Harriger M D, Supp A P, Warden G D, Holder I A
Department of Surgery, University of Cincinnati, Cincinnati, OH 45229, USA.
Wound Repair Regen. 1997 Apr-Jun;5(2):191-7. doi: 10.1046/j.1524-475X.1997.50212.x.
Cultured skin substitutes have become therapeutic alternatives for treatment of acute and chronic skin wounds, but all models of these substitutes are avascular and susceptible to microbial destruction during vascularization. To develop a practical management protocol for increased survival of skin substitutes, experimental wounds were contaminated with Pseudomonas aeruginosa and treated with a formulation of noncytotoxic antimicrobial agents (polymyxin B, neomycin, ciprofloxacin, mupirocin, amphotericin B) in a nutrient medium (vehicle). Cultured skin substitutes consisting of human keratinocytes and fibroblasts attached to collagen-glycosaminoglycan sponges were grafted to 2 x 2 cm full-thickness wounds on athymic mice that were contaminated with the strain SBI-N of P. aeruginosa at 1 x 10(4), 1 x 10(5), and 1 x 10(6) organisms/wound. Experimental wounds were irrigated with 1 ml/day topical antimicrobial solution for 10 days, and controls received vehicle only. Two, three, and four weeks after grafting, wounds were traced and swabbed for microbial culture and areas were measured with planimetry. At 4 weeks, biopsy samples were scored histochemically for immunoreactivity to HLA antigens. Data analysis by chi-square, analysis of variance, and Tukey's test shows that treatment of contaminated wounds with noncytotoxic topical antimicrobials is associated with an increased area of healed wounds, positive detection of HLA antigens, and negative cultures for P. aeruginosa. These results show that microbial contamination of cultured skin substitutes on full-thickness wounds may be managed effectively during graft vascularization. However, this formulation of antimicrobial agents is not currently approved for human use and is investigational only. Effective management of microbial contamination suggests that clinical efficacy of avascular tissue analogs may be increased by local application of noncytotoxic antimicrobial agents.
培养的皮肤替代物已成为治疗急慢性皮肤伤口的治疗选择，但这些替代物的所有模型都是无血管的，并且在血管化过程中易受微生物破坏。为了制定提高皮肤替代物存活率的实用管理方案，将实验伤口用铜绿假单胞菌污染，并用营养培养基（载体）中的一种无细胞毒性抗菌剂（多粘菌素B、新霉素、环丙沙星、莫匹罗星、两性霉素B）制剂进行处理。将由附着在胶原-糖胺聚糖海绵上的人角质形成细胞和成纤维细胞组成的培养皮肤替代物移植到无胸腺小鼠2×2cm的全层伤口上，这些伤口分别用1×10⁴、1×10⁵和1×10⁶个铜绿假单胞菌SBI-N菌株污染。实验伤口每天用1ml局部抗菌溶液冲洗10天，对照组仅接受载体。移植后2、3和4周，追踪伤口并擦拭进行微生物培养，并用平面测量法测量面积。在4周时，对活检样本进行组织化学评分，以检测对HLA抗原的免疫反应性。通过卡方检验、方差分析和Tukey检验进行数据分析表明，用无细胞毒性局部抗菌剂治疗污染伤口与愈合伤口面积增加、HLA抗原阳性检测以及铜绿假单胞菌培养阴性相关。这些结果表明，在移植物血管化过程中，可以有效管理全层伤口上培养皮肤替代物的微生物污染。然而，这种抗菌剂制剂目前尚未被批准用于人类，仅用于研究。微生物污染的有效管理表明，通过局部应用无细胞毒性抗菌剂可以提高无血管组织类似物的临床疗效。
