CN114618026B - Barrier membrane capable of promoting bone regeneration and preparation method thereof - Google Patents
Barrier membrane capable of promoting bone regeneration and preparation method thereof Download PDFInfo
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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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/005—Ingredients of undetermined constitution or reaction products thereof
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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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/146—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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/148—Materials at least partially resorbable by the body
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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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/16—Biologically active materials, e.g. therapeutic substances
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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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- 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
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
Abstract
The invention discloses a barrier membrane capable of promoting bone regeneration and a preparation method thereof, wherein the barrier membrane comprises a compact barrier layer and a loose collagen layer which is overlaid and paved on the barrier layer; the barrier layer is made of a dermal matrix membrane and/or a pericardium membrane of pigs and/or cattle, and the collagen layer is made of a peritoneal membrane and/or a small intestine submucosa of the pigs and/or the cattle. When in use, the barrier layer faces soft tissues and the collagen layer faces bone tissues, the barrier layer can prevent epithelial fiber cells with higher migration speed from growing into a bone defect area to interfere with bone formation, delay degradation time, better match the speed of bone formation and meet the requirement of bone growth time; the collagen layer has a special pore structure, allows osteoblasts with low migration speed to enter a bone defect area preferentially, promotes the growth of the osteoblasts, and has good practical and medical values.
Description
Technical Field
The present invention relates to a barrier membrane capable of promoting bone regeneration and a preparation method thereof.
Background
Leading to the generation of bone regeneration theory and technology, which is derived from regenerative treatment of periodontal disease, scholars such as Nyman use barrier membranes to block gingival connective tissue and epithelium from contacting the root surface so that damaged periodontal tissue is reconstructed. This technique of applying barrier membrane protection in periodontal soft and hard tissue regeneration is called guided tissue regeneration (guided bone regeneration, GTR). With the rapid development of the field of stomatology, a specific space is formed and maintained in a bone defect area by utilizing a barrier membrane, so that bone cells with a low growth rate are fixedly planted in the area, and simultaneously epidermis and connective tissue cells with a high tissue growth rate grow to the defect area, thereby realizing the regeneration of bone tissues, and the concept of guided bone regeneration (guided bone regeneration, GBR) is independent from the theory of guided tissue regeneration. At present, most of products on the market for guiding regeneration of oral tissues are collagen sponges extracted from bovine achilles tendons. Products for bone regeneration mainly utilize the formation of blood clots in the area of bone defects below the barrier membrane, followed by vascular ingrowth, granulation tissue formation, osteoblast migration recruitment and formation of woven bone, followed by lamellar bone formation and remodeling, the overall process having a very high similarity to the bone growth and development process.
Existing barrier films can be classified into non-absorbable and absorbable based on their degradation characteristics. The non-absorbable barrier membranes mainly include peptide omentum and polytetrafluoroethylene membranes, which have greater stiffness, higher space maintainability, and better clinical handling; however, the disadvantages are also prominent, and there is a risk of exposure due to the need for secondary surgery to remove the membrane for clinical use. Absorbable biofilms include synthetic polymer films which degrade at a slow rate and have processability, but do not have an active substance themselves, and thus do not have osteoinductive capacity and degradation products may trigger inflammatory reactions and rejection reactions, and thus absorbable biofilms are preferred.
Currently, the absorbable biological film on the market is mainly a collagen film, however, a large amount of clinical data show that the time required for complete regeneration of bone is about three months, and ideally, the degradation speed of the collagen film should be matched with the formation speed of bone; however, the collagen barrier film in the current market has shorter degradation time and can not meet the requirement of bone growth; the mechanical property is poor, collapse is easy to occur in the using process, the vertical bone increment is insufficient, and the bone repairing effect is not ideal; meanwhile, the material is animal-derived material, has higher immunogenicity, is easy to generate rejection reaction and inflammatory reaction, and increases the infection risk.
Disclosure of Invention
In order to solve the above problems, the present invention provides a barrier membrane for promoting bone regeneration, which is a double-layered membrane obtained from animal-derived tissues through a series of treatment methods, comprises a dense barrier layer and a relatively loose collagen layer, and can prolong degradation time and promote regeneration of osteoblasts better, and a method for preparing the same. The specific technical scheme is as follows:
first, the present invention provides a barrier membrane for promoting bone regeneration, comprising a dense barrier layer and a loose collagen layer overlaid and laid on the barrier layer; the barrier layer is made of dermis matrix membrane and/or pericardium membrane of pig and/or cattle,
the collagen layer is prepared from peritoneum and/or small intestine submucosa of pig and/or cattle.
