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	反复冻融配合超声振荡洗涤制备猪脱细胞真皮基质
	张国安 ; 宁方刚 ; 钟京鸣 ; 于东宁 ; 赵南明 ; Zhang GA ; Ning FG ; Zhong JM ; Yu DN ; Zhao NM
	2010-06-09 ; 2010-06-09
关键词	脱细胞真皮基质 制备 R318.0
其他题名	Swine acellular dermal matrix prepared by repeated freezing and thawing combined with ultrasonic surge
中文摘要	目的:改进猪脱细胞真皮基质制备方法,并与传统制备方法进行比较,为临床大面积烧伤患者治疗提供更安全、更廉价的异体皮源。方法:实验于2003-10/2007-03在北京积水潭医院烧伤研究所完成。①实验材料:实验动物为3月龄清洁中华香猪,由北京积水潭医院动物室提供。实验符合动物伦理学标准。②实验方法:取25kg实验室环境养殖的健康中华香猪,麻醉处死去毛后取躯干部位全层皮肤,仔细剃除毛发,去除脂肪。采用反复冻融,配合超声振荡和化学去污剂处理的复合方法,获得脱细胞猪真皮基质。③实验评估:利用常规病理检查和电子显微镜检查其有无明显细胞碎片残留,测试其机械强度、亲水性和细胞毒性,通过动物埋藏实验确定其组织相容性,并以戊二醛交联的脱细胞异体皮,戊二醛交联的脱细胞猪皮,以及新鲜人皮作对照,结果:①常规病理检查可见真皮结构完整,无明显破坏;上皮细胞清除彻底,无明显细胞碎片残留,电子显微镜检查见脱细胞猪皮的胶原结构完整,未发现破坏痕迹,无明显细胞残留。②各组中脱细胞猪皮基质的极限应力强度和新鲜人皮接近,无明显统计学差别(P>0.05);戊二醛交联的脱细胞猪皮强度最低,戊二醛交联的脱细胞异体皮的强度最高,且与新鲜人皮相比也有明显统计学差别(P<0.01)。③戊二醛交联的脱细胞异体皮亲水性最强,其余三者相似。④脱细胞猪皮基质浸提液中细胞生长旺盛,7d后脱细胞猪皮基质两次测试相对增殖度值分别为:182,169。按照6级分级标准,脱细胞猪皮基质毒性级别为0级,在国家标准允许范围内。⑤脱细胞猪皮基质植入1,3周,边界不清,炎性反应与吸收均不明显,新生血管与成纤维细胞长入,植入12周,植入物与组织融为一体,新生血管与成纤维细胞填充真皮支架。结论:所制备的猪脱细胞真皮基质产品真皮基质完整,细胞毒性低,物理性能与人皮接近。; AIM:The objective of the study was to improve the preparative method of swine acellular dermal matrix(SADM) ,compare with the traditional methods,and provide a safe,low-cost production from an abundant source of homogeneous skin in the treatment of large area burn patients.METHODS:The experiment was performed at Burn Institute of Ji Shui Tan Hospital from October 2003 to March 2007.①Chinese fragrant pigs aged 3 months of clean grade were selected from Animal Room of Ji Shui Tan Hospital.The experiment was approved by animal ethical standard.②Healthy Chinese fragrant pigs of 25 kg were killed after narcosis,shaved and fully peeled.The subcutaneous fat was removed.SADM was obtained by repeated freezing and thawing combined with ultrasonic surge and chemical subtraction.③The cell fragments and structure were tested by routine pathological method and electron microscope.The tensile strength,hydrophilicity and cytotoxicity were determined.Histocompatibility was measured by animal implantation experiment.Tests were compared with acellular human skin composite with glutaraldehyde,acellular swine skin composite with glutaraldehyde and fresh human skin.RESULTS:①Integral dermis was seen by routine pathological method.Endothelial cells were removed fully,and collagen structure of the SADM was integrity and no cell fragment was found under electron microscope.②Ultimate stress of SADM was similar to fresh human skin(P > 0.05) .The stress of acellular swine skin composite with glutaraldehyde was the lowest,while the stress of acellular human skin composite with glutaraldehyde was the highest,and significant differences were found compared with that of fresh human skin(P < 0.01) .③The hydrophilicity of acellular human skin composite with glutaraldehyde was the highest.Others were similar in hydrophilicity.④Cells grew productively in acellular swine matrix leaching liquor.Seven days later,proliferation rates of two measures were 182 and 169 respectively.According to 6-grading standards,cytotoxicity of acellular swine matrix was 0 grade,which was in the range of national standard.⑤Obscure boundary,insignificant inflammatory reaction and insignificant resorption were found at weeks 1 and 3 after acellular swine matrix implantation,and new vessels and fibroblasts appeared.At week 12,implant and tissues fused together,and dermal stent was filled with new vessels and fibroblasts.CONCLUSION:This SADM production is characterized by integral dermal matrix,low cytotoxicity and similar physical function as human skin.; 清华-裕元医学科学研究基金支持项目~~
语种	中文 ; 中文
内容类型	期刊论文
源URL	[http://hdl.handle.net/123456789/58324]
专题	清华大学
推荐引用方式 GB/T 7714	张国安,宁方刚,钟京鸣,等. 反复冻融配合超声振荡洗涤制备猪脱细胞真皮基质[J],2010, 2010.
APA	张国安.,宁方刚.,钟京鸣.,于东宁.,赵南明.,...&Zhao NM.(2010).反复冻融配合超声振荡洗涤制备猪脱细胞真皮基质..
MLA	张国安,et al."反复冻融配合超声振荡洗涤制备猪脱细胞真皮基质".(2010).

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