摘要
目的观察制备的组织工程皮肤基底膜的组织学特征。方法取门诊正常儿童包皮环切术之包皮,采用胰蛋白酶胶原酶顺序消化得到角质形成细胞(KC)和成纤维细胞(Fb)悬液。制备复方壳多糖组织工程皮肤,浸没培养3 d后,继续行气液界面培养。将培养7、10、15 d的复方壳多糖组织工程皮肤用中性甲醛溶液固定后石蜡包埋、切片,行HE及高碘酸-雪夫(PAS)染色,并用免疫组织化学染色法观察基底膜的重要成分:Ⅳ型胶原、Ⅶ型胶原及层黏连蛋白(LN)的存在情况。结果HE染色可见培养的组织工程皮肤表皮结构分化良好,大致可分为基底层、棘层和角质层,各层均有数量不等的扁平梭形细胞。PAS染色显示真皮表皮间有一均匀红染的条带。免疫组织化学染色结果显示,Ⅳ型胶原、Ⅶ型胶原及LN呈阳性表达。结论复方壳多糖组织工程皮肤基底膜构建良好。
Objective To observe the histological characteristics of constructed basement membrane in tissue-engineered skin. Methods Forskins from circumcision in normal children were obtained with informed consent of the parents, and then the epidermal keratinocytes （KC） and dermal fibroblasts（Fb） were isolated with trypsin and collagenase D digestion in sequence. Tissue engineered skin with composite chitosan was maintained in a submerged state for 3 days, and then at the air-liquid interface. The tissue-engineered skins were fixed in neutral formalin and then embedded in paraffin after culture for 7,10 and 15 days, respectively for immunohistological examination of the basement membrane component, including the condition of collagen type IV（COL-IV） , collagen type Ⅶ（COL-Ⅶ） , and laminin （LN）. Results HE staining showed that the keratinocytes formed a fine stratified squamous epithelium with the presence of basal, spinous, granular and corneous cell layers, and there was various amount of cells in flat and fusiform shape in each layer. It was found that a regular red staining strip situated at the dermal epidermal junction. Positive staining of collagen Ⅳ, collagen Ⅶ as well as LN was observed by immunohistological examination. Conclusion The results suggest that the composite chitosan tissue engineered skin has a good prospect for clinical use because it presents a perfect reconstruction of basement membrane.
出处
《中华烧伤杂志》
CAS
CSCD
北大核心
2007年第5期369-371,共3页
Chinese Journal of Burns
基金
国家高技术研究发展计划(2003AA205022)
重庆市科技计划项目(7897)
关键词
壳多糖
组织工程
皮肤
人工
基底膜
Chitin
Tissue engineering
Skin,artificial
Basilar membrane
