巴马香猪皮肤比较生物学特性及其无细胞真皮的研究
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摘要
为寻找人类皮肤研究合适的动物模型,科学家们研究了多种动物的皮肤,发现猪的皮肤在解剖、生理等方面与人类皮肤最为接近。但由于普通商品猪遗传背景不清晰,个体差异大,生长速度快,不适于用作实验动物。巴马香猪是我国独特的小型猪种之一,体型小,操作容易,遗传稳定,个体差异小,是一种理想的实验动物。但其缺乏与人的比较医学生物学系统基础资料,制约了巴马香猪在医学生物学中的应用。
无细胞真皮作为一种有效的临床用生物材料,具有重要的临床应用价值。但国内目前仅有人无细胞真皮产品面世,且价格昂贵。随着国家人体器官移植技术临床应用管理暂行规定的正式实施,人组织器官的应用将面临来源、伦理、法律等一系列问题,而且还具有传染HIV、HBV等血液传播性疾病的危险。因此,以标准化实验用小型猪无细胞真皮替代人无细胞真皮具有良好的应用前景。
为了推广巴马香猪在医学生物学中的应用,评估巴马香猪作为人皮肤模型动物在创伤修复领域的应用价值,同时为了开发巴马香猪皮肤来源的异种移植生物材料,本课题从比较医学的角度,对巴马香猪与人皮肤对比进行了组织学结构、免疫组化反应特点、超微结构、皮肤胶原蛋白理化特性等的系统研究;同时,制备了巴马香猪无细胞真皮,与人无细胞真皮对比进行了安全性有效性评价,以期利用巴马香猪无细胞真皮替代人无细胞真皮而应用于创伤修复领域,为其产品研发形成基础,同时也进一步比较巴马香猪皮肤与人皮肤在应用特性方面的相似性。全文主要结果及结论如下:
1.本课题对巴马香猪皮肤的组织学结构、免疫组化反应特点、超微结构、胶原蛋白理化特性等进行了系统研究,发现巴马香猪的皮肤结构除汗腺结构与人不同外,其它结构及其特点与人极为相似。其4月龄小型猪皮肤发育程度最接近于成人皮肤,各层厚度及结构都与成人皮肤相似。上述研究结果表明,巴马香猪可作为研究人类皮肤的模型动物。
巴马香猪在皮肤厚度、分层、真表皮的高低起伏的连接方式、发达的浅表血管系统以及皮肤的超微结构都与人类皮肤相似;皮肤中所含细胞种类和成纤维细胞、血管内皮朗格罕细胞、肥大细胞的密度也与人类皮肤相近;具有与人类Ⅰ型胶原、Ⅲ型胶原、层粘连蛋白、纤维连接蛋白、中间丝相关蛋白相似的抗原决定簇,可与抗人血清发生交叉反应,且分布特点与人类皮肤相似;真皮中主要的胞外基质构成成分-Ⅰ型胶原的结构及理化特性也几乎与人类Ⅰ型胶原相同。但巴马香猪皮肤中的汗腺为顶泌汗腺,而人类皮肤中主要为外泌汗腺;另外,巴马香猪皮肤中的色素细胞在身体的不同部位差异很大;巴马香猪皮肤缺乏与抗人CD34、ICAM-1、S-100等抗体对应的抗原决定簇。
2.优化了无细胞真皮的制备方法,制备了巴马香猪无细胞真皮,对其与人无细胞真皮对比进行了生物学特性与安全性有效性评价研究。对巴马香猪与人无细胞真皮对比进行了胞外基质的种类和分布、微生物学检测、致敏实验、淋巴细胞增殖实验、细胞毒性实验、组织学相容性实验、单纯无细胞真皮移植实验以及结合大鼠自体皮的复合移植实验等实验研究。实验结果表明,巴马香猪无细胞真皮与人无细胞真皮的形态结构、抗原性、生物相容性、血管化速度及动物实验效果无显著差异,显示巴马香猪无细胞真皮可望替代人无细胞真皮应用于创伤修复领域。
经0.25%胰酶4℃处理过夜,0.5%Triton X-100室温振荡处理12h制备而成的巴马香猪无细胞真皮,具有如下特点:成功去除了引起强烈免疫排斥反应的表皮和细胞成分,完整保留了基底膜,与抗原性有关的胞外基质结蛋白和波形蛋白也得到有效的去除;保留了Ⅰ型胶原、Ⅲ型胶原、Ⅳ型胶原、纤维连接蛋白等与细胞粘附、生长相关的胞外基质;具有与人无细胞真皮相似的无菌、无致敏反应、生物相容性好、抗原性低、血管化速度快等优良的生物学特性;将其与大鼠自体刃厚皮复合植入大鼠创面后,巴马香猪无细胞真皮与人无细胞真皮组的移植皮片存活率、创面收缩率相似,且与大鼠同种无细胞真皮组的移植皮片存活率和创面收缩率无明显差异,未引发明显的免疫排斥反应。表明巴马香猪无细胞真皮可望替代人无细胞真皮应用于创伤修复领域。
The skin of domestic animals has been investigated extensively in order to find a suitable experimental model to be used in research studies on human skin.Of all the animals examined pig skin is more similar to human skin anatomically and physiologically.But the domestic pig is not suitable for the biomedicine research for its unsharp genetic background,great individual difference and fast growth velocity.Bama miniature pig,one distinct breed of miniature pigs in China,is an ideal animal model for its high inbreeding degree,genetic stability,small body size.However basic research data of Bama miniature pig is absent,so its application in biomedicine is limited.
Acellular dermal matrix used as dermal substitute provides to splite-thickness skin grafts and to cultured epidermal autografts.A thick demal bed at the recepient site inhibits and hypertrophic scar formation of split-tickness skin grafts.The clinic applicationable product of acellular dermal matrix was only originated from human by now.But with the enforcement of laws and regulations about human organ transplantation,the use of tissue or organs originated from human may face the problems of source limitations,ethical issues,law restriction and the risk of infection of HIV or HBV.So utlizing pig acellular dermal matrix as the substitute of human acellular dermal matrix into clinical use will be a potential solution in the future.
To accumulate for Bama miniature pig as animal model used in study on human skin and form research basis for Bama minipig acellular dermal matrix in order to utilize it into xeno-transplant in clinical use,skin samples from Bama miniature pigs and human were excised and the morphology,ultrastructre,immnohistochemistry and physico-chemical property of collagen typeⅠextracted from Bama miniature pig skin were studied.Resemblance and disparity between Bama miniature pig skin and human skin were found and described.Then Bama miniature pig acellular dermal matrix was prepared.Its bionomics,safe and efficacy were studied compared to human acellular dermal matrix.The results and conclusions are summarized as follows:
1.Bama miniature pig skin were strikingly similar to human skin except it posesses apocrine glands instead of eccrine sweat glands in human skin.4 months Bama miniature pig skin was the most similar to human skin because all layers thickness were similar between Bama miniature pig skin and human skin.Our results indicated that Bama miniature pigs could be used as model for study on human skin,it also could be used as materials to prepare dermal substitute in wound repair.
