摘要
背景:研究证实,去表皮的真皮可以作为真皮替代物,在其上接种角质形成细胞后形成表皮结构。但有关真皮替代物细胞生物活性、组织结构特点及基底膜成分分析的研究报道较少。目的:观察人去表皮真皮细胞活性及组织结构特征。方法:将健康成人皮瓣用56℃PBS溶液处理以去除表皮,用液氮连续冻融处理去除真皮中细胞成分,获得去表皮真皮。以组织块培养法观察去表皮真皮细胞活性。以苏木精染色检测去表皮真皮细胞核,以波形蛋白免疫组化检测去表皮真皮成纤维细胞成分。以PAS染色及Ⅳ型胶原免疫组化检测基底膜及其成分。以VG染色检测去表皮真皮胶原纤维,Weigert染色检测弹力纤维,VG与Weigert双染色检测胶原纤维及弹力纤维,透射及扫描电镜观察去表皮真皮超微结构。结果与结论:用组织块培养方法培养的去表皮真皮2周无细胞生长。苏木精-伊红染色显示去表皮真皮中无细胞核、波形蛋白免疫组化显示去表皮真皮中无波形蛋白表达。VG染色显示去表皮真皮胶原纤维染成玫瑰红色,Weigert染色显示去表皮真皮弹力纤维染成紫黑色,双染色进一步显示胶原纤维与弹力纤维均匀排列。去表皮真皮表面及附属器残留部位PAS反应强阳性,Ⅳ型胶原表达明显。透射及扫描电镜下观察到去表皮真皮中胶原﹑弹力纤维交错排列,间有孔隙,相互交织成网。去表皮真皮无活细胞成分,真皮基质表面及附属器管腔壁仍保留糖原﹑Ⅳ型胶原等基底膜成分,真皮基质中富含胶原及弹力纤维,是一种类似在体真皮的三维胶原基质。
BACKGROUND： Study confirmed that the de-epidermized dermis （DED） can be used as dermal substitute and may form epidermal structure after incubating keratinocytes. However, the cell biological activity, tissue structure characteristics and the basement membrane component analysis of dermal substitute have been reported less. OBJECTIVE： To investigate the cell activity and the tissue structure characteristics of DED. METHODS： Skin flap was treated with 56 ℃ phosphate buffered solution to remove the epidermis, and the dermal cell components were deleted by freezing and thawing with liquid nitrogen to obtain DED. The DED cell activity was detected with tissue culture method, hematoxylin nuclear staining was used to determine the DED cell nuclei, and vimentin immunohistochemistry was applied for fibroblast determinations. The basement membrane and its components were detected using Periodic Acid-Schiff staining and collagen type IV immunohistochemistry. Van Gieson stain, Weigart stain and those double staining were respectively used to determine DED collagen fibers and elastic fibers. The DED ultrastructure was observed under transmission and scanning electron microscope. RESULTS AND CONCLUSION： Using tissue culture method, the cultured DED did not exhibit cell growth at 2 weeks. Hematoxylin-eosin staining showed no nuclear in DED, vimentin immunohistochemistry showed no vimentin expressed in DED. Van Gieson staining showed DED collagen fibers were stained as rose red, Weigert staining showed DED elastic fibers were stained as purplish black, double staining further demonstrated uniform arrangement of collagen fibers and elastic fibers. DED surface and the remaining appendages were strongly positive for Periodic Acid-Schiff staining, and type IV collagen expression was significant. Transmission and scanning electron microscope results showed that, the DED elastic fibers and collagen overlap arranged with pore intervals, they intercrossed into a network. There is no living cell component in DED, dermal ma
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第11期2076-2080,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
the National Natural Science Foundation of China, No. 30872283~~
