准曲线计算供试液的蛋白浓度，按照下式换算样品中的残留蛋白含量(mg/g)：
其中，C为样品中残留蛋白的含量(mg/g)；
Cts为供试液中残留蛋白的终浓度(mg/mL)；
V为匀质样品管中加入的裂解液的体积(公式中为1mL)；
M为样品的质量(g)。
6、浸提液pH
取样品，加入生理盐水作为浸提介质，当样品厚度≤1.0mm时样品表面积/浸提液体积＝3cm2/mL，样品厚度>1.0mm时样品表面积/浸提液体积＝1.25cm2/mL，在37℃下浸提72小时，制备供试液，测试pH值。
上述测试的结果如下表2所示：
表2
动物实验：
使用28只骨骼发育成熟的比格犬(雌雄不限，年龄12个月，重量15.6±0.5kg)。按照预定术后观察时间(0、6、12和24周)分为四组，每组随机分配7只动物。每只动物随机选择一侧前肢进行肩袖桥接手术，作为实验组；另一侧不做处理，作为正常对照组。
比格犬采取侧卧位，舒泰静脉麻醉，随机备皮左侧或右侧肩关节。非备皮侧为正常对照侧，不做任何处理；备皮侧为手术实验侧。手术过程如下：在肩膀上方的皮肤上切开一个4-5cm的切口，然后解剖到三角肌，与肌肉纤维平行分开该肌肉，以暴露冈下肌腱。将肌腱与大结节锐性切离，并在肌腱交界处横切，从肱骨大结节的肌腱连接处完全切除冈下肌腱，造成长度约20mm的缺损。肌腱的切除部分随囊一起移开，完全暴露肩胛盂关节。在大结节旁做两个5×5mm的骨隧道，相距5mm。取实施例1提供的肩袖补片(长：50mm，宽： 30mm，厚：1.6mm)一张，将其裁剪为图1所示形状，其中A、B为该肩袖补片相对的两侧。将该肩袖补片的A侧通过骨隧道固定于肱骨头，B侧缝合于肌肉组织，缝合方式如图2所示。用强生4-0vicryl可吸收缝线逐层间断缝合创口，用不可吸收缝合线连续缝合皮肤。术后每只比格犬肌肉注射头孢噻呋钠8mg/kg，连续5天。
达到预定术后观察时间后取样(取样部位如图3所示)，进行生物力学检测和组织学检测。所有时间点动物中，5只进行生物力学检测，2只进行组织学检测。
动物实验的大体检查结果如下：
1、健康状态
所有的动物都能在手术后立即走动并承重，无伤口感染，平均体重无统计学差异，无意外死亡，皮肤愈合良好。
2、肌腱横截面面积
肌腱横截面面积是通过将横截面视为矩形，并测量中间区域肌腱的宽度和厚度，计算得到，结果如下表2所述。
表2不同时间点肌腱横截面面积(标准差)
从表2可以看出，术后6周、12周和24周，手术侧肌腱均比正常侧显著增粗(P<0.01，有非常显著的统计学意义)。
3、肌腱断裂位置
将0周(正常侧和手术侧)、6周(手术侧)、12周(手术侧)和24周(手术侧)4个时间点所获得的25个样本在拉力机上测定生物力学指标。关注的断裂位置主要为肌肉处、肌肉肌腱连接处、肌腱 中段和腱骨界面。测试结果如下表3所示：
表3肌腱断裂位置统计
从表3可以看出，在术后0周，正常侧5根样本均在肌肉处断裂，手术侧均在腱骨界面处；术后6周，3根样本在肌腱中段断裂，2根样本在腱骨界面断裂；术后12周，4根样本在韧带中段断裂，1根样本在肌肉处断裂；24周时，所有样本的断裂处均与正常对照侧类似，在肌肉处断裂，表明再生后的肌腱的该项指标恢复到正常水平。
4、手术侧肱骨剖面
图4为术后6周(A)、12周(B)和24周(C)，手术侧肱骨的剖面图。可以看出，各个时间点，肌腱在骨隧道入口和周围皮质处形成腱骨界面(见图中星号标示处)。第6周和第12周，由于骨隧道内的机械张力较低，肩袖补片材料明显可见(箭头所示部分)。
5、拉断最大力
样本在拉力机上拉断时瞬间的拉力为拉断最大力(单位：N)。肩袖补片植入后6-24周，正常侧和手术侧样本最大力测定结果如图5所示。其中，术后6周，手术侧样本最大力为140.54±23.06N，是正常对照侧(354.96±79.30N)的40.15％；术后12周，手术侧样本最大力为188.04±51.31N，是正常侧(443.89±62.22N)的42.76％；而术后24周，手术侧样本最大力为506.72±129.36N，与正常侧(485.54±148.66N)相比无统计学差异(P>0.05)，是正常侧的 95％-100％。
6、抗拉强度
抗拉强度为样本单位面积的最大力(单位MPa)。肩袖补片植入后6-24周，正常侧和手术侧样本的抗拉强度测定结果如图6所示。从图6可以看出，由于手术侧肌腱横截面比正常对照侧增粗，因此相应的抗拉强度比正常对照侧低。其中，术后6周、12周和24周，手术侧样本的抗拉强度分别为正常对照侧的24.25％、15.28％和49.98％。可见，手术侧样本的抗拉强度在逐渐增大。
7、组织学检测
术后6周、12周和24周，每个组2只动物进行组织学评价。从肱骨上的肌腱连接处(包括大结节)的近端采集肌腱和再生肌腱；同样采集对侧正常对照标本，以检查其固有的冈下肌腱的组织学外观。将肩关节标本纵行切开，充分暴露修复的组织、肌肉和腱骨结合界面。用10％中性福尔马林固定，5％硝酸溶液进行脱钙处理。所有组织学切片经过HE和三色染色，在光学显微镜下观察。
1)图7为预定术后各观察时间关节腔内再生的肌腱组织的HE染色和Masson染色光学显微照片。从图7中可以看出，从术后第6周，所有手术侧肌腱组织已经形成。在临近肌肉侧的手术缝线附近可以看到炎性细胞的浸润。肩袖补片材料已经基本完全降解。
2)图8为预定术后各观察时间再生的腱骨愈合结构的HE染色光学显微照片，其中，术后6周：A(×20)和D(×40)；术后12周：B(×20)和E(×40)；术后24周：C(×20)和F(×40)。从图8中可以看出，在腱骨界面，腱骨愈合四层结构主要出现在骨隧道入口及其周围的皮质处。典型的腱骨愈合分别由四层组织组成循序排列的肌腱组织和Sharpy氏纤维，非钙化的纤维结构，钙化纤维状结构以及骨组织。从第6周，可以在显微镜下明确分辨出。术后6-12周，骨髓道内可见肩袖补片残留和慢性炎性介质；而第24周，骨隧 道内由排列无序的胶原蛋白样结构组成。
虽然，上文中已经用一般性说明、具体实施方式及试验，对本发明作了详尽的描述，但在本发明基础上，可以对之作一些修改或改进，这对本领域技术人员而言是显而易见的。因此，在不偏离本发明精神的基础上所做的这些修改或改进，均属于本发明要求保护的范围。

