非培养皮肤细胞喷雾移植技术用于小型猪创伤修复的研究  

Non-cultured skin cell-spray grafting technology for full-thickness skin wound repair in a minipig model

作　　者：岳亮 贠志敏 李素波 余群芳 任延玲 马瑞麟 崔宏图 钟鹏飞 张雪 宫锋 檀英霞 YUE Liang;YUN Zhi-min;LI Su-bo;YU Qun-fang;REN Yan-ling;MA Rui-lin;CUI Hong-tu;ZHONG Peng-fei;ZHANG Xue;GONG Feng;TAN Ying-xia(Institute of Health Service and Transfusion Medicine,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China;Experimental Animal Center,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100071,China)

机构地区：[1]军事科学院军事医学研究院卫生勤务与血液研究所,北京100850 [2]军事科学院军事医学研究院实验动物中心,北京100071

出　　处：《军事医学》2023年第2期104-110,共7页Military Medical Sciences

摘　　要：目的 探讨非培养皮肤细胞结合喷雾移植技术用于小型猪全层皮肤损伤的修复治疗效果。方法 巴马小型猪背部两侧创建6个直径3 cm的全层皮肤切除创面,分生理盐水组、脱细胞猪真皮基质粉组(ADM组)、多点注射细胞治疗组(细胞注射组)和喷洒细胞治疗组(细胞喷洒组)进行治疗。术后连续监测实验动物的生理状况及创面愈合情况,切取创面组织进行HE染色和细胞角蛋白-10(CK10)、CK14及血管性血友病因子(VWF)免疫组织化学染色,评估创面皮肤损伤修复效果。结果 术后14 d开始,细胞喷洒组的愈合速度更快;21 d时,细胞喷洒组创面的新生皮肤和毛发等皮肤附属器产生。HE染色结果证明,细胞治疗促进皮肤再生,且细胞喷洒组修复效率更高、新生皮肤更薄、更均一。免疫组化结果证明,术后7 d开始,细胞喷洒组中CK10阳性新生皮肤组织更长,修复速度更快;修复后期,细胞喷洒组CK10/CK14阳性新生皮肤更平整光滑,实现了美学修复。另外,细胞喷洒组VWF阳性细胞出现的时间更早,数量更多,排列更规律整齐,证明其新生皮肤富含更多的血管和毛囊,实现了功能修复。结论 非培养皮肤细胞结合喷雾移植技术的应用显著提高了小型猪全层皮肤损伤的愈合效率,更重要的是实现了创面的功能及美学修复,为后续临床转化奠定了理论和技术基础。该技术为平、战时创面的早期救治开辟了一条新的有效途径,有望成为未来烧伤治疗的新模式。Objective To investigate the efficacy of non-cultured skin cells combined with spray grafting technique against full-thickness skin lesions in minipigs. Methods Six full-thickness skin excision wounds of 3 cm in diameter were created on both sides of the backs of Bama minipigs,which were treated with saline,powdered porcine acellular dermal matrix(ADM),multisite injection of cell therapy,and sprayed cell therapy respectively. The physiological status and wound healing of the experimental animals were continuously monitored postoperatively,and wound tissues were excised for HE staining and immunohistochemical staining of cytokeratin-10(CK10),CK14,and von Willebrand factor(VWF)to evaluate the effect of wound repair. Results Healing was faster in the cell spraying group starting at 14 days after surgery. On day 21,skin appendages such as hair were formed in the regenerated skin of the cell spraying group. The results of HE staining proved that cell treatment promoted skin regeneration,and the repair efficiency was higher and the newly regenerated skin was thinner and more homogeneous in the cell spraying group. The immunohistochemical results showed the that the CK10 positive regenerated skin tissue was longer in the cell spraying group starting 7 days after surgery. At the later stage of repair,the CK10/CK14 positive regenerated skin in the cell spraying group was flatter and smoother,which was a sign of esthetic restoration. In addition,the VWF-positive cells in the cell spraying group appeared earlier,in greater numbers,and were arranged more regularly and neatly,suggesting that their new skin was rich in blood vessels and hair follicles and achieved functional repair. Conclusion Non-cultured skin cells combined with spray grafting technique can significantly improve the healing efficiency of full-thickness skin lesions in minipigs. What’s more,the technique can achieve the functional and aesthetic repair of wounds,which can be used as a potential means for future burn treatment.

关 键 词：非培养皮肤细胞 细胞喷雾移植 全层皮肤损伤 创面修复 毛囊再生 

分 类 号：R62[医药卫生—整形外科]