组织工程真皮替代物的实验研究
|
摘要
深度皮肤缺损创面的修复在烧伤、创伤、慢性皮肤溃疡、瘢痕整形等的治疗中具有重要意义。传统移植自体皮的治疗方法常常导致供皮区色素改变和瘢痕形成,而皮肤缺损面积较大时自体皮源不能满足移植的要求。皮肤组织工程技术的发展与应用为改善创面修复质量,解决皮源问题开辟了崭新的途径。其中真皮替代物的研制是皮肤组织工程的基础也是技术关键。国外研究者已成功地开发出了多种真皮替代物,并已初步应用于深度烧伤创面的修复及瘢痕整形等领域,取得了一定临床效果,但是制品的价格非常昂贵。因此,研制和开发适合我国国情和经济实用的高质量真皮替代物具有重要的意义。
本研究应用组织工程学的原理和方法,用本实验室提取的Ⅰ型胶原复合壳聚糖制备出双层胶原-壳聚糖海绵支架,并对其理化和生物学性能进行了系统研究;用从大鼠皮肤中分离出皮肤成纤维细胞,将其种植在支架上体外构建了人工真皮替代物,并将人工真皮替代物移植到SD大鼠全层皮肤缺损创面上,得出以下结果:
1.以胶原和壳聚糖为原料,戊二醛为交联剂,采用冷冻干燥的方法制备了双层胶原-壳聚糖海绵支架;制备的胶原-壳聚糖海绵支架具有较高的孔隙率、良好的吸水性能和一定的力学强度;生物学评价结果证明,该支架无细胞毒性,无明显的皮肤刺激性;皮下埋植28天仍能观察到支架存在、具有良好的组织相容性,支架可在体内缓慢降解吸收。
2.成功地从新生SD大鼠皮肤中分离出成纤维细胞;将成纤维细胞种植到胶原-壳聚糖海绵支架上,在体外构建出了具有一定功能活性的人工真皮替代物。
3.通过建立的SD大鼠皮肤缺损模型,用构建的人工真皮替代物在SD大鼠脊背两侧皮肤进行了移植实验,对人工真皮替代物促进组织再生能力进行了评价。实验结果表明,该人工真皮替代物组织附着性、吸液性好,能促进伤口愈合。组织学检查结果表明,该人工真皮替代物具有引导组织形成,促进组织细胞生长的效果。
总之,本研究构建的活性人工真皮替代物理化性能和生物相容性好,可以作为人工真皮替代物用于皮肤缺损的修复。
Closure of full-thickness skin defects is important for the treatment of patients with burn, traumatic injury, chronic skin ulcer or hyperthopic scar. The traditional approach is grafting autologous skin, which often results in heavy pigmentation and scarring in donor sites, and the amount of skin obtained by this method is limited for patients with large area skin injuries. The technique of skin tissue engineering has been a new method to provide adequate skin grafts and improve wound healing quality. The development of dermal replacement is particular crucial to skin tissue engineering. Various dermal analogs have been developed and used as dermal template to cover full-thickness wounds successfully. But it is too expensive to afford. Therefore, development of economical dermal replacement with good quality is necessary.
In this study, the bilayer collagen-chitosan sponge scaffold was fabricated with type I collagen and chitosan, the structures and properties of the scaffold were studied. Dermal fibroblasts were isolated from SD rat skin, then the cells were transplanted into the sponge scaffold to construct the tissue engineered dermal substitutes in vitro. The artificial skin was transplanted into full-thickness wounds of SD rats. The main results of the thesis are as follows:
1. A porous scaffold with suitable pore sizes and internconnected pore structure was prepared with collagen and chitosan by freeze-drying, and glutaraldehyde (GA) was added to the scaffold as cross-linking bridges. The test indicated the scaffold had good mechanical property and water-absorbing ability. The biological evaluation tests showed that the collagen-chitosan dermal scaffold had no cytotoxicity and no remarkable sensitization for rabbit skin. Subcutaneous implantation indicated the collagen-chitosan dermal scaffold had good tissue compatibility. However, the scaffold was still observed on 28th day, which demonstrated that it could be biodegradated and absorbed slowly.
2. Dermal fibroblasts were isolated from SD rat skin successfully. We transplanted the fibroblasts on the collagen-chitosan scaffold to construct the dermal substitute in vitro. The artificial dermal substitute with functionally vitality could be called a living artificial dermal substitute.
3. The skin defect model was established in SD rats. The capcity of the artificial dermal substitute promoting tissue regeneration was evaluated by transplanting it on the back of the SD rats. The results of the experiment showed that the artificial dermal substitute was good at adhering the tissue and absorbing liquid and it could promote the wounds healing. The result of the histological observation showed that the artificial dermal substitute could guide the ingrowth of fresh tissues and promote the growth of epidermal cells and fibroblasts.
According to the results of this study, we concluded that the living artificialdermal substitute had not only good properties of physics and chemistry but alsogood biocompatibility. So it could become an artificial dermal substitute applied inrepairing the skin defect.
本研究应用组织工程学的原理和方法,用本实验室提取的Ⅰ型胶原复合壳聚糖制备出双层胶原-壳聚糖海绵支架,并对其理化和生物学性能进行了系统研究;用从大鼠皮肤中分离出皮肤成纤维细胞,将其种植在支架上体外构建了人工真皮替代物,并将人工真皮替代物移植到SD大鼠全层皮肤缺损创面上,得出以下结果:
1.以胶原和壳聚糖为原料,戊二醛为交联剂,采用冷冻干燥的方法制备了双层胶原-壳聚糖海绵支架;制备的胶原-壳聚糖海绵支架具有较高的孔隙率、良好的吸水性能和一定的力学强度;生物学评价结果证明,该支架无细胞毒性,无明显的皮肤刺激性;皮下埋植28天仍能观察到支架存在、具有良好的组织相容性,支架可在体内缓慢降解吸收。
2.成功地从新生SD大鼠皮肤中分离出成纤维细胞;将成纤维细胞种植到胶原-壳聚糖海绵支架上,在体外构建出了具有一定功能活性的人工真皮替代物。
3.通过建立的SD大鼠皮肤缺损模型,用构建的人工真皮替代物在SD大鼠脊背两侧皮肤进行了移植实验,对人工真皮替代物促进组织再生能力进行了评价。实验结果表明,该人工真皮替代物组织附着性、吸液性好,能促进伤口愈合。组织学检查结果表明,该人工真皮替代物具有引导组织形成,促进组织细胞生长的效果。
总之,本研究构建的活性人工真皮替代物理化性能和生物相容性好,可以作为人工真皮替代物用于皮肤缺损的修复。
Closure of full-thickness skin defects is important for the treatment of patients with burn, traumatic injury, chronic skin ulcer or hyperthopic scar. The traditional approach is grafting autologous skin, which often results in heavy pigmentation and scarring in donor sites, and the amount of skin obtained by this method is limited for patients with large area skin injuries. The technique of skin tissue engineering has been a new method to provide adequate skin grafts and improve wound healing quality. The development of dermal replacement is particular crucial to skin tissue engineering. Various dermal analogs have been developed and used as dermal template to cover full-thickness wounds successfully. But it is too expensive to afford. Therefore, development of economical dermal replacement with good quality is necessary.
