CN114618026A - 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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Abstract
The invention discloses a barrier film capable of promoting bone regeneration and a preparation method thereof, wherein the barrier film comprises a compact barrier layer and a loose collagen layer which is overlaid and paved on the barrier layer; the barrier layer is prepared from porcine and/or bovine dermal matrix membrane and/or pericardium, and the collagen layer is prepared from porcine and/or bovine peritoneum and/or small intestine submucosa. When the bone growth barrier layer is used, the barrier layer faces soft tissues, the collagen layer faces bone tissues, the barrier layer can prevent epithelial fibrocytes with high migration speed from growing into a bone defect area to interfere 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 preferentially enter the bone defect area, promotes the growth of the osteoblasts, and has good practical and medical values.
Description
Technical Field
The 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, Nyman et al use the barrier membrane to block the contact of gingival connective tissue and epithelium with the root surface, so that the damaged periodontal tissue is reconstructed. This technique of applying barrier membrane protection in periodontal soft and hard tissue regeneration is called guided bone regeneration (GTR). Subsequently, with the rapid development of the field of oral medicine, a barrier membrane is utilized to form and maintain a specific space in a bone defect area, so that bone cells with a slow growth speed are planted in the area, and meanwhile, epidermal and connective tissue cells with a fast tissue growth speed grow to the defect area, so that the regeneration of bone tissues is realized, and therefore, the concept of Guided Bone Regeneration (GBR) is independent from the theory of guided tissue regeneration. At present, most of products for guiding the regeneration of oral tissues in the market are collagen sponges extracted from bovine achilles tendons. The product for bone regeneration mainly utilizes the formation of blood clots in the bone defect area below the barrier membrane, the subsequent growth of blood vessels, the formation of granulation tissues, the migration and recruitment of osteoblasts and the formation of braided bone, and then the formation and reconstruction of layered bone, and the whole process has extremely high similarity with the growth and development process of the bone.
The existing barrier film can be divided into two types of non-absorbable and absorbable according to different degradation characteristics. Non-absorbable barrier membranes mainly include peptide omentum and polytetrafluoroethylene membranes, which have greater stiffness, greater space maintenance, and better clinical operability; however, the disadvantages are significant, and the clinical application requires a second operation to remove the membrane and risks exposure. Absorbable biofilms include synthetic polymer membranes which are slow in degradation rate and processable but do not have active substances themselves and thus have no osteoinductive capacity and degradation products may induce inflammatory and body rejection reactions, and absorbable biofilms are preferred.
The absorbable biological membrane on the market is mainly a collagen membrane, however, a large amount of clinical data show that the time required for complete regeneration of bones is about three months, and ideally, the degradation speed of the collagen membrane is matched with the bone formation speed; however, the degradation time of the collagen barrier membrane on the current market is short, and the requirement of bone growth cannot be met; the mechanical property is poor, and the bone is easy to collapse in the using process, so that the vertical bone increment is insufficient, and the bone repairing effect is not ideal enough; meanwhile, as the vaccine is an animal-derived material, the vaccine 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-mentioned problems, the present invention provides a barrier membrane for promoting bone regeneration, which is a double-layered membrane obtained by subjecting animal-derived tissues to a series of treatments, and includes a dense barrier layer and a relatively loose collagen layer, and which can prolong the degradation time and can better promote the regeneration of osteoblasts, and a method for preparing the same. The specific technical scheme is as follows:
firstly, the invention provides a barrier film capable of promoting bone regeneration, which comprises a compact barrier layer and a loose collagen layer overlaid and laid on the barrier layer; the barrier layer is prepared from porcine and/or bovine dermal matrix membrane and/or pericardium,
the collagen layer is prepared from peritoneum and/or small intestine submucosa of pig and/or cattle.
Preferably, the barrier layer capable of promoting bone regeneration is made of 1-5 layers of porcine and/or bovine dermal matrix membranes and/or pericardium, and the structure of the barrier layer has no pores; the collagen layer is formed by stacking 1-5 layers of peritoneum and/or small intestine submucosa of pigs and/or cattle, and the pore sizes of the layers are sequentially increased along with the stacking sequence.
