不同年龄阶段人脱细胞真皮的制备及细胞渗透性研究

Preparation and infiltration analysis of human acellular dermal matrix from different age stage

目的 制备不同年龄阶段的人脱细胞真皮（ADM）,并进行体外细胞渗透性研究,为改良ADM支架提供新的思路.方法 利用氢氧化钠（NaOH）消蚀＋冻融法制备ADM,对比青年人与老年人皮肤制备的ADM孔隙率和细胞毒性,在两种真皮的网状层接种人包皮成纤维细胞（FB）,于接种后3、6、9d通过HE染色比较不同年龄阶段ADM的细胞渗透性.结果 成功获得ADM,两组ADM细胞毒性低,老年组ADM孔隙率较青年组更理想,老年组ADM更多见到细胞形成复层并长入基质内部.结论 NaOH消蚀＋冻融法制备的ADM性状优良,其中老年人ADM的细胞渗透性较青年人更为理想,是一种良好的组织工程皮肤支架.

Objective To study the preparation of acellular dermal matrix （ADM） from different age stages,and to analyze the infiltration of ADM.Methods ADM was prepared by means of combined treatments with sodium hydroxide solution and freeze thawing,and the porosity,cell toxicity were also analyzed.The fibroblasts were seeded on the reticular dermis in both groups,after seeded for 3,6,9 days,the composites were observed by HE staining,the infiltration of ADM between young people （20-30 years old） and the elderly population （60-80 years old） were compared.Results We acquired ADM successfully by sodium hydroxide solution plus freeze thawing.ADM showed low cytotoxicity,ADM from aged skin displayed better porosity than youth group.HE staining demonstrated fibroblast infiltrated into ADM in more samples of aged ADM than young group.Conclusions Sodium hydroxide solution plus freeze thawing is an ideal way to prepare ADM.Compared with young people,skins from elderly population have more ideal infiltration,and it is therefore suitable for building tissue engineering skin.