Derivative scaffold based on skin-derived acellular matrix and construction method ofderivative scaffold

A technology of acellular matrix and construction method, which is applied in the field of tissue engineering scaffold and its construction, can solve the problems that have not been reported, and achieve the effect of improving mechanical strength, broad-spectrum antibacterial performance, and increasing cell adhesion

Active Publication Date: 2021-07-09
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Gelatin and chitosan have been fully recognized and widely used in the field of tissue engineering, but the skin tissue engineering composite scaffold based on acellular matrix has not been reported yet

Method used

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  • Derivative scaffold based on skin-derived acellular matrix and construction method ofderivative scaffold
  • Derivative scaffold based on skin-derived acellular matrix and construction method ofderivative scaffold
  • Derivative scaffold based on skin-derived acellular matrix and construction method ofderivative scaffold

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Preparation of decellularized extracellular matrix scaffold (as a comparative example) and detection of decellularized efficiency

[0037] (1) Buy fresh pigskin from the market, wash it and cut it into 1.0cm 2 The small pieces were placed in a PBS solution containing 0.5wt% trypsin and 5mM ethylenediaminetetraacetic acid (EDTA), stirred in a water bath at 30°C for 8h for decellularization, and the stirring speed was 300rpm;

[0038] (2) Place the matrix obtained in step (1) in a PBS solution containing 5.0wt% Triton-X-100 and 20mM EDTA, stir at room temperature for 48h to decellularize, and the stirring speed is 300rpm;

[0039] (3) Place the matrix obtained in step (2) in an aqueous solution containing 20% ​​isopropanol, and stir at room temperature for 24 hours to elute excess fat at a stirring speed of 300 rpm;

[0040] (4) Wash the matrix obtained by step (3) with PBS solution at room temperature for 3 times, each time at an interval of 1h, and wash with ...

Embodiment 2

[0044] The preparation of embodiment 2 gelatin / chitosan (Gel / CS) composite support (as comparative example)

[0045] (1) Weigh 3 g of gelatin (Gel) and 1 g of chitosan (CS) into 100 mL of 0.5 mol / L acetic acid solution in turn, stir and dissolve under heating at 50°C.

[0046] (2) The Gel / CS solution in step (1) was slowly poured into a 24-well plate, frozen in a -20°C refrigerator for 12 hours, and then freeze-dried with a freeze dryer for 24 hours to obtain a porous composite scaffold.

[0047] (3) The composite scaffold in step (2) is cross-linked with a cross-linking agent for 24 hours at room temperature. The cross-linking agent is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), ethanesulfonic acid (MES ) and ethanol mixed solution. Among them, MES is used as the buffer solution of the cross-linking agent, the concentrations of EDC, NHS and MES are all 50mmol / L, and the volume fraction of the ethanol solution is 60%.

[00...

Embodiment 3

[0049] Example 3 Preparation of Decellularized Matrix / Gelatin / Chitosan (dECM / Gel / CS) Composite Scaffold

[0050] (1) Buy fresh pigskin from the market, wash it and cut it into 0.2cm 2 The small pieces of the mixture were placed in a PBS solution containing 0.1wt% trypsin and 1mM ethylenediaminetetraacetic acid (EDTA), stirred in a water bath at 10°C for 2h to decellularize, and the stirring speed was 100rpm;

[0051] (2) Place the matrix obtained in step (1) in a PBS solution containing 0.5wt% Triton-X-100 and 5mM EDTA, and stir at room temperature for 12h to decellularize, and the stirring speed is 100rpm;

[0052] (3) Place the substrate obtained in step (2) in an aqueous solution containing 5% isopropanol, stir for 12 hours at room temperature to elute excess fat, and the stirring speed is 100 rpm;

[0053] (4) Wash the substrate obtained in step (3) with PBS solution at room temperature for 3 to 4 times, with an interval of 1 hour each time, and wash with a solution conta...

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Abstract

The invention discloses a derived scaffold based on a skin-derived acellular matrix and a construction method of the derived scaffold. Through a decellularization technology, firstly, pigskin is subjected to decellularization treatment through a pancreatin method, a Triton-X-100 method and a DNase method in sequence, then a freeze-dried decellularization matrix is digested through pepsase, then the decellularization matrix is mixed with gelatin and chitosan in a matching mode, and the porous three-dimensional acellular matrix derived scaffold is obtained through freeze drying. The pigskin-derived acellular matrix is selected as the matrix material, and the gelatin and chitosan are compounded to prepare the composite porous scaffold, so that advantage complementation of the functions of the materials is realized. The pigskin is wide in source and low in cost, and meanwhile, the extracellular matrix is a substance synthesized and secreted in the growth process of cells and can provide a natural growth microenvironment of fibroblasts to the maximum extent; the gelatin is compounded to improve the mechanical strength of the scaffold and increase the cell adhesiveness of the scaffold; the chitosan is compounded to improve the mechanical strength and water absorption of the scaffold and endow the scaffold with broad-spectrum antibacterial performance. The scaffold can be used as an excellent biomedical material in the field of skin tissue engineering.

Description

technical field [0001] The invention belongs to the fields of tissue engineering, material science and biology, and provides a tissue engineering scaffold composed of skin-derived decellularized matrix, gelatin and chitosan and a construction method thereof. Background technique [0002] As the largest organ of the human body, the skin acts as a barrier to maintain the stability of the internal environment and prevent microbial invasion, and plays important functions such as temperature regulation, body fluid balance, vitamin D synthesis, and waste excretion. Skin defects caused by external injuries or diseases often lead to loss of body fluids and bacterial infection, causing secondary damage to patients and even threatening their lives. As a new skin repair technology, skin tissue engineering has the characteristics of high efficiency, wide range of material sources, no immune rejection, and low cost. [0003] The first step in skin tissue engineering repair technology is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/48A61L27/54A61L27/56A61L27/60
CPCA61L27/362A61L27/54A61L27/48A61L27/56A61L27/60A61L2300/404A61L2300/236C08L89/00C08L5/08
Inventor 宋克东徐杰聂毅李丽颖胡雪岩苏雅房换刘天庆
Owner DALIAN UNIV OF TECH
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