肝素化羊脱细胞真皮基质对大鼠深Ⅱ度创面愈合的影响

EFFECTS OF HEPARINIZED SHEEP ACELLULAR DERMIS MATRIX ON WOUND HEALING OF DEEP PARTIAL-THICKNESS BURN IN RATS

目的制备肝素化羊脱细胞真皮基质(ADM),观察其对大鼠深Ⅱ度烫伤创面愈合的影响。方法①制成异种(羊)ADM,再用低分子肝钠素注射液浸泡处理,合成肝素化羊ADM。对肝素化羊ADM进行组织学观察,测定吸附量,进行细菌、真菌培养。②96只Wistar大鼠制作背部深Ⅱ度烫伤模型,随机分为4组,A组(n=24)肝素化羊ADM覆盖创面,B组(n=24)异种(羊)ADM覆盖创面,C组(n=24)肝素钠乳膏涂沫创面,D组(n=24)为对照组。测算3,7,14,21d创面愈合率,于7,14d光镜观察创面组织学变化,于14d电镜观察成纤维细胞结构。观察各组创面、粪、尿有无出血现象。结果①肝素化羊ADM质感柔软有弹性,不易断裂。含有低分子肝素钠(4047.50±497.49)IU/g。光镜检查、电镜检查未见表皮和真皮细胞残留,胶原结构基本完整。细菌和真菌培养呈阴性。②伤后14,21d,A组创面愈合率优于B,C,D组(P<0.05)。伤后14d光镜观察,A组与B,C,D组比较表皮组织、肉芽组织、胶原纤维明显增多。伤后14d电镜观察,A组与B,C,D组比较成纤维细胞细胞器更丰富。各组动物创面、粪便及尿液均无出血现象。结论肝素化羊ADM能够促进大鼠深Ⅱ度创面愈合,适合用作生物敷料。

Objective To prepare the heparinized sheep acellular dermal matrix and investigate its effects on wound healing of deep partial-thickness burn in rats.Methods ①Sheep acellular dermal matrix was fabricated and immersed in heparin sodium injection to prepare the heparinized sheep acellular dermal.The histological characteristics were tested with optical microscope and electron microscope.Bacteria and fungal culture were tested.The heparin content was also calculated.②96 Wistar rats were subjected to deep partial thickness scalding and were randomly divided into A group（n=24,with heparinized sheepskin acellular dermal matrix on the wounds）,B group （n=24,with sheep acellular dermal matrix on the wounds）,C group （n=24,with heparin sodium cream on the wounds）,and D group （n=24,with gauze on the wounds as control）.The healing rates of the traumatic wounds were compared at 3,7,14 and 21 postburn days（PBD）.The growth of epidermis,granulation tissue and collagen fibers were evaluated under light microscope on 7 and 14 PBD.The growth of fibroblast was observed under transmission electron microscope on 14 PBD.To find out whether bleeding side effects were occurring,the traumatic wounds,fecal and urine were observed.Results ①The heparinized sheep acellular dermal matrix was soft,extensibility and undisrupted solid with hydrophilic quality.No cell fragment was found by routine histological methods and electron microscope.4047.50±497.49IU heparin sodium injection were contained in per gramme heparinized sheep acellular dermal matrix.Bacteria and fungal culture were negative.②The wound healing rate on 14 and 21 PBD in A group were higher when compared with that in B,C,D groups （P〈0.05）.Light microscope observation showed that the growth of epidermis,granulation tissue of A group were obviously better than those of B,C,D groups on 14 PBD.Transmission electron microscope observation showed that the fibroblast of the A group was better than those of B,C,D groups on 14 PBD.No bleeding side effects were observed.Conclusion The heparinized sheep acellular dermal matrix could significantly promote wound healing of deep partial thickness burn on Wistar rat.It meets the requirements for biological dressing.