In this study, the bilayer collagen-chitosan sponge scaffold was fabricated with type I collagen and chitosan, the structures and properties of the scaffold were studied. Dermal fibroblasts were isolated from SD rat skin, then the cells were transplanted into the sponge scaffold to construct the tissue engineered dermal substitutes in vitro. The artificial skin was transplanted into full-thickness wounds of SD rats. The main results of the thesis are as follows:
1. A porous scaffold with suitable pore sizes and internconnected pore structure was prepared with collagen and chitosan by freeze-drying, and glutaraldehyde (GA) was added to the scaffold as cross-linking bridges. The test indicated the scaffold had good mechanical property and water-absorbing ability. The biological evaluation tests showed that the collagen-chitosan dermal scaffold had no cytotoxicity and no remarkable sensitization for rabbit skin. Subcutaneous implantation indicated the collagen-chitosan dermal scaffold had good tissue compatibility. However, the scaffold was still observed on 28th day, which demonstrated that it could be biodegradated and absorbed slowly.
2. Dermal fibroblasts were isolated from SD rat skin successfully. We transplanted the fibroblasts on the collagen-chitosan scaffold to construct the dermal substitute in vitro. The artificial dermal substitute with functionally vitality could be called a living artificial dermal substitute.
3. The skin defect model was established in SD rats. The capcity of the artificial dermal substitute promoting tissue regeneration was evaluated by transplanting it on the back of the SD rats. The results of the experiment showed that the artificial dermal substitute was good at adhering the tissue and absorbing liquid and it could promote the wounds healing. The result of the histological observation showed that the artificial dermal substitute could guide the ingrowth of fresh tissues and promote the growth of epidermal cells and fibroblasts.
According to the results of this study, we concluded that the living artificialdermal substitute had not only good properties of physics and chemistry but alsogood biocompatibility. So it could become an artificial dermal substitute applied inrepairing the skin defect.
引文
[1]R.Langer,J.Vacanti.Tissue engineering[J].Science,1993,260:920-926.
[2]Brandt J,Nilsson A,Kanje M,et al.Acutely-dissociated Schwann cells used intendon autografts for bridging nerve defects in rats:A new principle for tissue engrneermg in nerve reconstruction[J].Scandinavian journal of plastic and reconstructive surgery and hand surgery,2005,39(6):321-325.
[3]杨志明.组织工程基础与临床[M].成都:四川科学技术出版社.2000.
[4]Marler JJ,Upton J,Langer R,et al.Transplantation of cells in matrices for tissue regeneration[J].Advanced Drug Delivery Reviews,1998,33(1-2):165-182.
[5]Saltzman WM,Parkhurst MR.,Parsons WP,et al.Three-dimensional cell cultures mimic tissues[J].Ann N YAcad Sci,1992,5:259.
[6]李雪盛,孙建军.组织工程种子细胞的研究进展[J].临床耳鼻咽喉科杂志,2004,18(5):313-314.
[7]Nerem RM.Cellular engineering[J].Ann Biomed Eng,1991,19:529-534.
[8]杨志明.组织工程[M].北京:化学工业出版社.2001.
[9]Watt FM,Hogan BL.Out of Eden:stem cells and their niches[J].Science,2000,287:1427-30.
[10]Weissman IL.Translating stem and progenitor cell biology to the clinic:barriers and opportunities[J].Science,2000,287(5457):1442-1446.
[11]Thomson JA.Embryonic stem cell lines derived from human blastocysts[J].Science,1998,282(5395):1827-1827.
[12]姚康德,尹玉姬.组织工程相关生物材料[M].北京:化学工业出版社.2003.
[13]Shin H,Jo S,Mikos AG.Biomimetic materials for tissue engineering[J].Biomaterials,2003,24(24):4353-4364.
[14]Maeda M,Tan S,Sano A,Fujioka K.Microstructure and release characteristics of the minipellet,a collagen based drug delivery system for controlled release of protein drugs[J].J.Controlled.Rel,1999,62:313-324.
[15]Jianbiao Ma,Hongyun Wang,He Binglin,et al.A preliminary in vitro study on the fabrication and tissue engineering applications of a novel chitosan bilayer material as a scaffold of human neofetal dermal fibroblasts[J].Biomaterials,2001,22(4):331.
[16]Kuen Yong Lee,David J.Mooney.Hydrogels for Tissue Engineering[J].Chemical Review,2001,101(7):1869-1877.
[17]Lie Ma,Changyou Gao,Zhengwei Mao,et al.Collagen/chitosan porous scaffolds with improved biostability for skin tissue engineering[J].Biomaterials,2003,24:4833-4841.
[18]王身国.组织工程细胞支架及其相关技术研究[J].现代康复.2001,5(8):16-18.
[19]W.T.Jodbey,A.Atala.In Vitro Systems for Tissue Engineering[J].Ann.N.Y.Acad Sci,2002,961:10-26.
[20]Saltzman WM.Weaving cartilage at zerog:the reality of tissue engineering in space[J].Proc Natl Acad Sci USA,1997,94(25):1338.
[21]胡江,陶祖莱.组织工程研究进展[J].生物医学工程学杂志.2000,17(1):75-79.
[22]金岩.组织工程学原理与技术[M].西安:第四军医大学出版社,2004.
[23]Yang S,Leong KF,Du Z,et al.The design of scaffolds for use in tissue engineering[J].Tissue Engineering,2002,8(1):1-11.
[24]Kim BS,Mooney DJ.Development of biocompatible synthetic extracellular matrices for tissue engineering[J].Trends Biotechnol.1998,16:224-230.
[25]Gao C;Y,Li A,Feng LX,et al.Factors controlling surface morpHology of porous polystyrene membranes prepared by thermally induced phase separation[J].Acta Polym Sin,2000,49(4):323-328.
[26]P.X.MA,J.W.Choi.Biodegradable polymer scaffolds with well-defined interconnected spherical pore network[J].Tissue Engineering,2001,7:23-33.
[27]Guoping Chen,Takashi Ushida,Tetsuya Tateishi.Development of biodegradable porous scaffolds for tissue engineering[J].Materials Science and Engineering,2001,17:63-69.
[28]曹谊林.组织工程学的研究进展[J].中国美容医学,2005,14(2):134-135.
[29]Matts P,Grosick T,Rust R.Human body skin:Structure,function,and correlation with perceived personal care needs and treatment choice[J].Journal of the American Academy of Dermatology,2005,52(3):91.
[30]Warner RR,Myers MC,Taylor DA.Electron-probe analysis of human-skin-element concentration profiles[J].Journal of Investigative Dermatology,1988,90(1):78-85.
[31]陈东.皮肤的结构和功能[J].中文轻工科技,2000,1:15.
[32]时宇.皮肤的结构与保健[J].生物学通报,1995,30(9):22.
[33]季红星.皮肤创伤愈合最新研究进展[J].中国美容医学,2001,10(5):440-442.
[34]Martin.P.Wound healing-Aiming for perfect skin regeneration[J].Science,1997,276(5309):75-81.
[35]杨友竹译.创伤愈合分子生物学的进展一粘附分子的调控[J].日本医学介绍,2001,22(4):173.
[36]Adzick NS,LorenzZ HE Cells,matrix,growth-factors,and the surgeon-the biology of scarless fetal wound repair[J].Annals of surgery,1994,220(1):10-18.
[37]查锡良.整合蛋白介导的信号传导研究进展[J].生命科学,1997,19(5):221-223.
