活性真皮基质的研制及生物学活性检测

Construction of living dermal matrix and detection of its bioactivity

摘要: 目的构建含有成纤维细胞(Fb)-胶原-猪去细胞真皮基质(PADM)的活性真皮基质。研究培养液中转化生长因子-β1(TGF-β1)和碱性成纤维细胞生长因子(bFGF)的分泌情况。方法将Fb与胶原混合种植于PADM的表面,收集第2、4、6、8、10d培养液样本,双抗体夹心ELISA法测定其中TGF-β1、bFGF的含量。结果 Fb在胶原内结构完整,与PADM形成复合真皮基质。TGF-β1和bFGF浓度培养第4天与第2天比较显著上升(P<0.01),并达到稳定水平。结论 Fb-胶原-PADM真皮替代物可作为构建组织工程全层皮肤的真皮支架。

Abstract: Objective To construct the living dermal matrix containing fibroblasts-collagen-PADM and to investigate the secretion of TGF-β1 and bFGF in culture medium. Methods Fibroblasts were mixed with collagen inoculated on the surface of PADM. Culture medium samples were collected after 2, 4, 6, 8, and 10 days. TGF-β1 and bFGF levels in the culture medium were measured by ELISA. Results The structure of fibroblasts located in collagen was intact. Composite dermal matrix was constructed with PADM. The TGF-β1 and bFGF levels were higher on day 4 than on day 2 （P＜0.01） and then remained stable. Conclusion Living dermal matrix containing fibroblasts-collagen-PADM can be used as a substrate for constructing dermal skin in tissue engineering.