Claims (21)

  1. 一种亲水性静电纺生物复合支架材料在制备治疗肩袖组织撕裂的植入物中的应用，其特征在于，所述复合支架材料是由纤维蛋白原、L-精氨酸或其盐酸盐的水溶液与P(LLA-CL)溶液共混，采用静电纺技术制备而得到的；其中，所述纤维蛋白原与L-精氨酸或其盐酸盐的质量比为1.2:1～12.5:1；
    所述纤维蛋白原、L-精氨酸或其盐酸盐的水溶液，其中的溶剂选自纯水、注射用水、盐溶液、缓冲液中的一种或多种；所述盐溶液选自氯化钠溶液、氯化钾溶液；所述缓冲液选自磷酸盐缓冲液、Tris-HCl缓冲液、甘氨酸缓冲液、D-Hank’s液。
  2. 根据权利要求1所述的应用，其特征在于，所述纤维蛋白原是来源于哺乳动物的纤维蛋白原。
  3. 根据权利要求2所述的应用，其特征在于，所述哺乳动物为人、猪、牛、羊或马。
  4. 根据权利要求1所述的应用，其特征在于，所述P(LLA-CL)中聚乳酸和聚己内酯的质量比为20:80～95:5。
  5. 根据权利要求1所述的应用，其特征在于，所述的P(LLA-CL)溶液中的溶剂选自六氟异丙醇、三氯甲烷、二甲基甲酰胺、四氢呋喃、氯仿或丙酮中的一种或者多种。
  6. 根据权利要求1所述的应用，其特征在于，所述纤维蛋白原、L-精氨酸或其盐酸盐的水溶液与P(LLA-CL)溶液共混后，其中纤维蛋白原:P(LLA-CL)的质量比为0.2:1～2.1:1。
  7. 根据权利要求1所述的应用，其特征在于，所述亲水性静电纺生物复合支架材料的平衡接触角小于55°。
  8. 根据权利要求1所述的应用，其特征在于，所述亲水性静电纺生物复合支架材料与水溶液接触后，总体积皱缩率不大于20％；孔隙率不低于30％。
  9. 根据权利要求1所述的应用，其特征在于，所述纤维蛋白原、 L-精氨酸或其盐酸盐的水溶液中，还进一步加载有抗菌物质，所述抗菌物质选自青霉素类、头孢菌素类、碳青酶烯类、氨基糖甙类、四环素类、大环内脂类、糖甙类、磺胺类、喹诺酮类、硝咪唑类、林克胺类、磷霉素、氯霉素、对粘菌素B、杆菌肽中的一种或多种。
  10. 根据权利要求9所述的应用，其特征在于，所述青霉素类选自青霉素、氨苄西林、羧苄西林；所述头孢菌素类选自头孢氨苄、头孢呋辛钠、头孢曲松、头孢匹罗；所述碳青酶烯类为硫霉素；所述氨基糖甙类选自庆大霉素、链霉素、卡那霉素；所述四环素类选自四环素、金霉素；所述大环内脂类选自红霉素、阿奇霉素；所述糖甙类为万古霉素；所述磺胺类选自磺胺嘧啶、甲氧苄啶；所述喹诺酮类选自吡哌酸、环丙沙星；所述硝咪唑类选自甲硝唑、替硝唑；所述林克胺类选自林可霉素、克林霉素。
  11. 根据权利要求9或10所述的应用，其特征在于，在该支架材料植入体内后15分钟内，所述抗菌物质的释放量不低于总加载量的30％。
  12. 一种肩袖补片，其特征在于，所述肩袖补片具有三维纳米网状结构，由包含纤维蛋白原复合物和聚乳酸-己内酯的原料制成。
  13. 根据权利要求12所述的肩袖补片，其特征在于，所述纤维蛋白原复合物包括如下重量份数的组分：纤维蛋白原0.1-20份、精氨酸盐酸盐0.1-10份、氯化钠0.01-10份和枸橼酸钠1-10份；
