摘要
目的:通过组织工程方法研究丝素蛋白/羟基磷灰石复合材料的细胞相容性及其修复包含性骨缺损的作用,探讨其临床应用的可能性。
方法:同种异体脂肪干细胞(Adipose-derived stromal cells,ADSCs)与丝素蛋白/羟基磷灰石共同培养构建组织工程骨。取新西兰大白兔36只,随机分为A、B、C 3组,每组12只,在兔股骨远端制造一4.5mm直径、10mm深度松质骨缺损。A组为实验组,植入复合ADSCs的丝素蛋白/羟基磷灰石;B组为实验对照组,单纯植入丝素蛋白/羟基磷灰石;C组为空白对照组,不作任何植入。
结果:ADSCs能在SF/HA材料上良好的黏附、增殖和生长。12周时X线、组织学检查显示A组完全修复了骨缺损区,B组部分修复了骨缺损区,A组的新生骨小梁多于B组(P <0.05)。C组未见骨修复。
结论:SF/HA具有良好的细胞相容性,并能成功修复兔股骨包含性骨缺损,有望成为骨组织工程理想的细胞外基质和骨移植替代材料。
Objective: To investigate the cellular compatibility of silk fibroin/hydroxyapatite (SF/HA) and its efficacy in repairing contained bone defects, and to explore the possibility of its clinical application.
Methods: SF/HA was co-cultured with Adipose-derived stromal cells(ADSCs)to repair bone defects.36 rabbits were randomly divided into 3 groups. Each group had 12 rabbits. The bone defects(4.5mm×4.5mm×10mm) in the condylus lateralis femoris of rabbits were implanted with ADSCs-SF/HA complex(group A),with simple SF/HA(group B) or left untreated(group C).
Result: Normal adherence, proliferation and growth of ADSCs on the SF/HA could be seen. The examination of X-ray and histology showed that the bone defects were repaired entirely in group A and partly in group B. The quantity of the newly formed bone in group A was more than that in group B (P <0.05). Repair showed no effect in group C.
Conclusion: SF/HA has good cellular compatibility, and can successfully repair contained bone defects of a rabbit femur. It can be used as the ideal alternative biomaterial for bone tissue engineering.
引文
1李玉宝.骨修复纳米生物材料及其发展前景.中国国际新材料产业研讨会,生物医用材料专业论坛, 2004: 4-11.
2 Akita S,Fukui M, Nakagawa H,et al. Cranial bone defect healing is accelerated by mesenchymal stem cells induced by coadministration of bone morphogenetic protein-2 and basic fibroblast growth factor[J].Wound Regen,2004,12(2):252-259.
3 Khan SN, Cammisa FP Jr, Sandhu HS, et al. The biology of bone grafting[J].Am Acad Orthop Surg,2005,13:77-86.
4 Zijderveld SA, ten Bruggenkate CM, van Den Bergh JP,et al. Fractures of the iliac crest after split-thickness bone grafting for preprosthetic surgery: report of 3 cases and review of the literature[J].Oral Maxillofac Surg,2004,62:781-786.
5 Lewandrowski K, Gresser JD, Wise DL, et al. Bioresorbable bone graft substitutes of different osteoconductivities: a histologic evaluation of osteointegration of poly (propylene glycol-co-fumaric acid)-based cement implants in rats[J].Biomaterials,2000,21:757-764.
6 Boone DW. Complications of iliac crest graft and bone grafting alternatives in foot and ankle surgery[J].Foot Ankle Clin,2003,8:1-14.
7 Lin AS, Barrows TH, Cartmell SH, et a1. Microarchitectural and mechanical characterization of oriented porous polymer scaffolds[J].Biomaterials, 2003,24:481-489.
8 Li S, De Wijn JR, Li J, et a1. Macroporous biphasic calcium phosphate scaffold with high permeability/porosity ratio[J].Tissue Eng,2003,90:535-548.
9 Proussaefs P, Lozada J, Valencia G,et al. Histologicevaluation of a hydroxyapatite onlay bone graft retrieved after 9 years: a clinical report[J].Prosthet Dent,2002,87:481-484.