Further preferably, the barrier membrane capable of promoting bone regeneration is made of three layers of porcine and/or bovine dermal matrix membranes and/or pericardium; the collagen layer is formed by overlapping three layers of porcine and/or bovine peritoneum and/or small intestine submucosa, and the pore sizes of the layers are 45-55 microns, 90-110 microns and 195-210 microns in sequence.
The invention further provides a preparation method of the barrier membrane capable of promoting bone regeneration, which comprises the following steps:
s1: and (3) barrier film treatment: cleaning porcine and/or bovine dermal matrix membrane and/or pericardium, defatting, removing virus, removing cells, and removing immunogen to obtain barrier membrane for preparing barrier layer;
s2: collagen membrane treatment: cleaning peritoneum and/or small intestine submucosa of pig and/or cattle, degreasing, removing hybrid protein and removing cells, and then removing immunogen to obtain a collagen membrane for preparing a collagen layer;
s3: film laying: firstly, stacking and paving the treated barrier film according to requirements and compacting the barrier film to form a compact barrier layer, then stacking and paving the collagen film on the barrier layer according to requirements to form a loose collagen layer, and sequentially increasing the pore size of each collagen film of the collagen layer according to the film laying sequence;
s4: and (3) dehydrating and drying: and after the membrane laying is finished, dehydrating and drying to bond the layers together, thus obtaining the barrier membrane capable of promoting bone regeneration.
In the preparation method of the barrier membrane capable of promoting bone regeneration, the barrier membrane treatment of step S1 specifically includes the following steps:
s1-1: washing porcine and/or bovine dermal matrix membrane and/or pericardium with clear water;
s1-2: soaking in a degreasing reagent for 10-25 h;
s1-3: soaking in a cell removing reagent for 3-5 h;
s1-4: the immunogen is removed and a barrier membrane for preparing the barrier layer is obtained.
Wherein the degreasing reagent is ethanol, sodium hydroxide or a mixed solution of chloroform and methanol; the cell removing 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 immunogen removing reagent is trypsin solution, the concentration of the trypsin solution is 0.1% -5%, and the pH range is 7-9.
Preferably, the concentration of the ethanol used by the degreasing reagent is 75% -95% ethanol and the concentration of a sodium hydroxide solution is 1-2 mol/L; the volume ratio of the chloroform to the methanol mixed solution is as follows: methanol =2 to 3: 1 to 2.
In the preparation method of the barrier membrane for promoting bone regeneration, the collagen membrane treatment in step S2 specifically includes the following steps:
s2-1: washing peritoneum and/or small intestine submucosa of pig and/or cattle with clear water;
s2-2: soaking in a degreasing reagent for 16-24 h;
s2-3: soaking in a cell removing reagent for 2-4 h;
s2-4: removing the immunogen to obtain the collagen membrane for preparing the collagen layer.
Wherein the degreasing reagent is n-hexane or acetone; the cell removal reagent is a hydrogen peroxide solution with the mass concentration of 0.1-2% or a sodium hypochlorite solution with the mass concentration of 0.01-1%; the immunogen removing reagent is a telopeptidase solution, the concentration of the telopeptidase solution is 0.1% -1%, and the pH range is 7-9.
In the preparation method of the barrier membrane capable of promoting bone regeneration, step S4 is performed by dehydration and drying, wherein the drying method is vacuum freeze drying or ethanol gradient dehydration and drying; the technological parameters of the vacuum freeze drying are as follows: drying for 30-45 h at-40-25 ℃; the technological parameters of the ethanol gradient dehydration drying are as follows: 25 to 100 percent of gradient dehydration and drying.
The beneficial effects of the invention are:
1) the barrier film is divided into a dense barrier layer and a loose collagen layer, when the barrier film is used, the barrier layer faces soft tissues, the collagen layer faces bone tissues, the barrier layer can prevent epithelial fibrocytes with high migration speed from growing into a bone defect area to interfere bone formation, the degradation time is delayed, the bone formation speed is better matched, and the bone growth time requirement is met.
