利用Collagen/PLCL复合纳米纤维电纺膜构建组织工程化气管补片的初步研究  

Preliminary study on construction of tissue-engineered tracheal patch by electrospun collagen/PLCL composite nanofibrous membranes

作　　者：何晓敏 付炜 王浩[1] 冯蓓[1,2] 刘珍伶 殷猛[1] 王伟 徐志伟[1] 郑景浩[1,2] HE Xiao-min;FU Wei;WANG Hao;FENG Bei;LIU Zhen-ling;YIN Meng;WANG Wei;XU Zhi-wei;ZHENG Jing-hao(Department of Cardiothoracic Surgery,Shanghai Children's Medical Center,Shanghai Jiaotong University School of Medicine,Shanghai 200127,China)

机构地区：[1]上海交通大学医学院附属上海儿童医学中心心胸外科,200127 [2]上海交通大学医学院附属上海儿童医学中心儿科转化医学研究所,200127

出　　处：《中华临床医师杂志（电子版）》2012年第22期7095-7099,共5页Chinese Journal of Clinicians(Electronic Edition)

基　　金：上海市级医院新兴前沿技术联合攻关项目课题(SHDC12010112);国家自然科学基金(31200735);上海市自然科学基金(12ZR1446500);上海交通大学医工交叉项目(YG2012MS36);上海市浦东新区科技发展基金(PKJ2012-Y48)

摘　　要：目的初步探讨以胶原/聚左旋乳酸-己内酯共聚物(Collagen/PLCL)复合纳米纤维电纺膜作为支架材料复合肋软骨细胞构建组织工程化气管补片的可行性。方法分离及培养2月龄新西兰兔肋软骨细胞,取第二代肋软骨细胞种植于Collagen/PLCL复合纳米纤维电纺膜,并进行扫描电镜观察,然后利用"三明治"法构建成细胞材料复合物,体外培养4周再植入裸鼠背部皮下,4周后取出进行大体观察、组织学染色以及Ⅱ型胶原免疫组织化学检测以评价组织成软骨效果。结果扫描电镜显示肋软骨细胞在该材料上黏附生长良好,细胞材料复合物在体内培养4周后已形成类软骨样组织,组织学检查HE染色可见有大量软骨陷窝形成,甲苯胺蓝、番红O染色可见有大量软骨基质分泌,Ⅱ型胶原免疫组织化学染色呈阳性表达提示该组织含有软骨组织特有的Ⅱ型胶原。结论 Collagen/PLCL复合纳米纤维电纺膜对于兔肋软骨细胞具有良好的生物相容性,复合肋软骨细胞形成的组织工程化软骨类似于正常透明软骨组织,适合于构建组织工程化气管补片。Objective To investigate the feasibility of electrospun collagen/PLCL composite nanofibrous membranes combined with costal chondrocytes to construct tissue-engineered tracheal patch.Methods Costal chondrocytes were isolated and expanded to two passages from New Zealand rabbits,seeded on the collagen/PLCL nanofibrous scaffolds for scanning electron microscopy.Then,the cell-scaffold compounds were constructed by a sandwich model,cultured in vitro for 4 weeks and subsequently implanted subcutaneously in nude mouse for additional 4 weeks.The cartilage-like tissues were evaluated by gross view,histology staining and typeⅡcollagen immunohistochemistry.Results We found that the scaffold facilitated the proliferation of costal chondrocytes from the results of scanning electron microscopy.The cell-scaffold compounds were similar to cartilage tissues after cultured in vivo for additional 4 weeks.HE staining proved amount cartilage lacuna formation,while the secretion of cartilage matrix were proved by toluidine blue and safranin O.Besides,immunohistochemistry proved the expression of typeⅡcollagen.Conclusions Electrospun collagen/PLCL composite nanofibrous membranes had the satisfactory biocompatibility and were considered as the suitable scaffolds for tissue-engineered tracheal patch.

关 键 词：胶原 组织工程 气管 软骨 静电纺 聚左旋乳酸-己内酯共聚物 

分 类 号：R318.08[医药卫生—生物医学工程]