腺病毒介导的mCbfa1基因治疗骨缺损的实验研究
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摘要
目的 以AdEasy腺病毒表达系统制备携带Cbfa1基因的高滴度腺病毒，感染小鼠成肌细胞C2C12，利用外源性基因编码的生长因子诱导其表达成骨细胞表型。再将感染的C2C12与高分子生物材料chronOS支架相复合，在体外构建组织工程化骨，移植修复小鼠股骨骨缺损，观察该方法的可行性和有效性，以期找到一种较好的修复骨缺损的方法。
方法 (1)将小鼠Cbfa1基因cDNA克隆到穿梭载体形成转移质粒pAdTrack-CMV-Cbfa1，经PmeⅠ酶切线性化后与骨架质粒感受态细胞AdEasier-1 Cells经电穿孔法同源重组得到腺病毒质粒pAdCbfa1并酶切鉴定，PacⅠ酶切线性化后感染293细胞包装后获得复制缺陷型重组腺病毒AdCbfa1。(2)以复制缺陷重组腺病毒介导小鼠Cbfa1基因感染C2C12后，以MTT法及流式细胞仪观察感染对细胞增殖活力及细胞周期的影响；用RT-PCR法、免疫组化染色以及Westernblot证实转基因C2C12的Cbfa1蛋白和Ⅰ型胶原的表达情况；通过改良Gomori钙钴法及ALP检测试剂盒检测两组细胞的ALP活性；通过放免分析试剂盒检测两组细胞的骨钙素(osteocalcin，OCN)含量。(3)制备孔径为150～300μm，孔隙率为90％的chronOS与腺病毒感染后的C2C12
Objective To clone human osteogenic sarcoma core binding factorα1 gene cDNA full length and generate high titers level recombinant adenoviruses containing hCbfal gene the replication defective recombinant adenoviruses AdEasy system was used. To induce expression of osteogenic phenotype by mCbfa1 gene the mouse C2C12cells were infected by recombinant adenovirus. Then the tissue-engineered bone was constructed by combination of infected C2C12 andchronOS and its feasibility and ability to enhance segmental bone defect healing of mouse femur was explored.
Methods (1) mCbfal gene was cloned into Shuttle vector to generate pAdTrack-CMV-Cbfal, which was linearized by PmeI and co-transformed into adenoviral backbone plasmid competent cells-AdEasier-1 Cells to obtain recombinant adenoviral plasmid- pAd-Cbfa1 by homologous recombination. Recombinants were selected and confirmed by restriction endonuclease analysis. After packed in 293 cells, the recombinant adenoviruses AdCbfal were generated. (2) The proliferation and cell cyle analysis of gene-modified C2C12 were assessed by MTT method and FCM after transfected with Ad-Cbfa1. The methods of RT-PCR, immunohistochemical staining and western blot were used to test the expression of Cbfa1 protein. Gomori stain and ALP testing kit were used to measure ALP activity and immunohistochemistry testing kit were used to measure OCN contents. In addition, the collagen type I were tested with the methods of RT-PCR and immunohistochemical staining. (3)
方法 (1)将小鼠Cbfa1基因cDNA克隆到穿梭载体形成转移质粒pAdTrack-CMV-Cbfa1，经PmeⅠ酶切线性化后与骨架质粒感受态细胞AdEasier-1 Cells经电穿孔法同源重组得到腺病毒质粒pAdCbfa1并酶切鉴定，PacⅠ酶切线性化后感染293细胞包装后获得复制缺陷型重组腺病毒AdCbfa1。(2)以复制缺陷重组腺病毒介导小鼠Cbfa1基因感染C2C12后，以MTT法及流式细胞仪观察感染对细胞增殖活力及细胞周期的影响；用RT-PCR法、免疫组化染色以及Westernblot证实转基因C2C12的Cbfa1蛋白和Ⅰ型胶原的表达情况；通过改良Gomori钙钴法及ALP检测试剂盒检测两组细胞的ALP活性；通过放免分析试剂盒检测两组细胞的骨钙素(osteocalcin，OCN)含量。(3)制备孔径为150～300μm，孔隙率为90％的chronOS与腺病毒感染后的C2C12
Objective To clone human osteogenic sarcoma core binding factorα1 gene cDNA full length and generate high titers level recombinant adenoviruses containing hCbfal gene the replication defective recombinant adenoviruses AdEasy system was used. To induce expression of osteogenic phenotype by mCbfa1 gene the mouse C2C12cells were infected by recombinant adenovirus. Then the tissue-engineered bone was constructed by combination of infected C2C12 andchronOS and its feasibility and ability to enhance segmental bone defect healing of mouse femur was explored.
Methods (1) mCbfal gene was cloned into Shuttle vector to generate pAdTrack-CMV-Cbfal, which was linearized by PmeI and co-transformed into adenoviral backbone plasmid competent cells-AdEasier-1 Cells to obtain recombinant adenoviral plasmid- pAd-Cbfa1 by homologous recombination. Recombinants were selected and confirmed by restriction endonuclease analysis. After packed in 293 cells, the recombinant adenoviruses AdCbfal were generated. (2) The proliferation and cell cyle analysis of gene-modified C2C12 were assessed by MTT method and FCM after transfected with Ad-Cbfa1. The methods of RT-PCR, immunohistochemical staining and western blot were used to test the expression of Cbfa1 protein. Gomori stain and ALP testing kit were used to measure ALP activity and immunohistochemistry testing kit were used to measure OCN contents. In addition, the collagen type I were tested with the methods of RT-PCR and immunohistochemical staining. (3)
引文
1. Cohn M J, Tickle C: Limbs: a model for pattern formation within thevertebrate body plan. Trends Genet 1996, 12: 253-258.
2. Hogan BLM: Bone morphogenetic proteins: multifunctionalregulators of vertebrate development. Genes Dev 1996, 10: 1580-1594.
3. Tabin C J: Retinoids, homeoboxes, and growth factors: towardmolecular models for limb development. Cell 1991, 66: 199-217.
4. Hall BK, Miyake T: The membranous skeleton: the role of cell condensations in vertebrate skeletogenesis. Anat Embryo/1992, 186: 107-124.
5. Horton WA: Morphology of connective tissue: cartilage.In Connective Tissue and Its Heritable Disorders. New York: Wiley-Liss, Inc; 1993: 73-84.