10 Meinela L, Fajardod R, Hofmanna S, et al. Silk implants for the healing of critical size bone defects[J].Bone,2005,37:688–698.
11 Lorenz M, Sandra H, Vassilis K et al. The inflammatory responses to silk films in vitro and in vivo[J].Biomaterials,2005,26:147-155.
12 Altmana GH,Horan RL,Lu HH,et al.Silk matrix for tissue engineered anterior cruciate ligments[J].Biomaterials,2002,23:4131-4141.
13 Zuk PA, ZhuM, MizunoH,et al.Multilineage cells from human adipose tissue: implications for cell-based therapies[J].Tissue Eng,2001,7:211-228.
1 Van Damme A, Vanden-Driessche T, Collen D, et a1. Bone marrow stromal cells astargets for gene therapy. Curr Gene Ther, 2002, 2: 195-209.
2 Pittenger MF, Mackay AM, Beck SC, et a1. Multlineage potential of adult human mesenchymal stem cells. Science, 1999, 284: 143-147.
3 Slack JM. Stem cells in epithelial tissue.Science 2000,287(5457):1431-1433.
4 Kaplan FS,Hahn GV,Zasloff MA et al.Heterotopic ossification:Two rare forms and what they can teach us.J Am Acad Orthop Surg.1994,2(5):288-96.
5 Zuk PA, Zhu M, Mizuno H, et al.Multilineage cells from human adipose tissue:implications for cell-based therapies.Tissue Eng,2001,7 (2):211-228.
6 Halvorsen YD,Franklin D,Bond Al,et al.Extracellular matrix mineralization and osteoblast gene expression by human adipose tissue-derived stromal cells.Tissue Eng,2001,7(6):729-741.
7 Dragoo JI, Samimi B,Zhu M,et al.Tissue-engineered cartilage and bone using stem cells from human infrapatellar fat pads. J Bone Joint Surg Am,2003,85(5):740-747.
8 Nakahara H, Goldberg VM, Caplan AI. Culture-expanded periosteal-derived cells exhibit osteochondrogenic potential in porous calcium phosphate ceramics in vivo. Clin Orthop, 1992, (276): 291-298.
9 Dennis V.H., Karsten H., Sevine H., et al. Comparison of viable cell yield from excised versus aspirated adipose tissue. Cells Tissues Organs. 2004, 178: 87-92.
10 Huss F.R., and Koatz G.. Adipose tissue processed for lipoinjection shows increased cellular survival in vitro when tissue engineering principles are applied. Scand J Plast Reconstr Hand Surg, 2002, 36: 166-171.
11 Halvorsen Y.D., Franklin D., Bond A.L., et al. Extracellular matrix mineralization and osteoblast gene expression by human adipose tissue-derived stromal cells.Tissue Eng,2001b, 7: 729-741.
12 Takamichi M, Yasuo K, Kazuhiro T, et al. Isolation of two human fibroblastic cell populations with multiple but distinct potential of mesenchymal differentiation by ceiling cylture of mature fat cells fro, subcutaneous adipose tissue. Differentiation,2005, 73: 69-78.
13 Sugihara H., Yonemitsu N., Miyabara S., et al. Primary cultures of uniocular fat cells:characteristics of growth in vitro and changes in differentiation properties. Differentiation, 1986, 31: 42-49.
14余方圆,卢世壁,袁玫等.脂肪干细胞向软骨方向诱导的初步研究.中国矫形外科杂志, 2004, 12(10): 762-764.
15 Hidemi H., Masato S., Kazunori M., er al. Osteogenic potential of human adipose tissue-derived stromal cells as an alternative stem cell source. Cell Tissue Organs,2004, 178: 2-12.
16 Bennett J.H., Joyner C.J., Triffitt J.T., et al.Adipocytic cells cultured from marrow have osteogenic potential. J Cell Sci, 1991, 99: 131-139.
17 Zuk PA,Zhu M,Ashjian P,et al. Human adipose tissue is a source of multipotent stem cell. Mol Biol Cell 2002; 13(12):4279-4295.
