脂肪间充质干细胞条件培养基制备温敏凝胶外用治疗皮肤烫伤
发布时间：2019-06-05 18:53
【摘要】：背景:有研究显示脂肪干细胞参与烫伤后皮肤组织的修复,但利用脂肪干细胞分泌物制备水凝胶,并治疗皮肤烫伤的研究尚未见报道。目的:分析人脂肪干细胞条件培养基水凝胶外用治疗小鼠皮肤烫伤模型的疗效。方法:采用酶解联合贴壁培养法,从脂肪组织培养获得脂肪干细胞并进行鉴定。取稳定增殖期细胞,收集其条件培养基,加入壳聚糖、甘露醇、β-甘油磷酸钠和透明质酸等制备温敏水凝胶,同时用新鲜脂肪干细胞完全培养基制备水凝胶。24只C57BL/6小鼠背部脱毛后用95℃铝块烫伤10 s快速建立3度皮肤烫伤模型,右侧以脂肪干细胞条件培养基水凝胶每天涂抹2次;左侧使用脂肪干细胞新鲜培养基水凝胶每天涂抹2次,连续涂抹7 d,观察愈合时间与愈合进程,计算愈合率,在烫伤后第4,14,28天取材进行苏木精-伊红染色观察组织病理学变化。结果与结论:①人脂肪间充质干细胞为成纤维状,增殖旺盛,平均倍增时间为55 h,可被诱导分化为成骨细胞、软骨细胞和脂肪细胞,高表达CD29、CD44、CD90和CD105,低表达CD31和CD34,证明其符合间充质干细胞的标准;②用人脂肪间充质干细胞条件培养基制备的温敏水凝胶在4-20℃呈现为清凉透明的黏稠液体,放入37℃15 min变为半固态凝胶;③95℃铝块烫伤小鼠表皮、真皮及皮下脂肪和肌肉组织的正常结构完全破坏,符合严重的3度烫伤标准,伤口面积均大致为3 cm~2;④在修复过程中发现,与新鲜培养基水凝胶对照组相比,脂肪干细胞条件培养基水凝胶组创口愈合时间短,伤口整洁,瘢痕增生少,创面组织结构恢复好;⑤脂肪干细胞条件培养基水凝胶组炎症浸润深度、炎症浸润细胞密度、肉芽组织厚度、成纤维细胞密度、新生血管密度和表皮厚度等指标显著好于新鲜培养基水凝胶组,差异有显著性意义(P0.05);⑥以上结果表明,人脂肪间充质干细胞条件培养基水凝胶可有效促进烫伤皮肤伤口愈合,提高再生皮肤质量。
[Abstract]:Background: some studies have shown that adipose stem cells are involved in the repair of skin tissue after scald, but the preparation of hydrogel from adipose stem cell secretions and the treatment of skin scald have not been reported. Objective: to analyze the efficacy of human adipose stem cells (HSC) conditioned culture medium hydrogel in the treatment of mouse skin scald model. Methods: adipose stem cells were obtained from adipose tissue by enzyme hydrolysis combined with adherent culture. The thermosensitive hydrogel was prepared by adding chitosan, mannitol, sodium 尾-glycerophosphate and hyaluronic acid into the conditioned medium for stable proliferation of cells. At the same time, hydrogel was prepared by fresh adipose stem cell complete medium. 24 C57BL/6 mice were scalded with 95 鈩，
本文编号：2493749
[Abstract]:Background: some studies have shown that adipose stem cells are involved in the repair of skin tissue after scald, but the preparation of hydrogel from adipose stem cell secretions and the treatment of skin scald have not been reported. Objective: to analyze the efficacy of human adipose stem cells (HSC) conditioned culture medium hydrogel in the treatment of mouse skin scald model. Methods: adipose stem cells were obtained from adipose tissue by enzyme hydrolysis combined with adherent culture. The thermosensitive hydrogel was prepared by adding chitosan, mannitol, sodium 尾-glycerophosphate and hyaluronic acid into the conditioned medium for stable proliferation of cells. At the same time, hydrogel was prepared by fresh adipose stem cell complete medium. 24 C57BL/6 mice were scalded with 95 鈩，
本文编号：2493749
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