The layer,the layer thickness,the heterogeneity of basal keratinocytes,the dermal-epidermal interface consisted of undulant downgrowths that interdigitated with dermal papillae,the structure and distribution of extracellular matrix,the network of vertically oriented capillary loops that extend into each dermal papilla emanate orom the superficial vascular plexus,were strikingly similar to that seen in humans.The ultrastructure and density of langerhans cell,fibroblast,vascular endothelial cell,mast cell were similar between Bama miniature pig skin and those of human.The reactivity and postive location of the Bama miniature pig skin with anti-human sera to lamilin,fibronectin,filaggrin,collagenⅠ,collagenⅢ,keratin were consistent with human skin.The HPLC chromatograph,UV spectroscopy, amino acids composition,electrophoresis of collgen typeⅠextracted from Bama miniature pig skin showed remarkable similarity to those of human collgen typeⅠ.A significant difference between Bama miniature pig and human was that Bama miniature pig possesses apocrine glands instead of the eccrine sweat glands in human skin.Melanocytes were the next-most prevalent cell type found in human skin,but in Bama miniature pig skin, melanocytes density were various on different areas.Another differences were that Bama miniature pig skin could not react with anti-human sera to CD34,ICAM-1,S-100.
2.Bama miniature pig acellular dermal matrix was prepared and the method of preparing was improved.There was no significant difference between Bama miniature pig acellular dermal matrix and human acellular dermal matrix in their histologic structure, antigenicity,biocompatibility,vascularization and efficacy.Our results indicated that Bama miniature pig acellular dermal matrix may be an ideal substitute of human acellular dermal matrix in wound repair.
Bama miniature pig acellular dermal matrix treated by 0.25 typsin at 4℃for one night,then treated by 0.5%triton X-100 succussed at room temperature for 12h had no epithelial components and dermal cell components,which could evoke an immune reaction when it is implanted.The prepared acellular dermal matrix remain several extracellular matrix may support cell attachment and proliferation,such as elastic fibre,collagenⅠ,collagneⅢ,collagenⅣ,fibronectin.It exhibited three important properties like human acellular dermal matrix:well biocompatibility,low antigenicity,vapid vascularization.After transplanted to rat's full thickness skin defects in combination with autologous thin split-thickness skin graft(STSG),Bama miniature pig acellular dermal matrix could significantly raise the rate of autologous thin STSG survival and significantly decreace the rate of wound contraction vs only transplant autologous thin STSG group.The effects were not significantly difference between Bama miniature pig acellular dermal matrix and human acellular dermal matrix.So it would be an ideal dermal substitute of human acellular dermal matrix for clinical use.
无细胞真皮作为一种有效的临床用生物材料,具有重要的临床应用价值。但国内目前仅有人无细胞真皮产品面世,且价格昂贵。随着国家人体器官移植技术临床应用管理暂行规定的正式实施,人组织器官的应用将面临来源、伦理、法律等一系列问题,而且还具有传染HIV、HBV等血液传播性疾病的危险。因此,以标准化实验用小型猪无细胞真皮替代人无细胞真皮具有良好的应用前景。
为了推广巴马香猪在医学生物学中的应用,评估巴马香猪作为人皮肤模型动物在创伤修复领域的应用价值,同时为了开发巴马香猪皮肤来源的异种移植生物材料,本课题从比较医学的角度,对巴马香猪与人皮肤对比进行了组织学结构、免疫组化反应特点、超微结构、皮肤胶原蛋白理化特性等的系统研究;同时,制备了巴马香猪无细胞真皮,与人无细胞真皮对比进行了安全性有效性评价,以期利用巴马香猪无细胞真皮替代人无细胞真皮而应用于创伤修复领域,为其产品研发形成基础,同时也进一步比较巴马香猪皮肤与人皮肤在应用特性方面的相似性。全文主要结果及结论如下:
1.本课题对巴马香猪皮肤的组织学结构、免疫组化反应特点、超微结构、胶原蛋白理化特性等进行了系统研究,发现巴马香猪的皮肤结构除汗腺结构与人不同外,其它结构及其特点与人极为相似。其4月龄小型猪皮肤发育程度最接近于成人皮肤,各层厚度及结构都与成人皮肤相似。上述研究结果表明,巴马香猪可作为研究人类皮肤的模型动物。
巴马香猪在皮肤厚度、分层、真表皮的高低起伏的连接方式、发达的浅表血管系统以及皮肤的超微结构都与人类皮肤相似;皮肤中所含细胞种类和成纤维细胞、血管内皮朗格罕细胞、肥大细胞的密度也与人类皮肤相近;具有与人类Ⅰ型胶原、Ⅲ型胶原、层粘连蛋白、纤维连接蛋白、中间丝相关蛋白相似的抗原决定簇,可与抗人血清发生交叉反应,且分布特点与人类皮肤相似;真皮中主要的胞外基质构成成分-Ⅰ型胶原的结构及理化特性也几乎与人类Ⅰ型胶原相同。但巴马香猪皮肤中的汗腺为顶泌汗腺,而人类皮肤中主要为外泌汗腺;另外,巴马香猪皮肤中的色素细胞在身体的不同部位差异很大;巴马香猪皮肤缺乏与抗人CD34、ICAM-1、S-100等抗体对应的抗原决定簇。
2.优化了无细胞真皮的制备方法,制备了巴马香猪无细胞真皮,对其与人无细胞真皮对比进行了生物学特性与安全性有效性评价研究。对巴马香猪与人无细胞真皮对比进行了胞外基质的种类和分布、微生物学检测、致敏实验、淋巴细胞增殖实验、细胞毒性实验、组织学相容性实验、单纯无细胞真皮移植实验以及结合大鼠自体皮的复合移植实验等实验研究。实验结果表明,巴马香猪无细胞真皮与人无细胞真皮的形态结构、抗原性、生物相容性、血管化速度及动物实验效果无显著差异,显示巴马香猪无细胞真皮可望替代人无细胞真皮应用于创伤修复领域。
经0.25%胰酶4℃处理过夜,0.5%Triton X-100室温振荡处理12h制备而成的巴马香猪无细胞真皮,具有如下特点:成功去除了引起强烈免疫排斥反应的表皮和细胞成分,完整保留了基底膜,与抗原性有关的胞外基质结蛋白和波形蛋白也得到有效的去除;保留了Ⅰ型胶原、Ⅲ型胶原、Ⅳ型胶原、纤维连接蛋白等与细胞粘附、生长相关的胞外基质;具有与人无细胞真皮相似的无菌、无致敏反应、生物相容性好、抗原性低、血管化速度快等优良的生物学特性;将其与大鼠自体刃厚皮复合植入大鼠创面后,巴马香猪无细胞真皮与人无细胞真皮组的移植皮片存活率、创面收缩率相似,且与大鼠同种无细胞真皮组的移植皮片存活率和创面收缩率无明显差异,未引发明显的免疫排斥反应。表明巴马香猪无细胞真皮可望替代人无细胞真皮应用于创伤修复领域。
The skin of domestic animals has been investigated extensively in order to find a suitable experimental model to be used in research studies on human skin.Of all the animals examined pig skin is more similar to human skin anatomically and physiologically.But the domestic pig is not suitable for the biomedicine research for its unsharp genetic background,great individual difference and fast growth velocity.Bama miniature pig,one distinct breed of miniature pigs in China,is an ideal animal model for its high inbreeding degree,genetic stability,small body size.However basic research data of Bama miniature pig is absent,so its application in biomedicine is limited.