18 De Ugarte DA,Alfonso Z,Zuk PA,et al. Differential expression of stem cell mobilization-associated molecules on multi-lineage cells from adipose tissue and bone marrow. Immunol Lett 2003; 89(2-3):267-270.
19 Gronthos S,Franklin DM,Leddy HA,et al.Surface protein characterization of human adipose tissue-derived stromal cells. J Cell Physiol 2001; 189(1):54-63.
20 Pittenger MF,Mackay AM,Beck SC,et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284(5411):143-147.
21 Aust L,Devlin B,Foster SJ,et al. Yield of human adipose-derived adult stem cells from liposuction aspirates. Cytotherapy 2004; 6(1):7-14.
22 Lee RH,Kim B,Choi I,et al. Characterization and expression analysis of mesenchymal stem cells from human bone marrow and adipose tissue.Cell Physiol Biochem 2004;14(4-6):311-324.
23 Roebuck KA,Finnegan A. Regulation of intercellular adhesion molecule-1(CD54)gene expression. J Leukoc Biol 1999; 66(6):876-888.
24 Gronthos S,Franklin DM,Leddy HA,et al.Surface protein characterization of human adipose tissue-derived stromal cells,J Cell Physiol,2001,189(1):54-63.
25 De Ugret DA,Alfonso Z,Zuk PA,et al.Defferendial expression of stem cells mobilization-assciated molecules on mutilineage cells from adipose tissue and bone marrow.Immunol Lett,2003,89(2-3):267-270.
1 Akita S,Fukui M, Nakagawa H,et al. Cranial bone defect healing is accelerated by mesenchymal stem cells induced by coadministration of bone morphogenetic protein-2 and basic fibroblast growth factor[J].Wound Regen,2004,12(2):252-259.
2 Khan SN, Cammisa FP Jr, Sandhu HS, et al. The biology of bone grafting[J].Am Acad Orthop Surg,2005,13:77-86.
3 Zijderveld SA, ten Bruggenkate CM, van Den Bergh JP,et al. Fractures of the iliac crest after split-thickness bone grafting for preprosthetic surgery: report of 3 cases and review of the literature[J].Oral Maxillofac Surg,2004,62:781-786.
4 Lewandrowski K, Gresser JD, Wise DL, et al. Bioresorbable bone graft substitutes of different osteoconductivities:a histologic evaluation of osteointegration of poly (propylene glycol-co-fumaric acid)-based cement implants in rats[J].Biomaterials, 2000,21:757-764.
5 Boone DW. Complications of iliac crest graft and bone grafting alternatives in foot and ankle surgery[J].Foot Ankle Clin,2003,8:1-14.
6曹谊林,周广东. 21世纪组织工程面临的机遇与挑战.中华医学杂志, 2005, 85: 2523-2525.
7 Lane JM, Sandhu HS.Current approaches to experimental bone grafting. Orthop Clin of North Am 1987; 18: 213-224.
8曹谊林,周广东. 21世纪组织工程面临的机遇与挑战.中华医学杂志, 2005, 85: 2523-2525.
9 Lane JM, Sandhu HS, Current approaches to experimental bone grafting. Orthop Clin of North Am, 1987, 18: 213-224.
10 Akita S,Fukui M, Nakagawa H,et al. Cranial bone defect healing is accelerated by mesenchymal stem cells induced by coadministration of bone morphogenetic protein-2 and basic fibroblast growth factor[J].Wound Regen,2004,12(2):252-259.
11 Wolter JR, Meyer RF. Sessile macreophages forming clear endothelium-like membrane on inside of successful keratoprosthesis.Trans Am Ophthalmol Soc, 1984, 82:187-202.
12 Langer R, Vacanti JP. Tissue Engineering. Sciences, 1993, 260(5110):920-6.
13 Casabona F, Martin I, Muraglia A, et al. Prefabricated engineered bone flaps: an experimental model of tissue reconstruction in plastic surgery. Plast Reconstr Surg, 1998,101(3):577-95.
14杨维东,毛天球.口腔领面骨替代材料与骨组织工程.实用口腔医学杂志.1999;15(6):469-71.