Preferably, the aforementioned barrier membrane capable of promoting bone regeneration is made of 1-5 layers of porcine and/or bovine dermal matrix membrane and/or pericardium membrane, and has no pores; the collagen layer is formed by laminating 1-5 layers of peritoneum and/or small intestine submucosa of pigs and/or cattle, and the pore sizes of all the layers are sequentially increased along with the lamination sequence.
Further preferably, the aforementioned barrier membrane capable of promoting bone regeneration is made of three layers of porcine and/or bovine dermal matrix membrane and/or pericardial membrane; the collagen layer is formed by stacking three layers of peritoneum and/or small intestine submucosa of pigs and/or cattle, and the pore size of each layer is 45-55 mu m, 90-110 mu m and 195-210 mu m in sequence.
Next, the present invention provides a method for preparing the aforementioned barrier membrane capable of promoting bone regeneration, comprising the steps of:
s1: barrier film treatment: washing dermis matrix membrane and/or pericardium membrane of pig and/or cattle, degreasing, removing virus, removing cell, and removing immunogen to obtain barrier membrane for preparing barrier layer;
s2: collagen membrane treatment: washing, degreasing, removing impurity protein and cell from the peritoneum and/or small intestine submucosa of the pig and/or the cow, and performing deimmunization to obtain a collagen film for preparing a collagen layer;
s3: film laying: firstly, superposing and tiling the treated barrier films according to requirements and compacting to form a compact barrier layer, then superposing and tiling the collagen films on the barrier layer according to requirements to form a loose collagen layer, and sequentially increasing the pore sizes of the collagen films of the collagen layer according to the sequence of tiling the films;
s4: dehydrating and drying: and (3) dehydrating and drying after the membrane is laid, and bonding all layers together to obtain the barrier membrane capable of promoting bone regeneration.
The aforementioned preparation method of the barrier membrane capable of promoting bone regeneration, wherein the step S1 of barrier membrane treatment specifically comprises the following steps:
s1-1: washing the dermis matrix membrane and/or pericardium membrane of pig and/or cattle with clear water;
s1-2: soaking in degreasing reagent for 10-25 h;
s1-3: soaking in cell eliminating reagent for 3-5 hr;
s1-4: the immunogen is removed to obtain a barrier membrane for preparing the barrier layer.
Wherein the degreasing reagent is ethanol, sodium hydroxide or mixed solution of chloroform and methanol; the decellularized reagent is hydrogen peroxide solution with the mass concentration of 1-5% or sodium hypochlorite solution with the mass concentration of 0.1-3%; the deimmunizing agent is trypsin solution, the concentration of which is 0.1-5% and the pH range is 7-9.
Preferably, the ethanol used by the degreasing reagent is 75% -95% ethanol, and the concentration of the sodium hydroxide solution is 1-2 mol/L; the volume ratio of the chloroform to the methanol mixed solution is: methanol=2 to 3:1 to 2.
The aforementioned preparation method of the barrier membrane capable of promoting bone regeneration, the step S2 of collagen membrane treatment specifically comprises the following steps:
s2-1: washing the peritoneum and/or small intestine submucosa of the pig and/or the cow with clear water;
s2-2: soaking in degreasing reagent for 16-24 h;
s2-3: soaking in cell eliminating reagent for 2-4 hr;
s2-4: removing the immunogen to obtain the collagen membrane for preparing the collagen layer.
Wherein the degreasing agent is n-hexane or acetone; the decellularized reagent is hydrogen peroxide solution with the mass concentration of 0.1-2% or sodium hypochlorite solution with the mass concentration of 0.01-1%; the deimmunizing agent is a solution of terminal peptidase, the concentration of which is 0.1-1% and the pH range is 7-9.
In the aforementioned preparation method of the barrier membrane capable of promoting bone regeneration, step S4 is dehydration drying, wherein the drying mode is vacuum freeze drying or ethanol gradient dehydration drying; the technological parameters of the vacuum freeze drying are as follows: -40-25 ℃ and drying time is 30-45 h; the technological parameters of ethanol gradient dehydration and drying are as follows: and (3) gradient dehydration and drying of 25-100%.