[38]Rudkin GH,Miller TA.Growth factor in surgery[J].Plas.Reconstr.Surg.,1996,97(2):469-476.
[39]Sundberg C,Ivarsson M,Gerdin B,et al.Pericytes as collagen-producing cells in excessive dermal scarring[J].Laboratory Investigation,1996,74(2):452-466.
[40]StepHen T.Semipermeable film dressing.In:Wound management and dressing[M].London:The PHarmaceutical Press,2000.
[41]Braun-Falco M.Gene therapy approaches in stimulating wound repair[J].Hautarzt,2002,53(4):238-243.
[42]Ravan.ti L,Kahari VM.Matrix metalloproteinases in wound repair[J].International journal of molecular medicine,2000,6(4):391-407.
[43]Green H,Kehinde O,Thomas J.Growth of cultured human epidermal cells intomultiple epithelia suitable for grafting[J].Proc Natl Acad Sci USA,1979,76(11):5665-5668.
[44]Naughton G.The Advanced Tissue Sciences Story[J].Sci Am,1999,280(4):84-85.
[45]Horch RE,Bannasch H,Stark GB.Culture human keratinocytes as a single cell suspension in fibrin glue combined with preserved dermal grafts enhance skinreconstruction in athymic mice full thickness wounds[J].Eur J Plast Surg,1999,22:209-213.
[46] Medalie DA, Eming SA, Tompkins Rq, et al. Evaluation of human skin, reconstituted from composite grafts of cultured keratinocytes and human acellular dermis transplanted to athymic mice[J]. J Invest Dermatol, 1996,107: 121-126.
[47].Butler CE, Orgill DP, Yannas IV, et al. Effect of keratinocytes seeding of collagen glycosaminoglycan membranes on the regeneration of skin in a porcinemodel[J]. Plast Reconstr Surg, 1998,101: 1572-1579.
[48] Horch RE, Debus M, Wagner Q, et al. Cultured human keratinocytes on type I collagen membranes to reconstitute the epidermis [J]. Tissue Engineering, 2000, 6: 53-58.
[49] Heck EL, Bergstresser PR, Baxter CR. Composite skin graft:frozen dermal allografts support the engarftment and expansion of autologous epidermis[J].J Trauma, 1985, 25(2):106-112.
[50] Cuono CB, Langdon R, Birchall N, et al. Composite autologous-allogeneic skin replacement:development and clinical application [J]. Plast Reconstr Surg, 1987, 80 (4):626-637.
[51] Cuono CB, Langdon R, McGuire J. Use of cultured epidermal autografts and dermal allografts as skin replacement after burn injury [J] .Lancet, 1986, l(8490):l 123-1124.
[52] Keis RW, Hoekstra MJ, Mackie DP et al. Historical appraisal of the use of skin allografts in the treatment of extensive full skin thickness burns at the red cross hospital burns center .Beverwijik,the Netherlands [J] .Burns, 1992,18 (2): 19-23.
[53] Schiozer, WA, Hartinger A, Henckel G, et al. Composite grafts of autogeneic cultured epidermis and glycerol-preserved allogeneic dermis for definitive coverage of full-thickness burn woundsxase reports[J]. Burns,1994,20 (6):503-507.
[54] Mizuno H, Takeda A, Uchinuma E. Creation of an acellular dermal matrix from frozen skin[J].Aesthetic Plast Surg, 1999, 23(5):316-322.
[55] Takami K, Matsuda T, Yoshitake M, et al. Dispase/detergent treated dermal matrix as a dermal substitute[J].Burns, 1996,22 (3):182-190.
[56] Wainwright DJ. Use of an acellular allograft dermal matrix (Alloderm) in the management of full-thickness burns[J].Burns,1995,21(4):243-248.
[57]Livesey SA,Herndon DN,Hollyoak MA,et al.Transplanted acellular allograft dermal matrix:potential as a template for the reconstruction of viable dermis[J].Transplantation,1995,60(1):1-9.
[58]姜笃银,陈壁,贾赤宇.戊二醛交联的异种/异体脱细胞真皮基质的制作及临床应用观察[J].第四军医大学学报,1999,20(5):371-374.
[59]Boyce ST,Kagan RJ,Meyer NA,et al.Cultured skin substitutes combined with Integra artificial skin to replace native skin autograft and allograft for the closure of excised full-thickness burns[J].J Burn Care Rehabil,1999,20(6):453-461.
[60]Supp AP,Wickett RR,Swope VB,et al.Incubation of cultured skin substitutes in reduced humidity promotes cornification in vitro and stable engraftment in athymic mice[J].Wound Repair Regen,1999,7(4):226-237.
[61]Harriger MD,Warden GD,Greenhalgh DG,et al.Pigmentation and micro anatomy of skin regenerated from composite grafts of cultured cells and biopolymers applied to full-thickness burn wounds[J].Transplantation,1995,59(5):702-707.
[62]Hansbrough JF,Cooper ML,Spievogel RS,et al.Evaluation of a biodegradable matrix containing cultured human fibroblasts as a dermal replacement beneath meshed skin grafts on athymic mice[J].Surgery,1992,111(4):438-446.
[63]Hansbrough JF,Dore C,Hasbrough WB.Clinical trials of a living dermal tissue replacement placed beneathed meshed,split-thickness skin grafts on excised burn wounds[J].J Burn Care Rehabil,1992,13(5):519-629.
[64]Choi YS,Hong SR,Lee YM,et al.Studies on gelatin-containing artificial skin:Ⅱ.Preparation and characterization of cross-linked gelatin-hyaluronate sponge[J].J Biomed Mater Res,1999,48(5):631-639.
[65]Falanga V,Margolis D,Alarez O,et al.Rapid healing of venous ulcer and lack of clinical rejection with an allogenic cultured human skin equivalent[J].Arch Dermatol.,1997,134:293-300.
[66]Boyce ST.Skin substitutes from cultured cells and collagen GAG polymers[J].Med Biol Eng Comput,1998,36:791-797.
[67]Zacchi V,Soranzo C,Cortivo R,et al.In vitro engineering of human skin like tissue[J].J Biomed mater Res,1998,40:187-192.
[68] Sabolinski M, Alvarez O, Auletta M, et al. Cultured skin as a"smart material"for healing wounds: experience in venous ulcers[J]. Biomaterials, 1996,17(3):311-318.
[69] Lukish JR, Eichelberger MR, Newman KD, et al. The use of a bioactive skin substitute decreases length of stay for pediatric burn patients[J]. Journal of Pediatric Surgery, 2001,36(8): 1121-1128.
[70] Wisser D. Skin replacement with a collagen based dermal substitute, autologous keratinocytes and fibroblasts in burn trauma[J]. Burns,2003,29:375-381.
[71] Hunt J. A. Initial experience of Integra in the treatment of post-burn anterior cervical neck contracture[J]. British Journal of Plastic Surger, 2000,53: 652-659.
[72] Falanga V, Margolis D, Alvarez O, et al. Rapid healing of venous ulcers and lack of clinical rejection with an allogeneic human skin equivalent[J]. Arch Dermatol, 1998,134:293-299.
[73] Susan M. Repair of cicatricial ectropion in an infant with harlequin ichthyosis using engineered human skin[J]. American Journal of OpHthalmology, 2002,134(3): 442-448.