15张琳,田杰谟.多孔HA人工骨的研究[J].透析与人工器官,2002,13(2):5-10.
16 Liu Dean-Mo.Influence of Solid Loading and Particle Size Distribution on the Porosity Development of Green Alumina Ceramic Mouldings[J].Ceramics Internatioa1, 1997,23:135-139.
17 Wei J, Li YB. Tissue engineering scaffold material of nano-apatite crystals and polyamide composite. Eur Polymer J, 2004, 40: 509-5l5.
18 Li Shipu, Zhang Shicheng, Chen Wenjie. Effects of hydroxya-patite ultrafine powder on colony formation and cytoskeletons of MGC-803 cell. Bioceramics 1996; 9(2):225-227.
19田莉,闵思佳,朱良均.丝素蛋白在软组织相容性材料方面的研究进展.蚕业科学2005; 31(2):187-190.
20 Meinela L, Fajardod R, Hofmanna S, et al. Silk implants for the healing of critical size bone defects. Bone 2005; 37: 688–698.
21 Landis WJ, Jacquet R, Hillyer J,et al.The potential of tissue engineering in orthopedics. Orthop Clin North Am. 2005,36(1):97-104.
22王莉,张亚峰,李春忠.羟基磷灰石/丝蛋白复合骨材料的生物相容性[J].华东理工大学学报,2007,33(1):47-51.
23 Kim BS, Mooney DJ. Development of biocompatible synthetic extracellular matrices for tissue engineering. Trends Biotechnol,1998,16(5):224-30.
24 ZeUin G, Gritli A, Linde A . Healing of mandibular defects with different biodegradable and non-biodegrable membranes: an experimental study in rats. Biomaterials, 1995, 16: 601-609.
25黄岚峰,刘建国,徐莘香.利用可生物降解共聚膜引导骨再生.中国修复重建外科杂志, 1999, 13: 321-325.
26易诚青,杜靖远,郑启新.转化生长因子-β1转基因生物衍生型移植体修复骨缺损.中华实验外科杂志, 2006, 23: 157-159.
27 Gazit D, Turgeman G, Kelley P, et al. Engineered pluripoten mesenchymal cells in integrate and differentiate in regenerating bone: a novel cell mediated gene therapy. J Gene Med, 1999,1(2):121-133.
28 Turksen K, Aubin犯.Positive and negative immunoselection for enrichment of two classes of osteoprogenitor cells. J Cell Biol,1991,114(2):373-84.
29 Ren L, Tsuru K, Hayakawa S, et a.l Novel approach to fabricate porous gelatin siloxane hybrids for bone tissue engineering[J]. Biomaterials. 2002, 23(24): 4765-4773.
30 Benedicte P, Corinne B, Philippe B,et al. Immuno-modulatory effect of human adipose tissue-derived adult stem cells: compareison with bone marrow mesenchymal stem cells. British Journal of Haematology, 2005, 129: 118-129.
31 Mcintosh K,Zvonic S,Grarrett S,et al.The immunogenicity of human adipose-derived stem cellstemporal changes in vitro[J].Stem Cells,2006,24(5):1246-1253.
1 Concannon MJ,Boschert MT,Fitzpatrick L,et al.The use of demineralized bone powder in an onlay graft moldel [J].Plastic&Recon Surg,1995,95(6): 1085.
2郑岳华,侯小妹,杨兆雄,等.多孔羟基磷灰石生物陶瓷的进展[J].硅酸盐通报,1995,3:20-24.
3张琳,田杰谟.多孔HA人工骨的研究[J].透析与人工器官,2002,13(2):5-10.
4 Liu Dean-Mo.Influence of Solid Loading and Particle Size Distribution on the Porosity Development of Green Alumina Ceramic Mouldings[J].Ceramics Internatioa1,1997,23:
35-139.
5王大章,费伟,刘松箔,等.高生物诱导活性复合人工骨整复骨缺损的实验研究[J].华西口腔医学杂志,1997,15(1):3.
6 Yamsaki H,Hegerotopic.Bone formation around porous hydroxyapatite ceramics in the subcutaneous of dogs[J]. Japan Oral Bio1,1990,32:190.
