干细胞培养物表面滴注对豚鼠皮肤重度缺损的修复作用

背景干细胞是动物和人的一种特殊功能的细胞,存在于多种组织之中,其大部分分化成各种特定组织器官,一部分保持干细胞状态,以备组织修复之用.有研究将间充质干细胞移植于烧伤创面,用以诱导皮肤干细胞的增生和活化,以达到治疗烧伤的目的.目的观察采用体外培养干细胞培养液滴注于豚鼠皮肤重度缺损局部对创面愈合时间及愈合情况的影响.设计随机分组,空白对照实验.单位吉林大学公共卫生学院毒理学教研室.材料成年健康豚鼠14只,体质量300～350 g,雌雄不限.方法实验于2003-03/09在吉林大学公共卫生学院毒理学教研室实验室完成.取10豚鼠,经颈部放血处死,提取骨髓,经过培养制成单细胞悬液备用.将14只豚鼠制成两侧重度皮肤损伤模型.随机选取10只豚鼠,其中一侧滴加干细胞培养物(干细胞组),而另一侧滴加培养液(培养液组);剩余4只动物不做任何治疗,作为空白对照组.3 d后,每两天用透明有机玻璃置创面上方,描画其形状,观察创面情况,并将其图形复制到透明玻璃纸上,在直角坐标纸上精确查出创面面积,计算创面愈合速度.主要观察指标大体观察各组豚鼠创面愈合情况及平均愈合时间和速度.结果14只豚鼠均进入结果分析.①各组豚鼠创面愈合情况3 d时,干细胞组创面干燥,无明显渗出,在创面底部有一层膜状物质,与纱布粘贴比较牢固,敷料不易脱离;培养液组创面情况与干细胞组基本一致,但无膜状物质形成,敷料易脱离;空白对照组动物创面炎症明显.②各组豚鼠创面愈合时间干细胞组明显少于培养液组和空白对照组(12.45±2.18)d比(26.29±1.38)d和＞30 d,P＜0.05].③各组豚鼠创面愈合速度干细胞组明显快于培养液组和空白对照组(40.42±2.14)mm2/d比(15.53±5.22)mm2/d和(10.27±4.57)mm2/d,P＜0.05].结论利用同种异体动物干细胞培养物进行皮肤表面滴注修复其损伤创面,干细胞能够在创面上生长,并向皮肤细胞转化,促进创面皮肤修复,用于皮肤重度缺损治疗是可行的.

Surface instillation of stem cell culture in repair of severe skin trauma in guinea pigs

BACKGROUND: Stem cells, cells with special function in animals and humans, exist in various tissues. Most of stem cells differentiate into special tissue organs and some of them remain in the status of stem cells for tissue repair. Mesenchymal stem cells were transplanted to burn wounds in some researches for inducing the proliferation and activation of skin stem cells so as to cure burn.OBJECTIVE: To observe the effect of stem cell culture medium cultured in vitro instilled locally into the severely traumatic skin in guinea pigs on healing time and healing degree of the wound.DESIGN: Random grouping and blank control trial.SETTING: Department of Toxicology, School of Public Health, Jilin University.MATERIALS: Totally 14 adult healthy guinea pigs of either gender weighing 300 to 350 g were recruited.METHODS: The experiment was conducted in the Laboratory of Toxicology Department, School of Public Health, Jilin University, from March to September 2003. Ten guinea pigs were put to death by bloodletting on the neck. The bone marrow was extracted and cultured in unicellular supematant fluid for use. The 14 guinea pigs were made into models of bilateral severe skin trauma.Ten of the guinea pigs were chosen randomly, stem cell culture was instilled into one side of the animals (stem cell group), while the culture medium was instilled into the other side of the animals (culture medium group). The remaining 4 guinea pigs that received no treatment were blank control group.Three days later, transparent lucite was put on the wound every other day for drawing the shape and observing the wound. After the shape was copied onto the transparent lucite, the wound area was worked out on the rectangular coordinate paper and the speed of wound healing was calculated.MAIN OUTCOME MEASURES: Gross observation was performed on the healing status of the wound and average healing time and speed of the guinea pigs in each group.wound healing status of the guinea pigs in each group: At day 3, the wound in stem cell group was dry without obvious exudation. There was a layer of membrane at the bottom of the wound and it stuck to the pledget closely so that the dressing could not easily be removed. The wound status in the culture medium was practically the same as that of the stem cells, but no membrane-ike material was formed and the dressing was easily removed. Inflammation appeared obviously in blank control group.nificantly less in stem cell group than in culture medium group and blank control group (12.45±2.18) days vs (26.29±1.38) days and ＞ 30It was obviously faster in stem cell group than in culture medium group and blank control group (40.42±2.14) mm2 per day vs (15.53±5.22) mm2 per day and(10.27±4.57) mm2 per day,P ＜ 0.05].CONCLUSION: The stem cells of homologous animals were instilled on the skin surface to repair injury wound. Stem cells can grow on the wound and transform into skin cells, which promotes the recovery of wounded skin. Therefore, it is feasible to treat severe skin defect with stem cells.