摘要
目的制备无细胞毒性猪去细胞真皮基质(PADM),评价其生物安全性,并观察基质在体内的转归情况。方法取0.3～0.4mm厚的猪断层皮片,用高渗盐水浸泡去除表皮,然后置于氢氧化钠溶液中,常温下水浴持续震荡去除真皮细胞,获得PADM,经交联和消毒后进行细菌学及猪病毒微生物学检测。将PADM植入SD大鼠皮下,于术后动态观察PADM大体及组织学形态变化。结果所获PADM柔软而有弹性,易于塑形,便于手术操作。PADM内无任何细胞成分,胶原纤维排列规则有序,弹力纤维丰富。细菌和病毒微生物学检测结果均为阴性。PADM植入SD大鼠皮下后未引发免疫排斥。术后3周,PADM与创面黏附牢固,不易从创面揭离;术后24周仍然可见其结构;早期PADM被大量炎性细胞浸润,随时间延长成纤维细胞逐渐增多,并出现新生的毛细血管,胶原纤维逐渐排列规则、致密。结论高渗盐水/氢氧化钠方法制备PADM过程简单,所得真皮基质无细胞毒性,生物安全性高,在体内可作为真皮支架长期存在。
Objective To prepare porcine acellular dermal matrix （PADM） without cytotoxicity, to evaluate its biological safety, and investigate its end result in vivo. Methods A piece of porcine split-thickness skin of 0. 3-0. 4mm was obtained, the epidermis was removed by hypertonic saline immersion, and then the skin was immersed in sodium hydroxide solution, and dermal cells were removed by shaking water bath at normal temperature to harvest PADM. After cross-linking and sterilization, bacteria and porcine virus were examined in the PADM. The PADM were embedded beneath the Sprague-Dawley （SD） rat＇s skin, and then the general and histological changes were observed after transplantation. Result The obtained PADM was soft and elastic, easy to be moulded, and convenient for operation. No cellular component was found in PADM, the collagen was regularly arranged and elastic fibers were abundant. Bacteria and virus examination of all specimens was negative. No immunologic rejection of PADM was found after being embedded beneath the skin of SD rat. The PADM adhered to the wound firmly, and it was difficult to detach 3 weeks after implantation, and the general structures of PADM beneath rat skin were discernible 24 weeks after implantation. The PADM was infiltrated by inflammatory cells in the early stage, fibroblasts and capillary vessels increased in number along with the time, and collagen fibers gradually gained a regular and compact arrangement in PADM. Conclusions The PADM prepared by hypertonic saline/sodium hydroxide method is a simple preparation process, and there is no cytotoxicity. It has a high biological safety and can be used as dermal scaffold with long-term existence in vivo.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2009年第9期1079-1081,共3页
Medical Journal of Chinese People's Liberation Army
基金
全军医学科研"十一五"计划专项课题(06Z054)
首都医学发展科研基金项目(2005-2040)
北京市科技计划研发攻关课题(H060920050830)
河北省2007年科学技术研究与发展指导计划项目(072761594)
关键词
皮肤移植
去细胞真皮基质
胶原
猪
skin transplantation
porcine acellular dermal matrix
collagen
swine