2) The collagen layer has a special pore structure, allows osteoblasts with low migration speed to preferentially enter a bone defect area, and promotes the growth of the osteoblasts.
3) The barrier film 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 pressure brought by wounds without collapse, and provides a basic guarantee for the effect of osteogenic repair.
4) The barrier layer is extruded by pressure, so that the layers are tightly connected, the degradation time in vivo is prolonged, and sufficient 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.
Drawings
FIG. 1 is a schematic structural diagram of a barrier membrane for promoting bone regeneration according to the present invention;
FIG. 2 is a pictorial view of a barrier membrane of the present invention that promotes bone regeneration;
fig. 3 is a photograph of a patient with an extracted tooth sutured with the barrier film 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 described in detail and completely with reference to the embodiments and the accompanying drawings.
Example 1
This example is to prepare a barrier membrane for promoting bone regeneration, which comprises a dense barrier layer and a loose collagen layer laid on the barrier layer; the barrier layer is prepared from 1-5 layers of porcine or bovine dermal matrix membranes or pericardium, and the collagen layer is prepared from 1-5 layers of porcine or bovine peritoneum or small intestine submucosa. In the embodiment, as shown in fig. 1, the barrier layer is formed by compacting three layers of bovine pericardium, and the structure has no pores; the collagen layer is formed by stacking three layers of small intestine submucosa of cattle, and the pore sizes of the layers are sequentially increased along with the stacking sequence and are respectively 50 microns, 100 microns and 200 microns; the preparation method of the barrier membrane capable of promoting bone regeneration specifically comprises the following steps:
s1: and (3) barrier film treatment: washing bovine pericardium with clear water, and soaking in a degreasing reagent for 20h for degreasing; and (3) removing viruses, soaking in a cell removing reagent for 4h for cell removing treatment, and finally performing immunogen removing by using a trypsin solution to obtain a barrier membrane for preparing a barrier layer. The degreasing reagent is a mixed solution of trichloromethane and methanol, and the volume ratio of the mixed solution is that the trichloromethane: methanol = 3: 2; the virus removing reagent is NaOH solution with the concentration of 1 mol/L; the cell removing reagent is a hydrogen peroxide solution with the mass concentration of 3%; the trypsin solution was 2% in concentration and had a pH of 8.
S2: collagen membrane treatment: washing the small intestine submucosa of the cattle with clear water, and soaking the small intestine submucosa in a degreasing reagent for 18h for degreasing and removing foreign protein; and then placing the membrane in a decellularization reagent for soaking for 3h for decellularization, and finally using a telopeptidase solution for immunogen removal to obtain a collagen membrane for preparing a collagen layer. The degreasing reagent is n-hexane; the protein removing reagent is a sodium dodecyl sulfate solution with the mass concentration of 0.5%; the cell removing reagent is a sodium hypochlorite solution with the mass concentration of 0.05%; the concentration of the solution of the de-terminative peptidase is 0.5 percent, and the pH value is 7; and then rolling by using rollers with different thicknesses to form different pores.
S3: film laying: firstly, the treated three layers of barrier films are overlapped, tiled and compacted to form a compact barrier layer according to requirements, then the three layers of collagen films are overlapped and tiled on the barrier layer according to requirements to form a loose collagen layer, and the pore sizes of the collagen films of the collagen layers are sequentially overlaid and tiled according to the film laying sequence of 50 microns, 100 microns and 200 microns.
S4: and (3) dehydrating and drying: after the membrane laying is finished, dehydrating and drying in an ethanol gradient dehydration drying mode to obtain a barrier membrane capable of promoting bone regeneration, 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 for 10 hr for dewatering to bond the layers together, and drying naturally.