    优选地，所述纤维蛋白原复合物包括如下重量份数的组分：纤维蛋白原3-15份、精氨酸盐酸盐0.5-5份、氯化钠0.3-5份和枸橼酸钠1-10份。
  14. 根据权利要求12或13所述的肩袖补片，其特征在于，所述聚乳酸-己内酯的Mn为30000-80000，Mw为80000-160000，Mw/Mn<3.0；
    优选地，所述聚乳酸-己内酯中聚乳酸链段与聚己内酯链段的质 量比为20:80～95:5。
  15. 根据权利要求12-14任一项所述的肩袖补片，其特征在于，所述肩袖补片的厚度为1.6～10mm，优选为2～3mm。
  16. 根据权利要求12-15任一项所述的肩袖补片，其特征在于，所述肩袖补片中的蛋白质含量为100-220mg/g，残留蛋白含量<12mg/g；
    优选地，所述肩袖补片的水浸提液的pH为6-8。
  17. 根据权利要求12-16任一项所述的肩袖补片，其特征在于，所述肩袖补片的孔隙率为41～100％，优选为41～80％；
    和/或，所述肩袖补片的吸水率为35～200％，优选为55～80％；
    和/或，所述肩袖补片的拉伸强度为2～5MPa；
    和/或，所述肩袖补片的断裂伸长率为100～200％。
  18. 根据权利要求12-17任一项所述的肩袖补片，其特征在于，所述肩袖补片还含有具有抗菌和/或消炎作用的药物。
  19. 一种如权利要求12--18任一项所述的肩袖补片的制备方法，其特征在于，所述制备方法包括如下步骤：
    将纤维蛋白原复合物溶液和聚乳酸-己内酯溶液混合后加入静电纺丝机的同一容量管中，进行静电纺丝；或者，将纤维蛋白原复合物溶液和聚乳酸-己内酯溶液分别加入到静电纺丝机的不同容量管中，同时进行静电纺丝；干燥后得到所述肩袖补片。
  20. 根据权利要求19所述的制备方法，其特征在于，所述聚乳酸-己内酯溶液的浓度为5～8g/100mL；
    优选地，所述聚乳酸-己内酯溶液的溶剂选自六氟异丙醇、三氯甲烷、二甲基甲酰胺、四氢呋喃和丙酮中的一种或至少两种的组合；
    优选地，所述纤维蛋白原复合物溶液的溶剂为水或水溶液；
    优选地，所述纤维蛋白原复合物溶液的浓度为8-29g/100mL；
    优选地，所述纤维蛋白原复合物溶液与聚乳酸-己内酯溶液中的 纤维蛋白原复合物与聚乳酸-己内酯的质量比为0.48～1.1:1。
  21. 根据权利要求19所述的制备方法，其特征在于，所述静电纺丝的条件为：电压差为15～140kV，溶液流速为3-399mL/h或401-960mL/h，针头与芯轴之间的距离为10-50cm；
    优选地，所述制备方法还包括：在所述肩袖补片表面涂覆具有抗菌和/或消炎作用的药物。
PCT/CN2023/073024 2022-01-30 2023-01-19 一种肩袖补片及其制备方法 WO2023143337A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202210114561.9A CN114432501A (zh) 2022-01-30 2022-01-30 一种肩袖补片及其制备方法
CN202210114561.9 2022-01-30