6. Lui VC, Ng L J, Nicholls J, Tam PP, Cheah KS: Tissue-specific and differential expression of alternatively spliced alpha 1 (11) collagenmRNAs in early human embryos. Dev Dyn 1995, 203: 198-211.
7. Ryan MC, Sandell L J: Differential expression of a cystein-dchdomain in the amino-terminal propeptide of type II (cartilage)procollagen by alternative splicing of mRNA, J Bio/Chem 1990, 265: 10334-10339.
8. Sandell U, Morris N, Robbins JR, Goldring MB: Alternatively splicedtype II procollagen mRNAs define distinct populations of cellsduring vertebral development: differential expression of theamino-propeptide. J Ce//Bio/1991, 114: 1307-1319.
9. Luo G, D'Souza R, Hogue D, Karsenty G: The matrix Gla proteingene is a marker of the chondrogenesis cell lineage during mouse development. J Bone Miner Res 1995, 10: 325-334.
10. Thorogood P: Differentiation and morphogenesis of cranial skeletal tissues. In The Skull. Edited by J Hanken, BK Hall. Chicago:University of Chicago Press; 1993.
11. Grigodadis AE, Heersche JNM, Aubin JE: Analysis of chondroprogenitor frequency and cartilage differentiation in anovelfamily of clonal chondrogenic rat cell lines. Differentiation 1996, 60: 299-307.
12. Suda T, Takaharashi N, Martin TJ: Modulation of osteoclast differentiation. Endocr Rev 1992, 13: 66-80.
13. Ng L J, Wheatley S, Muscat GE, Conway-Campbell J, Bowles J, o Wright E, Bell DM, Tam PP, Cheah KS, Koopman P: SOX9 binds DNA, activates transcription, and coexpresses with type II collagen during chondrogenesis in the mouse. Dev Biol 1997, 183: 108-121.
14. Zhao Q, Eberspaecher H, Lefebvre V, De Crombrugghe B: Parallelexpression of Sox9 and Col2al in cells undergoingchondrogenesis. Dev Oyn 1997, 209: 377-386.
15. Wagner T, Wirth J, Meyer J, Zabel B, Held M, Zimmer J, Pasantes J,Bricarelli FD, Keutel J, Hustert E: Autosomal sex reversal andcampomelic dysplasia are caused by mutations in and around theSRY-related gene SOX9. Cell 1994, 79: 1111-1120.
16. Lefebvre V, Huang W, Harley VR, Goodfellow PN, De Crombrugghe B: SOX9 is a potent activator of the chondroeyte-specific enhancerof the pro alphal (11) collagen gene. Mol Cell Biol 1997, 17: 2336-2346.
17. Bell DM, Leung KK, Wheatley SC, Ng L J, Zhou S, Ling KW,Do Sham MH, Koopman P, Tam PP, Cheah KS: SOX9 directly regulatesthe type-II collagen gene. Nat Genet 1997, 16: 174-178.
18. Suva U, Winslow GA, Wettenhall RE, Hammonds RG, Moseley JM,Diefenbach-Jagger H, Rodda CP, Kemp BE, Rodriguez H, Chen EYet aL: A parathyroid hormone-related protein implicated inmalignant hypercalcemia: cloning and expression. Science 1987, 237: 893-896.
19. Karaplis AC, Luz A, Glowacki J, Bronson RT, Tybulewicz VL,Kronenberg HM, Mulligan RC: Lethal skeletal dysplasia fromtargeted disruption of the parathyroid hormone-related peptidegene. Genes Dev 1994, 8: 277-289.
20. Lanske B, Karaplis AC, Lee K, Luz A, Vortkamp A, Pirro A,Karperien M, Defize LHK, Ho C, Mulligan RC et al.: PTH/PTHrP receptor in early development and Indian hedgehog-regulated bonegrowth. Science 1996, 273: 663-666.
21. Schipani E, Kruse K, Juppner H: A constitutively active mutant PTHPTHrP receptor in Jansen-type metaphyseal chondrodysplasia.Science 1995, 268: 98-100.
22. Bitgood M J, McMahon AP: Hedgehog and Bmp genes arecoexpressed at many diverse sites of cell-cell interaction in themouse embryo. Dev Biol 1993, 172: 126-138.
23. Vortkamp A, Lee K, Lanske B, Segre GV, Kronenberg HM, Tabin CJ:Regulation of rate of cartilage differentiation by Indian hedgehogand PTH-related protein. Science 1996, 273: 613-622.
24. Chiang C, Litingtung Y, Lee E, Young KE, Corden JL, Westphal H,Beachy PA: Cyclopia and defective axial patterning in mice lackingSonic hedgehog gene function. Nature 1998, 383: 407-413.
25. Naski MC, Wang Q, Xu J, Omitz DM: Graded activation of fibroblast growth factor receptor 3 by mutations causing achondroplasiaand thanatophoric dysplasia. Nat Genet 1996, 13: 233-237.
26. Colvin JS, Bohne BA, Harding GW, McEwen DG, Ornitz DM:Skeletal overgrowth and deafness in mice lacking fibroblastgrowth factor receptor 3. Nat Genet 1996, 12: 390-397.
27. Vu TH, Shipley JM, Bergers G, Berger JE, Helms JA, Hanahan D,te Shapiro SD, Senior RM, Werb Z: M MP-9/Gelatinase B is a keyregulator of growth plate angiogenesis and apoptosis ofhypertrophic chomdrocytes. Cell 1998, 93: 411-422.
28. Ducy P, Karsenty G: Two distinct osteoblast-specific cis-actingelements control expression of a mouse osteocalcin gene.Mo/Cell Biol 1995, 15: 1858-1869.
29. Geoffroy V, Duty P, Karsenty G: A PEBP2/AML-I-related factorincreases osteocalcin promoter activity through its binding to anosteoblast-specific cis-acUng elemenL J BiD/Chem 1995, 270: 30973-30979.
30. Merriman HL, van Wijnen A J, Hiebert S, Bidwell JP, Fey E, Lian J,Stein J, Stein GS: The tissue-specific nuclear matrix protein,NMP-2, is a member of the AM L/CBF/PEBP2/Runt domaintranscription factorfamily: interactions with the osteocalcin gene promoter. Biochemistry 1995, 34: 13125-13132.
31. Gergen JP, Wieschaus E: The localized requirements for a gene affecting segmentation in Drosophila: analysis of larvae mosaic for runL Dev Bio11985, 109: 321-335.
