异种脱细胞真皮基质制备方法的改进

An improved method for heterogenous acellular dermal matrix fabrication

背景:应用液氮进行大张羊脱细胞真皮基质制作时,复温过程中真皮基质易破碎,不易制出大张完整的脱细胞羊真皮基质。目的:探索更好的羊脱细胞真皮基质的制作方法。方法:改变低温条件,变液氮降温为应用-80℃低温冰箱进行异种皮肤的冷冻处理,破坏细胞成分,利用洗脱液振荡洗涤,去除细胞碎片,制备出完整的大张羊脱细胞真皮基质,并对其脱细胞程度和胶原三维支架结构的完整性进行测定。结果与结论:通过-80℃低温冰箱进行反复冻融制备的大张羊脱细胞真皮基质,较为完整,常规病理检查无明显细胞碎片残留;免疫组化检测无明显细胞碎片;电子显微镜检查胶原结构完整。应用改良-80℃低温冰箱和液氮进行反复冻融制作的大张羊脱细胞真皮基质对比,脱细胞程度和其他生物学性能无明显差异,但完整性前者明显优于后者,适合临床应用。

BACKGROUND： It is difficult to prepare complete and large piece of goat acellular dermal matrix using liquid nitrogen method due to breakability of dermal matrix during rewarming period. OBJECTIVE： To explore a better method for goat acellular dermal matrix fabrication. METHODS： In the course of freezing goat skin, a-80℃ refrigerator was used as a substitute for liquid nitrogen. Cell components of goat skin were removed by freezing and thawing repeatedly, and then cell fragments were washed off using a zwitterionic with ultrasonic machine. The cell fragments and structure were tested. RESULTS AND CONCLUSION： By strict tests, collagen structure of the heterogeneous dermal matrix fabricated here was integrity and no cell fragment was found. The biomechanical characteristics were similar with acellular dermal matrix fabricated by routine repeated frozen method. Compared with routine repeated frozen method, the acellular degree and biological properties of goat acellular dermal matrix prepared by improved method had no significant differences, but the integrity of latter was eminent.