The beneficial effects of the invention are as follows:
1) The barrier membrane is divided into a compact barrier layer and a loose collagen layer, when the barrier layer is used, the barrier layer faces soft tissues, the collagen layer faces bone tissues, the barrier layer can prevent epithelial fiber cells with high migration speed from growing into a bone defect area to interfere with bone formation, the degradation time is delayed, the speed of bone formation is better matched, and the requirement of bone growth time is met.
2) The collagen layer has a special pore structure, allows osteoblasts with low migration speed to enter a bone defect area preferentially, and promotes the growth of the osteoblasts.
3) The barrier membrane is prepared from six layers of animal-derived tissues, and the formed product has good ductility, good mechanical property and stiffness, has a good supporting effect, can bear the pressure brought by a wound without collapsing, and provides a basic guarantee for the effect of bone repair.
4) The barrier layer is extruded by pressure, so that the layers are tightly connected, the degradation time in the body is prolonged, and enough time and space are provided for bone growth; the collagen layer adopts milder reagent, retains active substances in the loose layer, and the active substances can promote bone growth to a certain extent.
Drawings
FIG. 1 is a schematic structural view of a barrier membrane for promoting bone regeneration according to the present invention;
FIG. 2 is a physical view of a barrier membrane for promoting bone regeneration according to the present invention;
fig. 3 is a photograph of a dental extraction patient stitched using a barrier membrane of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described below with reference to the embodiments and the accompanying drawings.
Example 1
This example is to prepare a barrier membrane that promotes bone regeneration, comprising a dense barrier layer and a loose collagen layer overlying the barrier layer; the barrier layer is prepared from 1-5 layers of pig or cow dermis matrix membranes or pericardium membranes, and the collagen layer is prepared from 1-5 layers of pig or cow peritoneum or small intestine submucosa. In this embodiment, as shown in fig. 1, the barrier layer is formed by compacting three layers of bovine pericardium, and the structure of the barrier layer has no pores; the collagen layer is formed by stacking three layers of calf intestinal submucosa, and the pore sizes of the layers are sequentially increased along with the stacking sequence and are respectively 50 mu m, 100 mu m and 200 mu m; the preparation method of the barrier membrane capable of promoting bone regeneration specifically comprises the following steps:
s1: barrier film treatment: washing bovine pericardium with clear water, soaking in degreasing agent for 20h for degreasing; removing virus, soaking in a decellularizing reagent for 4 hours to perform decellularizing treatment, and finally performing deimmunization by using a trypsin solution to obtain the barrier membrane for preparing the barrier layer. The degreasing reagent is a mixed solution of chloroform and methanol, and the volume ratio of the mixed solution is that the mixed solution is composed of chloroform: methanol=3: 2; the virus-removing reagent is NaOH solution with the concentration of 1 mol/L; the decellularization reagent is hydrogen peroxide solution with the mass concentration of 3%; the trypsin solution had a concentration of 2% and a pH of 8.
S2: collagen membrane treatment: washing the submucosa of the small intestine of the cattle with clear water, soaking in a degreasing reagent for 18h to degrease and remove impurity proteins; then placing the collagen membrane in a decellularization reagent for soaking for 3 hours to decellularize, and finally, performing deimmunization by using a terminal-removing peptidase solution to obtain the collagen membrane for preparing the collagen layer. The degreasing reagent is n-hexane; the impurity-removing protein reagent is a sodium dodecyl sulfate solution with the mass concentration of 0.5%; the decellularization reagent is sodium hypochlorite solution with the mass concentration of 0.05%; the concentration of the solution of the terminal peptidase is 0.5%, and the pH value is 7; then rolling with rollers of different thicknesses to form different pores.
S3: film laying: the treated three-layer barrier film is overlapped, tiled and compacted according to the requirement to form a compact barrier layer, then the three-layer collagen film is overlapped, tiled on the barrier layer according to the requirement to form a loose collagen layer, and the pore sizes of the collagen films of the collagen layer are sequentially tiled according to the film paving sequence of 50 mu m, 100 mu m and 200 mu m.
S4: dehydrating and drying: after the membrane is laid, dehydrating and drying by adopting an ethanol gradient dehydrating and drying mode, so that a barrier membrane capable of promoting bone regeneration can be obtained, as shown in figure 2; the ethanol gradient dehydration and drying process comprises the following steps: soaking in 25%, 45%, 65%, 85%, and 100% ethanol sequentially for 10 hr for dehydration to bond the layers together, and taking out and naturally drying after dehydration.
