【作者】 梁志为；

【导师】 谭谦；

【作者基本信息】 东南大学 ， 外科学， 2004， 硕士

【摘要】 【目的】体外构建一种含有角质形成细胞和血小板衍化生长因子(PDGF)基因转染的成纤维细胞的复合皮肤,并将此种复合皮移植到动物皮肤缺损模型,观察创面愈合的效果。【方法】在体外利用已制备的无细胞异种(猪)真皮基质、培养的角质形成细胞和PDGF基因转染的成纤维细胞构建三种类型复合皮:A 型,培养的角质形成细胞+无细胞猪真皮基质(CK+AX);B型,培养的角质形成细胞+无细胞猪真皮基质+培养的成纤维细胞(CK+AX+Fb);C型,培养的角质形成细胞+无细胞猪真皮基质+PDGF基因转染的成纤维细胞(CK+AX+PFb)。通过相差显微镜观察、组织切片HE染色等方法进行检测。分别将三型复合皮分成A、B、C三组,进行动物移植实验,观察皮片存活率、创面收缩率、真皮血管化程度等,并与单纯角质形成细胞膜片移植组(D组)和用凡士林纱布覆盖的对照组(E组)进行比较。【结果】体外培养后2w, C型复合皮HE染色显示有4层以上的角质形成细胞,有基底膜形成,并有轻度角质化。移植术后2w, 与其他各组相比,B、C两组成活率高,复合皮与创面边缘融合较好,与基底贴附紧密,尤以C组情况好;E组呈结痂愈合。移植后6w,C组创面愈合好,表面光滑,弹性较好,抗摩擦性强,收缩不明显。D组愈合后成活不稳定,皮片薄,质脆,表面不光滑,抗摩擦性差,有部分创面愈合后再度形成创面,各组均无毛发生长。创面收缩率:D、E两组收缩最为明显,A、B组次之,C组收缩最少(P<0.05)。存活率:除C、D两组间差异显著外(P<0.05),余各组间存活率无显著性差异。但与其他各组相比,C组复合皮完全存活数较多,未存活数少。血管化情况:移植后2w,C组真皮浅层内可见较多新生毛细血管长入,B组次之,A 组毛细血管长入较少(P<0.05);移植后4w各组真皮浅层内毛细血管数逐渐增加,但C组毛细血管数仍明显高于A、B两组;移植后6w,各复合皮内毛细血管数差异不显著(P﹥0.05)。【结论】用角质形成细胞和PDGF基因传染的成纤维细胞在无细胞异种?(猪)真皮上培养来构建复合皮,经组织学检查、动物移植实验,证实构建的复合皮具有良好的结构,能够迅速血管化,创面修复后具有良好的形态。更多还原

【Abstract】 【Objective】 To establish a composite skin with combining cultured keratinocytes, acellular pig dermis and fibroblasts transfected with PDGF gene in vitro and to observe the healing effects by transplanting the composite skin to the wounds of rat back. 【Methods】 Three types of composite skins were reconstructed by using cultured keratinocytes(CK), acellular xenodermis(AX) and the fibroblasts(Fb) with PDGF gene(PFb): Type A (CK+AX), type B (CK+AX+Fb) and type C (CK+AX+PFb). The growth states of keratinocytes and fibroblasts were observed under phase contrast microscope. The composite skins were detected by histological examination (HE staining). Three types of composite skins which were considered as three groups of A, B and C were transplanted to the wounds on rat back. The wounds of group A, B and C were compared with the wounds transplanted with cultured keratinocyte membrane (group D) and the wounds covered with Vaseline (group E) in skin takes, contraction of the wounds and the vescularization in acellular pig dermis after transplantation.【Results】 More than four-layer keratinocytes, the basement membrane and slight keratinization were observed after histological HE staining. The skin take rates of group B and group C in 2 weeks after skin grafting were much higher that those in any other groups. In 6 weeks the healed wounds in group C were smooth, elastic and wearable. The wound contraction rates in group D and group E were most obvious comparing with any other groups(P﹤0.05). The take rates had significant differences between group C and group D, and had no difference among other groups. In 2～4 weeks after skin grafting the vescularization rate in group C was higher than that of group B and group A(P﹤0.05). The vescularization rates in all groups had no significant differences in 6 weeks(P﹥0.05). 【Conclusions】 The composite skins are constructed with cultured keratinocytes and fibroblasts with PDGF gene on acellular xenodermis. It is concluded that the composite skins we had established have good structure and function and play an important role in wound healing.更多还原