[74] Barbora DvoRankova, MSc, Zuzana Holikovfi, MD.Reconstruction of epidermis by grafting of keratinocytes cultured on polymer support-clinical study. International Journal of Dermatology. 2003,42: 219-223.
[75] Veves A, Falanga MArmstrong Dg, et al. The Apligraf diabetic foot ulcer study:graftskin, a human skin equivalent, is effective in the management of noninfected neuropathic diabetic foot ulcers: a prospective randomized multicenter clinical trial. Diabetes Care. 2001, 24: 290-296.
[76] Maier JP, Lippitt C, Mooney EK, et al. Use of tissue-engineered skin in the dermal atropHy patient with traumatic avulsion injuries[J]. Ann Plast Surg, 2002,49(1 ):67-73.
[77] Susan M. Repair of cicatricial ectropion in an infant with harlequin ichthyosis using engineered human skin[J]. American Journal of OpHthalmology, 2002,134(3): 442-443.
[78] Meggan N, Banta MD, Robert S, et al. Modulating Diseased Skin with Tissue Engineering: Actinic Purpura Treated with Apligraf[J]. Dermatol Surg, 2002,28(12): 1103-1110.
[79] OpHof Rvan, Rheden RE, Von den HJ, et al Oral keratinocytes cultured on dermal matrices form a mucosa-like tissue[J]. Biomaterials, 2002, 23(17):3741-3478.
[80] Kearney JN. Clinical evaluation of skin substitutes [J]. Bums,2001,27:545-551.
[81]Gallico GG.Biologic skin substitutes[J].Clin Plast Surg,1990,17(3):519-526.
[82]Brain A,Purkis P,Coates P,et al.Survival of cultured allogeneic keratinocytes transplanted to deep dermal bed assessed with probe specific for Y chromosome[J].Br Med J,1989,298:917-919.
[83]Sheridan RL,Tomplcin RCS.Skin substitutes in bums[J].Bums,1999,2597:103-110.
[84]Black AF,Berthod F,L' heureux,et al.In vitro reconstruction of a human capillary like network in tissue engineering skin equivalent[J].FASEB,1998,12:13-16.
[85]李荟元.创伤动物研究模型—兔耳瘢痕模型的建立与应用[M].西安:第四军医大学出版社,2005.
[86]付小兵,王德文.创伤修复基础[M].北京:人民军医出版社,1997.
[87]付小兵,王亚平,常国友,等.碱性成纤维细胞生长因子加速小型猪背部创伤修复的实验研究[J].中华创伤杂志,1995,11:134-136.
[88]Davidson JM.Animal models for wound repair[J].Arch Dermatol Res,1998,290:1211.
[89]Livesey SA,Herndon DN,Hollyoak MA,et al.Transplanted acellular allograft dermal matrix:potentital as a temp late for the reconstruction of viable derm is[J].Transplantation,1995,60:1.
[90]Jameela SR,Jayakrishnan A.Glutaraldehyde crosslinked chitosan microspHeres as a long acting biodegradable drug delivery vehicle:studies on the in vitro release of mitoxantrone and in vivo degradation of microspHeres in rat muscle[J].Biomaterials,1995,16:769-775.
[91]Shanmugasundaram N,Ravichandran P,Reddy PN,et al.Collagen-chitosan polymeric scaffolds for the in vitro culture of human epidermoid carcinoma cells[J].Biomaterial,2001,22:1943-1951.
[92]Stone H,Wright T,Clarke R,et al.Healing at skin graft donor sites dressed with chitosan[J].Br J Plast Surg,2000,53:601-606.
[93]Hongyi Yang,Qiqing Zhang.Preparation and Characterization of Collagen-GAGs Bioactive Matrices for Tissue Engineering[J].J Mater Sci Technol,2001,17:495-500.
[1]Donate I,Stredanska S,Silvestrini G,et al.The aggregation of pig articular chondrocyte and synthesis of extracellular matrix by a lactose-modified chitosan[J].Biomaterials,2005,26:987-998.
[2]Tomihata K,Ilcada Y.In vitro and in vivo degradation of films of chitin and its deacetylated derivatives[J].Biomaterials,1997,18:567-573.
[3]Muzzarelli R,Tarsi R,Filippini O,et al..Antimicrobial properties of N-carboncybutyl chitosan[J].Antimicrob Agents Chemother,1990,34:2019-2023.
[4]HongyiYang,Qiqing Zhang.Preparation and Characterization of Collagen-GAGs Bioactive Matrices for Tissue Engineering[J].J Mater Sci.Technol,2001,17(5):495-500.
[5]关嫚,任磊,吴婷,等.新型双层皮肤组织工程支架构建[J].厦门大学学报,2006,45:379-382.
[6]Shapiro L,Cohen S.Novel alginate sponges for cell culture and transplantation[J].Biomaterials,1997,18:583-590.
[7]黄志海,董寅生,郭超,等.壳聚糖—胶原多孔支架制备研究[J].化工时刊,2004,18(10):36-39.
[8]Ma Jianbiao,Wang Hongjun,He Binglin,et al.A preliminary in vitro study on the fabrication and tissue engineering applications of a novel chitosan bilayer material as a scaffold of human neofetal dermal fibroblasts[J].Biomaterials,2001,22:331-336.
[9]Ye Fen,YinYuji,Sun Guangjie,et al.The applications of chitosan scaffolds for tissue engineering[J].Functional Materials,2002,33(5):359-461.
[10]中华人民共和国药典[M].北京:人民卫生出版社,1990.
[11]杨晓芳.生物材料生物相容性评价研究进展[J].生物医学工程学杂志,2001,18(1):123-128.
[12]王剑虹,黄庆科,陈民新.抑制肝癌细胞的核转录因子活性增强阿霉素的抗癌作用[J].中华肝脏病杂志,2004,6:375.
[13]陈耀富,印木泉,贺清玉.可吸收止血敷料的细胞毒性和致突性研究[J].第二军医大学学报,1997,18(2):166-169.
[14]宁丽,薛森,黄海宁等.皮肤再生膜的生物相容性系列研究[J].中国修复重建外科杂志,2000,14(1):44-48.
[15]Friess W.Collagen-biomaterial for drug delivery[J].Eur J PHarm.BiopHarm,1998,45:113-136.
[16]Maeda M,Tani S,Sano A,Fujioka K.Microstructure and release characteristics of the minipellet,a collagen based drug delivery system for controlled release of protein drugs[J].J Controlled Rel,1999,62:313-324.
[17]刘晋宇,侯立中,颜炜群,等.组织工程化人工复合皮的构建[J].中国修复重建外科杂志,2001,15:235-239.
[18]杨君,肖仕初,夏照帆.海绵状胶原膜真皮替代物的研制及实验研究[J].中华烧伤杂志,2002,18:23-25.
[19]郑化,杜予民,余家会,等.交联壳聚糖膜的制备及其性能的研究[J].高等学校化学学报,2000,21(5):809-812.
[20]叶勇,王坤余,廖隆理,等.胶原蛋白—壳聚糖—聚乙烯醇共混膜的特性研究[J].中国皮革,2003,32(23):1-4.
[21]Gao Changyou,Wang Dengyong,Shen Jiacong.Fabrication of porous collagen-chitosan scaffolds with controlling microstructure for dermal equivalent[J].Polymers for Advanced Technologies,2003,14(6):373-379.