Example 2
This example was also prepared by preparing a barrier membrane for promoting bone regeneration comprising a dense barrier layer and a loose collagen layer overlaid on the barrier layer; the barrier layer is prepared from porcine or bovine dermal matrix membrane or pericardium, and the collagen layer is prepared from porcine or bovine peritoneum or small intestine submucosa. In this example, the barrier layer was a three-layer compacted porcine dermal matrix film, which was not porous in structure. The collagen layer is formed by stacking three pig peritoneum layers, and the pore sizes of the layers are sequentially increased along with the stacking sequence; the pore sizes of the layers in this example were 45 μm, 95 μm, and 195 μm, respectively. The preparation method of the barrier membrane capable of promoting bone regeneration specifically comprises the following steps:
s1: and (3) barrier film treatment: washing the porcine dermal matrix membrane with clear water; then placing the mixture in a degreasing reagent for soaking 15 to degrease and remove viruses, and then placing the mixture in a cell removing reagent for soaking for 3.5 hours to remove cells; finally, the immunogen is removed by using a trypsin solution to obtain a barrier membrane for preparing a barrier layer. The degreasing reagent is a mixed solution of ethanol and methanol; the volume ratio of the ethanol: methanol = 2: 1; the virus removing reagent is NaOH solution with the concentration of 1 mol/L; the cell removing reagent is a 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 a degreasing reagent for 12h for degreasing and removing impure protein, then soaking in a cell removal reagent for 2h for cell removal treatment, and finally removing immunogen by adopting a telopeptidase removing solution to obtain a collagen membrane for preparing a collagen layer; the degreasing reagent is acetone; the protein removing reagent is a sodium dodecyl sulfate solution with the mass concentration of 0.5%; the cell removing reagent is a hydrogen peroxide solution with the mass concentration of 0.5%; the concentration of the solution of the de-termining peptidase is 0.2 percent, and the pH range is 7; and then rolling by using rollers with different thicknesses to form different pores.
S3: film laying: firstly, the three layers of treated barrier films are overlapped, tiled and compacted to form a compact barrier layer according to requirements, then the treated collagen films are overlapped and tiled on the barrier layer according to requirements to form a loose collagen layer, and the pore sizes of the collagen films of the collagen layers are sequentially overlapped according to the film laying sequence of 45 mu m, 95 mu m and 195 mu m.
S4: and (3) dehydrating and drying: after the membrane laying is finished, dehydrating and drying in a vacuum freeze drying mode to enable the layers to be bonded together, and obtaining the barrier membrane capable of promoting bone regeneration. The technological parameters of the vacuum freeze drying are as follows: drying at-40 ℃ for 45 h.
Example 3
This example was also prepared by preparing a barrier membrane for promoting bone regeneration comprising a dense barrier layer and a loose collagen layer overlaid on the barrier layer; the barrier layer is prepared from porcine or bovine dermal matrix membrane or pericardium, and the collagen layer is prepared from porcine or bovine peritoneum or small intestine submucosa. In the embodiment, the barrier layer is formed by compacting two pig heart envelopes and one bovine dermal matrix film, and the structure of the barrier layer has no pores; the collagen layer is formed by overlapping two layers of porcine small intestine mucosa and one layer of bovine peritoneum, the pore sizes of the layers are sequentially increased along with the overlapping sequence, and the pore sizes of the layers are respectively 55 microns, 100 microns and 210 microns. The preparation method of the barrier membrane capable of promoting bone regeneration specifically comprises the following steps:
s1: and (3) barrier film treatment: washing pig heart envelope and bovine dermal matrix membrane with clear water, soaking in degreasing reagent for 24h for degreasing and virus removal, soaking in decellularization reagent for 5h for decellularization treatment, and finally adding trypsin solution for immunogen removal 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 degreasing to mixing is as follows: methanol = 3: 1; the virus removing reagent is NaOH solution with the concentration of 1 mol/L; the cell removing reagent is a hydrogen peroxide solution with the mass concentration of 5%; the trypsin solution was 5% in concentration and pH 9.