32. Kagoshima H, Shigesada K, Satake M, Ito Y, Miyoshi H, Ohki M,Pepling M, Gergen P: The Runt domain identifies a new family ofheteromeric transcriptional regulators. Trends Genet 1993, 9: 338-341.
33. Ito Y, Bae S-C: The runt domain transcription factor PEBP2/CBF,and its involvment in human leukemia. In Oncogenes asTranscriptional Regulators, vol 2. Edited by Yaniv M, Ghysdael J.Basel: Birkhauser Verlag; 1997: 107-132.
34. Satake M, Nomura S, Yamagushi-lwai Y, Takahama Y, Hashimoto Y,Niki M, Kitamura Y, Ito Y: Expression of the runt domain-encodingPEBP'2a genes in T cells during thymic developmenL Mo/Cell BiD/1995, 15: 1662-1670.
35. D,,cy P, Zhang R, Geoffroy V, Ridall A, Karsenty G: Osf2/Cbfal: a transcriptional activator of ostaoblast differentiation. Cell 1997, 89: 747-754.
36. Otto F, Thornell AP, Crompton T, Denzel A, Gilmour KC, Rosewell IR, Stamp GWH, Beddington RSP, Mundlos S, Olsen B: Cbfal, a candidate gene for cleidocranial dysplasia syndrome, is essentialfor osteoblast differentiation and bone development. Cell 1997, 89: 765-771.
37. Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K,Shimizu Y, Bronson RT, GaD Y-H, Inada Met a/.: Targeted disruptionof Cbfal results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell 1997, 89: 755-764.
38. Mundlos S, Otto F, Mundlos C, Mulliken JB, Aylsworth AS, Albright S, Lindhout D, Cole WG, Henri W, Knoll JHM et aL: Mutations involvingthe transcription factor CBFA1 cause cleidocranial dysplasia. Ce111997, 89: 773-779.
39. Lee B, Thirunavukkarasu K, Zhou L, Pastore L, Baldini A, Geoffroy V, Duty P, Karsenty G: Missense mutations abolishing DNA bindingOSF2/CBFA1 in patients affected with cleidocranial dysplasia. Nat Genet 199'7, 16: 307-311.
40. Tondravi MM, McKercher SR, Anderson K, Erdmann JM, Ouiroz M,Maki R, Teitelbaum SL: Osteopetrosis in mice lackinghaematopoietic transcription factor PU.1. Nature 1997, 386: 81-84.
41. Yoshida H, Hayashi S, Kunisada T, Ogawa M, Nishikawa S,Okamura H, Sudo T, Shultz LD, Nishikawa S: The murine mutationosteopetrosis is in the coding region of the macrophage colonystimulating factor gene. Nature 1990, 345: 442-444.
42. Simonet WS, Lacey DL, Dunstan CR, Kelley M, Chang MS, Luthy R,Lee Nguyen HQ, Wooden S, Bennett L, Boone T et al.: Osteoprotegerin:a novel secreted protein involved in the regulation of bone density. Cell 1997, 89: 309-319.
43. Bucay N, Sarosi I, Dunstan CR, Morony S, Tarpley J, Capparelli C,Scully S, Tan HL, Xu W, Lacey DL eta/.: Osteoprotegedn-deflcient mice develop early onset osteoporosis and arterial calcification.Genes Dev 1998, 12: 1260-1268.
44. Lacey DL, Timms E, Tan HL, Kelley M J, Dunstan CR, Brugess T, Elliot R, Colombero A, Elliot G, Scully S, Hsu Her.:Osteoprotegerin ligand is a cytokine that regulates osteoclastdifferentiation and activation. Cell 1998, 93: 165-168.
45. Yasuda H, Shima N, Nakagawa N, Yamaguchi K, Kinosaki M, Mochizuki S, Tomoyasu A, Yano K, Goto M, Murakami A:Osteoclast differentiation factor is a ligand for osteoprotegerin/osteclastogenesis-inhibitory factor and isidentical to TRANCE/RANKL Proc Nat/Acad Sci USA 1998, 95: 3597-35602.
46. Wang ZQ, Ovitt C, Grigoriadis AE, Mohle-Steinlein U, Ruther U,Wagner EF: Bone and haeroatopoiatic defects in mice lackingc-los. Nature 1999, 360: 741-745.
47. Grigoriadis AE, Wang ZCI, Cecchini MG, Hofstetter W, FelixR,Fleisch HA, Wagner EF: c-Fos: a key regulator of osteoclastroacrephagelineage determination and bone remodeling.Science 1994, 266: 443-448.
48. Johnson RS, Spiegelman BM, Papaioannou V: Pleiotropic effects of a null mutation in the c-los proto-oncogene. Cell 1992, 71: 577-586.
49. Franzoso G, Carlson L, Xing L, Poljak L, Shores EW, Brown KD, Leonardi A, Tran T, Boyce BF, Siebenlist U: Requirement forNF-kappa B in osteoclast and B-cell development. Genes Dev1997, 11: 3482-3496.
50. Hodgkinson CA, Moore KJ, Nakayama A, Steingrimsson E,Copeland NG, Jenkins NA, Arnheiter H: Mutations at the mouseroicrophthalroia locus are associated with defects in a geneencoding a novel basic-helix-loop-helix-zipper protein. Cell 1993, 74: 395-404.
51. Satake M, Nomura S, Yamaguchi-lwai Y, Brown KD. Expression of the Runt domain-encoding PEBP2 alpha genes in T cells during thymic development. Mol Cell Biol. 1995; 15(3): 1662-70.
52. Mundlos S, Otto F, Mundlos C, Xing L. Mutations involving the transcription factor CBFA1 cause cleidocranial dysplasia. Cell. 1997; 89(5): 773-9.
53. Xiao ZS,Thomas R.,Hinson TK.. Genomic structure and isoform expression of the mouse, rat and human Cbfal/Osf2 transcription factor. Gene. 1998; 214(1-2): 187-97.
54. Harada H, Tagashira S, Fujiwara M, McLarren KW. Cbfal isoforms exert functional differences in osteoblast differentiation. J Biol Chem. 1999; 274(11): 6972-8.
55. Marks SC, Hermey DC: The structure and development of bone. In Principles of Bone Biology. Edtted by Bllezikian JP, Ralsz LG, RodanGA. London: Academic Press; 1996: 3-14.
56. Suds T, Udagawa N, Takahashl N: Cells of bone: osteoclastgeneration. in Prinuples of Bone Biology. Edited by Bilezikian JP,Ralsz LG, Rodan GA. London: Academic Press; 1996: 87-102.
