微孔化猪脱细胞真皮基质与大鼠骨髓间充质细胞对裸鼠皮肤附件细胞再生的作用  

Effects of microporous porcine acellular dermal matrix combined with bone marrow mesenchymalcells of rats on the regeneration of cutaneous appendages cells in nude mice

作　　者：罗旭[1] 辛国华[2] 曾逃方 林才[1] 曾元临[2] 李郁葱[4] 邱泽亮 

机构地区：[1]浙江省温州医科大学附属第一医院烧伤·伤口中心,325000 [2]南昌大学第一附属医院烧伤科 [3]宜春市人民医院烧伤科 [4]赣州市人民医院烧伤整形科 [5]上海市浦东新区周浦医院重症医学科

出　　处：《中华烧伤杂志》2013年第6期541-547,共7页Chinese Journal of Burns

基　　金：浙江省自然科学基金重点项目（Z2080985）;浙江省“十二五高校重点学科”危重病与灾害救援医学;温州市科技计划项目（Y20080101）

摘　　要：目的观察微孔化猪ADM复合含有骨髓间充质干细胞（BMSC）的大鼠骨髓间充质细胞群对裸鼠部分皮肤附件细胞再生的作用。方法将1只清洁级健康小白猪处死，切取制作面积约20cm×10Cll、0．3mm厚断层真皮片，通过激光打孔、高渗盐溶液脱细胞、交联等处理制作激光微孔化猪ADM（LPADM），无孔猪ADM仅行脱细胞、交联等处理，进行外观、组织学、扫描电镜观察。将1只SD大鼠处死，取股骨和胫骨，分离培养骨来源的骨髓间充质细胞群，取第3代贴擘细胞进行成骨、成脂肪细胞分化实验。之后将其接种于LPADM、无孔猪ADM上，构建骨髓间充质细胞一LPADM和骨髓间充质细胞一无孔猪ADM。取21只健康裸鼠随机区组分为骨髓间充质细胞一LPADM＋无孔猪ADM组（简称A组，6只）、LPADM＋刃厚皮片组（简称B组，6只）、骨髓间充质细胞一I。PADM＋刃厚皮片组（简称C组，6只）、骨髓间允质细胞一无孔猪ADM＋刃厚皮片组（简称D组，3只），麻醉后在其背部正中做一2cin×2cm的全层皮肤缺损创面，达深筋膜，同时切取相同大小刃厚皮片备用，并分别移植相应材料覆盖创面．．于移植术后5、7、14d观察裸鼠局部情况及不良反应，各组分别处死1只裸鼠切取全部移植物，HE染色观察其组织结构；移植术后7、14d透射电镜下观察相应复合物中新生皮肤附件情况。结果（1）LPADM、无孔猪ADM呈瓷白色，柔软、有弹性，组织学观察显示真皮基质末见细胞成分；扫描电镜示孔径中胶原纤维排列有序；LPADM还有微孔结构。（2）细胞传至第3代时，形态趋于一致，呈Fb样，生长较快。，（3）诱导分化实验表明，细胞可向成骨细胞、成脂肪细胞分化。（4）移植术后5d，A组无孔猪ADM局部干燥，I）组皮片局部干燥坏死，A、D组均未见感染及炎症反应；B、c组移植皮片成活。移植术后7、14d，A组表面的覆�Objective To observe the effects of microporous porcine acellular dermal matrix （ADM） combined with bone marrow mesenchymal cells （BMMCs） population containing bone mesenchymal stem cells （BMSCs） of rats on the regeneration of cutaneous appendages cells in nude mice. Methods Split-thickness dermal grafts, 20 cmx 10 em in size and 0.3 mm in thickness, were prepared from a healthy pig which was sacrificed under sanitary condition. Laser microporous porcine ADM （LPADM） was produced by laser punching, hypertonic saline solution acellular method, and crosslinking treatment, and nonporous porcine ADM （NPADM） was produced by the latter two procedures. Then the appearance observation, his- tological examination and scanning electron microscope observation were conducted. BMMCs were isolated and cultured from tibia and femur after sacrifice of an SD rat. Osteogenic and adipogenic differentiation ex- periments were conducted among the adherent cells in the third passage. Then they were inoculated to LPADM and NPADM to construct BMMCs-LPADM and BMMCs-NPADM materials. Twenty-one healthy nude mice were divided into BMMCs-LPADM ＋ NPADM group （ A, n = 6） , LPADM ＋ split-thickness skin graft group （ B, n = 6） , BMMCs-LPADM ＋ split-thickness skin graft group （ C, n = 6） , BMMCs-NPADM ＋ split-thickness skin graft group （ D, n = 3 ） according to randomized block. After anesthesia, a 2 cmx 2 cm full-thickness skin defect reaching deep fascia was reproduced in the middle of the back of each nude mouse, and a split-thickness skin graft of the same size was obtained, and then prepared skin grafts were transplan- ted to cover the wounds respectively. On post transplantation day （PTD） 5, 7, and 14, local condition and adverse effects observation was conducted; one nude mouse was sacrificed each time to harvest all the trans- plant for tissue structure observation with HE staining. On PTD 7 and 14, neonatal skin appendages in corre- sponding composite materials were observed with transmissio

关 键 词：伤口愈合 皮肤 真皮 再生 骨髓间充质干细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]