S2: collagen membrane treatment: washing pig small intestine mucosa and cattle peritoneum with clear water, soaking in a degreasing reagent for 20h for degreasing and removing foreign protein, soaking in a cell removing reagent for 4h for cell removal, and removing immunogen with a telopeptidase solution to obtain a collagen membrane for preparing a collagen layer. The degreasing reagent is acetone; the protein removing reagent is a sodium dodecyl sulfate solution with the mass concentration of 0.5%; the cell removing reagent is a sodium hypochlorite solution with the mass concentration of 1%; the concentration of the solution of the de-termining peptidase is 0.8 percent, and the pH range is 8; and then rolling by using rollers with different thicknesses to form different pores.
S3: film laying: firstly, two layers of processed pig heart envelopes and a layer of bovine dermal matrix are overlapped, tiled and compacted to form a compact barrier layer according to requirements, then two layers of pig small intestine mucous membranes and a layer of bovine peritoneum are overlapped and tiled on the barrier layer according to requirements to form a loose collagen layer, and the pore sizes of the collagen films of the collagen layer are sequentially tiled according to the membrane-laying sequence of 55 microns, 100 microns and 210 microns.
S4: and (3) dehydrating and drying: after the membrane laying is finished, dehydrating and drying in a freeze-drying mode to obtain the barrier membrane capable of promoting bone regeneration. The technological parameters of dehydration and drying are as follows: drying at-40 deg.C for 40 h.
Example 4 application example
The embodiment is used for verifying the using effect of the product. After the patient completes the tooth extraction, the bone repair material is filled in the tooth extraction socket, and the barrier film of the present invention is used for suturing, as shown in fig. 3. The physical isolation function of the barrier membrane is utilized to separate different periodontal tissues, the dense barrier layer faces to soft tissues during sewing, epithelial fibrocytes with high migration speed are prevented from growing into a bone defect area to interfere bone formation, the loose collagen layer faces to the bone tissues, enough space is provided for osteoblasts with low migration speed, the growth of osteoblasts is promoted, and the normal connection relation between the periodontal tissues and the root surface is reestablished.
In 100 patients tracked and investigated, the bone repair effect after tooth extraction is good, the time for complete absorption of the used barrier membrane is 110-128 days, the time required for complete regeneration of bones is met, and the phenomenon of wound surface collapse caused by insufficient vertical bone increment does not occur in 100 patients; and the serious rejection or inflammatory reaction is basically not generated, and the using experience effect is good.
Generally, the barrier film is divided into a dense barrier layer and a loose collagen layer, the barrier layer faces soft tissues and the collagen layer faces bone tissues in use, the barrier layer can prevent epithelial fibrocytes with high migration speed from growing into a bone defect area to interfere 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 preferentially enter a bone defect area, and promotes the growth of the osteoblasts; the barrier film is prepared from animal-derived tissues, and the formed product has good ductility, good mechanical properties and stiffness, good supporting effect, capability of bearing pressure brought by wounds without collapse and capability of providing basic guarantee for the effect of osteogenic repair. The barrier layer is extruded by pressure, so that the layers are tightly connected, the degradation time in vivo is prolonged, and enough time and space are provided for bone growth; the collagen layer adopts a milder reagent, retains active substances in the loose layer, 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 attributes thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive. In addition, it should be understood that, although the present specification describes embodiments, one embodiment is not included, and the description is only for clarity, and those skilled in the art should be able to take the description as a whole, and the embodiments may be appropriately combined to form other embodiments understood by those skilled in the art.
Claims (10)
1. A barrier membrane for promoting bone regeneration, comprising: the device comprises a dense barrier layer and a loose collagen layer which is overlaid and laid on the barrier layer;
the barrier layer is prepared from porcine and/or bovine dermal matrix membrane and/or pericardium,
the collagen layer is prepared from peritoneum and/or small intestine submucosa of pig and/or cattle.
2. A barrier membrane for promoting bone regeneration according to claim 1, wherein: the barrier layer is made of 1-5 layers of porcine and/or bovine dermal matrix films and/or pericardium, and pores do not exist in the structure; the collagen layer is formed by overlapping 1-5 layers of porcine and/or bovine peritoneum and/or small intestine submucosa, and the pore sizes of the layers are sequentially increased along with the overlapping sequence.