Acellular dermal matrix used as dermal substitute provides to splite-thickness skin grafts and to cultured epidermal autografts.A thick demal bed at the recepient site inhibits and hypertrophic scar formation of split-tickness skin grafts.The clinic applicationable product of acellular dermal matrix was only originated from human by now.But with the enforcement of laws and regulations about human organ transplantation,the use of tissue or organs originated from human may face the problems of source limitations,ethical issues,law restriction and the risk of infection of HIV or HBV.So utlizing pig acellular dermal matrix as the substitute of human acellular dermal matrix into clinical use will be a potential solution in the future.
To accumulate for Bama miniature pig as animal model used in study on human skin and form research basis for Bama minipig acellular dermal matrix in order to utilize it into xeno-transplant in clinical use,skin samples from Bama miniature pigs and human were excised and the morphology,ultrastructre,immnohistochemistry and physico-chemical property of collagen typeⅠextracted from Bama miniature pig skin were studied.Resemblance and disparity between Bama miniature pig skin and human skin were found and described.Then Bama miniature pig acellular dermal matrix was prepared.Its bionomics,safe and efficacy were studied compared to human acellular dermal matrix.The results and conclusions are summarized as follows:
1.Bama miniature pig skin were strikingly similar to human skin except it posesses apocrine glands instead of eccrine sweat glands in human skin.4 months Bama miniature pig skin was the most similar to human skin because all layers thickness were similar between Bama miniature pig skin and human skin.Our results indicated that Bama miniature pigs could be used as model for study on human skin,it also could be used as materials to prepare dermal substitute in wound repair.
The layer,the layer thickness,the heterogeneity of basal keratinocytes,the dermal-epidermal interface consisted of undulant downgrowths that interdigitated with dermal papillae,the structure and distribution of extracellular matrix,the network of vertically oriented capillary loops that extend into each dermal papilla emanate orom the superficial vascular plexus,were strikingly similar to that seen in humans.The ultrastructure and density of langerhans cell,fibroblast,vascular endothelial cell,mast cell were similar between Bama miniature pig skin and those of human.The reactivity and postive location of the Bama miniature pig skin with anti-human sera to lamilin,fibronectin,filaggrin,collagenⅠ,collagenⅢ,keratin were consistent with human skin.The HPLC chromatograph,UV spectroscopy, amino acids composition,electrophoresis of collgen typeⅠextracted from Bama miniature pig skin showed remarkable similarity to those of human collgen typeⅠ.A significant difference between Bama miniature pig and human was that Bama miniature pig possesses apocrine glands instead of the eccrine sweat glands in human skin.Melanocytes were the next-most prevalent cell type found in human skin,but in Bama miniature pig skin, melanocytes density were various on different areas.Another differences were that Bama miniature pig skin could not react with anti-human sera to CD34,ICAM-1,S-100.
2.Bama miniature pig acellular dermal matrix was prepared and the method of preparing was improved.There was no significant difference between Bama miniature pig acellular dermal matrix and human acellular dermal matrix in their histologic structure, antigenicity,biocompatibility,vascularization and efficacy.Our results indicated that Bama miniature pig acellular dermal matrix may be an ideal substitute of human acellular dermal matrix in wound repair.
Bama miniature pig acellular dermal matrix treated by 0.25 typsin at 4℃for one night,then treated by 0.5%triton X-100 succussed at room temperature for 12h had no epithelial components and dermal cell components,which could evoke an immune reaction when it is implanted.The prepared acellular dermal matrix remain several extracellular matrix may support cell attachment and proliferation,such as elastic fibre,collagenⅠ,collagneⅢ,collagenⅣ,fibronectin.It exhibited three important properties like human acellular dermal matrix:well biocompatibility,low antigenicity,vapid vascularization.After transplanted to rat's full thickness skin defects in combination with autologous thin split-thickness skin graft(STSG),Bama miniature pig acellular dermal matrix could significantly raise the rate of autologous thin STSG survival and significantly decreace the rate of wound contraction vs only transplant autologous thin STSG group.The effects were not significantly difference between Bama miniature pig acellular dermal matrix and human acellular dermal matrix.So it would be an ideal dermal substitute of human acellular dermal matrix for clinical use.
引文
1.Lavker RM,Kwong F,Kligman AM.Changes in skin surface patterms withage[J].Jgerontol,1980,35:348-354
2.Braverman IM.Ultrastructure and organization of the cutaneous microvasculature in normal and pathologic states[J].J Invest Dermatol,1989,93:2S-9S
3.Gray GM,Yardely HJ.Lipid composition of cell isolated from pig,human,and rat epidermis[J].J Lipid Res,1975,16:434-440
4.Vardaxis NJ.Brans TA,Boon ME,et al.Confocal laser scaning microscopy of porcine skin:implications for human wound healing studies[J].J Anat,1997,190:601-611
5.Morris GM,Hopewell JW.Epidermal cell kinetics of the pig:a review[J].Cell Tissue Kinetics,1990,23:271-282
6.Marcarian HQ,Calhoun ML.Mocroscopic anatomy of the integument of adult swine[J].Am J Vet Res,1966,27:765-772
7.Forbes PD.Vascular supply of the skin and hair in swine.In:Montagna W,Dobson RL.editors.Advances in the biology of skin:Hair growth.Vol.9.Oxford:Pergamon,1969:419-432
8.Nicolaides N,Fu HC,Rice GR.The skin surface lipids of man compared with those of eighteen species of animals[J].J Invest Derm,1968,51:83-89
9.Gray GM,White RJ,Williams RH,et al.Lipid composition of the superficiall stratum corneum of pig epidermis[J].Br J Dermatol,1982,106:59-63
10.Wollina U,Berger U,Mahrle G.Immunohistochemistry of porcine skin[J].Acta Histochem,1991,90:87-91
11.Qvist MH,Hoeck U,Kreilgaard B,et al.Evaluation of Gottingen minipig skin for transdermal in vitro permeation studies[J].European Journal of Pharmaceutical Sciences,2000,11:59-68
12.Meyer W,Scharz R,Neurand K.The skin of domestic mammals as a model for the human skin with special reference to the domestic pig[J].Curr Probl Dermatol,1978,7:39-52
13.潘玉英,白文霞,苏宁.兔阴道黏膜组织学与阴道黏膜刺激性形态学改变[J].中国比较医学杂志,2005,15(4):227-228
14.中华医学会.临床技术操作规范-病理学分册[M].北京:人民军医出版社,2004.130-136
15.谢宝松,施作霖,李强.非小细胞肺癌Ki-67的表达及其意义[J].福建医药杂志,2002,24(2):1-2
16.Morimura S,Nagata H,Uemura Y,et al.Development of an effective process for utilization of collagen from livestock and fish waste[J].Process Biochemistry,2002(37):1403-1412
17.王韦,孙家玲,刘俊龙.可溶性胶原的制作与临床应用[J].生物医学工程学杂志,1998,5:224-225
18.Tory P,Sullivan MD,William H,et al.The pig as a model for human wound healing[J].Wound Rep Reg,2001,9:66-76.
