CN113633827A - Silk woven meniscus implant and preparation method thereof - Google Patents
Silk woven meniscus implant and preparation method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
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- 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
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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- 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
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
- A61L27/3608—Bone, e.g. demineralised bone matrix [DBM], bone powder
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- 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
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- 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
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/30—Compounds of undetermined constitution extracted from natural sources, e.g. Aloe Vera
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- 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
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
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- 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
- A61L2430/06—Materials or treatment for tissue regeneration for cartilage reconstruction, e.g. meniscus
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Abstract
The invention discloses a silk woven meniscus implant and a preparation method thereof. The silk is adopted as a weaving material and is prepared by bone glue infusion molding. The invention adopts the three-dimensional weaving technology to realize the mechanical integrated weaving of the silk, so that the precise forming of the woven body becomes possible, and the industrial production is easy; the allogenic bone glue is used as a molding perfusion material, so that the material has better biocompatibility; the bone glue is prepared from the decalcified bone meal, and the formed meniscus has multiple pores, is favorable for the attachment of surrounding nutrient substances, and is favorable for wound healing.
Description
Technical Field
The invention belongs to the technical field of preparation of meniscus repairing materials, and particularly relates to a silk woven meniscus implant and a preparation method thereof.
Background
The physician website statistics in 2018 show that meniscus injury has become the most common knee joint injury, and the prevalence rate of acute meniscus tear reaches 61 cases/100000 people. Meniscal reconstruction has become one of the research hotspots of researchers in the field of injury repair, the main modes of which include allogeneic meniscus transplantation and artificial substitute transplantation. Allogeneic meniscus transplantation can effectively relieve pain of patients and improve knee joint function, but has the problems of insufficient graft source, difficult tissue preservation and the like. Artificial meniscal implant materials are divided into two classes of natural and synthetic materials.
Collagen meniscal graft (CMI) by vy Sports Medicine, USA
Conducting commercial investment and research and development. The product can generate new tissues without toxic and side effects, and effectively improve clinical symptoms of patients and clinical scores of knee joints; but the mechanical property can not meet the requirement of maintaining the normal function of the knee joint, and the commodity is only suitable for the case that the peripheral edge of the meniscus is not damaged. The British Qrteq company develops further research and development to prepare a polycaprolactone/polyurethane meniscus scaffold with the pore diameter of 150-335 mu m and the porosity of 80%
The product can reduce pressure on tibial plateau, promote tissue growth and protect articular cartilage, and improve clinical score of knee joint of patients; however, polyurethanes may produce small amounts of degradable toxic diamines.
The American company Active Implants developed a commercial, long-standing alternative polycarbonate Polyurethane (PCU) meniscal implant
The product has good qualityDue to the composite structure and design, the artificial meniscus does not need to be fixed to bone or soft tissue. An implantable cartilaginous tissue repair device (200680038582.6) is disclosed by oxford biomaterials ltd.a three-dimensional fibrous layer is formed from silk by one or more of winding or weaving or compressing a felt or twisting or knitting or braiding or stitching or embroidering or bonding a cloth layer, and a hydrogel is cast, the hydrogel comprising gelatin, fibrin, hyaluronic acid, alginate, etc. Affiliated drum building hospital of Nanjing university's department of medicine discloses a medical knee joint meniscus (200720131799.3), the designed meniscus is formed by crocheting silk and wrapping type I collagen. Beijing aerospace university's application discloses an artificial meniscus scaffold based on a fiber weaving structure and a weaving method (201710999938.2), the meniscus scaffold is formed by stacking and connecting silk in series after being woven in a single layer by a traditional weaving method, and is solidified by a pouring mode and wrapped with solid materials such as collagen, polyvinyl alcohol hydrogel and polyurethane. The materials such as hydrogel, type I collagen and collagen have poor histocompatibility and poor mechanical properties.
Disclosure of Invention
The invention aims to provide a silk woven meniscus implant and a preparation method thereof.
A silk woven meniscus implant is prepared by using silk as a weaving material and carrying out bone glue infusion forming.