3. A barrier membrane for promoting bone regeneration according to claim 2, wherein: the barrier layer is formed by compacting three layers of porcine and/or bovine dermal matrix membranes and/or pericardium; the collagen layer is formed by overlapping three layers of peritoneum and/or small intestine submucosa of pig and/or cattle, and 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.
4. A method for preparing a barrier membrane for promoting bone regeneration according to any one of claims 1 to 3, wherein: the method comprises the following steps:
s1: and (3) barrier film treatment: cleaning porcine and/or bovine dermal matrix membrane and/or pericardium, defatting, removing virus, removing cells, and removing immunogen to obtain barrier membrane for preparing barrier layer;
s2: collagen membrane treatment: cleaning peritoneum and/or small intestine submucosa of pig and/or cattle, degreasing, removing hybrid protein and removing cells, and then removing immunogen to obtain a collagen membrane for preparing a collagen layer;
s3: film laying: firstly, stacking and paving the treated barrier film according to requirements and compacting the barrier film to form a compact barrier layer, then stacking and paving the collagen film on the barrier layer according to requirements to form a loose collagen layer, and sequentially increasing the pore size of each collagen film of the collagen layer according to the film laying sequence;
s4: and (3) dehydrating and drying: after the membrane laying is finished, dehydration and drying are carried out, so that the layers are bonded together, and the barrier membrane capable of promoting bone regeneration can be obtained.
5. The method for preparing a barrier membrane for promoting bone regeneration according to claim 4, wherein: the barrier film treatment of step S1 specifically includes the steps of:
s1-1: washing porcine and/or bovine dermal matrix membrane and/or pericardium with clear water;
s1-2: soaking in a degreasing reagent for 10-25 h;
s1-3: soaking in a cell removing reagent for 3-5 h;
s1-4: the immunogen is removed and a barrier membrane for preparing the barrier layer is obtained.
6. The method for preparing a barrier membrane for promoting bone regeneration according to claim 5, wherein:
the degreasing reagent is ethanol, sodium hydroxide or a mixed solution of trichloromethane and methanol;
the cell removing 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 immunogen removing reagent is a trypsin solution, the concentration of the trypsin solution is 0.1% -5%, and the pH range is 7-9.
7. The method for preparing a barrier membrane for promoting bone regeneration according to claim 6, wherein: the concentration of ethanol used by the degreasing reagent is 75-95% ethanol and sodium hydroxide solution is 1-2 mol/L; the volume ratio of the chloroform to the methanol mixed solution is chloroform: methanol =2 to 3: 1 to 2.
8. The method for preparing a barrier membrane for promoting bone regeneration according to claim 4, wherein: the collagen membrane treatment of step S2 specifically includes the steps of:
s2-1: washing peritoneum and/or small intestine submucosa of pig and/or cattle with clear water;
s2-2: soaking in a degreasing reagent for 16-24 h;
s2-3: soaking in a cell removing reagent for 2-4 h;
s2-4: removing the immunogen to obtain a collagen membrane for preparing a collagen layer.
9. The method for preparing a barrier membrane for promoting bone regeneration according to claim 8, wherein:
the degreasing reagent is n-hexane or acetone;
the cell removal reagent is a hydrogen peroxide solution with the mass concentration of 0.1-2% or a sodium hypochlorite solution with the mass concentration of 0.01-1%;
the immunogen removing reagent is a telopeptidase solution, the concentration of the telopeptidase solution is 0.1% -1%, and the pH range is 7-9.
10. The method for preparing 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 dehydration drying;
the technological parameters of the vacuum freeze drying are as follows: drying for 30-45 h at-40-25 ℃;
the technological parameters of the ethanol gradient dehydration drying are as follows: 25 to 100 percent of gradient dehydration and drying.
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| CN115429939A (en) * | 2022-09-23 | 2022-12-06 | 博纳格科技(天津)有限公司 | Preparation method of oral biomembrane with controllable product performance |
| 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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