19.姜笃银,张红,周爱珍等.免疫组化方法检测猪与人皮肤通用抗原标志[J].中国冶金工业医学杂志,2003,20(4):236-239
20.Weinstein GD.Autoradiographic studies of turnover time and protein synthesis in pig epidermis[J].Journal of Investigative Dermatology,1965,44:413-419
21.Vardaxis NJ,Brans TA,Boon ME,et al.Confocal laser scanning microscopy of porcine skin:implications for human wound healing studies[J].J Anat,1997,190:601-611
22.Stadelmann WK,Digenis AG,Tobin GR.Physiology and healing dynamics of chroniv cutaneous wounds[J].Am J S urg,1998;176(2A Suppl):26S-38S
23.Cook H,Davies KJ,Harding KG,et al.Defective extracellular matrix reorganization by chronic wound fibroblasts is associated with alterations in TIMP-1,TIMP-2,and MMP-2activity[J].J Invest Dermatol,2000,115(2):225-233
24.Iyer VR,Eisen MB,Ross DT,et al.The transcriptional program in the response of human fibroblasts to serum[see comments][J].Science,1999,283(5398):83-87
25.程天民主编。创伤战伤病理学[M]。北京,解放军出版社,第1版。1992:28-42,398-426
26.Kimura I,Honda R,Okai H,et al.Vasclar endothelial growth factor promotes cell-cycle transition from G0-to-G1 phase in aubcultured endothelial cells of dabetic rat thoracc aorta[J].Jpn-J-Phrmacol.2000,83(1):47-55
27.Wang DI,Gotlieb AI.Fibroblast growth factor 2 enhances early stages of in vitro endothelial repair by microfilament bundle reorganization and cell elongation[J].Exp-Mol-Pathol.1999,66(3):179-190
28.Trautmann A,Toksoy A,Engelhardt E,et al.Mast cell involvement in normal human skin wound healing:expression of monocyte chemoattractant protein-lis correlated with recruitment of mastcells which synthesize inerleukin-4 in vivo[J].J Pathol,2000,190(1):100-156
29.Berton A,Levi Schaffer F,Emonard-H,et al.Activation of fibroblasts in collagen lattices by mast cell extract:amodel of fibrosis[see comments][J].Clin Exp Allergy,2000,30(4):485-492
30.Hermes B,Feldmann BI,Welker P,et al.Altered expression of mast cell cymase and tryptase and of c-kit in human cutaneous scar tissue[J].J Invest Dermatol,2000,114(1):51-55
31.Moresi JM,Hom TD1 Distribution of Langerhans cells in human hair follicle[J].J Cutan Pathol,1997,24(10):638-640
32.康克非,朱学骏,赵辨主编.免疫皮肤病学[M].北京:人民卫生出版社,1992:36-43
33.姜笃银,陈壁,贾赤宇等.异种脱细胞真皮基质抗原性的实验研究[J].中华烧伤杂志,2003,19(3):155-158.
34.Heinrich W,Lange PM,Stirz T,Iancu C,Heidermann E.Isolation and characterization of the large cyanogens bromide peptides from the al-and α2-chains in pig skin collagen[J].FEBS Letters,1971,16:63-67
35.颜汉卿,徐国风,生物医学材料学[M].天津:天津科技翻译出版公司。1983:327-328
36.陈金辉,齐顺贞,贺占国,等.不同种属脱细胞真皮与自体皮复合移植的比较性研究[J].中华烧伤杂志,2003,19(5):300-302
37.司徒镇强,吴军正.细胞培养[M].西安:世界图书出版公司,1996,183-184
38.陈瑞铭主编.动物组织培养技术及其应用[M].北京:科学出版社,1991:1-100.
39.刘鼎新,吕证宝主编.细胞生物学研究方法与技术[M].北京:北京医科大学、中国协和医科大学联合出版社,1990:156-190
40.Srivastava S,Gorham S D,Courtney JM..The attachment and growth of an established cell lines on collagen,chemically modified collagen,and collagen composite surfaces[J].Biomaterials,1990,11(4):162-168
41.梁卫东,石应康.细胞培养法评价生物材料生物相容性研究进展[J].生物医学工程学杂志,1999,16(1):86-90
42.Takami Y,Matsuda T,Yoshitake M,et al.Dispase/detergent treated dermal matrix as a dermal substitute[J].Burns,1996,22(3):182-190
43.Wang HJ,Chen TM,Cheng T Y.Use of a porcine dermis template to enhance widely expanded mesh autologous split—thickness skin graft growth:Preliminary report[J].J Trauma,1997,42(2):177-182
44.Walter RJ,Jennings LJ,Matsuda T,et al.Dispase/triton treated acellular dermal matrix as a dermal substitute in rats[J].Curr Surg,1997,54(5):371-374
45.Munster AM,Smith M,Shalom A.Acellular allograft dermal matrix:immediate or delayed epidermal coverage[J].Burns,2001,27(2):150-153
46.Walter RJ,Matsuda T,Reyes HM,et al.Charactefization of acellular dermal matrices (ADMs) prepared by two different methods[J].Burns,1998,24(2):104-113
47.廖立新,陈刚泉,李国辉,等.异种无细胞真皮的制备[J].江西医学院学报.2004,44(1):88-89.
48.杨燕妮,彭代智,伍素华,等异种无细胞真皮制备方法比较及其在移植中的意义[J].中国临床康复,2004,8(23):4772-4773
49.Van Luyn MJ,VanWachem PB,Olde Damink LH,et al.Secondary cyto-toxicity of cross-linked dermal sheep collagens during repeated exposure to human fibroblasts[J].Biomaterials,1992,13(4):1017-1022.
50.Medalie,DA,Tompkins RG,Morgan JR.Evaluation of acellular human dermis as a dermal analog in a composite skin graft[J].ASAIO J,1996,42:M455-462
51.Medalie DA,Eming SA,Tompkins RG,et al.Evaluation of human skin reconstituted from composite grafts of cultured keratinocytes and human acellular dermis transplanted to ahymic mice[J].J.Invest.Dermatol,1996,107:121-127
52.Fitzgerald AMP,Kirkpatrick JR,Foo ITH,et al.Human skin histology as demonstrated by Herovici's stain:a guide for the improvement of dermal substitutes for use with cultured keratinocytes[J].Burns,1996,22:200-202
53.Rennekampff HO,Hansbrough JF,Woods V,et al.Integrin and matrix molecule expression in cultured skin replacements[J].J.Burn Care Rehab,1996,17:213-221
54.Harry E.Nutrition needs of mammalian cells in tissue culture[J].Science,1955,122:3168-3171
55.Yourtee MD,Tong PY,Ro se LA,et al.Effect of spiroortho carbonate volume modifier comonomers on the in vitro toxicology of rial non—shrinking dental epoxycopolymers[J].Res Commun Mol Pathol Pharmacol,1994,86(3):347-351.