A preparation method of a silk woven meniscus implant comprises the following steps:
(1) selecting silk as a weaving material, based on a stitch-bonding technology, flatly laying a plurality of layers of continuous silk fibers and sewing in a Z direction to form an oriented three-dimensional fabric, wherein the weaving body is characterized by a semi-ring C shape or O shape, and the longitudinal section of the weaving body is a wedge-shaped structure with a high outer side and a low inner side;
(2) immersing the silk woven scaffold prepared in the step (1) in a sodium carbonate aqueous solution, degumming for 10-30 minutes at 80-120 ℃, washing for 1-3 times by warm water, and repeating for 1-6 times;
(3) taking allogeneic cortical bone or cancellous bone, and preparing the allogeneic cortical bone or cancellous bone into decalcified bone powder after powdering, decalcifying, cleaning and freeze-drying;
(4) mixing the decalcified bone meal with purified water, and steaming in an autoclave at the temperature of 100-;
(5) and (3) placing the silk braid prepared in the step (2) into a mold with a meniscus-shaped groove, pouring bone glue, infiltrating in vacuum, and performing freeze drying and molding.
The diameter of the silk in the step (1) is 0.2-0.8 mm.
The number of the layers is 2-20.
The concentration of the sodium carbonate aqueous solution is 0.01-0.05M.
The decalcified bone meal in the step (4): the proportion of purified water is 1: (1-5).
The invention has the beneficial effects that: the invention adopts the three-dimensional weaving technology to realize the mechanical integrated weaving of the silk, so that the precise forming of the woven body becomes possible, and the industrial production is easy. The invention adopts the allogeneic bone glue as the molding perfusion material, and has better biocompatibility. The bone glue is prepared from the decalcified bone meal, and the formed meniscus has multiple pores, is favorable for the attachment of surrounding nutrient substances, and is favorable for wound healing.
Drawings
FIG. 1 shows the results of cytotoxicity test.
FIG. 2 shows the observation results under a paraffin section microscope.
Fig. 3 is a graph of the stiffness and elastic modulus of the braid stent and human meniscus.
Detailed Description
In order that the invention may be more fully understood, reference will now be made to the following description. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
EXAMPLE 1 preparation of Silk woven meniscus implants
1. Silk weaving
Weaving: silk with the fiber diameter of 0.5mm is selected as a weaving material, based on the stitch-bonding technology, continuous silk fibers are flatly laid in 5 layers and sewn in the Z direction to form a directional three-dimensional fabric, the weaving body is characterized by being semi-ring C-shaped, and the longitudinal section of the weaving body is of a wedge-shaped structure with the high outer side and the low inner side.
Degumming: degumming the silk woven scaffold in 0.02M sodium carbonate aqueous solution at 100 ℃ for 20 minutes, washing with warm water for 2 times, and repeating for 3 times.
2. Preparation of bone glue
Preparing the decalcified bone: the allogeneic cortical bone is prepared into decalcified bone powder after powdering, decalcifying, cleaning and freeze-drying;
preparing bone glue: according to the calcium-removed bone meal: the proportion of purified water is 1: 3.5, steaming and boiling the mixture for 5 hours at 105 ℃ in an autoclave to form bone glue.
3. Composite molding
And (3) placing the braided stent into a mold with a meniscus-shaped groove, pouring bone glue, infiltrating in vacuum, and performing freeze drying and molding.
Example 2
1. Silk weaving
Weaving: silk with the fiber diameter of 0.5mm is selected as a weaving material, based on the stitch-bonding technology, continuous silk fibers are flatly laid in 5 layers and sewn in the Z direction to form an oriented three-dimensional fabric, the weaving body is characterized by being O-shaped, and the longitudinal section of the weaving body is of a wedge-shaped structure with the high outer side and the low inner side.
Degumming: degumming the silk woven scaffold in 0.02M sodium carbonate aqueous solution at 100 ℃ for 20 minutes, washing with warm water for 2 times, and repeating for 3 times.
2. Preparation of bone glue
Preparing the decalcified bone: the allogeneic cancellous bone is prepared into decalcified bone powder after powdering, decalcifying, cleaning and freeze-drying;
preparing bone glue: according to the calcium-removed bone meal: the proportion of purified water is 1: 3.5, steaming and boiling the mixture for 5 hours at 105 ℃ in an autoclave to form bone glue.
3. Composite molding
And (3) placing the braided stent into a mold with a meniscus-shaped groove, pouring bone glue, infiltrating in vacuum, and performing freeze drying and molding.
Experimental example 1: material cytotoxicity test:
the experimental method comprises the following steps: the braided body stent prepared in example 1 was prepared into a square body, and after the material was sterilized, it was extracted at a ratio of 3cm2Perml (surface area/volume) was added to high sugar DMEM medium and soaked for 24 hours. The L929 cells were arranged in 4X 104One well was inoculated in a 96-well plate, and after 24 hours, 100ul of the leaching solution was added to each well of the experimental group, and the normal medium was added to the control group. After 24 hours, the activity of each group of cells was examined by the CCK8 method.