57. Aubln JE, Turksen K, Heersche JNM: Osteoblastic lineage. In Cellularand Molecular Bology of Bone. Edited by Noda M. London:Academic Press; 1993: 1-45.
58. Welntraub H: The MyoD family and myogenesis: redundancy,networks, and thresholds. Cell 1993, 75: 1241-I 244.
59. Tontonoz P, Hu E, SpIegelman BM: Stimulation of adipogenesis infibroblasts by PPARr2, a lipid-activated transcription factor. Cell 1994, 79: 1147-1156.
60. Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K,Shimlzu Y, Bronson RT, Gao YH, lnada Met a/.: Targeted disruption of Cbfal results in a complete lack of bone formation owi& tomaturational arrest of osteoblasts. Cell 1997, 89: 755-764.
61. Otto F, Thornell AP, Crompton T, Denzel A, Gllmour KC, Rosewell IR, Stamo GWH. Beddlnaton RSP. Mundlos S. Olsen BR et al.: Cbfal. acandidate g&e for cieidocranial dysplasia syndrome, is essentialfor osteoblast differentiation and bone development. Cell 1997, 89: 765-771.
62. Duty P, Zhang R, Geoffroy V, Rldall AL, Karsenty G: Osf2/Cbfal: a transcriptional activator of osteoblast differentiation. Cell 1997, 89: 747-754.
63. Ogawa E, Maruyama M, Kagoshlma H, lnuzuka M, Lu J, Satake M,Shiaesada K. Ito Y: PEBP2IPEA2 represents a new familv of traiscription factor homologous toihe products of the drosophila runt and the human AMLf gene. Proc Nat/ Acad So USA 1993, 90: 6859-6863.
64. Bae SC, Yamaguchl-lwai Y, Ogawa E, Maruyama M, lnuzuka M,Kaaoshima H. Shiaesada K. Satake M. Ito Y: Isolation of PEBP2aB cDiA repre&ntiig the m&se homolog of human acute myeloidleukemia gene, AMLI. Oncogene 1993, 8: 809-814.
65. Bae SC, Takahashi E, Zhang VW, Ogawa E, Shigesada K, Namba Y,Satake M, Ito Y: Cloning, mapping and expression of PEBP2c&, athird gene encoding the mammalian Runt domain. Gene 1995, 159: 245-248.
66. Kanla MA, Banner AS, Duffy JB, Gergen JP: The Drosophilasegmentation gene runt encodes a novel nuclear regulatoryprotein that is also expressed in the developing nervous system.GenesDev1990, 4: 1701-1713.
67. Ogawa E, lnuzaka M, Maruyama M, Satake M, Naito-Fqmoto M, Ito Y,ShIResada K: Molecular cloning and characterization of PEBPSP,theheterodimeric partner of a novel Drosophila runt-related DNA binding protein PEBP2a V/rology 1993, 194: 314-331.
68. Vaillant F, Blyth K, Terry A. A full-length Cbfal gene product perturbs T-cell development and promotes lymphomagenesis in synergy with myc. Oncogene. 199925; 18(50): 7124-34.
69. Wang S, Wang a, Crute BE, Melnlkova IN, Keller SR, Speck NA:Cloning and characterization of subunits of the T-cell receptorand murine leukemia virus enhancer core-binding factor. MO/Cell Biol 1993, 13: 3324-3339.
70. Zeng C, Wtjnen AJV, Stein JL, Meyers S, Sun W, Shopland L,Lawrence JB, Penman S, Llan JB, Stein GS, Hiebert SW:Identification of a nuclear matrix targeting signal in the leukemiaand bone-related AML/CBF-a transcription factors. Proc Nat/ Acad SC; USA 1997, 94: 6746-6751
71. Kamachl Y, Ogawa E, Asano M, lshlda S, Murakami Y, Satake M, Ito Y,Shigesada K: Purification of a mouse nuclear factor that binds to both the A and B cores of the polyomavirus enhancer. J Viral 1990, 64: 4808-4819.
72. Drissi H,Lue Q, Shakoori R, Gibert L. Transcriptional autoregulation of the bone related CBFA 1/RUNX2gene.J Cell Physiol,2000, 1284(3): 341-350.
73. Wang S, Speck NA: Purification of core-binding factor, a protein that binds the conserved core site in murine leukemia virus enhancers. MO/ Cell B/o/1992, 12: 89-102.
74. Okuda T, van Deursen J, Hlebert SW, Grosveld G, Downing JR:AMLI, the target of multiple chromosomal translocation in humanleukemia, is essential for normal fetal liver hematopoiesis. Cell 1996, 84: 321-330.
75. Wang a, Stancy T, Binder M, Marin-PadlIla M, Sharpe AH, SpeckNA:Disruption of the Cbfa2 gene causes necrosis and hemorrh agingin the central nervous system and blocks definitive hematopoiesis. Proc Nat/Acad Sci USA 1996, 93:3444-3449.
76. Sasakl K, Yagl H, Bronson RT, Tomlnaga K, Matsunashl T, Deguchl K,Tani Y, Klshlmoto T, Komorl T: Absence of fetal liver hematopoiesis in mice deficient in transcriptional coactivator core binding factor. Proc Nat/AcadSo USA 1996, 93:i 2359-Ⅰ 2363.
77. Marzia M,Sims NA, Voit A. Decteaed c-Src expression enhances osteoblast differentiation and bone formation.J Cell Biol,2000,151 (2):311-320.
78. Wang Q, Stacy T, Miller JD, Lewis AF, Gu TL, Huang X, Bushweller JH, Bodes JC, Alt FW, Ryan Get al.: The CBFb subunit is essential for CBFa2 (AMLI) function in viva. Cell 1996, 87:697-708
79. Satake M, Nomura S, Yamaguchl-lwal Y, Takahama Y, Hashlmoto Y.Nikl M, Kltamura Y, Ito Y: Expression of the runt domain-encoding PEBP2a genes in T cells during thymic development. MO/Cell Bfol 1995, 15:1662-1670.
80. Wargnler A, Legros-Malda S, Bosselut R, Bourge JF, Lafaurle C, Ghysdael J, Sasportes M, Paul P: Identification of human granzyme B promoter regulatory elements interacting with activated T-cellspecific proteins: implication of Ikaros and CBF binding sites in promoter activation, froc Nat/AcadSo USA 1995, 92:6930-6934.
