In vivo biocompatibility of whole-kidney acellular matrix scaffolds prepared by perfusion method

doi:10.3969/j.issn.2095-4344.2015.16.013

Abstract

Abstract:

BACKGROUND: The whole-kidney acellular matrix scaffold in rats prepared by perfusion method is proved to have good in vitro biocompatibility, but in vivo biocompatibility is still unclear.
OBJECTIVE: To produce a whole-kidney acellular matrix scaffold in rats by perfusion method and to evaluate the in vivo biocompatibility of the scaffold.
METHODS: The whole-kidney acellular matrix scaffold in Wistar rats was prepared by perfusion method and evaluated with the following tests. (1) Acute toxicity test: mice were subject to intraperitoneal injection of whole-kidney acellular matrix scaffold extract liquid, normal saline and phenol. (2) Hemolytic test: Anticoagulant blood samples from New Zealand rabbits were mixed with whole-kidney acellular matrix scaffold extract liquid,
normal saline and distilled water, respectively. (3) Pyrogen test: Whole-kidney acellular matrix scaffold extract liquid was injected into the ear vein of New Zealand rabbits. (4) Intracutaneous stimulation test: Whole-kidney acellular matrix scaffold extract liquid was injected subcutaneously into New Zealand rabbits for observing whether there was a skin stimulus response. (5) Subcutaneous implantation test: The whole-kidney acellular matrix scaffold was implanted subcutaneously into the back of New Zealand rabbits.
RESULTS AND CONCLUSION: There was no cell residual in the whole-kidney acellular matrix scaffold prepared by perfusion method, and no acute systemic toxicity, hemolytic reaction, pyrogen response, and skin stimulus respons, indicating the scaffold has a good histocompatibility in the rabbits. These findings suggest that the whole-kidney acellular matrix scaffolds in Wistar rats prepared by perfusion method have good in vivo biocompatibility.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: Tissue Engineering, Kidney, Acute Toxicity Tests

CLC Number:

R318

Chen Jie, Yang Jing-qiu, Liu Chun-xiao. In vivo biocompatibility of whole-kidney acellular matrix scaffolds prepared by perfusion method[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(16): 2529-2533.

Figures/Tables 3

2.4 溶血实验结果实验组与阴性对照组标本离心后均有红细胞沉积，上清液接近无色，未见溶血表现。阳性对照组未见红细胞沉积，上层液体颜色为淡红色。实验组、阴性对照组、阳性对照组的A值分别为0.042±0.004、0.042±0.001及0.117±0.001，实验组、阴性对照组A值低于阳性对照组(P < 0.05)，前两组间比较差异无显著性意义(P > 0.05)。经溶血率公式计算，肾脏脱细胞基质材料的相对溶血率为0.58%，符合医用材料溶血率规定要求。 2.5 热源实验结果实验前监测新西兰兔体温1周，均在38.2-38.6 ℃之间，日夜体温波动平稳且最高与最低体温相差均不超过0.4 ℃。经耳缘静脉缓慢注入肾脏脱细胞基质浸提液后，新西兰兔活动自如，未出现烦躁不安、精神萎靡等不适表现，体温升高均在0.6 ℃以下且3只新西兰兔体温升高总和< 1.4℃，符合热源实验的评价标准。 2.6 皮内刺激实验结果所有新西兰兔皮内注射后观察24，48，72 h，以生理盐水为浸提介质肾脏脱细胞基质浸提液注射区及生理盐水注射区均无红斑、水肿及溃烂等明显皮肤刺激症状，内皮反应记分分别为0.083、0.083；以植物油为浸提介质肾脏脱细胞基质浸提液及植物油注射区均可见不同程度的水肿、破溃，内皮反应记分分别为2.250、2.083；实验组与各对照组的平均记分之差均小于1，符合生物材料标准。 2.7 皮下植入实验结果植入物植入后所有新西兰兔状态良好，进食可，伤口均无感染、红肿，渗液，伤口1期愈合。2，4周时植入部位皮肤无异常，无植入物排出。取出植入物后行苏木精-伊红染色，2周时可见在脱细胞基质纤维组织之间有少量多形核细胞、淋巴细胞及成纤维细胞浸润，脱细胞基质支架仍清晰可见；4周时脱细胞基质已基本降解，较难辨认，移植物周围炎性细胞浸润减轻，移植物原有红色胶原纤维与新生淡红色胶原纤维相互交织，纤维之间可见纤细的血管和大量成纤维细胞，周围组织结构正常(图3)。"

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