[22]顾震宇,李文俊.壳聚糖溶液冷冻诱导相分离—一种制备高孔隙率膜的新方法[J].科学通报,1999,44(9):933-937.
[23]Chen Guoping,Ushida T,Tateishi T.Development of biodegradable porous scaffolds for tissue engineering[J].Materials Science and Engineering,2001,17(1):63-69.
[24]Yannas IV,Burke JF,Gordon PL,et al.Design of an artificial skin Ⅱ.Control of chemical composition[J].J Biomed Mater Res,1980,14(4):107-131.
[25]Lie Ma,Changyou Gao,Zhengwei Mao,et al.Collagen/chitosan porous scaffolds with improved biostability for skin tissue engineering[J].Biomaterials,2003,24:4833-4841
[1]Harshad A.Navsaria,Simon R.Myers,Irene M.Leigh,et al.Culturing skin in vitro for wound therapy[J].Tibtech,1995,13:91-100.
[2]Gallico GG.Biologic skin substitutes[J].ClinPlast Surg,1990,17:519-526.
[3]Hongyi Yang,Qiqing Zhang.Preparation and Characterization of Collagen-GAGs Bioactive Matrices for Tissue Engineering[J].J Mater Sci.Technol,2001,17(5):495-500.
[4]R.I.弗雷谢尼.动物细胞培养[M].北京:科学出版社,2004.
[5]马刚,王胜,陈世义,等.皮肤成纤维细胞收缩功能的研究[J],白求恩医科大学学报,1996,22(3):272-273.
[6]C.FA.Culling.组织病理学与组织化学技术手册[M].北京:科学出版社,1982.
[7]曹谊林,刘伟,崔磊.组织工程学理论与实践[M].上海:上海科学技术出版社,2004.
[8]Garner WL.Epidermal regulation of dermal fibroblast activity[J].Plast Reconstr surg,1998,102(1):135-139.
[9]Demarchez M,Hartmarm D J,Regnier M,et al.The role of fibroblasts in dermal vascularization and remodeling of reconstructed human skin after transplantation onto the nude mouse[J].Transplantation,1992,54(2):317-326.
[10]Coulomb B,Friteau L,Baruch J,et al.Advantage of the presence of living fibroblasts within in vitro reconstructed skin for grafting in human[J].Plast Reconstr Surg,1998,101(7):1891-1903.
[11]Berthod F,Sahuc F,Hayek D,et al.Deposition of collagen fibril bundles by long-term culture of fibroblasts in a collagen sponge[J].Biomed Mater Res,1996,32(1):87-93.
[12]Lamme EN,Van Leeuwen RT,Brandsma K,et al.Higher numbers of autologous fibroblasts in an artificial dermal substitute improve tissue regeneration and modulate scar tissue formation[J].J Pathos,2000,190(5):595-603.
[13]陈光,周平,潘銮凤,等.天然生物材料壳聚糖支架上人胚肺成纤维细胞的生长[J].化学学报,2004,62(10):992-997.
[1]Steven T,Boyce.Fabrication,quality assurance,and assessment of cultured skin substitutes for treatment of skin wounds[J].Biochemical Engineering Journal,2004,20:107-112.
[2]Belasubramani M,Kumar TR,Babu M.Skin substitutes:a review[J].Bums,2001,27:534-544.
[3]Hunt JA,Van Der Laan JS,Schakenraad J,et al.Quantitative in vivo assessment of tissue response to dermal sheep collagen in abdominal wall defects[J].Biomaterials,1993,14:378-382.
[4]Livesey SA,Hemdon DN,Hollyoak MA,et al.Transplanted acellular allograft dermal matrix:potentital as a temp late for the reconstruction of viable derm is[J].Transplantation,1995,60:1.
[5]HongyiYang,Qiqing Zhang.Preparation and Characterization of Collagen-GAGs Bioactive Matrices for Tissue Engineering[J].J Mater Sci.Technol,2001,17(5):495-500.
[6]郑军,黄晓元,韦星.重组人表皮生长因子促进大鼠皮肤创面愈合的研究[J].中华整形外科杂志,2005,21:379-383.
[7]付小兵,王德文.现代创伤修复学[M].北京:人民军医出版社,1999.
[8]郝光荣.实验动物学[M].上海:第二军医大学出版社,1999.
[9]付小兵,孙同柱,盛志勇.几种用于创伤修复研究的动物模型[J].中华实验外科杂志,2001,12:40-43.
[10]Shahabeddin L,Berthod F,Damour O,et al.Characterization of skin reconstructed on chitosan cross linked collagen glycosam inoglycan matrix[J].Skin PHarmacol,1990,3:107.
[2]Brandt J,Nilsson A,Kanje M,et al.Acutely-dissociated Schwann cells used intendon autografts for bridging nerve defects in rats:A new principle for tissue engrneermg in nerve reconstruction[J].Scandinavian journal of plastic and reconstructive surgery and hand surgery,2005,39(6):321-325.
[3]杨志明.组织工程基础与临床[M].成都:四川科学技术出版社.2000.
[4]Marler JJ,Upton J,Langer R,et al.Transplantation of cells in matrices for tissue regeneration[J].Advanced Drug Delivery Reviews,1998,33(1-2):165-182.
[5]Saltzman WM,Parkhurst MR.,Parsons WP,et al.Three-dimensional cell cultures mimic tissues[J].Ann N YAcad Sci,1992,5:259.
[6]李雪盛,孙建军.组织工程种子细胞的研究进展[J].临床耳鼻咽喉科杂志,2004,18(5):313-314.
[7]Nerem RM.Cellular engineering[J].Ann Biomed Eng,1991,19:529-534.
[8]杨志明.组织工程[M].北京:化学工业出版社.2001.
[9]Watt FM,Hogan BL.Out of Eden:stem cells and their niches[J].Science,2000,287:1427-30.
[10]Weissman IL.Translating stem and progenitor cell biology to the clinic:barriers and opportunities[J].Science,2000,287(5457):1442-1446.
[11]Thomson JA.Embryonic stem cell lines derived from human blastocysts[J].Science,1998,282(5395):1827-1827.
[12]姚康德,尹玉姬.组织工程相关生物材料[M].北京:化学工业出版社.2003.
[13]Shin H,Jo S,Mikos AG.Biomimetic materials for tissue engineering[J].Biomaterials,2003,24(24):4353-4364.
[14]Maeda M,Tan S,Sano A,Fujioka K.Microstructure and release characteristics of the minipellet,a collagen based drug delivery system for controlled release of protein drugs[J].J.Controlled.Rel,1999,62:313-324.
[15]Jianbiao Ma,Hongyun Wang,He Binglin,et al.A preliminary in vitro study on the fabrication and tissue engineering applications of a novel chitosan bilayer material as a scaffold of human neofetal dermal fibroblasts[J].Biomaterials,2001,22(4):331.
[16]Kuen Yong Lee,David J.Mooney.Hydrogels for Tissue Engineering[J].Chemical Review,2001,101(7):1869-1877.
[17]Lie Ma,Changyou Gao,Zhengwei Mao,et al.Collagen/chitosan porous scaffolds with improved biostability for skin tissue engineering[J].Biomaterials,2003,24:4833-4841.
[18]王身国.组织工程细胞支架及其相关技术研究[J].现代康复.2001,5(8):16-18.
