Preparation method and application of acellular collagen matrix membrane
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A collagen matrix and decellularization technology, applied in the field of preparation of acellular collagen matrix membrane, can solve the problems of destroying the biocompatibility of collagen membrane, affecting the biological effect of collagen membrane, easy residue of detergents and enzymes, etc. Barrier function and space maintenance function, avoid biocompatibility damage, promote cell adhesion effect
Pending Publication Date: 2022-02-11
HOSPITAL OF STOMATOLOGY SUN YAT SEN UNIV
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Problems solved by technology
The existing decellularization schemes mostly use detergents and enzymes, which have high decellularization efficiency, but may cause damage to the structure of collagen fibers, thereby affecting the biological effects of the collagen membrane. Moreover, detergents and enzymes are easy to remain and severely damage the collagen membrane. In addition, there are still some problems that need to be improved when the acellular collagen matrix membrane is used for the regeneration of soft and hard tissues: ①Degradation speed is too fast;② Insufficient mechanical strength; ③High risk of soft tissue complications
[0005] Therefore, it is necessary to optimize the decellularization scheme of the extracellular matrix, reduce the residue of decellularized reagents, use the decellularization method to regulate the physical and chemical properties of the decellularized collagen matrix membrane, overcome the shortcomings of its fast degradation speed and insufficient mechanical strength, and improve the strength of the collagen matrix material. Soft Tissue Healing Efficacy
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Embodiment 1
[0050] A preparation method of decellularized collagen matrix membrane, such as figure 1 shown, including the following steps:
[0051] S1. Take the pig peritoneum, pretreat the pig peritoneum, wash the blood stains with pure water first, use scissors without tension to remove as much fat and muscle tissue as possible, keep the biofilm thickness of about 0.5mm-2mm, and remove the biofilm Cut the film into 30mm×30mm size, soak it in pure water for 12 hours, and obtain the first intermediate product—biofilm, which is ready for use;
[0052] S2,
[0053] S2-1, repeated freeze-thaw treatment
[0054] Soak the biofilm obtained in step S1 in liquid nitrogen at -80°C, the volume ratio of biofilm to liquid nitrogen is 1:10, let it stand for 5 minutes, and melt it in flowing pure water at 25°C for 10 minutes, repeat 3 times; Wash with water, soak in pure water for 15 minutes, repeat 5 times;
[0055] S2-2. Alkali treatment
[0056] Soak the biofilm after S2-1 treatment in 2g / L sod...
Embodiment 2
[0073] A preparation method of decellularized collagen matrix membrane, such as figure 1 shown, including the following steps:
[0074] S1. Take the pig peritoneum, pretreat the pig peritoneum, wash the blood stains with pure water first, use scissors without tension to remove as much fat and muscle tissue as possible, keep the biofilm thickness of about 0.5mm-2mm, and remove the biofilm Cut the film into 30mm×30mm size, soak it in pure water for 12 hours, and obtain the first intermediate product—biofilm, which is ready for use;
[0075] S2,
[0076] S2-1, repeated freeze-thaw treatment
[0077] Soak the biofilm obtained in step S1 in liquid nitrogen at -80°C, the volume ratio of biofilm to liquid nitrogen is 1:10, let it stand for 5 minutes, and melt it in flowing pure water at 25°C for 10 minutes, repeat 7 times; Wash with water, soak in pure water for 15 minutes, repeat 5 times;
[0078] S2-2. Alkali treatment
[0079] Soak the biofilm after S2-1 treatment in 2g / L sod...
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Abstract
The invention provides a preparation method of an acellular collagen matrix membrane, and the method comprises the following steps: performing pretreating to obtain a mammalian membrane tissue, and then carrying out repeated freeze-thawing treatment, acid-base treatment, gradient dehydration treatment, organic matter degreasing treatment and hypotonic / hypertonic treatment on the mammalian membrane tissue, thereby effectively removing tissue immunogenicity, retaining an ordered collagen fiber structure, and improving the decellularized collagen matrix membrane; then performing drying and sterilizing to obtain an acellular collagen matrix membrane, wherein the decellularized collagen matrix membrane has excellent flexibility, biocompatibility and soft and hard tissue regeneration promoting performance, has a smooth-surface and rough-surface double-layer bionic structure, can retain a natural three-dimensional pore scaffold structure similar to the tissue, retains a collagen structure, and has good water absorption and thermal denaturation stability; source materials are easy to obtain, the cost is low, the decellularized collagen matrix membrane has a stronger barrier function and a space maintaining function, and the surface appearance, the elastic modulus, the porosity and the pore size of the decellularized collagen matrix membrane can be regulated and controlled through different freeze-thaw treatment modes.
Description
【Technical field】 [0001] The invention relates to the fields of chemical engineering, bioengineering, biomechanics, medicine and material science, and in particular to a preparation method and application of an acellular collagen matrix membrane. 【Background technique】 [0002] Collagen membrane is a common soft and hard tissue regeneration repair material in clinic because of its good biocompatibility, tissue integration performance, biodegradability and operability. Collagen membrane materials can be divided into acellular collagen matrix membrane and purified collagen membrane according to the process. The decellularized collagen matrix membrane obtained by decellularization of whole tissues or organs (such as dermis, heart valve, bladder, small intestine, pericardium, peritoneum, etc.), because of its bionic structure and tissue-specific proteins that are conducive to regeneration, is compatible with Synthetic collagen is more biocompatible and more widely used in regen...
Claims
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