明胶海绵支架体内构建组织工程化颌下腺的研究  

The experiment research on reconstituting tissue engineering submandibular gland with collagen sponge scaffold in vivo.

作　　者：朱耀旻[1] 王洋[2] 周青[3] 王玉新[3] 郑苍尚[1] 

机构地区：[1]深圳市第二人民医院口腔科,广东深圳518035 [2]深圳市第二人民医院神经内科,广东深圳518035 [3]中国医科大学口腔医学院口腔外科,辽宁沈阳110002

出　　处：《临床口腔医学杂志》2010年第11期645-648,共4页Journal of Clinical Stomatology

基　　金：深圳市科技计划项目(200902043)

摘　　要：目的:研究采用组织工程技术通过颌下腺细胞与明胶海绵复合物的体内移植,探讨在同种异体动物体内构建具有生命活力的组织工程化颌下腺的可行性。方法:选用Wistar大鼠18只,分三组。A组:明胶海绵材料与鼠颌下腺细胞复合培养物;B组:单纯明胶海绵材料;C组:空白切开缝合对照。材料植入大鼠背部,术后3、7和15 d分期观察植入物的大体形态、组织学和超微结构,BrdU标记表达,及免疫排斥反应变化。结果:A组植入体内后颌下腺细胞在材料生长,形成的组织表面白色且有光泽、致密,组织学可见材料排列较为规则,15 d细胞仍成活并大量增殖,BrdU表达为阳性;B组植入体内7 d后材料逐渐变细,15 d后材料被逐渐降解吸收出现中断;A组分别于3、7、15 d进行扫描电镜观察可见颌下腺细胞数目逐渐增多,黏附在材料表面上生长,伸出突起,并可见明显的细胞分裂增殖,细胞形成功能性团块;A、B两组与C组相比3、5、7 d时白细胞数量增多明显,但逐步降低,第15天三组白细胞数量趋于一致。三组淋巴细胞数量变化不大,无显著性差异。结论:同种异体颌下腺细胞与明胶海绵材料复合构建的组织工程颌下腺,植入免疫功能正常的动物体内,细胞具有生命力,能够在材料上黏附生长、增殖,免疫反应随时间增加逐渐趋于正常。Objective：To reconstitute tissue engineering submandibular gland by implant RSGCs combined with collagen sponge scaffold in vivo.Method：18 Wistar rats were divided into three groups according to inserts（group A： RSGCs seeded on collagen sponge;group B：collagen sponge;group Cs：ham-operated controls）.RSGCs labeled by Brdu were seeded on collagen sponge.Inserts of group A and B were implanted either beneath the skin on the back or adjacent to the neck.At 3,7and 15days post-implantation,implant sites were examined local and systemic responses were assessed by conventional clinical chemistry and hematology analyses.HE and brdu immunohistochemical examined the responses of tissue.Scanning electron microscope was used to observe the growth behavior of RSGCs on collagen sponge scaffolds.Result：In group A,RSGCs can adhere and keep proliferating and differentiating on collagen fiber.The results of Brdu labelled cells were positive by immunohistochemical methods at each time point.At the 15th day RSGCs proliferate on the collagen sponge scaffold and form functional lumen unit.In group Bi,t is similar to group A at the 3rd and 7th day,but at the 15 th day after implantation the insert is not completely in shape.Postoperative inflammatory reaction of either group A or group B is strong until the 7 th day.Neutrophil levels were elevated initially postimplantation of implant groupsa,bove those seen in the shamoperated rat,but returned to similar values by 15 days.lymph cell levels has no significance change.Conclusion：Implantation of allogenous RSGCs can adhere and keep proliferating and differentiating on collagen fiber in vivo.There happen acute and temple Immunological rejection.But this rejection has no effect on proliferation and differentiation of RSGCs.This method can used to reconstitute tissue engineering submandibular gland in vivo.

关 键 词：组织工程 颌下腺细胞 明胶海绵支架 细胞培养 

分 类 号：R782[医药卫生—口腔医学] Q813.11[医药卫生—临床医学]