摘要
目的 为皮肤缺失的移植提供具有生物活性的表皮、真皮的组织工程化人工复合皮肤。方法 将体外传代培养的表皮角朊细胞和真皮成纤维细胞分别接种在经冷冻干燥及戊二醛交联的 I型胶原基质网架的两侧 ,在液面下培养 1周后 ,改为气 -液界面培养 ,动态观测人工皮肤光镜下组织学形态及电镜下超微结构。结果 人工复合皮肤具有表皮、真皮双层结构。培养过程中表皮逐步增殖、分化、发育 ,形成基底层、棘细胞层、颗粒层和角质层 ;真皮基质网架渐渐降解 ,并逐步被增殖的成纤维细胞及其所分泌的细胞外基质所取代 ,完成真皮构建。结论 利用组织工程技术体外构建人工复合皮肤 ,可用于皮肤缺失者的自体移植治疗 ,体外构建的复合人工皮肤可从根本上解决自体供皮不足 ,且细胞源于自体皮肤 ,排除了发生免疫排斥反应和疾病传播的风险 ,具有安全、有效。
Objective To fabricate artificial human skin with the tissue engineering methods. Methods The artificial epidermis and dermis were fabricated based on the successful achievements of culturing human keratinocytes(Kc) and fibroblasts (Fb) as well as fabrication of collagen lattice. It included: ① Culture of epidermal keratinocytes and dermal fibroblasts: Kc isolated from adult foreskin by digestion of trypsin dispase. Followed by comparison from aspects of proliferation, differentiation of the Kc, overgrowth of Fb and cost benefits. ② Fabrication of extracellular matrix sponge: collagen was extracted from skin by limited pepsin digestion, purified with primary and step salt fraction, and identified by SDS PAGE. The matrix lattice was fabricated by freeze dryer and cross linked with glutaraldehyde, in which the collagen appeared white, fibrous, connected and formed pores with average dimension of 180 to 260 μm. ③ Fabrication artificial human skin: The artificial skin was fabricated by plating subcultured Kc and Fb separately into the lattice with certain cell density, cultured for one week or so under culture medium, then changed to air liquid interface, and cultured for intervals. Results The artificial skin was composed of dermis and epidermis under light microscope. Epidemis of the skin consisted of Kc at various proliferation and differentiation stages, which proliferated and differentiated into basal cell layer, prickle cell layer, granular layer, and cornified layer. Conifilament not only increased in number, but also gathered into bundles. Keratohyalin granules at different development stages increased and became typical. The kinetic process of biochemistry of the skin was coincide with the changes on morphology. Conclusion Tissue engineered skin epuivalent has potential prospects in application of repairing skin defect with advantages of safe, effective and practical alternatives.
出处
《中国修复重建外科杂志》
CAS
CSCD
2001年第4期235-239,共5页
Chinese Journal of Reparative and Reconstructive Surgery
基金
吉林省科委资助项目 (95 3470 )
关键词
组织工程
人工皮肤
角朊细胞
成纤维细胞
Tissue engineering Artificial human skin Keratinocytes Fibroblasts