81. Geoffroy V, Corral DA, Zhou L, Lee B, Karsenty G: Genomic organization, expression of the human CBFAI gene, and evidence for an alternative splicing event affecting protein function. Mamm Genome 1998, 9:54-57.
82. Ahn MY, Bae SC, Maruyama M, Ito Y: Comparison of the humangenomic structure of the Runt domain-encoding PEBP2/CBFagene family. Gene 1996, 168:279-280.
83. Mundlos S. Otto F. Mundlos C. Mulliken JB. Avlsworth AS. Albrlaht S. Lindhout D, Cole WG, Henri W, Knoll JI-IM'etal.: Mutations invilvingthe transcription factor CBFAI cause eleidoeranial dysplasia. Cell 1997, 89:?73-779.84. Banerjee C, McCabe LR, Chow J, Hiebert SW, Stetn JL, Stein GS, Llan JB: Runt homology domain proteins in osteoblast differentiation: AMLI/CBFA1 is a maior component of a bonespecific complex. J Cell Biochem 1967, 66: 1-8.
85. Jarvls JL, Keats TE: Cleidocranial dysostosis, a review of 40 new cases. Am J Radio/1974, 121: 5-16.
86. Mundlos S, Mulllken JB, Abramson DL, Warman ML, Knoll JHM, Olsen BR: Genetic mapping of cleidocranial dysplasia and evidence of a microdeletion in one family. Hum MO/Genet 1995, 4: 71-75.
87. Mundlos S, Huang LF, Selby P, Olsen BR: Cleidocranial dysplasia in mice. Ann NY Acad So 1996, 785: 301-302.
88. Kvrishman V, Ma YL,Moseley JM. Bone anabolic effects of sonic/Indian hedgehog are mediated by BMP-2/4-dependent pathways in the neonatal rat metatarsal model. Endocrinology, 2001, 142(2): 940-947.
89. Fagenholz PJ,Warren SM,Greenwald JA, Llan JB. Osteoblast gene expression is differentially regulated by TGF-β isoforms.J Craniofac Surg,2001, 12(2): 183-190.
90. Gallea S,Lallemand F, Atfi A. Activiation of mitogen-activated protein kinase cascades is involved in regulation of bone morphogenetic protein 2-induced osteoblast differentiation in pluripotent C2C 12 cells.Bone,2001, 28(5): 491498.
91. Gibert L, He X,Farmer P. Expression of the osteoblast differentiation factor RUNX2(Cbfal/AML3/Pebp2alphaA) is inhibited by tumor necrosis factor-alpha.J Biol Chem,2002, 277: 2695-2701.
92. Levanon D, Negreanu V, Bernstein Y, Bar-Am I, AVIVI L, Groner Y:AMLI, AML2, and AML3, the human members of the runt domain gene-family: cDNA structure, expression, and chromosomal localization. Genomlcs 1994, 23: 425-432.
93. Wotton D, Ghysdael J, Wang S, Speck NA, Owen M J: Cooperative binding of ets-1 and core binding factor to DNA. MO/Cell Biol 1994, 14: 840-850.
94. Zhang Y, Bae S, Takahashi E, Ito Y: The cDNA cloning of thetranscripts of human PEBP2aA/CBFAI mapped to 6p12.3--21.I,the locus for cleidocranial dysplasia. Oncogene 1997, 15: 367-371.
95. Lomri A,Lemonnier J,Delannoy P. Increased expression of protein kinase Calpha, interleukin-Ialpha, and RhoA guanosine 5'-triphosphatase in osteoblasts expressing the Ser252Trp fibroblast growth factor 2 Apert mutation:identifieation by analysis of complementary DNA microarray.J Bone Miner Res,2001, 16(4): 705-712
96. Abe, E, Yamamoto M., Taguchi Y, Lecka-Czernik B, O'Brien CA.,Economides A. N, Stahl N, Jilka R L, and Manolagas SC. Essential requirement of BMPs-2/4 for both osteoblast and osteoclast formation in murine bone marrow cultures from adult mice: Antagonism by noggin. J Bone Miner Res 15: 663-673; 2000.
97. Anderson D M., Maraskovsky E, Billingsley WL, Dougall WC, Tometsko ME, Roux ER., Teepe MC, DuBose RF, Cosman D, and Galibert L.A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function. Nature 390: 175-179; 1997.
98. Ashton BA, Allen TD, Howlett CR, Eaglesom CC, Hattori A,and Owen M. Formation of bone and cartilage by marrow stromal cells in diffusion chambers in vivo. Clin Orthopaed Rel Res 151: 294-307; 1980.
99. Benson MD, Aubin JE, Xiao G, Thomas PE, and Franceschi RT.Cloning of a 2.5 kb murine bone sialoprotein promoter fragment and functional analysis of putative Osf2 binding sites. J Bone Miner Res 14: 396-405; 1999.
100. Corral DA., Amling M., Priemel M, Loyer E, Fuchs S, Duty P, Baron R, and Karsenty G. Dissociation between bone resorption and bone formation in osteopenic transgenic mice. Proc Natl Acad Sci USA 95: 13835-13840; 1998.
101. Ducy P, Amling M., Takeda S, Priemel M., Schilling A F, Beil FT, Shen J, Vinson C, Rueger J M., and Karsenty G. Leptin inhibits bone formation through a hypothalamic relay: A central control of bone mass. Cell 100: 197-207; 2000.
102.Zhou YX,Xu X,Ch L1. A Pro250 Arg substitution in mouse Fgfrl causes increased expression of cbfal and premature fusion of calvarial sutures.Hum Mol Genet, 2000, 9(13): 2001-2008.
103.Kannan S, Lasmoles F, Oyajobi BO, Chen H. Reciproal control of osteoblast/chondroblast and osteoblast/adipocyte differentiation of multipotential clonal human marrow stromal F/STRO-1 +cells.J Cell Biochem,2001, 81(1): 23-38.
104.Mernman HL, van Wljnen A J, Hiebert S, BIdwell JP, Fey E, Llan J,Stein J, Stein GS: The tissue-specific nuclear matrix protein. NMP-2. is a member of the AML/CBF/PEBP2/Runt domain transcription factor family: interaction with the osteocalcin gene promoter. Bochemistry 1995, 34: 13125-13132.
105.Geoffroy V, Duty P, Karsenty G: A PEBP2cL/AML-I -related factor increases osteocalcin promoter activity through its binding to an osteoblast-specific cis-acting element. J Viol Chem 1995, 270: 30973-30979.