Example 2
This example also shows the preparation of a barrier membrane for promoting bone regeneration comprising a dense barrier layer and a loose collagen layer overlying the barrier layer; the barrier layer is made of a dermal matrix membrane or a pericardium membrane of a pig or a cow, and the collagen layer is made of a peritoneum or small intestine submucosa of the pig or the cow. In this embodiment, the barrier layer is formed by compacting three layers of porcine dermal matrix membranes, and the structure of the barrier layer is free of pores. The collagen layer is formed by overlapping three layers of pig peritoneum, and the pore sizes of all layers are sequentially increased along with the overlapping sequence; the pore sizes of the layers in this example were 45 μm, 95 μm and 195. Mu.m, respectively. The preparation method of the barrier membrane capable of promoting bone regeneration specifically comprises the following steps:
s1: barrier film treatment: washing the pig dermis matrix membrane with clear water; then soaking in a degreasing reagent for 15 hours to degrease, removing viruses, and then soaking in a decellularizing reagent for 3.5 hours to decellularize; finally, the trypsin solution is used for deimmunization, and a barrier membrane for preparing the barrier layer is obtained. The degreasing reagent is a mixed solution of ethanol and methanol; the volume ratio of the ethanol is as follows: methanol=2: 1, a step of; the virus-removing reagent is NaOH solution with the concentration of 1 mol/L; the decellularization reagent is hydrogen peroxide solution with the mass concentration of 2%; the trypsin solution had a concentration of 1.5% and a pH of 7.5.
S2: collagen membrane treatment: washing pig peritoneum with clear water, soaking in degreasing agent for 12h to degrease, removing impurity protein, soaking in decellularizing agent for 2h to decellularize, and finally performing deimmunization by using terminal-removing peptidase solution to obtain collagen film for preparing collagen layer; the degreasing reagent is acetone; the impurity-removing protein reagent is a sodium dodecyl sulfate solution with the mass concentration of 0.5%; the decellularization reagent is hydrogen peroxide solution with the mass concentration of 0.5%; the concentration of the solution of the terminal peptidase is 0.2%, and the pH range is 7; then rolling with rollers of different thicknesses to form different pores.
S3: film laying: firstly, stacking and tiling three layers of treated barrier films according to requirements and compacting to form a compact barrier layer, then stacking and tiling the treated collagen films on the barrier layer according to requirements to form a loose collagen layer, and stacking the pore sizes of the collagen films of the collagen layer according to the order of 45 mu m, 95 mu m and 195 mu m of the stacked films.
S4: dehydrating and drying: and (3) dehydrating and drying the membrane by adopting a vacuum freeze drying mode after membrane laying is finished, and bonding all layers together to obtain the barrier membrane capable of promoting bone regeneration. The technological parameters of the vacuum freeze drying are as follows: -40 ℃ and drying time is 45h.
Example 3
This example also shows the preparation of a barrier membrane for promoting bone regeneration comprising a dense barrier layer and a loose collagen layer overlying the barrier layer; the barrier layer is made of a dermal matrix membrane or a pericardium membrane of a pig or a cow, and the collagen layer is made of a peritoneum or small intestine submucosa of the pig or the cow. In the embodiment, the barrier layer is formed by compacting two layers of pig pericardium and one layer of bovine dermis matrix membrane, and the structure of the barrier layer is free of pores; the collagen layer is formed by superposing two layers of pig small intestine mucosa and one layer of bovine peritoneum, and the pore sizes of the layers are sequentially increased along with the superposition sequence, and the pore sizes of the layers in the embodiment are 55 mu m, 100 mu m and 210 mu m respectively. The preparation method of the barrier membrane capable of promoting bone regeneration specifically comprises the following steps:
s1: barrier film treatment: washing the porcine pericardium membrane and the bovine dermis matrix membrane with clear water, soaking in a degreasing agent for 24 hours for degreasing, removing viruses, soaking in a decellularizing agent for 5 hours for decellularizing treatment, and finally adding a trypsin solution for deimmunizing to obtain the barrier membrane for preparing the barrier layer. The degreasing reagent is a mixed solution of sodium hydroxide and methanol, and the volume ratio of the de-mixed solution is sodium hydroxide: methanol=3: 1, a step of; the virus-removing reagent is NaOH solution with the concentration of 1 mol/L; the decellularization reagent is hydrogen peroxide solution with the mass concentration of 5%; the trypsin solution had a concentration of 5% and a pH of 9.