7 Ripamonti U,Crooks J,Kirkbride AN,et al.Sintered porous hydroxyapatites with intrinsic osteoinductive activity:geometric induction of bone formation[J].South African Journal of Science,1999,95(8):335.
8 Yuan H, Kurashina K, de Bruijn JD,et al.A preliminary study on osteoinduction of two kind of calcium phosphate ceramic[J]Biomaterials,1999,20:1799-1806.
9黄立业,徐可.纳米针状羟基磷灰石涂层的制备及其性能的研究[J].硅酸盐学报,1999,27(3):351-356.
10 Mizuno M, Shindo M, Kobayashi D,et al.Osteogenesis by bone marrow stromal cells maintained on type I collagen matrix gels in vivo[J]. Bone,1997,20:101-107.
11 Trudy McKee, James R. McKee,et al. Biochemistry: an introduction. The McGraw- Hill companies, Inc. 1999.
12 Kevor S,Tenhuisen, Roger I,et al.Formation and properties of a synthetic bone composite: Hydroxyapatite-collagen[J]. J Biomed Mater Res, 1995, 29(7):803-810.
13 Du C, Cui Fz, Feng Ql,et al.Tissue response to nano-hydroxyapatite/collagen composite implants in marrow cavity[J]. J Biomed Mater Res, 1998, 42(4):540-548.
14冯庆玲,崔福斋.纳米羟基磷灰石/胶原骨修复材料[J].中国医学科学院学报,2002,24(2):124-128.
15廖素三,崔福斋,张伟,等.组织工程中胶原基纳米骨复合材料的研制[J].中国医学科学院学报,2003,25(1):36-38.
16姚晖,杜昶,杨韶华,等.纳米羟晶/胶原仿生骨修复家兔颅颌骨缺损的实验研究[J].透析与人工器官,2000,11(2):5-8.
17凌翔,陶学金,陈卫民.胶原/纳米磷酸三钙复合人工骨修复骨缺损的实验研究[J].临床口腔医学杂志,2003,19(2):74-76.
18王韦,牟贤龙,赵霞,等.胶原、羟基磷灰石(C/HA)组织埋藏[J].生物医学工程学杂志,1995,12(3):197-200.
19 Chang MC. Preparation of a porous hydroxyapatite/collagen nanlocomposite using glutaraldehyde as a crosslinkage agent [J]. J Mater Sci, 2001, 20: 1199-1201.
20 Altman GH, Diaz F, Jakuba C,et al.Silk-based biomaterials[J].Biomaterials, 2003,24(3):401-416.
21 Furuzono T, Taguchi T, Kishida A,et al.Preparation and characterization of apatite deposited on silk fabric using an alternate soaking process[J]. J Biomed Mater Res,2000,50(3):344-352.
22卢神州,李明忠,白伦,等.羟基磷灰石/丝素蛋白纳米复合颗粒的制备[J].丝绸,2006,(2):17-19.
23 Wang L,Nemoto R,Senna M,et al.Changes in microstructure and physico-chemical properties of hydroxyapatite-silk fibroin nanocomposite with varying silk fibroin content[J].Journal of the European Ceramic Society,2004,24(9):2707-2715.
24王莉,张亚峰,李春忠.羟基磷灰石/丝蛋白复合骨材料的生物相容性[J].华东理工大学学报,2007,33(1):47-51.
25刘芳,贾德民.聚乳酸及其复合材料在骨组织工程方面的研究进展[J].生物医学工程学杂志,2001,18(3):448-450.
26 Zhang Ruiyun, Peter X. Ma.Poly(α-hydroxyl acids)/hydroxyapatite porous composites for bone-tissue engineering[J]. J Biomed Mater Res, 1999,44(4):446-455.
27 Deng XM,Hao JY,Wang CS.Preparation and mechanical properties of nanocomposites of poly (D,L-lactide) with Ca-deficient hydroxyapatite nanocrystals[J].Biomaterials, 2001,22(21):2867-2873.
28吕彩霞,姚子华.羟基磷灰石/壳聚糖复合材料研究进展[J].化工进展,2006,25(7):755-759.