106.Banerlee C, Hlebert SW, Stein JL, Liar- JB, Stein GS: An AML-1 consensus sequence binds an osteoblast-specific complex and transcriptionally activates the osteocalcin gene. Proc Nail Acad SC/ USA 1996, 93: 4968-4973.
107.Mlyazakl Y. Setoguchl M, Yoshlda S, Hlguchl Y, Aklzukl S, Yamamoto S:The mouse osteopontin gene. J B/o/ Chem 1990, 265: 14432-14438.
108.Yang R, Gerstenfeld LC: Structural analysis and characterization of tissue and hormonal responsive expression of the avian bone sialoprotein (BSP) gene. J Cell Bwchem 1996, 64: 128-139.
109.Rossert J, Eberspaecher H, Crombrugghe BD: Separate cis-acting DNA elements of the mouse pro-al (I) collagen promoter direct expression of reporter genes to different type I collagen-producingcells in transgenic mice. J Cell.&o/1995, 129: 142 1-1432.
110.Bedalov A, Salvatorl R, Dodlg M, Kronenberg MS, Kapural B,Bogdanovlc Z, Kream BE, Woody CO, Clark SH, Mack K:Regulation of collal expression in type I collagen producing tissues:identification of a 49 base pair region which is required for transgene expression in bone of transgenic mice. J Bone Miner Res 1995. 10: 1443-1451.
111.TSUJI K, Ito Y, Noda M: Expression of the PEBP2aA/AML3/CBFAl gene is regulated by BMP4/7 heterodimer and its overexpressionsuppresses type I collagen and osteocalcin gene expression inosteoblastic and nonosteoblastic mesenchymal cells. Bone 1998, 22. 87-92.
112.Chang DJ, JI C, Kim KK, Casmghlno S, McCarthy TL, Centrella M:Reduction in transforming growth factor p receptor I expression and transcription factor CBFal on bone cells by glucocorticoid. J 6101 Chem 1998, 273: 4892-4896.
113.Lee B, Thirunavukkarasu K, Zhou L, Pastore L, BaldIn A, Hecht J, Geoffroy V, Duty P, Karsenty G: Missense mutations abolishing DNA binding of the osteoblast-specific transcription factor OSF2/CBFAi in cleidocranial dysplasia. Nat Genet 1997, 16: 307-309
114.Stewart M, Terry A, Hu M, O'hara M, Blyth K, Baxter E, Cameron E,Onions DE, Neil JC: Proviral insertions induce the expression of bone-specific isoforms of PEBP2aA (CBFAI): evidence for a newmyc collaborating oncogene. Proc Nat/Acad So USA 1997, 94: 8646-8651.
115.Pend& AM, Balbin M, Llano E, Jimenez MG, Lbpez-Ottn C: Structural analysis and promoter characterization of the human collagenase-3 (MMPI3). Genomtcs 1997, 40: 222-233.
116.Mc Cauley LK.Kolr Paige AJ.Chen H. Parath.yroid hormone stimulates fra-2 expression in osteoblastic cells in vitro and in vivo.Endocrinology, 2001, 142(5): 1975-1981.
117.Naot D,Callon KE,Grey A. A potential role for adrenomedullin as a local regulator of bone growth[J ]. Endocrinology,2001, 142(5): 1849-1857.
118.Hay E.Lemonnier J,Modrowski D. N- and E-cadherin mediated early human calvaria osteoblast differentiation promoted by bone morphgenetie protein-2.J Cell Physiol,2000, 183(1): 117-128.
119.Zalman AL, Lenz J: Transcriptional activation of a retrovirus enhancerby CBF (AMLI) requires a second factor: evidence for cooperativity with c-Myb. J I/iro/1996, 70: 5618-5629.
120.Schorpp M, Mattel M-G, Herr I, Gack S, Schaper J, Angel P:Structural organization and chromosomal localization of the mouse collagenase type I gene. B--ochem J 1995, 308: 211-217.
121.Zhang D, Hethermgton C J, Meyers S, Rhoades KL, Larson CJ,Chen H, Hlebert SW, Tenen DG: CCAAT enhancer-binding protein (C/EBP) and AML1 (CBFa2) synergistically activate the macrophage colony-stimulating factor receptor promoter. Mol Cell Biol 1996, 16: 1231-1240.
122.Tardlf G, Pelletier J-P, Dupuis M, Hambor JE, Mar&-Pelletler J:Cloning, sequencing and characterization of the 5' -flankingregion of the human collagenase-3 gene. B/o&em J 1997, 323: 13-16.
123.Otto T, Sugimoto T, Saijoh K, FUJII M. Chihara K: Cloning and characterization of the 5' -flanking region of the mouse diastrophic dysplasia sulphate transpoter gene. Bochem Bmphys Res Commun 1997, 238: 738-743.
124.Bronckers AL,Sasguri K, Engeles MH.Transcription and immunolocalization of Runx2/Cbfal/PebpzαA in developing osteoblasts phagocytose osteocytes.Microsc Res Tech,2003; 61(6): 540-548.
125.Djouad F, Plence P, Bony C, Schaper J. ImmunosuPpressive elect of ruesenchym stem cells favors tumor growth in allogeneic animals. Blood, 2003, 102(10): 3837--3844.
126.Lazarus HM, Koco N, Devine SM, Hart Q. Cotransplantation of HLA. identical sibling culture, expanded mesenchymal stem cells and hemato. poietic stem cells in hematologic malignancy patients. Biol BloodMarrow Transplant. 2005, 11(5): 389--398.
127.Bartholomew A, Sturgeon C, Siatskas M. Mesenchymal stem cells suppress lymphocyte proliferation in vitro and prolong skin survival in vivo. Exp Hematol, 2002, 30(1): 42-48.
128.Deng W, Han Q, Liao L, Le Blanc K. Long--term distribution of adultstem cells in diferent tissues after transplantation. Stem Cell and CellularTherapy, 2003, 1(1): 62--65.
129.Devine S M, Cobbs C, Jennings M, Mevorach D. Mesenchymal stem cells distribute to a wide range of tissues following systemic infusion into nonhuman primates. Blood, 2003, 101(8): 2999--3001.
130.Aggarwal S, Pittenger M F. Human mesenchymal stem cels modulate allogeneic immune cell responses. Blood, 2005, 105(4): 1815--1822.
