Yang Bin, Qiu Ri-Sheng, Hong Qing-Qi, Ji Chen-Yang
Department of Plastic Surgery, the First Affiliated Hospital, Guangzhou Medical College, Guangzhou 510120, China.
Zhonghua Zheng Xing Wai Ke Za Zhi. 2009 May;25(3):200-4.
To study the effect of tissue-engineered skin loaded with keratinocyte growth factor (KGF) nanocapsules for skin defect on athymic mice.
The acellular dermal matrix (ADM) loaded with KGF-ADM was constructed by means of phacoemulsification solvent evaporation and low temperature drying. The human epidermal stem cells and fibroblasts were captured and identified, then cultivated on the surface of the KGF-ADM. The cell growth was observed. The tissue-engineered skin without KGF was used as sham group. The autogenous skin graft was used as control group. 2 and 6 weeks after the skin was transplanted to the back of athymic mice, the contraction and histological healing of the transplanted skins were observed respectively. Then the immunofluorescence examination with anti-human K10-FITC and beta1-integrin-Cy3 were applied to detect the origin, growth and differentiation of epidermal and dermal cells in tissue-engineered skin.
The epidermal stem cells grew well and attached tightly on KGF-ADM. There were small round stem cells and polygonal terminally-differentiated cells, which appeared a partly cloning growth and a tendency of merging. The tissue-engineered skin with KGF nanocapsules gained better result in repairing the skin defects as compared with the blank group and the control group 2 and 6 weeks after transplantation. The regenerative skin cells could connect and mix closely with the athymic mouse skin cells on the border of skin defect. Meanwhile, the regenerative skin existed some contraction. The histological observation with HE staining showed that the regenerative skin possessed intact epidermis with several cell layers and normal keratose stratum, among which there were still some beta1-integrin (+) cells which represented epidermal stem cells or transient amplifying cells when they were tested by immunofluorescence after 6 weeks of transplantation.
The tissue-engineered skin loaded with KGF nanocapsules had a better result in repairing athymic mice skin defects than common tissue-engineered skin without KGF nanocapsules or skin auto-graft.
研究负载角质形成细胞生长因子（KGF）纳米胶囊的组织工程皮肤对无胸腺小鼠皮肤缺损的修复作用。
采用超声乳化溶剂蒸发法和低温干燥法构建负载KGF的脱细胞真皮基质（KGF-ADM）。采集并鉴定人表皮干细胞和成纤维细胞，然后接种于KGF-ADM表面，观察细胞生长情况。以未负载KGF的组织工程皮肤作为假手术组，自体皮肤移植作为对照组。将皮肤移植到无胸腺小鼠背部2周和6周后，分别观察移植皮肤的收缩情况和组织学愈合情况。然后采用抗人K10-FITC和β1整合素-Cy3免疫荧光检测组织工程皮肤中表皮和真皮细胞的来源、生长及分化情况。
表皮干细胞在KGF-ADM上生长良好，贴附紧密。可见圆形小干细胞和多边形终末分化细胞，呈部分克隆生长及融合趋势。移植2周和6周后，负载KGF纳米胶囊的组织工程皮肤在修复皮肤缺损方面比空白组和对照组效果更好。再生皮肤细胞在皮肤缺损边缘能与无胸腺小鼠皮肤细胞紧密连接并融合。同时，再生皮肤存在一定程度的收缩。HE染色组织学观察显示，再生皮肤表皮完整，有多层细胞及正常角质层，移植6周后免疫荧光检测仍有一些β1整合素（+）细胞，代表表皮干细胞或短暂增殖细胞。
负载KGF纳米胶囊的组织工程皮肤在修复无胸腺小鼠皮肤缺损方面比普通未负载KGF纳米胶囊的组织工程皮肤及自体皮肤移植效果更好。
