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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 of the material. Insufficient thermal stability and poor biocompatibility, therefore, the decellularization method that maintains the stability of the three-dimensional structure of the dermal matrix to the greatest extent and retains bioactive components such as proteoglycans and growth factors is a hot and difficult point in current research

Method used

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

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Embodiment 1

[0017] The present invention will be further described below in conjunction with accompanying drawings and Examples, and the protection scope of the present invention is not limited to the following: Embodiment 1: Preparation of decellularized dermal matrix material

[0018] A preparation method of decellularized dermal matrix material, which comprises the following steps:

[0019] S1. Peeling: take the discarded pigskin, scrape off the hair on the surface, cut out the dermal matrix with a thickness of 0.1-3mm and contain the epidermis, and remove the fat tissue at the same time;

[0020] S2. Soaking: Soak the peeled dermal matrix in an acetaldehyde solution containing glycine for 1-2 hours, the mass percent concentration of glycine is 4-5%, and the ratio of the volume of the solution to the mass of the dermal matrix is ​​2:1;

[0021] S3. Decellularization: the soaked dermal matrix is ​​placed in the eluent for elution, the solid-liquid ratio of the eluent to the dermal matri...

Embodiment 2

[0023] Embodiment 2: scanning electron microscope observation (SEM)

[0024] (1) Method: Cut the acellular dermal matrix material prepared in Example 1 into 5×5mm 2 Size, use tweezers to place the bracket sample on the stage with conductive glue, press it lightly and blow it with a dust blower to fix it; spray gold on the bracket sample; after the gold spray is completed, put the stage with the sample into in a scanning electron microscope. The structure of the sample was observed with a voltage of 5.0KeV and a current of 10mA.

[0025] (2) Results: results such as figure 1 As shown, by SEM observation, the porcine acellular dermal matrix material has a honeycomb porous network structure inside after being freeze-dried by a freeze dryer, the connectivity between the pores is good, and the fibers in the matrix are clustered, indicating that the material is The collagen fiber structure is not damaged during the preparation process and is well preserved. This porous structure ...

Embodiment 3

[0026] Embodiment 3: cell proliferation experiment:

[0027] (1) Method: The porcine acellular dermal matrix material prepared in Example 1 was cut into small pieces and put into 24-well plates. The samples were immersed in the culture medium for 2h. Trypsinize L929 cells with 0.25% trypsin in a culture flask, and prepare cell suspension at a concentration of 1×10 5 / mL. Carefully aspirate a 100 µL aliquot of the cell suspension and add it evenly to the surface of the sample, then place in a constant temperature incubator for 4 h. Subsequently, 900 μL of medium was added to each well to continue culturing, and the DMEM medium was replaced every 3 days. Cultures were aspirated on days 1, 3, 5, 7 and 9. Add the same amount of fresh DMEM medium, followed by 50 μL of CCK-8 reagent to each well, and place in 37 °C CO 2 Shake slowly for another 4 h in the incubator. Take a 200 µL aliquot from each well and transfer to a 96-well plate. Read the absorbance at 450 nm with a micr...

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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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