中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (32): 6010-6014.doi: 10.3969/j.issn.1673-8225.2010.32.026

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

羊膜负载表皮干细胞促进糖尿病大鼠创面的愈合

钟清玲1,2，刘德伍2，刘繁荣1，彭  燕2，于  玫1，肖鲁良1   

  1. 1南昌大学医学院，江西省南昌市 330006；2南昌大学第一附属医院烧伤研究所，江西省南昌市 330006
  • 出版日期:2010-08-06 发布日期:2010-08-06
  • 通讯作者: 刘德伍，博士，教授，主任医师，博士生导师，南昌大学第一附属医院烧伤中心，江西省南昌市 330006 dewuliu@126. com
  • 作者简介:钟清玲☆，女，1971年生，江西省宜春市人，汉族，南昌大学医学院在读博士，副教授，主要从事创面修复和组织工程方面的研究。 Qingling5218@ 163.com
  • 基金资助:

    国家自然科学基金资助项目(30560058)；江西省教育厅科技项目(GJJ09115)；江西省卫生厅科技项目(20092021)。

Amniotic membrane loading epidermal stem cells accelerates wound healing in diabetic rats

Zhong Qing-ling 1,2, Liu De-wu2, Liu Fan-rong1, Peng Yan2, Yu Mei1, Xiao Lu-liang1   

  1. 1 Medical College of Nanchang University, Nanchang  330006, Jiangxi Province, China; 2 Institute of Burn Surgery, First Affiliated Hospital of Nanchang University, Nanchang  330006, Jiangxi Province, China
  • Online:2010-08-06 Published:2010-08-06
  • Contact: Liu De-wu, Doctor, Professor, Chief physician, Doctoral supervisor, Institute of Burn Surgery, First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China dewuliu@126.com
  • About author:Zhong Qing-ling☆, Studying for doctorate, Associate professor, Medical College of Nanchang University, Nanchang 330006, Jiangxi Province, China; Institute of Burn Surgery, First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China Qingling5218@163. com
  • Supported by:

    the National Natural Science Foundation of China, No. 30560058*; the Science and Technology Project of Education Department in Jiangxi Province, No. GJJ09115*; the Science and Technology Project of Health Department in Jiangxi Province, No. 20092021*

PDF

686

被引次数

26

摘要：

背景：表皮干细胞作为皮肤组织的特异性干细胞，具有强大增殖及多向分化潜能，与创面修复紧密相关。近期研究表明糖尿病皮肤创面愈合过程中表皮干细胞数量减少、活性降低是导致其创面难愈的重要原因。
目的：观察表皮干细胞在糖尿病大鼠创面愈合中的作用。
方法：分离培养及鉴定SD大鼠表皮干细胞，并以BrdU标记。建立糖尿病SD大鼠创面模型，抽签法随机分为3组：表皮干细胞组创面移植羊膜负载BrdU标记的表皮干细胞；羊膜组创面移植羊膜；空白对照组创面未给予干预。 观察创面愈合情况、计算创面愈合率，苏木精-伊红及免疫组织化学SP法检测创面愈合组织中BrdU及增殖细胞核抗原表达。用图像分析软件测量阳性细胞积分吸光度平均值。
结果与结论：表皮干细胞组治疗后7d创面缩小明显，治疗后14 d创面基本愈合，创面愈合率明显高于羊膜组、空白对照组 (P < 0.01)。表皮干细胞组创面及新生表皮中可见BrdU阳性细胞，而另两组皮肤创面组织中始终未见BrdU阳性细胞。各组创面组织中可见增殖细胞核抗原阳性细胞表达，但表皮干细胞组的阳性细胞积分吸光度平均值与羊膜组、空白对照组比较差异有显著性意义(P < 0.01)。结果证实糖尿病大鼠创面愈合过程中表皮干细胞与创缘表皮移行、创面的上皮化有直接关联，可有效促进其创面愈合。

关键词: 表皮干细胞, 糖尿病, 创面愈合, 羊膜, 干细胞移植

Abstract:

BACKGROUND: As the specific stem cells in skin tissues, epidermal stem cells (ESCs) which posses high proliferative potential and multiple-directional potential play a key role in repairing and rebuilding of the skin. Recent studies showed that the decreased amount and the reduced proliferation and differentiation capacity of ESCs may be one of the important mechanisms of difficult wound recovery in diabetes mellitus.
OBJECTIVE: To investigate the effects of ESCs on wound healing of diabetic rats. 
METHODS: ESCs of Sprague Dawley rats were isolated, cultured, identified and labeled with 5-bromo-2’-deoxyuridine (BrdU). The wound models of diabetic SD rats were established, and then divided into ESCs, HAM, blank control groups. Human amniotic membrane (HAM) loading labeled BrdU ESCs were implanted to ESCs group. HAM was implanted to HAM group, but blank control group was not intervened. General situation of wound healing was observed, and the healing rate of wound was calculated. Hematoxylin and eosin staining and immunohistochemical staining (SP method) were used to detect BrdU and expression of proliferating cell nuclear antigen in healing tissues. Mean value of integral absorbance of positive cells was measured with image analysis software.
RESULTS AND CONCLUSION: At 7 days after treatment, the wounds of ESCs group reduced significantly. At 14 days after treatment, wounds of ESCs group were almost completely healed. Compared with HAM and blank control groups, the wound healing rate of ESCs group was significantly higher (P < 0.01). BrdU-positive cells in the wounds and newborn epidermis of ESCs group were visible, while wound tissue of HAM and blank control groups had no BrdU-positive cells. Proliferating cell nuclear antigen-positive cells in wound tissue of each group could be seen, but mean value of integral absorbance of positive cells showed significantly differences in ESCs group compared to HAM and blank control groups (P < 0.01). The results have confirmed that ESCs have a direct correlation with epidermal migration of wound margin and wound epithelialization in diabetic rats, may contribute to healing of diabetic skin wound.

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