[19]W.T.Jodbey,A.Atala.In Vitro Systems for Tissue Engineering[J].Ann.N.Y.Acad Sci,2002,961:10-26.
[20]Saltzman WM.Weaving cartilage at zerog:the reality of tissue engineering in space[J].Proc Natl Acad Sci USA,1997,94(25):1338.
[21]胡江,陶祖莱.组织工程研究进展[J].生物医学工程学杂志.2000,17(1):75-79.
[22]金岩.组织工程学原理与技术[M].西安:第四军医大学出版社,2004.
[23]Yang S,Leong KF,Du Z,et al.The design of scaffolds for use in tissue engineering[J].Tissue Engineering,2002,8(1):1-11.
[24]Kim BS,Mooney DJ.Development of biocompatible synthetic extracellular matrices for tissue engineering[J].Trends Biotechnol.1998,16:224-230.
[25]Gao C;Y,Li A,Feng LX,et al.Factors controlling surface morpHology of porous polystyrene membranes prepared by thermally induced phase separation[J].Acta Polym Sin,2000,49(4):323-328.
[26]P.X.MA,J.W.Choi.Biodegradable polymer scaffolds with well-defined interconnected spherical pore network[J].Tissue Engineering,2001,7:23-33.
[27]Guoping Chen,Takashi Ushida,Tetsuya Tateishi.Development of biodegradable porous scaffolds for tissue engineering[J].Materials Science and Engineering,2001,17:63-69.
[28]曹谊林.组织工程学的研究进展[J].中国美容医学,2005,14(2):134-135.
[29]Matts P,Grosick T,Rust R.Human body skin:Structure,function,and correlation with perceived personal care needs and treatment choice[J].Journal of the American Academy of Dermatology,2005,52(3):91.
[30]Warner RR,Myers MC,Taylor DA.Electron-probe analysis of human-skin-element concentration profiles[J].Journal of Investigative Dermatology,1988,90(1):78-85.
[31]陈东.皮肤的结构和功能[J].中文轻工科技,2000,1:15.
[32]时宇.皮肤的结构与保健[J].生物学通报,1995,30(9):22.
[33]季红星.皮肤创伤愈合最新研究进展[J].中国美容医学,2001,10(5):440-442.
[34]Martin.P.Wound healing-Aiming for perfect skin regeneration[J].Science,1997,276(5309):75-81.
[35]杨友竹译.创伤愈合分子生物学的进展一粘附分子的调控[J].日本医学介绍,2001,22(4):173.
[36]Adzick NS,LorenzZ HE Cells,matrix,growth-factors,and the surgeon-the biology of scarless fetal wound repair[J].Annals of surgery,1994,220(1):10-18.
[37]查锡良.整合蛋白介导的信号传导研究进展[J].生命科学,1997,19(5):221-223.
[38]Rudkin GH,Miller TA.Growth factor in surgery[J].Plas.Reconstr.Surg.,1996,97(2):469-476.
[39]Sundberg C,Ivarsson M,Gerdin B,et al.Pericytes as collagen-producing cells in excessive dermal scarring[J].Laboratory Investigation,1996,74(2):452-466.
[40]StepHen T.Semipermeable film dressing.In:Wound management and dressing[M].London:The PHarmaceutical Press,2000.
[41]Braun-Falco M.Gene therapy approaches in stimulating wound repair[J].Hautarzt,2002,53(4):238-243.
[42]Ravan.ti L,Kahari VM.Matrix metalloproteinases in wound repair[J].International journal of molecular medicine,2000,6(4):391-407.
[43]Green H,Kehinde O,Thomas J.Growth of cultured human epidermal cells intomultiple epithelia suitable for grafting[J].Proc Natl Acad Sci USA,1979,76(11):5665-5668.
[44]Naughton G.The Advanced Tissue Sciences Story[J].Sci Am,1999,280(4):84-85.
[45]Horch RE,Bannasch H,Stark GB.Culture human keratinocytes as a single cell suspension in fibrin glue combined with preserved dermal grafts enhance skinreconstruction in athymic mice full thickness wounds[J].Eur J Plast Surg,1999,22:209-213.
[46] Medalie DA, Eming SA, Tompkins Rq, et al. Evaluation of human skin, reconstituted from composite grafts of cultured keratinocytes and human acellular dermis transplanted to athymic mice[J]. J Invest Dermatol, 1996,107: 121-126.
[47].Butler CE, Orgill DP, Yannas IV, et al. Effect of keratinocytes seeding of collagen glycosaminoglycan membranes on the regeneration of skin in a porcinemodel[J]. Plast Reconstr Surg, 1998,101: 1572-1579.
[48] Horch RE, Debus M, Wagner Q, et al. Cultured human keratinocytes on type I collagen membranes to reconstitute the epidermis [J]. Tissue Engineering, 2000, 6: 53-58.
[49] Heck EL, Bergstresser PR, Baxter CR. Composite skin graft:frozen dermal allografts support the engarftment and expansion of autologous epidermis[J].J Trauma, 1985, 25(2):106-112.
[50] Cuono CB, Langdon R, Birchall N, et al. Composite autologous-allogeneic skin replacement:development and clinical application [J]. Plast Reconstr Surg, 1987, 80 (4):626-637.
[51] Cuono CB, Langdon R, McGuire J. Use of cultured epidermal autografts and dermal allografts as skin replacement after burn injury [J] .Lancet, 1986, l(8490):l 123-1124.
[52] Keis RW, Hoekstra MJ, Mackie DP et al. Historical appraisal of the use of skin allografts in the treatment of extensive full skin thickness burns at the red cross hospital burns center .Beverwijik,the Netherlands [J] .Burns, 1992,18 (2): 19-23.
[53] Schiozer, WA, Hartinger A, Henckel G, et al. Composite grafts of autogeneic cultured epidermis and glycerol-preserved allogeneic dermis for definitive coverage of full-thickness burn woundsxase reports[J]. Burns,1994,20 (6):503-507.
[54] Mizuno H, Takeda A, Uchinuma E. Creation of an acellular dermal matrix from frozen skin[J].Aesthetic Plast Surg, 1999, 23(5):316-322.
[55] Takami K, Matsuda T, Yoshitake M, et al. Dispase/detergent treated dermal matrix as a dermal substitute[J].Burns, 1996,22 (3):182-190.
[56] Wainwright DJ. Use of an acellular allograft dermal matrix (Alloderm) in the management of full-thickness burns[J].Burns,1995,21(4):243-248.
[57]Livesey SA,Herndon DN,Hollyoak MA,et al.Transplanted acellular allograft dermal matrix:potential as a template for the reconstruction of viable dermis[J].Transplantation,1995,60(1):1-9.
[58]姜笃银,陈壁,贾赤宇.戊二醛交联的异种/异体脱细胞真皮基质的制作及临床应用观察[J].第四军医大学学报,1999,20(5):371-374.
[59]Boyce ST,Kagan RJ,Meyer NA,et al.Cultured skin substitutes combined with Integra artificial skin to replace native skin autograft and allograft for the closure of excised full-thickness burns[J].J Burn Care Rehabil,1999,20(6):453-461.
[60]Supp AP,Wickett RR,Swope VB,et al.Incubation of cultured skin substitutes in reduced humidity promotes cornification in vitro and stable engraftment in athymic mice[J].Wound Repair Regen,1999,7(4):226-237.