Publications (1)

Publication Number Publication Date
WO2023143337A1 true WO2023143337A1 (zh) 2023-08-03

Family

ID=81371229

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2023/073024 WO2023143337A1 (zh) 2022-01-30 2023-01-19 一种肩袖补片及其制备方法

Country Status (2)

Country Link
CN (1) CN114432501A (zh)
WO (1) WO2023143337A1 (zh)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114432501A (zh) * 2022-01-30 2022-05-06 上海松力生物技术有限公司 一种肩袖补片及其制备方法
CN116370708B (zh) * 2023-01-15 2024-07-19 北京化工大学 一种具有压电效应的肩袖补片及其制备方法
CN117883216B (zh) * 2024-01-19 2024-08-09 常州集硕医疗器械有限公司 一种生物可吸收肩袖补片及其制备方法

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011035017A2 (en) * 2009-09-18 2011-03-24 The Trustees Of The University Of Pennsylvania Artificial meniscal implants
CN105705172A (zh) * 2013-11-19 2016-06-22 上海松力生物技术有限公司 用于组织再生的亲水性静电纺生物复合支架材料及其制法与应用
US20160175487A1 (en) * 2013-06-28 2016-06-23 Medprin Regenerative Medical Technologies Co., Ltd. Tissue repair scaffold and preparation method and purpose thereof
CN107510520A (zh) * 2017-09-21 2017-12-26 上海松力生物技术有限公司 十字交叉韧带再生性植入物及其制备方法与应用
US20190054205A1 (en) * 2017-05-16 2019-02-21 Embody Llc Biopolymer compositions, scaffolds and devices
CN113842498A (zh) * 2021-08-31 2021-12-28 上海市第六人民医院 一种促进肩袖腱骨愈合和抑制肌肉脂肪浸润的电纺膜支架及其制备方法
CN114432501A (zh) * 2022-01-30 2022-05-06 上海松力生物技术有限公司 一种肩袖补片及其制备方法

