激光微孔化猪脱细胞真皮基质的设计、制备及同步移植实验

Design,preparation and synchronous transplantation experiments of laser micropore porcine acellular dermal matrix

目的构建一种新型真皮替代物——激光微孔化猪脱细胞真皮基质材料(LPADM)并验证其修复全层皮肤缺损的效果。方法取新鲜健康小白猪断层真皮,一部分经高钠-SDS法脱细胞获得脱细胞真皮基质(ADM),另一部分则先行激光打孔再经脱细胞获得LPADM;检测LPADM的物理性状。在SD大鼠(n=30)背部制作2.0 cm×2.0 cm的全层皮肤缺损创面(n=4),分别采用LPADM自体中厚皮片(L组)和无孔ADM自体中厚皮片(A组)复合移植及自体全厚皮片(F组)和中厚皮片原位移植(C组)修复。分别于术后第3、5、7、10、14天,每组随机选取6只大鼠,观察创面皮片成活情况,处死动物后采集创面组织行组织学检查。结果制备的LPADM呈瓷白色,柔软有光泽;显微镜下观察未见细胞残留。移植术后第3天,组织学检查发现L组创面组织有内皮细胞围成的空腔样结构;术后第3、5、7、10天,L组和C组创面移植皮片成活良好,L组血管样结构中已可见红细胞;A组皮片起泡甚至发黑坏死;F组皮片部分起小泡,但泡皮下基底红润。术后第14天,L组真皮微孔中可见大量血管形成,L组、C组和F组移植皮片全部或基本存活,创面愈合率分别为(99.10±0.66)%、(99.25±0.23)%、(97.07±1.32)%,差异无统计学意义(P>0.05);A组皮片全部坏死脱落,创面愈合率为(27.46±2.05)%,显著低于其他各组(P<0.01)。结论构建的新型激光微孔化猪脱细胞真皮基质,同步复合自体皮移植,移植成功率高,较快实现早期血管化,为一种较理想的真皮替代物。

Objective To prepare laser micropore porcine acellular dermal matrix（LPADM）,a new type of dermal substitute,and to verify the healing effect on full-thickness skin defect.Methods Split-thickness dermis of the healthy white pigs was harvested and treated by high-sodium-SDS method,and the porcine acellular dermal matrix（ADM） was obtained.Besides,LPADM was prepared using laser micropore technology on the dermis,followed by the procedure of high-sodium-SDS acellular treatment.The physical properties of LPADM were evaluated.Four full-thickness skin wounds（2.0 cm×2.0cm） were created on each dorsum of 30 SD rats.Group L（LPADM grafted with the split-thickness autograft）,group A（nonporous ADM grafted with the split-thickness autograft）,group F（full-thickness autograft） and group C（split-thickness autograft） were divided on the basis of different grafts used through one-step method.Six rats were randomly selected on day 3,5,7,10 and 14 after surgery,the survival of skin graft was observed,and histological examination was conducted after sacrifice.Results The well-prepared micropore LPADM was characterized by its porcelain white color,softness and good elasticity.Histological examination revealed that LPADM was totally devoid of epidermis and cellular components.On day 3 after surgery,the cavity-like structure formed by surrounding endothelial cells was observed in group L.On day 3,5,7 and 10 after surgery,the grafts of group L and C survived well,and new vessels were generated in group L.However,epidermal necrosis with black and blistering tissues occurred in group A,while some grafts blistering with ruddy epidermal basement were observed in group F.On day 14 after surgery,abundant blood vessels formed in group L,and all the grafts survived in group L,C and F,with the wound healing rates of（99.10±0.66）%,（99.25±0.23）%,and（97.07±1.32）% respectively,and there was no significant difference among groups（P0.05）.The wound healing rate of group A was（27.46±2.05）%,which was significantly lower than those of the other groups（P0.01）.Conclusion The new type of micropore porcine acellular dermis matrix can be used as an ideal dermal substitute due to high survival rate when grafted with the split-thickness autograft and the ability of initiating early revascularization.