131.Le Blanc K, Rasmusson I, Gotherstrem C. Mesenchymal stem cells inhibit the expression of CD25 (interleukin-2 receptor) and CD38 on phytohaemagglutinin--activated lymphocytes. Scand JImmunol, 2004, 60(3): 307--315.
132.Beyth S, Borovsky z, Mevorach D. Human mesenchymal stem cells alter antigen presenting cell maturation an d induce (?). Blood, 2005, 105(5): 2214--2219.
133.Arinzeh T L, Peter S J, Arehambault M P, et al. Allogeneic II chymal stem cells regenerate bone in a critical--sized canine segmental defect. J Bone Joint Surg Am, 2003, 85-A(10): 1927--1935.
134.Vacanti CA, Bonassar LJ. An overview of tissue engineered bone. Clin Orthop, 1999, (367Supp 1): 375.
135.Chen JL, Hunt P, McElvain M. Osteoblast precursor cells are found in CD34+ cells from human bone marrow. Stem Cells, 1997, 15(5): 368-375
136.Vogel W, Grunebach F, Messam CA, et al. Heterogeneity among human bone marrow---derived mesenchymal stem cells and neural progenitor cells. Haematologica, 2003, 88(2): 126-131
137.Nuttall ME, Patton A J, Olivera DL, Hart Q. Human trabecar bone cells are able to expres bo th osteoblastic an d adipo cytic phenotype: implications for osteogenic disorders. J Bone Miner Res, 1998, 13(3): 371-376
138.Khor E, Lim LY. Implantable applications ofchitin andchitosan.Biomaterials 2003; 24: 2339-2349.
139.Domish M, Kaplan D, Skaugrud O. Standards and guidelines for biopolymers in tissue-engineered medical products. Ann N Y Acad Sci 2001; 944: 388-397.
140.Athanasiou KA, Shah AR, Hernandez RJ, LeBaron RG. Basic science ofarticular cartilage repair. Clin Sports Med 2001; 20: 223-247.
141.Madihally SV, Matthew HWT. Porous chitosan scaffolds for tissue engineering. Biomaterials 1999; 20: 1133-1142.
142.Mao JS, Cui YL, Wang XH, Sun Y, Yin YJ, Zhao HM, De Yao K. A preliminary study on chitosan and gelatin polyelectrolyte complex cytocompatibility by cell cycle and apoptosis analysis.Biomaterials 2004; 25: 3973-3981.
143.VandeVord PJ, Matthew HW, DeSilva SP, Mayton L, Wu B,Wooley PH. Evaluation ofthe biocompatibility of a chitosanscaffold in mice. J Biomed Mater Res 2002; 59: 585-590.
144.Suh JKF, Matthew HWT. Application ofchitosan-base d polysaccharide biomaterials in cartilage tissue engineering: a review.Biomaterials 2000; 21: 2589-2598.
145.Hu Q, Li B, Wang M, Shen J. Preparation and characterization of biodegradable chitosan/hydroxyapatite nanocomposite rods via in situ hybridization: a potential material as internal fixation of bone fracture. Biomaterials 2004; 25: 779-785.
146.Risbud MV, Bhonde RR. Polyacrylamide-chitosan hydrogels: in vitro biocompatibility and sustained antibiotic release studies.Drug Deliv 2000; 7: 69-75.
147.Chow KS, Khor E. Novel fabrication of open-pore chitin matrixes. Biomacromolecules 2000; 1: 61--67
148.Geng L, Feng W, Hutmacher DW, Wong Y, Loh H, Fuh JYH.Direct writing of chitosan scaffolds using a robotic system. Rapid Prototyping J 2005; 11: 90-97.
149.Aimin C, Chunlin H, Juliang B, Tinyin Z, Zhichao D. Antibiotic loaded chitosan bar. An in vitro, in vivo study of a possible treatment for osteomyelitis. Clin Orthop 1999; 366: 239-247.
150. Hu SG, Jou CH, Yang MC. Protein adsorption, fibroblast activity and antibacterial properties ofpoly(3-hyd roxybutyric acid-co-3-hydroxyvaleric acid) grafted with chitosan and chitooligosaccharide after immobilized with hyaluronic acid. Biomaterials2003 ;24:2685-2693.
151. Buranapanitkit B, Srinilta V, Ingviga N, Oungbho K, Geater A, Ovatlarnpom C. The efficacy of a hydroxyapatite composite as a biodegradable antibiotic delivery system. Clin Orthop 2004;424:244-252.
152. Chenite A, Chaput C, Wang D, Combes C, Buschmann MD,Hoemann CD, Leroux JC, Atkinson BL, Binette F, Selmani A.Novel injectable neutral solutions ofchitosan form biodegradable gels in situ. Biomaterials 2000;21:2155-2161.
153. Tuzlakoglu K, Alves CM, Mano JF, Reis RL. Production and characterization of chitosan fibers and 3-D fiber mesh scaffolds for tissue engineering applications. Macromol Biosci 2004;4:811-819.
154. Xia W, Liu W, Cui L, Liu Y, Zhong W, Liu D, Wu J, Chua K,Cao Y. Tissue engineering ofcartilage with the use ofchitosan-gelatin complex scaffolds. J Biomed Mater Res 2004;71 B:373-380.
155. Risbud M, Ringe J, Bhonde R, Sittinger M. In vitro expression of cartilage-specific markers by chondrocytes on a biocompatible hydrogel: implications for engineering cartilage tissue. Cell Transplant 2001;10:755-763.
156. Sittinger M, Hutmacher DW, Risbud MV. Current strategies for cell delivery in cartilage and bone regeneration. Curr Opin Biotechnol 2004;15:411-418
157．秦辉，许建中，周强，胡光运．两种构建方法对组织工程骨种子细胞粘附和增殖的影响．第三军医大学学报，2004，26(11)：990—992．
158．赵敏，许建中，周强，李新民．脉冲电磁场促进人骨髓问充质干细胞成骨的研究．中国矫形外科杂志，2004，12(6)：43-45
159．朱灏，许建中，周强，李新民．不同年龄人骨髓问充质干细胞体??外增殖及成骨分化的研究．中国矫形外科杂志，2004．12(10)：759—761．
160. Abdallah B M, Haack Sorensen M, Bums J S. Maintenance of diferentiation potential of human bo ne manDw mesenchymal stem tens immortalized by human telomerase reverse transcdptase gene despite of extensive proliferation. Biochem Biophys Res Commun, 2005. 326(3): 527—538.