[61]Harriger MD,Warden GD,Greenhalgh DG,et al.Pigmentation and micro anatomy of skin regenerated from composite grafts of cultured cells and biopolymers applied to full-thickness burn wounds[J].Transplantation,1995,59(5):702-707.
[62]Hansbrough JF,Cooper ML,Spievogel RS,et al.Evaluation of a biodegradable matrix containing cultured human fibroblasts as a dermal replacement beneath meshed skin grafts on athymic mice[J].Surgery,1992,111(4):438-446.
[63]Hansbrough JF,Dore C,Hasbrough WB.Clinical trials of a living dermal tissue replacement placed beneathed meshed,split-thickness skin grafts on excised burn wounds[J].J Burn Care Rehabil,1992,13(5):519-629.
[64]Choi YS,Hong SR,Lee YM,et al.Studies on gelatin-containing artificial skin:Ⅱ.Preparation and characterization of cross-linked gelatin-hyaluronate sponge[J].J Biomed Mater Res,1999,48(5):631-639.
[65]Falanga V,Margolis D,Alarez O,et al.Rapid healing of venous ulcer and lack of clinical rejection with an allogenic cultured human skin equivalent[J].Arch Dermatol.,1997,134:293-300.
[66]Boyce ST.Skin substitutes from cultured cells and collagen GAG polymers[J].Med Biol Eng Comput,1998,36:791-797.
[67]Zacchi V,Soranzo C,Cortivo R,et al.In vitro engineering of human skin like tissue[J].J Biomed mater Res,1998,40:187-192.
[68] Sabolinski M, Alvarez O, Auletta M, et al. Cultured skin as a"smart material"for healing wounds: experience in venous ulcers[J]. Biomaterials, 1996,17(3):311-318.
[69] Lukish JR, Eichelberger MR, Newman KD, et al. The use of a bioactive skin substitute decreases length of stay for pediatric burn patients[J]. Journal of Pediatric Surgery, 2001,36(8): 1121-1128.
[70] Wisser D. Skin replacement with a collagen based dermal substitute, autologous keratinocytes and fibroblasts in burn trauma[J]. Burns,2003,29:375-381.
[71] Hunt J. A. Initial experience of Integra in the treatment of post-burn anterior cervical neck contracture[J]. British Journal of Plastic Surger, 2000,53: 652-659.
[72] Falanga V, Margolis D, Alvarez O, et al. Rapid healing of venous ulcers and lack of clinical rejection with an allogeneic human skin equivalent[J]. Arch Dermatol, 1998,134:293-299.
[73] Susan M. Repair of cicatricial ectropion in an infant with harlequin ichthyosis using engineered human skin[J]. American Journal of OpHthalmology, 2002,134(3): 442-448.
[74] Barbora DvoRankova, MSc, Zuzana Holikovfi, MD.Reconstruction of epidermis by grafting of keratinocytes cultured on polymer support-clinical study. International Journal of Dermatology. 2003,42: 219-223.
[75] Veves A, Falanga MArmstrong Dg, et al. The Apligraf diabetic foot ulcer study:graftskin, a human skin equivalent, is effective in the management of noninfected neuropathic diabetic foot ulcers: a prospective randomized multicenter clinical trial. Diabetes Care. 2001, 24: 290-296.
[76] Maier JP, Lippitt C, Mooney EK, et al. Use of tissue-engineered skin in the dermal atropHy patient with traumatic avulsion injuries[J]. Ann Plast Surg, 2002,49(1 ):67-73.
[77] Susan M. Repair of cicatricial ectropion in an infant with harlequin ichthyosis using engineered human skin[J]. American Journal of OpHthalmology, 2002,134(3): 442-443.
[78] Meggan N, Banta MD, Robert S, et al. Modulating Diseased Skin with Tissue Engineering: Actinic Purpura Treated with Apligraf[J]. Dermatol Surg, 2002,28(12): 1103-1110.
[79] OpHof Rvan, Rheden RE, Von den HJ, et al Oral keratinocytes cultured on dermal matrices form a mucosa-like tissue[J]. Biomaterials, 2002, 23(17):3741-3478.
[80] Kearney JN. Clinical evaluation of skin substitutes [J]. Bums,2001,27:545-551.
[81]Gallico GG.Biologic skin substitutes[J].Clin Plast Surg,1990,17(3):519-526.
[82]Brain A,Purkis P,Coates P,et al.Survival of cultured allogeneic keratinocytes transplanted to deep dermal bed assessed with probe specific for Y chromosome[J].Br Med J,1989,298:917-919.
[83]Sheridan RL,Tomplcin RCS.Skin substitutes in bums[J].Bums,1999,2597:103-110.
[84]Black AF,Berthod F,L' heureux,et al.In vitro reconstruction of a human capillary like network in tissue engineering skin equivalent[J].FASEB,1998,12:13-16.
[85]李荟元.创伤动物研究模型—兔耳瘢痕模型的建立与应用[M].西安:第四军医大学出版社,2005.
[86]付小兵,王德文.创伤修复基础[M].北京:人民军医出版社,1997.
[87]付小兵,王亚平,常国友,等.碱性成纤维细胞生长因子加速小型猪背部创伤修复的实验研究[J].中华创伤杂志,1995,11:134-136.
[88]Davidson JM.Animal models for wound repair[J].Arch Dermatol Res,1998,290:1211.
[89]Livesey SA,Herndon DN,Hollyoak MA,et al.Transplanted acellular allograft dermal matrix:potentital as a temp late for the reconstruction of viable derm is[J].Transplantation,1995,60:1.
[90]Jameela SR,Jayakrishnan A.Glutaraldehyde crosslinked chitosan microspHeres as a long acting biodegradable drug delivery vehicle:studies on the in vitro release of mitoxantrone and in vivo degradation of microspHeres in rat muscle[J].Biomaterials,1995,16:769-775.
[91]Shanmugasundaram N,Ravichandran P,Reddy PN,et al.Collagen-chitosan polymeric scaffolds for the in vitro culture of human epidermoid carcinoma cells[J].Biomaterial,2001,22:1943-1951.
[92]Stone H,Wright T,Clarke R,et al.Healing at skin graft donor sites dressed with chitosan[J].Br J Plast Surg,2000,53:601-606.
[93]Hongyi Yang,Qiqing Zhang.Preparation and Characterization of Collagen-GAGs Bioactive Matrices for Tissue Engineering[J].J Mater Sci Technol,2001,17:495-500.
[1]Donate I,Stredanska S,Silvestrini G,et al.The aggregation of pig articular chondrocyte and synthesis of extracellular matrix by a lactose-modified chitosan[J].Biomaterials,2005,26:987-998.
[2]Tomihata K,Ilcada Y.In vitro and in vivo degradation of films of chitin and its deacetylated derivatives[J].Biomaterials,1997,18:567-573.
[3]Muzzarelli R,Tarsi R,Filippini O,et al..Antimicrobial properties of N-carboncybutyl chitosan[J].Antimicrob Agents Chemother,1990,34:2019-2023.
[4]HongyiYang,Qiqing Zhang.Preparation and Characterization of Collagen-GAGs Bioactive Matrices for Tissue Engineering[J].J Mater Sci.Technol,2001,17(5):495-500.
[5]关嫚,任磊,吴婷,等.新型双层皮肤组织工程支架构建[J].厦门大学学报,2006,45:379-382.