29赵峰,郭善一,尹玉姬.壳聚糖—明胶网络/羟基磷灰石复合材料支架的研究—制备及形貌[J].中国修复重建外科杂志,2001,15(5):276-279.
30 Lind M. Growth factor stimulation of bone healing. Effects on osteoblasts, osteomies, and implants fixation [J]. Acta Orthop Scand, 1998, 283 (Supp1): 32-37.
31 Worch Hartmut, Scharnweber Dieter.Biologization of titanium materials:Opportunities of materials science at the interface of life and inert matter.Z Metallkd,1998,89(2):153.
32 Jin QM, Takita H, Kohgo T, et al. Effects of geometry of hydroxyapatite as a cell substratum in BMP-induced ectopic bone formation [J]. J Biomed Mater Res, 2000, 52: 491-499.
33 Chistolini P, Ruspantini I, Bianco P, et al. Biomechanical evaluation of cell-loaded and cell-free hydroxyapatite implants for the reconstruction of segmental bone defects [J]. J Mater Sci Mater Med, 1999, 10: 739-743.
34 Flautre B, Ansehne K, Delecourt C, et al. Histological aspects in bone regeneration of an association with porous hydroxyapaite and bone marrow cells [J]. J Mater Sci, Mater Med, 1999, 10: 811-814.
35 Guo L, Guo X, Leng Y, et al. Nanoindentation study of interfaces between calciumphosphate and bone in an animal spinal fusion mode1 [J]. J Biomed Mater Res, 2001, 54: 554-559.
36李爱民,孙康宁,尹衍升,等.生物材料的发展应用评价与展望[J].山东大学学报, 2002, 6, 287-293.
1 Zuk PA, ZhuM, MizunoH, et a.l Multilineage cells from human adipose tissue: implications for cell-based therapies [J].Tissue Eng, 2001, 7:211-228.
2 Dennis VH, Karsten H, Sevine H, et al. Comparison of viable cell yield from excised versus aspirated adipose tissue[J]. Cells Tissues Organs. 2004, 178: 87-92.
3 Lalikos JF, Li YQ, Roth TP,et al.Biochemical assessment of cellular damage after adipocyte harvest[J].J Surg Res, 1997, 70: 95-100.
4 Huss FR, and Koatz G.Adipose tissue processed for lipoinjection shows increased cellular survival in vitro when tissue engineering principles are applied[J].Scand J Plast Reconstr Hand Surg, 2002,36:166-171.
5 Harmelen Von V, Skurk T, Rohrig K, et al. Effect of BMI and age on adipose tissuecellularity and differentiation capacity in women[J].Int J Obes, 2003,27:889-895.
6 Halvorsen YD,Franklin D,Bond AL,et al.Extracellular matrix mineralization and osteoblast gene expression by human adipose tissue-derived stromal cells[J].Tissye Eng,2001b,7:729-741.
7 Aust LB, Masato S, Kazynori M, et al.Yield of human adipose-derived adult stemcells from liposution aspirates.Cytotherapy,2004,6:7-14.
8 Prunet Marcassus B, Cousin B, Caton D,et al. From heterogeneity to plasticity in adipose tissues: site-specific differences [J]. Exp Cell Res, 2006,312(6):727-736.
9 Qu CQ, Zhang G H,Zhang L J,et al. Osteogenic and adipogenic potential of porcine adipose mesenchymal stem cells[J]. In Vitro Cell Dev Biol Anim, 2007,43(2):95-100.
10张国华,屈长青,杨公社,等.猪脂肪间充质干细胞的分离培养及其成脂分化[J].动物学报( Zhang G H, Qu C Q, Yang G S.Culture and adipogenesis differentiation of porcine adipose mesenchymal stem cells [J]. Acta Zool Sin), 2006,52(5):934-941.
11 De Ugarte D A, Morizono K, Elbarbary A,et al. Comparison of multi-lineage cells from human adipose tissue and bone marrow[J].CellsTissues Organs, 2003,174(3):101-109.
12 Pittenger MF,Mackay AM,Beck SC,et al. Multilineage potential of adult humanmesenchymal stem cells[J]. Science,1999,284(5411):143-147.
