摘要
目的 观察成纤维细胞与人工真皮共同移植对移植床及真皮海绵内微血管形成、胶原沉积的影响 ,促进人工真皮海绵内类真皮组织形成 ,缩短“二期植皮”的间隔时间。方法 Wistar大鼠 18只 ,背部两侧各制作 2 .5 cm×2 .5 cm全层皮肤缺损创面 ,在移植人工真皮之前分为两组 ,即成纤维细胞移植组 (A组 ) :向创面喷洒混入密度为 1.0×10 5/cm2 的同种真皮成纤维细胞的纤维蛋白胶 0 .5 ml;对照组 (B组 ) :只喷洒纤维蛋白胶 0 .5 ml。于移植后 5和 10天分别切取移植的真皮及周围组织 ,进行 HE染色、Masson染色、VEGF免疫抗体染色和墨汁灌注染色 ,观察移植床及真皮海绵内微血管新生及胶原沉积状态 ;于移植后 5天行伊文蓝组织灌注 ,分光光度计测定真皮海绵内染料含量 ,定量检测真皮海绵内循环渗透情况。 结果 移植后 5天 ,新生血管主要集中在真皮下移植床 ,A组新生血管较 B组明显增多 ,A组 (9.6 4± 2 .36 )个 /高倍视野 ,B组 (3.88± 1.6 2 )个 /高倍视野 ,差异有统计学意义 (P<0 .0 5 ) ;VEGF阳性细胞亦明显增多。移植后 10天 ,移植床及真皮内均有微血管形成 ,A组血管数量和管径均较 B组增加 ,胶原沉积量也明显增加。A组(46 .0 4± 8.90 )个 /高倍视野 ,B组 (30 .0 8± 7.76 )个 /高倍视野 ,有统计学意义
Objective To evaluate the effects of cryopreserved cultured allogenic dermal fibroblasts on angiogenesis and fibroplasia while artificial dermis grafting by spraying the cells on the graft bed.Methods Full thickness skin defect was made on the back of Wistar rat, fibroblasts mixed into fibrin glue (fibroblast group) and same amount fibrin glue (control group) were sprayed separately between the wound bed and artificial dermis in cell density of 1.0×10 5 cells/cm 2 before the artificial dermis was grafted. On day 5 after grafting, the graft and surrounding tissue were examined histologically for angiogenesis and fibroplasia in the dermis and wound bed with hematoxylin eosin stain, VEGF antibody stain, Masson's trichrome stain and India ink stain. Evans blue perfusion method was also used for detecting the angiogenesis quantitatively.Results In the fibroblast group, the angiogenesis of graft bed was significantly accelerated on day 5 after grafting; the numbers of the newly formed capillaries were 9.64±2.36/HP in the fibroblast group and 3.88±1.62/HP in the control group (P<0.05). And on day 10 after grafting the angiogenesis was accelerated not only in graft bed but also in the artificial dermis when compared with control group, the newly formed capillaries network was clearly observed in the artificial dermis. Otherwise, the synthesis of collagen was increased in the dermis on day 10 after grafting in the fibroblast group when compared with control group. The immunoreactivity of VEGF antibody in the fibroblast group also showed a stronger expression than that in control group on day 5 after grafting, the numbers of positive cells were 46.04±8.90/HP in the fibroblast group and 30.08±7.76/HP in the control group(P<0.05).Conclusion Transplantation of cryopreserved dermal fibroblasts while artificial dermis grafting can accelerate the angiogenesis and fibroplasia in the artificial dermis and graft bed, thereby accelerate the formation of dermal-like tissue in the artificial dermis.
出处
《中国修复重建外科杂志》
CAS
CSCD
2004年第3期205-208,共4页
Chinese Journal of Reparative and Reconstructive Surgery
