CAJ | 학술논문

目的研究荷载角质细胞生长因子(keratinocyte growth factor,KGF)纳米微囊的新型组织工程皮肤对裸鼠皮肤缺损的修复效果及特点.方法采用超声乳化一溶剂挥发法及低温干燥法,制备KGF纳米微囊,并构建KGF-脱细胞真皮基质(acellular dermal matrix,ADM);分离培养和鉴定人表皮干细胞群和成纤维细胞;接种表皮干细胞群于KGF-ADM之上,观察其生长情况;将荷载KGF纳米微囊的组织工程皮肤移植于裸鼠皮肤缺损处,以无KGF纳米微囊的组织工程皮肤为空白组,以其自体皮肤移植作对照组.于术后2,6周时分别观察修复区组织学愈合及皮片挛缩情况,并应用抗人角蛋白10及β1-整合素免疫荧光检测修复区表皮和真皮层细胞来源、分化及生长情况.结果表皮干细胞群在KGF-ADM表面生长良好,粘贴紧密,可见到多角形的终末表皮细胞及小圆形的表皮干细胞,活性良好,有连接成片的趋势,部分形成克隆团块.以荷载KGF纳米微囊组织工程皮肤修复裸鼠皮肤缺损,2、6周时修复效果均优于空白组及对照组,移植的组织工程皮肤边缘可与邻近皮肤完全融合,但存在一定的挛缩.镜下可见修复区组织工程皮肤表皮细胞分层良好,与ADM紧密结合,能产生正常角质层.6周时实验组修复区组织工程皮肤切片免疫荧光检测,基底层仍存有少量β1-整合素阳性的表皮干细胞或短暂扩充细胞.结论所构建的荷载KGF纳米微囊组织工程皮肤修复裸鼠皮肤缺损的效果,优于无KGF纳米微囊的普通组织工程皮肤及裸鼠自体全厚皮片移植修复效果.
목적 연구하재각질세포생장인자(keratinocyte growth factor,KGF)납미미낭적신형조직공정피부대라서피부결손적수복효과급특점.방법 채용초성유화일용제휘발법급저온간조법,제비KGF납미미낭,병구건KGF-탈세포진피기질(acellular dermal matrix,ADM);분리배양화감정인표피간세포군화성섬유세포;접충표피간세포군우KGF-ADM지상,관찰기생장정황;장하재KGF납미미낭적조직공정피부이식우라서피부결손처,이무KGF납미미낭적조직공정피부위공백조,이기자체피부이식작대조조.우술후2,6주시분별관찰수복구조직학유합급피편련축정황,병응용항인각단백10급β1-정합소면역형광검측수복구표피화진피층세포래원、분화급생장정황.결과 표피간세포군재KGF-ADM표면생장량호,점첩긴밀,가견도다각형적종말표피세포급소원형적표피간세포,활성량호,유련접성편적추세,부분형성극륭단괴.이하재KGF납미미낭조직공정피부수복라서피부결손,2、6주시수복효과균우우공백조급대조조,이식적조직공정피부변연가여린근피부완전융합,단존재일정적련축.경하가견수복구조직공정피부표피세포분층량호,여ADM긴밀결합,능산생정상각질층.6주시실험조수복구조직공정피부절편면역형광검측,기저층잉존유소량β1-정합소양성적표피간세포혹단잠확충세포.결론 소구건적하재KGF납미미낭조직공정피부수복라서피부결손적효과,우우무KGF납미미낭적보통조직공정피부급라서자체전후피편이식수복효과.
Objective To study the effect of tissue-engineered skin loaded with keratinocyte growth factor (KGF) nanocapsules for skin defect on athymic mice. Methods The acellular dermal matrix(ADM) loaded with KGF-ADM was constructed by means of phaceemulsification solvent evaporation and low temperature drying. The human epidermal stem cells and fibroblasts were captured and identified, then cultivated on the surface of the KGF-ADM. The cell growth was observed. The tissue-engineered skin without KGF was used as sham group. The autogenous skin graft was used as control group. 2 and 6 weeks after the skin was transplanted to the back of athymic mice, the contraction and histological healing of the transplanted skins were observed respectively. Then the immunofluorescence examination with anti-human KI0-FITC and β1-integrin-Cy3 were applied to detect the origin, growth and differentiation of epidermal and dermal ceils in tissue-engineered skin. Results The epidermal stem ceils grew well and attached tightly on KGF-ADM. There were small round stem cells and polygonal terminally-dlfferentiated cells, which appeared a partly cloning growth and a tendency of merging. The tissue-engineerred skin with KGF nanocapsules gained better result in repairing the skin defects as compared with the blank group and the control group 2 and 6 weeks after transplantation. The regenerative skin cells could connect and mix closely with the athymic mouse skin cells on the border of skin defect. Meanwhile, the regenerative skin existed some contraction. The histological observation with HE staining showed that the regenerative skin possessed intact epidermis with several cell layers and normal keratose stratum, among which there were still some β1-integrin(+) cells which represented epidermal stem cells or transient amplifying cells when they were tested by immunofluoreseence after 6 weeks of transplantation. Conclusions The tissue-engineered skin loaded with KGF naneeepsules had a better result in repairing athymic mice skin defects than common tissue-engineered skin without KGF nanocapsules or skin auto-graft.