56.Takami Y,Matsuda T,Yoshitake M,et al.Dispase/detergent treated dermal matrix as a dermal substitute[J].Burns,1996,22(3):182-190
57.孙红,车鹏和,陈铎等.一种新型无细胞基质生物相容性的实验研究[J].中国美容医学,2004,13(2):138-139
58.Desagun EZ,Botts JL,Srivastava A,et al.Long-term outcome of xenogenyic dermal matrix implantation in immuneocompetent rats[J],J Surg Res 2001:96(1):96-106
59.Srivastva A,DeSagun EZ,Jennings LJ,et al.Use of porcine acellular dermal matrix as a dermal substitute in rats[J].Ann Surg,2001,233(3):400-408.
60.Walter RJ,Matsuda T,Reyes HM,et al.Characterization of acellular dermal matrices (ADMs) prepared by two different methods[J].Burns,1998,24(2):104-113
61.Boyce ST,Hansbrough JH.Biologic attachment,growth and differentiation of cultured human epidermal keratinocytes on a graftable collagenand chondroitin-6-sulfate substrate[J].Surgery,1988,103(4):421-431
62.Hansbrough JF,Cooper ML,Cohen R,et al.Evaluation of a biodegratable matrix containing cultured human fibroblasts as a dermal replacement beneath meshed skin grafts on athymic mice[J].Surgery,1992,111(4):438-446
63.Wang HJ,Chen TM,Cheng T Y.Use of a porcine dermis template to enhance widely expanded mesh autologous split—thickness skin graft growth:Preliminary report[J].J Trauma,1997,42(2):177-182
64.Richters CD,Hoekstra MJ,Pont JS,et al.Immunology of skin transplantation[J].Clincs in Dermatology,2005,23:338-342
1.Takami Y,Matsuda T,Yoshitake M,et al.Dispase/detergent treated dermal matrix as a dermal substitute[J].Burns,1996,22(3):182-190
2.Wang HJ,Chen TM,Cheng T Y.Use of a porcine dermis template to enhance widely expanded mesh autologous split—thickness skin graft growth:Preliminary report[J].J Trauma,1997,42(2):177-182
3.Walter RJ,Jennings LJ,Matsuda T,et al.DispasePtriton treated acellular dermal matrix as a dermal substitute in rats[J].Curr Surg,1997,54(5):371-374
4.Munster AM,Smith M,Shalom A.Acellular allograft dermal matrix:immediate or delayed epidermal coverage[J].Burns,2001,27(2):150-153
5.Walter RJ,Matsuda T,Reyes HM,et al.Characterization of acellular dermal matrices (ADMs) prepared by two different methods[J].Burns,1998,24(2):104-113
6.贾生贤,廖镇江,黄伯高.无细胞真皮基质与自体皮复合移植愈合过程的观察[J].实用美容整形外科杂志,2000,11:136-139
7.孙红,冯光珍,黑砚等.同种异体人脱细胞真皮基质的细胞相容性[J].中国临床康复,2004,8(23):4770-4771
8.廖立新,陈刚泉,李国辉等.异种无细胞真皮的制备[J].江西医学院学报,2004,44(1):88-90
9.Desagun EZ,Botts JL,Srivastava A,et al.Long-term outcome of xenogenyic dermal matrix implantation in immuneocompetent rats[J],J Surg Res 2001:96(1):96-106
10.杨燕妮,彭代智,伍素华等.异种无细胞真皮制备方法比较及其在移植中的意义[J].中国临床康复,2004,8(23):4772-4773
11.Abenavoli FM,Corelli R.About AlloDerm[J].Plast Reconstr Surg,2001,108(7):2175-2176
12.Sclafani AP,Romo T,Jacono AA et al.Evaluation of acellular dermal graft(AlloDerm)sheet for soft tissue augmentation:a 12 year follow-up of clinical observations and histological findings[J].Arch Facial Plast Surg 2001,3(2):101-103
13.姜笃银,陈壁,贾赤宇等.异种脱细胞真皮基质抗原性的实验研究[J]。中华烧伤杂志,2003,19(3):155-158.
14.Srivastva A,DeSagun EZ,Jennings LJ,et al.Use of porcine acellular dermal matrix as a dermal substitute in rats[J].Ann Surg,2001,233(3):400-408.
15.吴秋合,姚敏,潘银根等.异种(猪)脱细胞真皮基质移植后炎症免疫反应的实验研究[J].中华整形外科杂志,2002,18(5):266-268
16.刘强,柴家科,杨红明等.两种去细胞处理猪皮与微粒皮复合移植对创面愈合的影响[J].中国医师杂志,2004,6(3):291-294
17.姜笃银,陈壁,汤朝武等.猪与人致敏的淋巴细胞混合培养及其细胞因子检测[J].西北国防医学杂志,2002,23(2):90-93
18.Van Luyn MJ,VanWachem PB,Olde Damink LH,et al.Secondary cyto-toxicity of cross-linked dermal sheep collagens during repeated exposure to human fibroblasts[J].Biomaterials,1992,13(4):1017-1022
19.Oshima H.Nakamura M.A study on reference standard for cytotoxicity assay of biomaterials[J].Biomed Mater,1994,4(4):327-333
20.Tsuchiya T.Study on the standardization of cyto toxicity tests and new standard referencematerials useful for evaluating the safety of biomaterials[J].J Biomater Appl,1994;9(2):138-142
21.Harry E.Nutrition needs of mammalian cells in tissue culture[J]Science,1955,122:3168-3171
22.Richardson RJ,Miller JA,Reichert WM.Polyimides as biomaterials Preliminary biocompatibility testing[J].Biomaterials,1993,14(9):627-632
23.Gretzer C,Eriksson AS,Allden B,et al.Monocyte activation on titanium sputtered polystyrene surfaces in vitro,the effect of culture conditions of interleuk in 21release.Biomaterials,1996;17(9):851-855
24.石应康,程述森,魏蜀亮等.不同交联改性牛心包材料的细胞毒性研究[J].生物医学工程学杂志,1995,12(4):350-352
25.Wataha JC,Hanks CT,Craig RG.In vitro effects of metal ions on cellular metabolism and the co relation between these effects and the uptake of the ions[J].J Biomed Mater Res,1994,28:427-430
26.Sgouras D.The tetrazo liumbased assay(MTT) as a preliminary screen for evaluation of in vitro cytotoxicity[J].J Mater Sci,1990,1:61-65
27.Yourtee MD,Tong PY,Ro se LA,et al.Effect of spiroorthocarbonate volume modifier comonomers on the in vitro toxicology of trial non shrinking dental epoxycopolymers [J].Res Commun Mol Pathol Pharmacol,1994,86(3):347-351
28.孙红,车鹏和,陈铎等.一种新型无细胞基质生物相容性的实验研究[J].中国美容医学,2004,13(2):138-139
29.冯祥生,潘银根,谭家驹等.异种(猪)脱细胞真皮与自体表皮复合移植研究[J].中华整形外科杂志,2000,16(1):40-42
30.Burke JF,Yannas IV,Quinby WC,et al.Successful use of a physiologically acceptable artificial shin in the treatment of extensive burn injury[J].Ann Surg,1981,194(4):413-427
31.Pruitt Jr BA,Levine NS.Characteristics and uses of biologic dressingand skin substitutes [J].Arch Surg,1984,119(3):312-322