S2: collagen membrane treatment: washing the mucous membrane of the small intestine of the pig and the peritoneum of the cow with clear water, soaking in a degreasing reagent for 20 hours to degrease and remove impurity proteins, then soaking in a decellularizing reagent for 4 hours to decellularize, and finally performing deimmunization by using a terminal peptidase solution to obtain the collagen membrane for preparing the collagen layer. The degreasing reagent is acetone; the impurity-removing protein reagent is a sodium dodecyl sulfate solution with the mass concentration of 0.5%; the decellularization reagent is sodium hypochlorite solution with the mass concentration of 1%; the concentration of the solution of the terminal peptidase is 0.8%, and the pH range is 8; then rolling with rollers of different thicknesses to form different pores.
S3: film laying: firstly, superposing and tiling the treated two layers of pig pericardium and one layer of cow dermis matrix according to the requirement to form a compact barrier layer, then superposing and tiling the two layers of pig small intestine mucosa and one layer of cow peritoneum on the barrier layer according to the requirement to form a loose collagen layer, and sequentially tiling the pore sizes of the collagen films of the collagen layer according to the membrane laying sequence of 55 mu m, 100 mu m and 210 mu m.
S4: dehydrating and drying: and (5) dehydrating and drying by adopting a freeze-drying mode after membrane laying is finished, so that the barrier membrane capable of promoting bone regeneration can be obtained. The technological parameters of dehydration and drying are as follows: drying at-40deg.C for 40 hr.
Example 4 application example
The embodiment verifies the use effect of the product. After the completion of tooth extraction, the patient fills the tooth extraction socket with bone repair material and uses the barrier membrane of the present invention for suturing, as shown in fig. 3. The physical isolation function of the barrier membrane is utilized to separate different periodontal tissues, a compact barrier layer faces soft tissues during suturing, epithelial fibroblasts with high migration speed are prevented from growing into a bone defect area to interfere bone formation, a loose collagen layer faces bone tissues, enough space is provided for osteoblasts with low migration speed, growth of the osteoblasts is promoted, and normal connection relation between periodontal and root surfaces is reestablished.
In 100 patients in follow-up investigation, the bone repair effect after tooth extraction is good, the time of complete absorption of the used barrier film is 110-128 days, the time required by complete regeneration of bone is met, and the phenomenon of wound surface collapse caused by insufficient vertical bone increment does not occur in 100 patients; and no serious rejection or inflammatory reaction occurs basically, and the use experience effect is good.
In general, the barrier membrane is divided into a dense barrier layer and a loose collagen layer, when the barrier layer is used, the barrier layer faces soft tissues, the collagen layer faces bone tissues, the barrier layer can prevent epithelial fiber cells with higher migration speed from growing into a bone defect area to interfere with bone formation, the degradation time is delayed, the bone formation speed is better matched, and the bone growth time requirement is met.
The collagen layer has a special pore structure, allows osteoblasts with low migration speed to enter a bone defect area preferentially, and promotes the growth of the osteoblasts; the barrier membrane is prepared from animal-derived tissues, and the formed product has good ductility, good mechanical property and stiffness, has a good supporting effect, can bear the pressure brought by a wound without collapsing, and provides a basic guarantee for the effect of bone repair. The barrier layer is extruded by pressure, so that the layers are tightly connected, the degradation time in the body is prolonged, and enough time and space are provided for bone growth; the collagen layer adopts a milder reagent, active substances in the loose layer are reserved, and the active substances can promote bone growth to a certain extent, so that the collagen layer has good practical and medical values.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. Accordingly, the embodiments are to be considered in all respects as illustrative and not restrictive. Furthermore, it should be understood that, although the present disclosure describes embodiments, this description is not intended to include only one embodiment, and those skilled in the art should understand that the present disclosure is not limited to the embodiments described herein, and that the embodiments described in the examples may be combined appropriately to form other embodiments that will be understood by those skilled in the art.
