苦味酸-天狼猩红偏振光法对瘢痕动物模型的鉴定

背景:在瘢痕研究中急需稳定可靠的实验动物模型。目的:运用组织学方法对裸鼠瘢痕动物模型进行鉴定,确定最佳使用时机。设计:随机对照的重复测量设计。单位:中山大学附属第一医院烧伤科。材料:实验于2004-01/03在中山大学医学院动物实验中心完成。15只4～6周龄裸鼠(性别随机,体质量15～25g)由中山大学医学院实验动物中心提供;增生性瘢痕来自烧伤愈后、瘢痕增生半年之患者手术切除标本。方法:于裸鼠背侧移植人增生性瘢痕,建立疤痕动物模型;移植后4周起,每周处死5只实验动物,取移植物,100g/L甲醛固定标本,持续3周。苦味酸-天狼猩红染色偏振光检测移植物及临床取材,观察组织特点。主要观察指标:①苦味酸-天狼猩红染色偏振光法读片结果。②计算机图像分析结果。结果:①苦味酸-天狼猩红染色偏振光法读片结果:各时段组移植物在偏振光下呈现相同的以黄、红粗大纤维为主、疏网状绿色纤细纤维散在分布的特点。②计算机图像分析结果:临床病理性增生性瘢痕中,Ⅰ型胶原约占74%,Ⅲ型胶原约占26%;4～6周移植物中,Ⅰ型胶原含量分别为(74.52±0.47)%,(74.43±0.53)%,(74.69±0.63)%;Ⅲ型胶原分别约为(25.48±0.47)%,(25.57±0.53)%,(25.31±0.63)%,差异不显著(P>0.05)。结论:在实验所设计时段中,移植物与临床取材特性一致,符合增生性瘢痕特点。苦味酸-天狼猩红染色偏振光检测瘢痕组织胶原是对增生性瘢痕组织测定的简便、有效手段;运用裸鼠建立瘢痕动物模型是行之有效的、稳定的。

Identification of scar animal models with method of picric-sirius red polarized light

BACKGROUND: Stable reliable experimental animal models are needed urgently in scar research.OBJECTIVE: Scar animal models of nude mice are evaluated with histological method to define optimal opportunity for using.DESIGN: Randomizly controlled and repetitively measured design.SETTING: Department of Burn, First Affiliated Hospital, Sun Yat-sen University.MATERIALS: The experiment was conducted at the Center for Animal Experiment, Medical College, Sun Yat-sen University between January 2004 and March 2004. Fifteen nude mice aged 4-6 weeks were provided by Center for Animal Experiment, Medical College, Sun Yat-sen University (of either gender with body mass of 15-25 g). Hyperplastic scar was gained from samples of exairesis in patients with burn after healing which is hyperplastic scar for half a year.METHODS: Human hyperplastic scar was grafted at dorsa of nude mice to establish scar animal models. After graft for four weeks, 5 experimental animals were killed every week, and grafts were gained. 100 g/L formalin was used to fix samples for 3 weeks. Picric-sirius red polarized light method was used to detect the graft and clinical materials, and histological feature was observed.MAIN OUTCOME MEASURES: ①Results of film reading of picric-sirius red polarized light method. ②Analytic result of computer image.RESULTS: ①Results of film reading of picric-sirius red polarized light method: The grafts showed the same feature of diffused distribution of mainly yellow and red thick fiber with thin-mesh green fiber under polarized light in every time segment group. ②Analytic result of computer image: In clinicopathological hyperplastic scar, type Ⅰ collagen was about 74%; type Ⅲ collagen accounted for about 26%. In the graft from 4-6 weeks, the contents of type Ⅰ collagen were (74.52 ±0.47)% , (74.43 ±0.53)% ,(74.69±0.63)%, respectively; The contents of type Ⅲ were (25.48±0.47)%, (25.57±0.53)%, (25.31±0.63)%, respectively, which had insignificant difference (P ＞ 0.05 ).CONCLUSION: In the time segment designed by experiment, the feature of graft and clinical material is coincident, which is accorded with the characteristics of hyperplastic scar. The detection of collagen of scar tissue with picric-sirius red polarized light method is a simple effective method for assessing the tissue of hyperplastic scar. Establishing scar models with nude mice is effective and stable.