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101780292B (zh) * 2010-02-09 2013-10-16 复旦大学附属中山医院 以纤维蛋白原为基础的三维多孔纳米支架及其制备方法
JP2012205720A (ja) * 2011-03-29 2012-10-25 Gunze Ltd 腱板及び肩関節内構成体再生材料
CN103800097B (zh) * 2012-11-14 2016-12-21 深圳迈普再生医学科技有限公司 一种组织修复用纤维膜及其制造方法和应用
CN104841022A (zh) * 2014-02-14 2015-08-19 赵金忠 一种纳米纤维膜在制备治疗肩袖损伤的材料中的应用

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011035017A2 (en) * 2009-09-18 2011-03-24 The Trustees Of The University Of Pennsylvania Artificial meniscal implants
US20160175487A1 (en) * 2013-06-28 2016-06-23 Medprin Regenerative Medical Technologies Co., Ltd. Tissue repair scaffold and preparation method and purpose thereof
CN105705172A (zh) * 2013-11-19 2016-06-22 上海松力生物技术有限公司 用于组织再生的亲水性静电纺生物复合支架材料及其制法与应用
US20190054205A1 (en) * 2017-05-16 2019-02-21 Embody Llc Biopolymer compositions, scaffolds and devices
CN107510520A (zh) * 2017-09-21 2017-12-26 上海松力生物技术有限公司 十字交叉韧带再生性植入物及其制备方法与应用
CN113842498A (zh) * 2021-08-31 2021-12-28 上海市第六人民医院 一种促进肩袖腱骨愈合和抑制肌肉脂肪浸润的电纺膜支架及其制备方法
CN114432501A (zh) * 2022-01-30 2022-05-06 上海松力生物技术有限公司 一种肩袖补片及其制备方法

Also Published As

Publication number Publication date
CN114432501A (zh) 2022-05-06

Similar Documents

Publication Publication Date Title
WO2023143337A1 (zh) 一种肩袖补片及其制备方法
AU2017291837B2 (en) Indirect method of articular tissue repair
CN105705172B (zh) 用于组织再生的亲水性静电纺生物复合支架材料及其制法与应用
Catto et al. Small diameter electrospun silk fibroin vascular grafts: Mechanical properties, in vitro biodegradability, and in vivo biocompatibility
Yoo et al. Augmented peripheral nerve regeneration through elastic nerve guidance conduits prepared using a porous PLCL membrane with a 3D printed collagen hydrogel
ES2936715T3 (es) Métodos y productos de colágeno para reparación de tejidos
CN102231992B (zh) 用于治疗肌腱和韧带损伤的血小板衍生生长因子的组合物和方法
JP2019520900A5 (zh)
US9168326B2 (en) Collagen materials and methods for obtaining same
KR101151358B1 (ko) 실크 및 콜라겐을 포함하는 복합 지지체 및 그 제조방법
WO2009102967A2 (en) Molecular enhancement of extracellular matrix and methods of use
JP6648056B2 (ja) コラーゲン膜を生成するための方法およびその使用
WO1998030141A2 (en) Devices for tissue repair and methods for preparation and use thereof
Yoshii et al. Functional restoration of rabbit spinal cord using collagen‐filament scaffold
JP2022524714A (ja) マイクロフルイディック押出
CN114732948A (zh) 肩袖补片及其制备方法
KR101536134B1 (ko) 연부조직 수복용 매트릭스의 제조방법
WO2023143335A1 (zh) 一种诱导皮肤组织再生的亲水性静电纺植入物
US9468708B2 (en) Composite matrix
Meimandi-Parizi et al. Novel application of a tissue-engineered collagen-based three-dimensional bio-implant in a large tendon defect model: A broad-based study with high value in translational medicine
CN111346262B (zh) 一种用于促进腱骨愈合的可注射钙磷陶瓷及其制备方法与应用
Li et al. Enhancing neuroinduction activity of PLCL-based nerve conduits through native epineurium integration
Zhang et al. Evaluation of extracellular matrix-based nerve conduits for long-gap peripheral nerve repair in a goat model
He et al. Silk Fibroin Scaffolds Facilitating the Repair of Rat Abdominal Wall Defect
Chiera A model scaffold for Anterior Cruciate Ligament tissue engineering

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23746218

Country of ref document: EP

Kind code of ref document: A1