13 Strem B M, Hicok K C, ZhuM,et al. Multipotential differentiation of adipose tissue-derived stem cells[J]. Keio J Med,2005,54(3):132-141.
14 Zuk PA,Zhu M,Ashjian P,et al. Human adipose tissue is a source of multipotent stem cell[J]. Mol Biol Cell,2002,13(12):4279-4295.
15 De Ugarte DA,Alfonso Z,Zuk PA,et al. Differential expression of stem cell mobilization-associated molecules on multi-lineage cells from adipose tissue and bone marrow[J]. Immunol Lett,2003,89(2-3):267-270.
16 Gronthos S,Franklin DM,Leddy HA,et al.Surface protein characterization of human adipose tissue-derived stromal cells[J].Cell Physiol,2001,1089(1):54-63.
17 Roebuck KA,Finnegan A.Regulation of intercellular adhesion molecule1(CD54)gene expression[J].Leukoc Biol,1999,66(6):876-888.
18 Aust L,Devlin B,Foster SJ,et al. Yield of human adipose-derived adult stem cells from liposuction aspirates[J].Cytotherapy,2004,6(1):7-14.
19 Lee RH,Kim B,Choi I,et al.Characterization and expression analysis of mesenchymal stem cells from human bone marrow and adipose tissue[J].Cell Physiol Biochem,2004,14(4-6):311-324.
20 Dragoo JL,Samimi B,Zhu M,et al. Tissue-engineered cartilage and bone using stem cells from human infrapatellar fat pads[J].Bone Joint Surg Br,2003, 85(5):740-747.
21 Tholpady SS,Katz AJ,Ogle RC,et al. Mesenchymal stem cells from rat visceral fat exhibit multipotential differentiation in vitro[J].Anat Rec,2003,272(1):398-402.
22 Halvorsen YD,Franklin D,Bond AL,et al.Extracellular matrix mineralization and osteoblast gene expression by human adipose tissue-derived stromal cells[J].Tissue Eng,2001,7(6):729-741.
23 Lendeckel S,Jodicke A,Christophis P,et al. Autologous stem cells(adipose) and fibrin glue used to treat widespread traumatic calvarial defects:case report[J]. Craniomaxillofac Surg,2004,32(6):370-373.
24 Huang JI,Zuk PA,Jones NF,et al.Chondroenic potential of multipotential cells from human adipose tissue. Plast Reconstr Surg.2004,113:585-594.
25 Denker AE,Nicoll SB,Tuan RS,et al.Formation of cartilage-like spheroids by micromass cultures of murine C3H10T1/2 cells upon treatment with transforming growth factor-1. Differentiation.1995, 59: 25.
26 Ahrens PB,Solursh M,Reiter RS,et al. Stage-related capacity for limbchondrogenesis in cell culture. Dev. Biol.1977, 60: 69.
27 Tavella S,Raffo P,Tacchetti C,et al.N-CAM and N-cadherin expression during in vitro chondrogenesis. Exp. Cell Res.1994, 215: 354.
28 Tacchetti C,Tavella S,Dozin B,et al. Cell condensation in chondrogenic differentiation. Exp. Cell Res.1992 ,200: 26.
29 Stott NS,Jiang TX,Chuong CM. Successive formative stages of precartilaginous mesenchymal condensations in vitro: Modulation of cell adhesion by Wnt-7A and BMP-2. J. Cell Physiol.1999, 180: 314.
30 Kirsch T,Mark K.Remodelling of collagen ypes I, II and X and calcification of human fetal artilage. Bone Miner.1992,2:107.
31 Price PA.Gla-containing proteins of bone. Connect. issue Res. 1989,21: 51.
32 Price PA,Urist MR,Otawara Y.Matrix Gla protein, a new arboxyglutamic acid- containing protein hich is associated with the organic matrix of one. Biochem. Biophys. Res. Commun. 1983,117: 765.
33余方圆,卢世壁,袁玫等.脂肪干细胞向软骨方向诱导的初步研究.中国矫形外科杂志.2004,12(10):762-764.