32.Boyce ST,Hansbrough JH.Biologic attachment,growth and differentiation of cultured human epidermal keratinocytes on a graftable collagenand chondroitin-6-sulfate substrate [J].Surgery,1988,103(4):421-431
33.Hansbrough JF,Cooper ML,Cohen R,et al.Evaluation of a biodegratable matrix containing cultured human fibroblasts as a dermal replacement beneath meshed skin grafts on athymic mice[J].Surgery,1992,111(4):438-446
34.肖仕初,夏照帆,杨君等.成纤维细胞-无细胞真皮替代物的生物学活性及移植试验[J].中华烧伤杂志,2001,17(4):231-232
35.Sheridan R,Choucair R,Donelan M,et al.Acellular allodermis in bums surgery:1-year resuils of a pilot trial[J].J Burn Care Rehabil,1998,19(6):528-530
36.Wagshall E,Lewis Z,Babich SB,et al.Acelluar dermal matrix allograft in the treatment of mucogingival defects in children:illustrativecase report[J].J Dent Child,2002,69(1):39-43
37.Hansbrough JF,Morgan J,Greenleaf G,et al.Development of a temporary living skin replacement composed of human neonatal fibroblasts cultured in biobrane,a synthetic dressing material[J].Surgery,1994,115(5):633-637
38.肖仕初,夏照帆,杨君等.成纤维细胞促进真皮替代物血管化的作用机制[J].中国修复重建杂志,2003,17(2):100-102
39.王军琳,刘源,金岩等.构建组织工程皮肤种子细胞的人血管内皮细胞的培养[J].中国临床康复,2003,17(8):1243-1244
2.Braverman IM.Ultrastructure and organization of the cutaneous microvasculature in normal and pathologic states[J].J Invest Dermatol,1989,93:2S-9S
3.Gray GM,Yardely HJ.Lipid composition of cell isolated from pig,human,and rat epidermis[J].J Lipid Res,1975,16:434-440
4.Vardaxis NJ.Brans TA,Boon ME,et al.Confocal laser scaning microscopy of porcine skin:implications for human wound healing studies[J].J Anat,1997,190:601-611
5.Morris GM,Hopewell JW.Epidermal cell kinetics of the pig:a review[J].Cell Tissue Kinetics,1990,23:271-282
6.Marcarian HQ,Calhoun ML.Mocroscopic anatomy of the integument of adult swine[J].Am J Vet Res,1966,27:765-772
7.Forbes PD.Vascular supply of the skin and hair in swine.In:Montagna W,Dobson RL.editors.Advances in the biology of skin:Hair growth.Vol.9.Oxford:Pergamon,1969:419-432
8.Nicolaides N,Fu HC,Rice GR.The skin surface lipids of man compared with those of eighteen species of animals[J].J Invest Derm,1968,51:83-89
9.Gray GM,White RJ,Williams RH,et al.Lipid composition of the superficiall stratum corneum of pig epidermis[J].Br J Dermatol,1982,106:59-63
10.Wollina U,Berger U,Mahrle G.Immunohistochemistry of porcine skin[J].Acta Histochem,1991,90:87-91
11.Qvist MH,Hoeck U,Kreilgaard B,et al.Evaluation of Gottingen minipig skin for transdermal in vitro permeation studies[J].European Journal of Pharmaceutical Sciences,2000,11:59-68
12.Meyer W,Scharz R,Neurand K.The skin of domestic mammals as a model for the human skin with special reference to the domestic pig[J].Curr Probl Dermatol,1978,7:39-52
13.潘玉英,白文霞,苏宁.兔阴道黏膜组织学与阴道黏膜刺激性形态学改变[J].中国比较医学杂志,2005,15(4):227-228
14.中华医学会.临床技术操作规范-病理学分册[M].北京:人民军医出版社,2004.130-136
15.谢宝松,施作霖,李强.非小细胞肺癌Ki-67的表达及其意义[J].福建医药杂志,2002,24(2):1-2
16.Morimura S,Nagata H,Uemura Y,et al.Development of an effective process for utilization of collagen from livestock and fish waste[J].Process Biochemistry,2002(37):1403-1412
17.王韦,孙家玲,刘俊龙.可溶性胶原的制作与临床应用[J].生物医学工程学杂志,1998,5:224-225
18.Tory P,Sullivan MD,William H,et al.The pig as a model for human wound healing[J].Wound Rep Reg,2001,9:66-76.
19.姜笃银,张红,周爱珍等.免疫组化方法检测猪与人皮肤通用抗原标志[J].中国冶金工业医学杂志,2003,20(4):236-239
20.Weinstein GD.Autoradiographic studies of turnover time and protein synthesis in pig epidermis[J].Journal of Investigative Dermatology,1965,44:413-419
21.Vardaxis NJ,Brans TA,Boon ME,et al.Confocal laser scanning microscopy of porcine skin:implications for human wound healing studies[J].J Anat,1997,190:601-611
22.Stadelmann WK,Digenis AG,Tobin GR.Physiology and healing dynamics of chroniv cutaneous wounds[J].Am J S urg,1998;176(2A Suppl):26S-38S
23.Cook H,Davies KJ,Harding KG,et al.Defective extracellular matrix reorganization by chronic wound fibroblasts is associated with alterations in TIMP-1,TIMP-2,and MMP-2activity[J].J Invest Dermatol,2000,115(2):225-233
24.Iyer VR,Eisen MB,Ross DT,et al.The transcriptional program in the response of human fibroblasts to serum[see comments][J].Science,1999,283(5398):83-87
25.程天民主编。创伤战伤病理学[M]。北京,解放军出版社,第1版。1992:28-42,398-426
26.Kimura I,Honda R,Okai H,et al.Vasclar endothelial growth factor promotes cell-cycle transition from G0-to-G1 phase in aubcultured endothelial cells of dabetic rat thoracc aorta[J].Jpn-J-Phrmacol.2000,83(1):47-55
27.Wang DI,Gotlieb AI.Fibroblast growth factor 2 enhances early stages of in vitro endothelial repair by microfilament bundle reorganization and cell elongation[J].Exp-Mol-Pathol.1999,66(3):179-190
28.Trautmann A,Toksoy A,Engelhardt E,et al.Mast cell involvement in normal human skin wound healing:expression of monocyte chemoattractant protein-lis correlated with recruitment of mastcells which synthesize inerleukin-4 in vivo[J].J Pathol,2000,190(1):100-156
29.Berton A,Levi Schaffer F,Emonard-H,et al.Activation of fibroblasts in collagen lattices by mast cell extract:amodel of fibrosis[see comments][J].Clin Exp Allergy,2000,30(4):485-492
30.Hermes B,Feldmann BI,Welker P,et al.Altered expression of mast cell cymase and tryptase and of c-kit in human cutaneous scar tissue[J].J Invest Dermatol,2000,114(1):51-55
31.Moresi JM,Hom TD1 Distribution of Langerhans cells in human hair follicle[J].J Cutan Pathol,1997,24(10):638-640
32.康克非,朱学骏,赵辨主编.免疫皮肤病学[M].北京:人民卫生出版社,1992:36-43
33.姜笃银,陈壁,贾赤宇等.异种脱细胞真皮基质抗原性的实验研究[J].中华烧伤杂志,2003,19(3):155-158.
