应用组织工程方法构建瘢痕疙瘩模型的实验研究

THE RESEARCH OF ASSEMBLYING ANIMAL MODELS OF KELOID EMPLOYING THE METHOD OF TISSUE ENGINEERING

目的 应用组织工程的原理和方法 ,构建人瘢痕疙瘩 (keloid,K)动物模型 ,探讨采用这一模型进行 K临床和实验室研究的可行性。 方法 取人体 K组织中成纤维细胞 (fibroblasts,FB)进行原代培养 ,将体外培养第 6～ 8代 FB,接种到培养液预湿的聚乳酸 -乙醇酸共聚物 (the copolymers of polylactic acid and polyglycolie acid,PL GA)支架上 ,形成体外复合体 ,将复合体转移至旋转式细胞培养系统容器内培养 ;1周后种植在 2 0只雌性裸鼠皮下 ,同体两侧对照 ;第 4、8周取材 ,对获得的 K组织进行组织学评价。 结果 术后裸鼠全部成活。复合体移植 8周后 ,在裸鼠皮下形成的 K保持了其原有的胶原形态 ,在电镜下可观察到植入物内同时存在纤维细胞和 FB,并可见丰富的粗面内质网 ,FB仍保持合成和分泌胶原的能力 ,细胞特性保持不变。 结论 PL GA与 K组织中 FB具有较好的亲和性 ,复合后在裸鼠体内可形成 K,值得进一步开发研制 ,以便为进行 K的研究提供更为合适的动物模型。

Objective To build animal models of keloid by method of tissue engineering and to discuss the feasibility of using it in clinical and lab researches. Methods Fibroblasts(FB) were isolated from keloids and cultured. The seventh and eighth generation of the cultured FBs were inoculated into the copolymers of polylactic acid and polyglycolic PLGA. After being cultured in rotatory cell culture system (RCCS)for 1 week,the FB was transplanted into athymic mice. The specimens were obtained 4 weeks and 8 weeks and examined histologically. Results All mice survived.The collagen patterns of all keloids were pressed in every specimen obtained 8 weeks. Fibrocytes and FB were observed in specimens by electronic microscope. There were abundent rough endoplasmic reticulum (RER) in FB, which indicated that FB's capability of synthesizing and secreting collagen was preserved and the cellular characteristic was remained. Conclusion There is a good affinity between PLGA and FB. The composition of PLGA and FB can form keloids in athymic mice,so that it deserves further researching and developing.