[6]Shapiro L,Cohen S.Novel alginate sponges for cell culture and transplantation[J].Biomaterials,1997,18:583-590.
[7]黄志海,董寅生,郭超,等.壳聚糖—胶原多孔支架制备研究[J].化工时刊,2004,18(10):36-39.
[8]Ma Jianbiao,Wang Hongjun,He Binglin,et al.A preliminary in vitro study on the fabrication and tissue engineering applications of a novel chitosan bilayer material as a scaffold of human neofetal dermal fibroblasts[J].Biomaterials,2001,22:331-336.
[9]Ye Fen,YinYuji,Sun Guangjie,et al.The applications of chitosan scaffolds for tissue engineering[J].Functional Materials,2002,33(5):359-461.
[10]中华人民共和国药典[M].北京:人民卫生出版社,1990.
[11]杨晓芳.生物材料生物相容性评价研究进展[J].生物医学工程学杂志,2001,18(1):123-128.
[12]王剑虹,黄庆科,陈民新.抑制肝癌细胞的核转录因子活性增强阿霉素的抗癌作用[J].中华肝脏病杂志,2004,6:375.
[13]陈耀富,印木泉,贺清玉.可吸收止血敷料的细胞毒性和致突性研究[J].第二军医大学学报,1997,18(2):166-169.
[14]宁丽,薛森,黄海宁等.皮肤再生膜的生物相容性系列研究[J].中国修复重建外科杂志,2000,14(1):44-48.
[15]Friess W.Collagen-biomaterial for drug delivery[J].Eur J PHarm.BiopHarm,1998,45:113-136.
[16]Maeda M,Tani S,Sano A,Fujioka K.Microstructure and release characteristics of the minipellet,a collagen based drug delivery system for controlled release of protein drugs[J].J Controlled Rel,1999,62:313-324.
[17]刘晋宇,侯立中,颜炜群,等.组织工程化人工复合皮的构建[J].中国修复重建外科杂志,2001,15:235-239.
[18]杨君,肖仕初,夏照帆.海绵状胶原膜真皮替代物的研制及实验研究[J].中华烧伤杂志,2002,18:23-25.
[19]郑化,杜予民,余家会,等.交联壳聚糖膜的制备及其性能的研究[J].高等学校化学学报,2000,21(5):809-812.
[20]叶勇,王坤余,廖隆理,等.胶原蛋白—壳聚糖—聚乙烯醇共混膜的特性研究[J].中国皮革,2003,32(23):1-4.
[21]Gao Changyou,Wang Dengyong,Shen Jiacong.Fabrication of porous collagen-chitosan scaffolds with controlling microstructure for dermal equivalent[J].Polymers for Advanced Technologies,2003,14(6):373-379.
[22]顾震宇,李文俊.壳聚糖溶液冷冻诱导相分离—一种制备高孔隙率膜的新方法[J].科学通报,1999,44(9):933-937.
[23]Chen Guoping,Ushida T,Tateishi T.Development of biodegradable porous scaffolds for tissue engineering[J].Materials Science and Engineering,2001,17(1):63-69.
[24]Yannas IV,Burke JF,Gordon PL,et al.Design of an artificial skin Ⅱ.Control of chemical composition[J].J Biomed Mater Res,1980,14(4):107-131.
[25]Lie Ma,Changyou Gao,Zhengwei Mao,et al.Collagen/chitosan porous scaffolds with improved biostability for skin tissue engineering[J].Biomaterials,2003,24:4833-4841
[1]Harshad A.Navsaria,Simon R.Myers,Irene M.Leigh,et al.Culturing skin in vitro for wound therapy[J].Tibtech,1995,13:91-100.
[2]Gallico GG.Biologic skin substitutes[J].ClinPlast Surg,1990,17:519-526.
[3]Hongyi Yang,Qiqing Zhang.Preparation and Characterization of Collagen-GAGs Bioactive Matrices for Tissue Engineering[J].J Mater Sci.Technol,2001,17(5):495-500.
[4]R.I.弗雷谢尼.动物细胞培养[M].北京:科学出版社,2004.
[5]马刚,王胜,陈世义,等.皮肤成纤维细胞收缩功能的研究[J],白求恩医科大学学报,1996,22(3):272-273.
[6]C.FA.Culling.组织病理学与组织化学技术手册[M].北京:科学出版社,1982.
[7]曹谊林,刘伟,崔磊.组织工程学理论与实践[M].上海:上海科学技术出版社,2004.
[8]Garner WL.Epidermal regulation of dermal fibroblast activity[J].Plast Reconstr surg,1998,102(1):135-139.
[9]Demarchez M,Hartmarm D J,Regnier M,et al.The role of fibroblasts in dermal vascularization and remodeling of reconstructed human skin after transplantation onto the nude mouse[J].Transplantation,1992,54(2):317-326.
[10]Coulomb B,Friteau L,Baruch J,et al.Advantage of the presence of living fibroblasts within in vitro reconstructed skin for grafting in human[J].Plast Reconstr Surg,1998,101(7):1891-1903.
[11]Berthod F,Sahuc F,Hayek D,et al.Deposition of collagen fibril bundles by long-term culture of fibroblasts in a collagen sponge[J].Biomed Mater Res,1996,32(1):87-93.
[12]Lamme EN,Van Leeuwen RT,Brandsma K,et al.Higher numbers of autologous fibroblasts in an artificial dermal substitute improve tissue regeneration and modulate scar tissue formation[J].J Pathos,2000,190(5):595-603.
[13]陈光,周平,潘銮凤,等.天然生物材料壳聚糖支架上人胚肺成纤维细胞的生长[J].化学学报,2004,62(10):992-997.
[1]Steven T,Boyce.Fabrication,quality assurance,and assessment of cultured skin substitutes for treatment of skin wounds[J].Biochemical Engineering Journal,2004,20:107-112.
[2]Belasubramani M,Kumar TR,Babu M.Skin substitutes:a review[J].Bums,2001,27:534-544.
[3]Hunt JA,Van Der Laan JS,Schakenraad J,et al.Quantitative in vivo assessment of tissue response to dermal sheep collagen in abdominal wall defects[J].Biomaterials,1993,14:378-382.
[4]Livesey SA,Hemdon DN,Hollyoak MA,et al.Transplanted acellular allograft dermal matrix:potentital as a temp late for the reconstruction of viable derm is[J].Transplantation,1995,60:1.
[5]HongyiYang,Qiqing Zhang.Preparation and Characterization of Collagen-GAGs Bioactive Matrices for Tissue Engineering[J].J Mater Sci.Technol,2001,17(5):495-500.
[6]郑军,黄晓元,韦星.重组人表皮生长因子促进大鼠皮肤创面愈合的研究[J].中华整形外科杂志,2005,21:379-383.
[7]付小兵,王德文.现代创伤修复学[M].北京:人民军医出版社,1999.
[8]郝光荣.实验动物学[M].上海:第二军医大学出版社,1999.
[9]付小兵,孙同柱,盛志勇.几种用于创伤修复研究的动物模型[J].中华实验外科杂志,2001,12:40-43.
[10]Shahabeddin L,Berthod F,Damour O,et al.Characterization of skin reconstructed on chitosan cross linked collagen glycosam inoglycan matrix[J].Skin PHarmacol,1990,3:107.