The detection results are shown in fig. 1, the cell viability of the control group is equivalent to that of the experimental group, and the difference is not statistically significant.
Experimental example 2: in vivo implantation experiment and histological observation of stent
The experimental method comprises the following steps:
the knitted meniscus prepared in example 1 was prepared in a rabbit meniscus shape, and the stent was sterilized by irradiation for use.
Establishing a complete defect model of the meniscus at the inner side of the rabbit and implanting a stent:
chloral hydrate 10% was injected intravenously into New Zealand big ear rabbits at 1.5ml/kg of ear margin. Fixing the rabbit in supine position on the dissecting table after anesthesia, sterilizing the skin of the left back leg, wrapping the rabbit feet with sterile gloves, and laying sterile sheets. Taking the front inner side incision of the knee joint, incising the skin and subcutaneous tissues layer by layer, opening the joint capsule, turning the patella outwards, and dissociating the soft tissues around the meniscus. The medial meniscus was exposed and dissociated and completely excised, and a model of complete defect of rabbit medial meniscus was constructed. Implanting a half-moon plate of the braided body, sewing front and rear angles of the braided body on the crossed ligament, and sewing the periphery on peripheral soft tissues. The subcutaneous tissue and the skin were sutured in order.
(iii) sacrifice of experimental animals 2 months after implantation in vivo, and removal of the knitted meniscal stent and the normal medial meniscus on the opposite side. The experimental specimen is fixed, dehydrated, embedded in paraffin, sliced, and the specimen slice is subjected to HE staining and observed under a microscope.
The observation result is shown in fig. 2, the growth of the fibroblast-like cells on the silk fiber can be seen in the stent body, the formation of new blood vessels can be seen, no obvious inflammatory cell distribution can be seen, and the degradation of the material is not obvious.
Experimental example 3: mechanical property detection and result of material
The experimental method comprises the following steps: the human meniscus and the knitted scaffold prepared in example 1 were taken, and excess tissues such as fat and the like around the specimen were peeled off and placed in a universal mechanical testing machine for a compression test.
The test results are shown in fig. 3, the rigidity and the elastic modulus of the braided body stent and the human meniscus are equivalent, and the difference has no statistical significance.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (6)
1. The silk woven meniscus implant is characterized in that silk is used as a woven material and is prepared by bone glue infusion forming.
2. A preparation method of a silk woven meniscus implant is characterized by comprising the following steps:
(1) selecting silk as a weaving material, based on a stitch-bonding technology, flatly laying a plurality of layers of continuous silk fibers and sewing in a Z direction to form an oriented three-dimensional fabric, wherein the weaving body is characterized by a semi-ring C shape or O shape, and the longitudinal section of the weaving body is a wedge-shaped structure with a high outer side and a low inner side;
(2) immersing the silk woven scaffold prepared in the step (1) in a sodium carbonate aqueous solution, degumming for 10-30 minutes at 80-120 ℃, washing for 1-3 times by warm water, and repeating for 1-6 times;
(3) taking allogeneic cortical bone or cancellous bone, and preparing the allogeneic cortical bone or cancellous bone into decalcified bone powder after powdering, decalcifying, cleaning and freeze-drying;
(4) mixing the decalcified bone meal with purified water, and steaming in an autoclave at the temperature of 100-;
(5) and (3) placing the silk braid prepared in the step (2) into a mold with a meniscus-shaped groove, pouring bone glue, infiltrating in vacuum, and performing freeze drying and molding.
3. The method for preparing the silk woven meniscus implant according to claim 2, wherein the diameter of the silk of step (1) is 0.2-0.8 mm.
4. The method for preparing the silk woven meniscus implant according to claim 2, wherein the plurality of layers is 2 to 20 layers.
5. The method for preparing the silk woven meniscus implant according to claim 2, wherein the concentration of the aqueous sodium carbonate solution is 0.01 to 0.05M.
6. The method for preparing the silk woven meniscus implant according to claim 2, wherein the decalcified bone meal of step (4): the proportion of purified water is 1: (1-5).
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| CN202110939707.9A CN113633827A (en) | 2021-08-16 | 2021-08-16 | Silk woven meniscus implant and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114214838A (en) * | 2021-12-13 | 2022-03-22 | 宜兴市新立织造有限公司 | Silk/gel multidirectional fiber sandwich meniscus and preparation method thereof |
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