Experimental study on genipin crosslinking improved the biological performance of rat kidney decellularized matrices
LIU Dan, DU Ao-ling, WANG Xin-wang, ZHANG De-ming, TANG Hui-li, WANG Yu-hang, CHEN Sheng-hua
Chinese Journal of Clinical Anatomy ›› 2019, Vol. 37 ›› Issue (1) : 71-76.
Experimental study on genipin crosslinking improved the biological performance of rat kidney decellularized matrices
Objective To improve the biological properties of the rat kidney acellular biological scaffold by genipin crosslinking. Methods Eighty healthy SD rats were divided into a normal group, an uncrosslinking group, a glutaraldehyde crosslinking group and a genipin crosslinking group. Rat kidneys and renal arteries were isolated and connected with peristaltic pumps. Rat kidneys were successively infused with heparinized PBS solution, 1% Triton X-100, 1% sodium dodecyl sulfate (SDS), deionized water to complete the preparation of rat kidney acellular biological scaffolds. The glutaraldehyde crosslinking group was infused with 1500 ml 0.625% glutaraldehyde (GA),and the genipin crosslinking group was immersed in 0.5% genipin solution at 37 ℃ for 24 hours. Non-glutaraldehyde and non-genipincrosslinking renal acellular biological scaffolds served as the uncrosslinking group. Four groups of kidneys were stained with HE, Masson and immunofluorescence, respectively. The morphology and ultrastructure of scaffolds were observed by immunofluorescence and electron microscopy, and mechanical properties were tested by tensile test. Results After crosslinking with genipin, HE and Masson staining showed that collagen fibers arranged more tightly and orderly, glomerular fibers were aggregated, Collagen I and Collagen IV fluorescence staining enhanced, and the honeycomb-like pore structure in the cross-linked acellular scaffold was more stereoscopic and typical. The structure of glomerular niche was clearer, and the elastic modulus of kidney in crosslinking group was significantly higher than uncrosslinking group. Conclusion Genipin crosslinking rat renal decellularized scaffold can greatly improve the biological properties of the scaffold and lay a foundation for cell implantation and organ regeneration.
Genipin; Glutaraldehyde / Biological scaffold; Kidney; Kidney regeneration
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