34.Heinrich W,Lange PM,Stirz T,Iancu C,Heidermann E.Isolation and characterization of the large cyanogens bromide peptides from the al-and α2-chains in pig skin collagen[J].FEBS Letters,1971,16:63-67
35.颜汉卿,徐国风,生物医学材料学[M].天津:天津科技翻译出版公司。1983:327-328
36.陈金辉,齐顺贞,贺占国,等.不同种属脱细胞真皮与自体皮复合移植的比较性研究[J].中华烧伤杂志,2003,19(5):300-302
37.司徒镇强,吴军正.细胞培养[M].西安:世界图书出版公司,1996,183-184
38.陈瑞铭主编.动物组织培养技术及其应用[M].北京:科学出版社,1991:1-100.
39.刘鼎新,吕证宝主编.细胞生物学研究方法与技术[M].北京:北京医科大学、中国协和医科大学联合出版社,1990:156-190
40.Srivastava S,Gorham S D,Courtney JM..The attachment and growth of an established cell lines on collagen,chemically modified collagen,and collagen composite surfaces[J].Biomaterials,1990,11(4):162-168
41.梁卫东,石应康.细胞培养法评价生物材料生物相容性研究进展[J].生物医学工程学杂志,1999,16(1):86-90
42.Takami Y,Matsuda T,Yoshitake M,et al.Dispase/detergent treated dermal matrix as a dermal substitute[J].Burns,1996,22(3):182-190
43.Wang HJ,Chen TM,Cheng T Y.Use of a porcine dermis template to enhance widely expanded mesh autologous split—thickness skin graft growth:Preliminary report[J].J Trauma,1997,42(2):177-182
44.Walter RJ,Jennings LJ,Matsuda T,et al.Dispase/triton treated acellular dermal matrix as a dermal substitute in rats[J].Curr Surg,1997,54(5):371-374
45.Munster AM,Smith M,Shalom A.Acellular allograft dermal matrix:immediate or delayed epidermal coverage[J].Burns,2001,27(2):150-153
46.Walter RJ,Matsuda T,Reyes HM,et al.Charactefization of acellular dermal matrices (ADMs) prepared by two different methods[J].Burns,1998,24(2):104-113
47.廖立新,陈刚泉,李国辉,等.异种无细胞真皮的制备[J].江西医学院学报.2004,44(1):88-89.
48.杨燕妮,彭代智,伍素华,等异种无细胞真皮制备方法比较及其在移植中的意义[J].中国临床康复,2004,8(23):4772-4773
49.Van Luyn MJ,VanWachem PB,Olde Damink LH,et al.Secondary cyto-toxicity of cross-linked dermal sheep collagens during repeated exposure to human fibroblasts[J].Biomaterials,1992,13(4):1017-1022.
50.Medalie,DA,Tompkins RG,Morgan JR.Evaluation of acellular human dermis as a dermal analog in a composite skin graft[J].ASAIO J,1996,42:M455-462
51.Medalie DA,Eming SA,Tompkins RG,et al.Evaluation of human skin reconstituted from composite grafts of cultured keratinocytes and human acellular dermis transplanted to ahymic mice[J].J.Invest.Dermatol,1996,107:121-127
52.Fitzgerald AMP,Kirkpatrick JR,Foo ITH,et al.Human skin histology as demonstrated by Herovici's stain:a guide for the improvement of dermal substitutes for use with cultured keratinocytes[J].Burns,1996,22:200-202
53.Rennekampff HO,Hansbrough JF,Woods V,et al.Integrin and matrix molecule expression in cultured skin replacements[J].J.Burn Care Rehab,1996,17:213-221
54.Harry E.Nutrition needs of mammalian cells in tissue culture[J].Science,1955,122:3168-3171
55.Yourtee MD,Tong PY,Ro se LA,et al.Effect of spiroortho carbonate volume modifier comonomers on the in vitro toxicology of rial non—shrinking dental epoxycopolymers[J].Res Commun Mol Pathol Pharmacol,1994,86(3):347-351.
56.Takami Y,Matsuda T,Yoshitake M,et al.Dispase/detergent treated dermal matrix as a dermal substitute[J].Burns,1996,22(3):182-190
57.孙红,车鹏和,陈铎等.一种新型无细胞基质生物相容性的实验研究[J].中国美容医学,2004,13(2):138-139
58.Desagun EZ,Botts JL,Srivastava A,et al.Long-term outcome of xenogenyic dermal matrix implantation in immuneocompetent rats[J],J Surg Res 2001:96(1):96-106
59.Srivastva A,DeSagun EZ,Jennings LJ,et al.Use of porcine acellular dermal matrix as a dermal substitute in rats[J].Ann Surg,2001,233(3):400-408.
