Optimized preparation method for decellularized skin flap
FENG Yulu1, ZHOU Lebin 1, 2, XUE Xiangyang1, MEI Jin 1, 3.
1.Anatomy Department, School of Basic Medicine, Wenzhou Medical University, Wenzhou 325035, China; 2.Department of Emergency, Yueqing People’s Hospital, Wenzhou 325600, China; 3.Scientific Research Center, Ningbo First Hospital, Ningbo 315000, China
Abstract：Objective: To prepare decellularized skin flaps by auxiliary shaking method and test their biocompatibility. Methods: After taking the skin flap of the lower abdominal wall of adult rats to remove blood, a magnetic stirrer was used to perform auxiliary shaking to remove cells. The biological characteristics of the harvested scaffolds were obtained by quantitative analysis of DNA, detection of SDS residues, and scanning electron microscopy. Then take the decellularized skin flap specimens embedded subcutaneously on the back of adult SD rats, count the number of new blood vessels to detect its biocompatibility, and the proportion of regulatory T lymph in the peripheral blood. Results: Compared with the control group, the DNA content of the decellularized group decreased by more than 95%; no obvious nucleus was seen in DAPI staining; scanning electron microscope and HE staining showed that a large amount of extracellular matrix remained in the decellularized group; SDS residues were far below the biological safety level; angiography showed that the branches of blood vessels were complete and clear, and the number of new blood vessels was significantly higher than that in the control group (P<0.05). The results of animal experiments showed that in decellularized group the number of blood vessels was regenerated and the proportion of regulatory T lymphocytes increased (P<0.05). Conclusion: A protocol for preparing decellularized skin flaps with good biocompatibility and complete stent structure is provided.
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