医药学论文：脱细胞真皮基质ADM制备前后的变化

【摘要】 目的 探讨脱细胞真皮基质制备前后出现的各种变化。方法 将人全厚皮切制成厚0.5mm的中厚皮片，先后经1M NaCl溶液、0.1％戊二醛、0.5％SDS、0.25％胰蛋白酶、冷冻干燥及水化处理，制得脱细胞真皮基质（ADM）。观察制备前后出现的各种变化。结果 皮片经处理后，ADM具有良好的弹性和柔韧性。皮片中所有细胞成分消失，胶原及弹力纤维结构完整、排列规则，基底膜存在。处理前后弹力/胶原纤维面密度分别为0.25±0.08和0.26±0.08、胶原含量分别为69.18±12.51和85.17±28.93、Ⅲ/Ⅰ型胶原比值分别为0.85±0.22和0.71±0.14，均无明显变化。结论 脱细胞真皮基质制备过程可有效去除皮肤中的细胞成分，完整保留真皮基质和基底膜，形成相对无免疫活性的脱细胞真皮基质。

【关键词】 脱细胞真皮基质 制备

Studies on the changes during preparation of acellular dermal matrix (ADM)

【Abstract】 Objective To investigate the changes during preparation of ADM.Methods Split-thickness allograft， 0.5mm thick， were obtained from full thickness human's skins with a dermatome. Donor skins were treated with 1 M NaCL， 0.1% glutaraldehyde， 0.5 % SDS and 0.25 % trypsin successively. The resulting acellular dermal matrices were then freeze-dried in a freeze-dryer and hydrated in PBS. The processing technique resulted in acellular dermal matrices (ADM). The dynamic changes during processing were researched.Results All the cellular components in the donor skins were removed and basement membrane was retained. Regular orientation and integral framework of collagen bundles and elastin was present in elastic and flexible ADM. The area-density of elastin/ collagen， gross and Ⅲ/Ⅰ type ratio of collagen were 0.25±0.08， 69.18±12.51， 0.85±0.22 in the donor skins and 0.26±0.08， 85.17±28.93， 0.71±0.14 in the ADM respectively. There was no change after donor skins were treated.Conclusion The processing technique removed all the cellular components from the donor skins， retained dermal matrix and basement membrane， and acquired acellular dermal matrix that exhibited no immunocompetence.

【Key words】 acellular dermal matrix preparation

脱细胞真皮基质 (acellular dermal matrix， ADM) 是一种良好的基质，可作为真皮替代品与自体表皮复合移植，亦可作为软组织填充材料单独使用，临床上应用愈来愈广泛。最早由Liversey研制成功，并申请了美国专利［1］；国内报道的制备方法差别较大，对此我们进行了研究，移植后亦获得了成功。现就其制备过程中出现的各种变化报道如下。

1 材料与方法

1.1 异体皮肤来源 异体皮肤来源于我院整形手术患者，术前查体正常，免疫学检查HBsAg、HCV、HIV、TP-Anti均为阴性。切下的健康皮肤制成厚约0.5mm的中厚皮片。

1.2 实验条件 整个实验过程均需在严格无菌条件下进行。

1.3 实验步骤 中厚皮片入1M NaCl溶液，37℃，浸泡24h→0.1%戍二醛缓冲液，37℃，浸泡5min→0.5%SDS溶液，37℃，浸泡2h→0.25%胰蛋白酶溶液，消化20min→大量的PBS溶液反复多次漂洗，至漂洗液无泡沫为止→冷冻干燥机，-60℃，ATM 56，持续12h→密封保存。

1.4 ADM的水化 室温下冷冻干燥的ADM置于PBS溶液中浸泡30min以上，即可使用。

2 结果

2.1 大体观察 皮片经高渗盐水处理后，表皮易于从真皮上呈片状完整剥离，有时在表皮和真皮之间形成水泡样间隙，毛发与表皮亦一起脱落。真皮呈瓷白色，富有弹性，形成基底膜和真皮两个面，其中基底膜面有光泽，比真皮面光滑（图1～4）。