Claims (5)
1. A method for preparing a barrier membrane capable of promoting bone regeneration, which is characterized in that: comprises a dense barrier layer and a loose collagen layer which is overlapped and paved on the barrier layer;
the barrier layer is formed by compacting 3 layers of pig and/or cow dermis matrix membranes, and the structure of the barrier layer is free of pores;
the collagen layer is formed by overlapping 3 layers of peritoneum of pigs and/or cattle, and the pore sizes of all layers are sequentially increased along with the overlapping sequence; the pore sizes of the layers are 45-55 mu m, 90-110 mu m and 195-210 mu m from bottom to top in sequence;
the preparation method of the barrier film comprises the following steps:
s1: barrier film treatment: washing the dermis matrix membrane of the pig and/or the cow, degreasing, removing virus, removing cells, and then performing deimmunization to obtain a barrier membrane for preparing a barrier layer; the degreasing reagent is ethanol, sodium hydroxide or mixed solution of chloroform and methanol; the reagent for cell removal is hydrogen peroxide solution with the mass concentration of 1-5% or sodium hypochlorite solution with the mass concentration of 0.1-3%; the reagent for removing immunogen is trypsin solution, the concentration of the reagent is 0.1% -5%, and the pH range is 7-9;
s2: collagen membrane treatment: washing, degreasing, removing impurity protein and cell from peritoneum of pig and/or cattle, and then performing deimmunization to obtain a collagen film for preparing a collagen layer; the degreasing reagent is n-hexane or acetone; the reagent for cell removal is hydrogen peroxide solution with the mass concentration of 0.1-2% or sodium hypochlorite solution with the mass concentration of 0.01-1%; the reagent for removing immunogen is a solution of terminal peptidase, the concentration of the reagent is 0.1% -1%, and the pH range is 7-9;
s3: film laying: firstly, superposing and tiling the treated barrier films according to requirements and compacting to form a compact barrier layer, then superposing and tiling the collagen films on the barrier layer according to requirements to form a loose collagen layer, and sequentially increasing the pore sizes of the collagen films of the collagen layer according to the sequence of tiling the films;
s4: dehydrating and drying: and (3) dehydrating and drying after the membrane is laid, and bonding all layers together to obtain the barrier membrane capable of promoting bone regeneration.
2. The method for producing a barrier membrane for promoting bone regeneration according to claim 1, wherein: step S1 the barrier film treatment specifically comprises the steps of:
s1-1: washing the dermis matrix membrane of pig and/or cattle with clear water;
s1-2: soaking in degreasing reagent for 10-25 hr;
s1-3: soaking in the cell removing reagent for 3-5 h;
s1-4: the immunogen is removed to obtain a barrier membrane for preparing the barrier layer.
3. The method for preparing a barrier membrane for promoting bone regeneration according to claim 2, wherein: when the degreasing reagent is ethanol and methanol mixed solution, the concentration of the ethanol is 75-95%, and the volume ratio of the ethanol to the methanol mixed solution is 2:1;
when the degreasing reagent is a mixed solution of sodium hydroxide and methanol, the concentration of the sodium hydroxide solution is 1-2 mol/L, and the volume ratio of the sodium hydroxide solution to the methanol is 3:1;
when the degreasing reagent is a mixed solution of chloroform and methanol, the volume ratio of the mixed solution of chloroform and methanol is 2-3:1-2.
4. A method for preparing a barrier membrane for promoting bone regeneration according to claim 3, wherein: the step S2 of collagen membrane treatment specifically comprises the following steps:
s2-1: washing the peritoneum of pig and/or cattle with clear water;
s2-2: soaking in degreasing reagent for 16-24 hr;
s2-3: soaking in the cell removing reagent for 2-4 h;
s2-4: removing the immunogen to obtain the collagen membrane for preparing the collagen layer.
5. The method for producing a barrier membrane for promoting bone regeneration according to claim 4, wherein: step S4, dehydrating and drying, wherein the drying mode is vacuum freeze drying or ethanol gradient dehydrating and drying;
the technological parameters of the vacuum freeze drying are as follows: -40-25 ℃ and drying time is 30-45 h;
the technological parameters of ethanol gradient dehydration and drying are as follows: and (3) gradient dehydration and drying of 25-100%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210262309.2A CN114618026B (en) | 2022-03-17 | 2022-03-17 | Barrier membrane capable of promoting bone regeneration and preparation method thereof |
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| CN115429939B (en) * | 2022-09-23 | 2023-06-16 | 博纳格科技(天津)有限公司 | Preparation method of oral biological film with controllable product performance |
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