60.Walter RJ,Matsuda T,Reyes HM,et al.Characterization of acellular dermal matrices (ADMs) prepared by two different methods[J].Burns,1998,24(2):104-113
61.Boyce ST,Hansbrough JH.Biologic attachment,growth and differentiation of cultured human epidermal keratinocytes on a graftable collagenand chondroitin-6-sulfate substrate[J].Surgery,1988,103(4):421-431
62.Hansbrough JF,Cooper ML,Cohen R,et al.Evaluation of a biodegratable matrix containing cultured human fibroblasts as a dermal replacement beneath meshed skin grafts on athymic mice[J].Surgery,1992,111(4):438-446
63.Wang HJ,Chen TM,Cheng T Y.Use of a porcine dermis template to enhance widely expanded mesh autologous split—thickness skin graft growth:Preliminary report[J].J Trauma,1997,42(2):177-182
64.Richters CD,Hoekstra MJ,Pont JS,et al.Immunology of skin transplantation[J].Clincs in Dermatology,2005,23:338-342
1.Takami Y,Matsuda T,Yoshitake M,et al.Dispase/detergent treated dermal matrix as a dermal substitute[J].Burns,1996,22(3):182-190
2.Wang HJ,Chen TM,Cheng T Y.Use of a porcine dermis template to enhance widely expanded mesh autologous split—thickness skin graft growth:Preliminary report[J].J Trauma,1997,42(2):177-182
3.Walter RJ,Jennings LJ,Matsuda T,et al.DispasePtriton treated acellular dermal matrix as a dermal substitute in rats[J].Curr Surg,1997,54(5):371-374
4.Munster AM,Smith M,Shalom A.Acellular allograft dermal matrix:immediate or delayed epidermal coverage[J].Burns,2001,27(2):150-153
5.Walter RJ,Matsuda T,Reyes HM,et al.Characterization of acellular dermal matrices (ADMs) prepared by two different methods[J].Burns,1998,24(2):104-113
6.贾生贤,廖镇江,黄伯高.无细胞真皮基质与自体皮复合移植愈合过程的观察[J].实用美容整形外科杂志,2000,11:136-139
7.孙红,冯光珍,黑砚等.同种异体人脱细胞真皮基质的细胞相容性[J].中国临床康复,2004,8(23):4770-4771
8.廖立新,陈刚泉,李国辉等.异种无细胞真皮的制备[J].江西医学院学报,2004,44(1):88-90
9.Desagun EZ,Botts JL,Srivastava A,et al.Long-term outcome of xenogenyic dermal matrix implantation in immuneocompetent rats[J],J Surg Res 2001:96(1):96-106
10.杨燕妮,彭代智,伍素华等.异种无细胞真皮制备方法比较及其在移植中的意义[J].中国临床康复,2004,8(23):4772-4773
11.Abenavoli FM,Corelli R.About AlloDerm[J].Plast Reconstr Surg,2001,108(7):2175-2176
12.Sclafani AP,Romo T,Jacono AA et al.Evaluation of acellular dermal graft(AlloDerm)sheet for soft tissue augmentation:a 12 year follow-up of clinical observations and histological findings[J].Arch Facial Plast Surg 2001,3(2):101-103
13.姜笃银,陈壁,贾赤宇等.异种脱细胞真皮基质抗原性的实验研究[J]。中华烧伤杂志,2003,19(3):155-158.
14.Srivastva A,DeSagun EZ,Jennings LJ,et al.Use of porcine acellular dermal matrix as a dermal substitute in rats[J].Ann Surg,2001,233(3):400-408.
15.吴秋合,姚敏,潘银根等.异种(猪)脱细胞真皮基质移植后炎症免疫反应的实验研究[J].中华整形外科杂志,2002,18(5):266-268
16.刘强,柴家科,杨红明等.两种去细胞处理猪皮与微粒皮复合移植对创面愈合的影响[J].中国医师杂志,2004,6(3):291-294
17.姜笃银,陈壁,汤朝武等.猪与人致敏的淋巴细胞混合培养及其细胞因子检测[J].西北国防医学杂志,2002,23(2):90-93
18.Van Luyn MJ,VanWachem PB,Olde Damink LH,et al.Secondary cyto-toxicity of cross-linked dermal sheep collagens during repeated exposure to human fibroblasts[J].Biomaterials,1992,13(4):1017-1022
19.Oshima H.Nakamura M.A study on reference standard for cytotoxicity assay of biomaterials[J].Biomed Mater,1994,4(4):327-333
20.Tsuchiya T.Study on the standardization of cyto toxicity tests and new standard referencematerials useful for evaluating the safety of biomaterials[J].J Biomater Appl,1994;9(2):138-142
21.Harry E.Nutrition needs of mammalian cells in tissue culture[J]Science,1955,122:3168-3171
22.Richardson RJ,Miller JA,Reichert WM.Polyimides as biomaterials Preliminary biocompatibility testing[J].Biomaterials,1993,14(9):627-632
23.Gretzer C,Eriksson AS,Allden B,et al.Monocyte activation on titanium sputtered polystyrene surfaces in vitro,the effect of culture conditions of interleuk in 21release.Biomaterials,1996;17(9):851-855
24.石应康,程述森,魏蜀亮等.不同交联改性牛心包材料的细胞毒性研究[J].生物医学工程学杂志,1995,12(4):350-352
25.Wataha JC,Hanks CT,Craig RG.In vitro effects of metal ions on cellular metabolism and the co relation between these effects and the uptake of the ions[J].J Biomed Mater Res,1994,28:427-430
26.Sgouras D.The tetrazo liumbased assay(MTT) as a preliminary screen for evaluation of in vitro cytotoxicity[J].J Mater Sci,1990,1:61-65
27.Yourtee MD,Tong PY,Ro se LA,et al.Effect of spiroorthocarbonate volume modifier comonomers on the in vitro toxicology of trial non shrinking dental epoxycopolymers [J].Res Commun Mol Pathol Pharmacol,1994,86(3):347-351
28.孙红,车鹏和,陈铎等.一种新型无细胞基质生物相容性的实验研究[J].中国美容医学,2004,13(2):138-139
29.冯祥生,潘银根,谭家驹等.异种(猪)脱细胞真皮与自体表皮复合移植研究[J].中华整形外科杂志,2000,16(1):40-42
30.Burke JF,Yannas IV,Quinby WC,et al.Successful use of a physiologically acceptable artificial shin in the treatment of extensive burn injury[J].Ann Surg,1981,194(4):413-427
31.Pruitt Jr BA,Levine NS.Characteristics and uses of biologic dressingand skin substitutes [J].Arch Surg,1984,119(3):312-322
32.Boyce ST,Hansbrough JH.Biologic attachment,growth and differentiation of cultured human epidermal keratinocytes on a graftable collagenand chondroitin-6-sulfate substrate [J].Surgery,1988,103(4):421-431
33.Hansbrough JF,Cooper ML,Cohen R,et al.Evaluation of a biodegratable matrix containing cultured human fibroblasts as a dermal replacement beneath meshed skin grafts on athymic mice[J].Surgery,1992,111(4):438-446
34.肖仕初,夏照帆,杨君等.成纤维细胞-无细胞真皮替代物的生物学活性及移植试验[J].中华烧伤杂志,2001,17(4):231-232
35.Sheridan R,Choucair R,Donelan M,et al.Acellular allodermis in bums surgery:1-year resuils of a pilot trial[J].J Burn Care Rehabil,1998,19(6):528-530
36.Wagshall E,Lewis Z,Babich SB,et al.Acelluar dermal matrix allograft in the treatment of mucogingival defects in children:illustrativecase report[J].J Dent Child,2002,69(1):39-43
37.Hansbrough JF,Morgan J,Greenleaf G,et al.Development of a temporary living skin replacement composed of human neonatal fibroblasts cultured in biobrane,a synthetic dressing material[J].Surgery,1994,115(5):633-637
38.肖仕初,夏照帆,杨君等.成纤维细胞促进真皮替代物血管化的作用机制[J].中国修复重建杂志,2003,17(2):100-102
39.王军琳,刘源,金岩等.构建组织工程皮肤种子细胞的人血管内皮细胞的培养[J].中国临床康复,2003,17(8):1243-1244