Preparation method of acellular dermal matrix material

An acellular dermis and matrix material technology, which is applied in the field of the preparation of acellular dermal matrix materials, can solve the problems of dermal matrix structural damage, poor material mechanical strength, insufficient thermal stability, etc., and achieves the promotion of cell growth and reproduction, and the improvement of stability. effect on cell adhesion and nutrient exchange

Inactive Publication Date: 2021-06-15
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the commonly used decellularization methods include four methods: physical method, chemical method, enzymatic method, and organic solvent. During the treatment process, they will cause varying degrees of damage to the structure of the dermal matrix and degrade the active factors in the matrix, resulting in poor mechanical strength

Method used

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  • Preparation method of acellular dermal matrix material
  • Preparation method of acellular dermal matrix material

Examples

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Example Embodiment

[0017] The present invention is further described below with reference to the accompanying drawings and examples, and the scope of the present invention is not limited to the following: Example 1: Preparation of decellular leather matrix materials

[0018] A method for preparing a degraded leather matrix material, which includes the following steps:

[0019] S1. Peeling: Take the discarded pig skin, scrape off the hair, cut the thickness of 0.1 to 3 mm and the skin of the leather substrate, while removing fat tissue;

[0020] Soak: Soak the lateral leather matrix in a acetaldehyde solution containing glycine, the mass percentage concentration of glycine is 4-5%, the ratio of the solution volume and the dermis matrix is ​​2: 1;

[0021] S3. Decentral: The soaked leather base is placed in the eluate, the water ratio of the washing solution and the dermis matrix is ​​10 mg: 1 ml, the mass percentage concentration of NaOH is 2%, Tritonx-100 The mass percentage concentration is 2%, and...

Example Embodiment

[0023] Example 2: Scanning Electron Microscopy (SEM)

[0024] (1) Method: Decamination leather matrix material prepared in Example 1 was cut into 5 × 5 mm 2 Size, placing the stent sample on a stage with a conductive glue, gently press and blow it with a blown ball; a spray gold for the stent sample; put the sample-containing stage after completion of the sample Scanning an electron microscope. Samples were subjected to the sample with a voltage of 5.0 keV and 10 mA.

[0025] (2) Result: figure 1 As shown, after SEM observation, the pig derivative leather matrix material is lyophilized to lyophilize, and the internal communication between the honeycomb-shaped porous network structure, the hole and the hole is better, and the fiber is concentrated in the matrix, indicating that the material is The collagen fiber structure is not damaged during the preparation process, which is kept well, which is conducive to the adhesion proliferation of cells and the nutritional exchange process ...

Example Embodiment

[0026] Example 3: Cell proliferation experiment:

[0027] (1) Method: The porcine dermal matrix material prepared in Example 1 was cut into a small piece and was placed in a 24-well plate. The sample was immersed in the culture solution for 2 h. The l929 cells were digestred with 0.25% trypsin in the culture flask, and the cell suspension was prepared, the concentration was 1 × 10 5 / ml. Carefully draw a cell suspension of 100 μl of the aliquot of the sample and evenly add it to the surface of the sample, then placed in a constant temperature incubator for 4 h. Subsequently, 900 μl of medium is added to each well, and the DMEM medium is replaced every 3 days. The culture fluid was absorbed in 1, 3, 5, 7 and 9 days. Add the same amount of fresh DMEM medium, then add 50 μl of CCK-8 reagent to each well and placed in CO of 37 ° C 2 The incubator was slowly shaken for 4 h. 200 μl aliquots were removed from each well and then transferred to a 96-well plate. The absorbance at 450 nm wa...

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Abstract

The invention discloses a preparation method of an acellular dermal matrix material, and belongs to the technical field of biological material preparation. The preparation method is characterized in that the acellular dermal matrix material with good stability and biocompatibility is prepared by decellularizing waste pigskin produced and processed. The prepared pig ADM material is of a honeycomb-shaped porous structure, and main components of pigskin collagen are well reserved; the tensile strength of the obtained pig ADM material can reach 2.06+/-0.48 MPa, and is obviously superior to the research value 0.28 MPa of previous people, so that the pig ADM material can meet more requirements of actual use; and the prepared pig ADM material has no cytotoxicity, has high porosity, is beneficial to cell adhesion and nutrient exchange, and shows an obvious promotion effect on growth and reproduction of cells.

Description

technical field [0001] The invention belongs to the technical field of biological material preparation, and in particular relates to a preparation method of an acellular dermal matrix material. Background technique [0002] Over the past 30 years, many advanced wound dressings have been developed globally, especially the use of natural polymers, including rubber, cellulose, and collagen, which have good biophasic properties, has been extensively studied. capacitive, degradable, and water-retaining properties. Among them, collagen is the most abundant animal protein, which provides mechanical strength to biological tissues and accelerates cell proliferation. Currently commercial collagen dressings are mostly prepared from bovine and porcine tissues, and collagen scaffolds can be prepared by decellularization, commonly known as acellular dermal matrix. Acellular dermal matrix is ​​a special type of biomaterial in which the highly immunogenic cellular components from the skin...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/56
CPCA61L27/362A61L27/3687A61L27/56
Inventor 白新鹏王国锭
Owner HAINAN UNIVERSITY
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