应用双光子显微镜观察自体成纤维细胞填充效果  

Filling effect of autologous skin fibroblasts： a study of two-photon fluorescence microscopy

作　　者：熊舒原[1] 曹宁[2] 察鹏飞[1] 卓双木[3] 陈建新[3] 

机构地区：[1]福建医科大学附属第一医院整形外科,福州350005 [2]福建省漳州市医院整形外科 [3]福建师范大学医学光电科学与技术教育部重点实验室

出　　处：《中华医学美学美容杂志》2008年第3期193-196,共4页Chinese Journal of Medical Aesthetics and Cosmetology

摘　　要：目的通过双光子显微镜观察体外培养的小鼠皮肤成纤维细胞皮内注射移植后的长期存活情况，了解胶原纤维等基质成分的变化。方法将增强型绿色荧光蛋白（EGFP）慢病毒液转染成功的成纤维细胞注射到小鼠皮内，分别于注射1、2个月后取材，行连续冰冻切片，双光子显微镜观察，对胶原的分布面积和真皮厚度做图像分析，并对所得数据进行统计学处理。结果双光子显微镜对注射移植细胞及胶原清晰成像，注射1个月时，胶原分布面积及真皮厚度与对照组比较差异无统计学意义（P〉0.05）；2个月后胶原分布面积及真皮厚度与对照组比较差异有统计学意义（P〈0.01）。结论成纤维细胞注射移植到小鼠皮内可以长期存活，能够分泌胶原并增加真皮厚度，为临床上应用自体真皮成纤维细胞进行软组织填充注射治疗的可行性，提供了理论基础。Objective To investigate the survival profile of the intradermally injected mouse autologous skin fibroblasts and the changes of the collagen fibers by using green fluorescent protein labeling and two-photon fluorescence microscopy. Methods The cultured cells were transfected by EGFP lentivirus, and then the cells were injected into the corresponding mouse skin. Biopsy was taken from the animals after 1 and 2 months. The specimens made serial frozen sections, the survival profile of the injected cells and the changes of the collagen fibers were observed by two-photon fluorescence microscopy. The collagenic area and dermal thickness were measured with image analysis software, and statistical analysis was also carried out. Results Two-photon fluorescence microscopy showed clear images of the injected cells and collagen fibers. Both the area of collagen fibers and the dermal thickness were significantly increased in injected cells after 2 months （P 〈0.05）, however, there were no difference between injected cells and control after 1 mouth （P 〉 0.05）. Conclusions Autologous cultured fibroblasts could survive in a long time after transplantating into the skin, and collagen could be newly produced, the depth of dermis increases, which provides a possibility to treat mini-defects of the tissue.

关 键 词：成纤维细胞 细胞移植 双光子显微镜 

分 类 号：R[医药卫生]