摘要
目的通过对培养系统中层黏蛋白(LN)合成表达的影响,观察垂直应力对组织工程皮肤生长及基底膜构建的作用。方法常规制备复方壳多糖组织工程皮肤,并根据应力作用与否分成两组培养。应力作用组每天接受垂直应力作用15 min,其他与对照组相同。在浸没培养3 d后,继续进行气-液界面三维培养。培养10、15 d后,行HE和免疫组化染色,观察组织工程皮肤的基底膜构建情况。采用Western Blot方法测定陈旧培养基中游离LN的含量。结果实验组在垂直应力作用下,复方壳多糖组织工程皮肤表皮分层分化较对照组的组织工程皮肤好,免疫组化染色结果显示实验组LN染色良好,且陈旧培养基中游离型LN-5的含量也高于对照组。结论垂直应力作用有助于组织工程皮肤系统中LN的表达,并最终促进体外基底膜结构的构建。
Objective To evaluate the effect of vertical stress on the development of tissue engineering skin and construction of basement membrane（BM） by influencing the laminin synthesis in vitro.Methods According to the stress exerting or not,composite chitosan tissue engineering skin were divided into two groups.The test group received vertical stress for 15 minutes every day.They were both carried out submerged culture for 3 days followed by air-liquid interface culture.After 10 or 15 days culture,the histological manifestation of BM in tissue engineering skin was observed by hematoxylin eosin staining,while laminin（LN）,the important component in BM,was detected immunohistochemically at the dermal-epidermal junction.Soluble LN-5 in conditioned culture medium was analyzed by Western Blot to investigate the role of vertical stress on BM formation in tissue engineering skin.Results In model of stress on tissue engineering skin,vertical stress applied for 15 minutes every day was helpful to restruct BM in vitro.The epithelium in test group differentiated more well than another group.It was found that LN,the major BM component,was positive immunohistochemically at the dermal-epidermal junction in the tissue-engineering skin.Soluble LN-5 in conditioned medium was higher in test group than the control group.Conclusion It is revealed that the vertical stress applied on tissue engineering skin is helpful for synthesis of laminin and promote the reconstruction of BM in vitro.
出处
《重庆医学》
CAS
CSCD
北大核心
2010年第20期2704-2706,2709,共4页
Chongqing medicine
基金
国家自然科学基金资助项目(30700793)
重庆市自然科学基金资助项目(CSTC2007BB5283)
关键词
组织工程皮肤
基底膜
器官型培养
垂直应力
层黏蛋白
tissue engineered skin
basement membrane
organotypic culture
vertical stress
laminin