34 Winter A,Breit S,Parsch D,et al. Cartilage-like gene expression in differentiated human stem cell spheroids:a comparison of bone marrow-derived and adipose tissue-derived stromal cells[J]. Arthritis Rheum,2003,48(2):418-429.
35 Sen A. Adipogenic potential of human adipose derived stromal cells from multiple donorsis heterogeneous[J].CellBiochem,2001,81:312-319.
36 Halbleib M, Skurk T, de Luca C, et al. Tissue engineering of white adipose tissue using hyaluronic acid- based scaffolds I: in vitro differentiation of human adipocyte precursor cells on scaffolds[J].Biomaterials,2003,24:3125-3132.
37 von Heimburg D, Zachariah S, Schwarz ,et al. Human preadipocytes seeded on freeze-dried collagen scaffolds investigated in vitro and in vivo[J].Biomaterials, 2001, 22(5):429-438.
38 Woodbury D,Schwarz EJ,Prockop DJ,et al. Adult rat and human bone marrow stromal cells differentiate into neurons[J].Neurosci Res,2000, 61(4):364-370.
39 Sanchez-Ramos J,Song S,Carsozo-Pelaez F,et al. Adult bone marrow stromal cells differentiate into neural cells in vitro[J].Exp Neuro,2000,164(2):247-256.
40 Ning H, Lin G, Lue TF, et al.Neuron-like differentiation of adipose tissue-derived stromal cells and vascular smooth muscle cells[J].Differentiation,2006, 74(9-10):510- 518.
41 Yang L,Zheng J,Wang C,et al. The in vitro study of the human adipose tissue-derived stromal cells differentiating into the neuron-like cells[J].Zhongguo Xiufu Chongjian Waike Zazhi,2006,20(8):783-786.
42 Dezawa M,Kanno H,Hoshino M,et al. Specific induction of neuronal cells from bone marrow stromal cells and application for autologous transplantation[J]. Clin Invest, 2004,113(12):1701-1710.
43 Safford KM,Hicok KC,Safford SD,et al. Neurogenic differentiation of murine and human adipose-derived stromal cells[J].Biochem Biophys Res Commun,2002,294(2): 371-379.
44 Ashjian PH,Elbarbary AS,Edmonds B,et al. In vitro differentiation of human processed lipoaspirate cells into early neural progenitors[J].Plast Reconstr Surg,2003, 111(6): 1922-1931.
45 Planat-Benard V,Menard C,Andre M,et al. Spontaneous cardiomyocyte differentiation from adipose tissue stroma cells[J].Circ Res,2004,94(2):223-229.
46 Gaustad KG,Boquest AC,Anderson BE,et al. Differentiation of human adipose tissue stem cells using extracts of rat cardiomyocytes[J].Biochem Biophys Res Commun, 2004,314(2):420-427.
47 Makino S, Fukuda K, Miyoshi S, et al. Cardiomyoctes can be generated from marrow stromal cells in vitro[J]. J Clin Invest, 1999, 103(5): 697-705.
48 Rangappa S,Fen C,Lee EH,et al. Transformation of adult mesenchymal stem cells isolated from the fatty tissue into cardiomyocytes[J].Ann Thorac Surg,2003,75(3): 775-779.
49 Mizuno H,Zuk PA,Zhu M,et al. Myogenic differentiation by human processed lipoaspirate cells[J].Plast Reconstr Surg,2002,109(1):199-209.
50 Bacou F,Andalousi RB,Daussin PA,et al. Transplantation of adipose tissue-derived stromal cells increases mass and functional capacity of damaged skeletal muscle[J]. Cell Transplant,2004,13(2):101-111.
51 Benedicte P, Corinne B, Philippe B,et al. Immuno-modulatory effect of human adipose tissue-derived adult stem cells: compareison with bone marrow mesenchymal stemcells. British Journal of Haematology, 2005, 129: 118-129.
52 Mcintosh K,Zvonic S,Grarrett S,et al.The immunogenicity of human adipose-derived stem cellstemporal changes in vitro[J].Stem Cells,2006,24(5):1246-1253.