161．裴国献，金丹．骨组织工程学种子细胞研究进展．中华创伤骨科杂志，2003，5(1)：55-57
162. Lahiji A, Sohrabi A, Hungerford DS, Frondoza CG, Lahiji A,Sohrabi A, Hungerford DS, Frondoza CG. J Biomed Mater Res2000;51(4):586-595;
163. Iwasaki N, Yamane ST, Majima T, Kasahara Y, Minami A,Harada K, Nonaka S, Maekawa N, Tamura H, Tokura S, Shiono M, Monde K, Nishimura S. Feasibility ofpolysacc haride hybrid materials for scaffolds in cartilage tissue engineering: evaluation ofchondrocyte adhesion to polyion complex fibers prepared from alginate and chitosan. Biomacromolecules 2004;5:828-833.
164. Cui YL, Qi AD, Liu WG, Wang XH, Wang H, Ma DM, Yao KD. Biomimetic surface modification of poly(L-lactic acid) with chitosan and its effects on articular chondrocytes in vitro.Biomaterials 2003;24:3859-3868.
165. Yamane S, Iwasaki N, Majima T, Funakoshi T, Masuko T,Harada K, Minami A, Monde K, Nishimura S. Feasibility of chitosan-based hyaluronic acid hybrid biomaterial for a novel scaffold in cartilage tissue engineering. Biomaterials 2005;26:611-619.
166. Niemeyer P, Secldnger A, Simank H G, Helm GA. Allogenic transplantation of human mesenchymal stem cells for tissue engineering purposes: fluirtro study. Orthopade, 2004, 33(12): 1346—1353.
167. Lee JE, Kim KE, Kwon IC, Aim HJ, Lee SH, Cho H, Kim HJ,Seong SC, Lee MC. Effects of the controlled-released TGF-beta 1 from??chitosan microspheres on chondrocytes cultured in a collagen/chitosan/glycosaminoglycan scaffold. Biomaterials 2004; 25: 4163--4173. 168.Taravel MN, Domard A. Collagen and its interactions with chitosan, III some biological and mechanical properties. Biomaterials1996; 17: 451-455.
169.Kim SE, Park JH, Cho YW, Chung H, Jeong SY, Lee EB, Kwon IC. Porous chitosan scaffold containing microspheres loaded with transforming growth factor-betal: implications for cartilage tissue engineering. J Control Release 2003; 91: 365-374.
170.Mattioli-Belmonte M, Gigante A, Muzzarelli RA, Politano R, De Benedittis A, Specchia N, Buffa A, Biagini G, Greco F. N,Ndicarboxymethyl chitosan as delivery agent for bone morphogenetic protein in the repair ofarticular cartilage. Med Biol Eng Comput 1999; 37: 130-134.
171.Lu JX, Prudhommeaux F, Meunier A, Sedel L, Guillemin G.Effects of chitosan on rat knee cartilages. Biomaterials 1999; 20: 1937-1944.
172.Boden SD. Overview ofthe biology of lumbar spine fusion and principles for selecting a bone graft substitute. Spine 2002; 27(Suppl 16): S26-31.
173.Hankey DP, McCabeRE, DohertyMJ, etal. Enhancementof human osteoblast proliferation and phenotypic expression when cultured in hurnfln senl(?)1. Acta()nhp Scand, 2001, 72(4): 395-401
174.Cooper LF, Masuda T, Whitson SW. et al. Formation of miner alizing osteoblast cultures on machined, titan ium oxide grit--blasted, and plasma--sprayed titanium surfaces. Int J Oral Maxillo. fac Implants, 1999, 14(1): 37-41
175.Saito N, Takaoka K. New synthetic biodegradable polymers as BMP carders for bone tissue engineering. Biomaterials 2003; 24: 2287-2293.
176.Alden TD, Varady P, Kallmes DF, Jane Jr JA, Helm GA. Bone morphogenetic protein gene therapy. Spine 2002, 27(Suppl 16): S87-93.
177. Yoon ST, Boden SD. Spine fusion by gene therapy. Gene Ther 2004; 11(4):360-367.
178. Wang J, de Boer J, de Groot K. Preparation and characterization ofelectrodepo sited calcium phosphate/chitosan coating onTi6A14V plates. J Dent Res 2004;83:296-301.
179. Bonadio J. Tissue engineering via local gene delivery: update and future prospects for enhancing the technology. Adv Drug Deliv Rev 2000;44:185-194.
180. Cha CW, Boden SD. Gene therapy applications for spine fusion.Spine 2003;28(Suppl 15):S74-84.
181. Lee KY, Kwon IC, Kim YH, Jo WH, Jeong SY. Preparation of chitosan self-aggregates as a gene delivery system. J Control Release 1998 ;51:213-220.
182. Koping-Hoggard M, Varum KM, Issa M, Danielsen S,Christensen BE, Stokke BT, Artursson P. Improved chitosanmediated gene delivery based on easily dissociated chitosan polyplexes ofhighly defined chitosan oligomers. Gene Ther 2004; 11:1441-1452.
183. Gan JC, Ducheyne P, Vresilovic EJ, Swaim W, Shapiro IM.Intervertebral disc tissue engineering I: characterization of the nucleus pulposus. Clin Orthop 2003;411:305-314.
184. Yung Lee J, Hall R, Pelinkovic D, Cassinelli E, Usas A,Gilbertson L, Huard J, Kang J. New use of a three-dimensional pellet culture system for human intervertebral disc cells: initial characterization and potential use for tissue engineering. Spine 2001 ;26:231 6-2322.
185．王常勇，王永红，郭希民，等．应用旋转生物反应器和微载体技术体外大规模扩增骨髓间充质干细胞．中华实验外科杂志，2002，19(4)：378-380
186. An HS, Thonar EJ, Masuda K. Biological repair ofinterverteb ral disc. Spine 2003;28(Suppl 15):S86-92.
187. Alini M, Roughley PJ, Antoniou J, Stoll T, Aebi M. A biological approach to treating disc degeneration: not for today, but maybe for tomorrow. Eur Spine J 2002;11(Suppl 2):S215-220.
188. Seol YJ, Lee JY, Park YJ, Lee YM, Young-Ku, Rhyu IC, Lee SJ,Han??SB, Chung CP. Chitosan sponges as tissue engineering scaffolds for bone formation. Biotechnol Lett 2004; 26: 1037-1041.
189.Seeherman H, Li R, Wozney J. A review ofpreclinic al program development for evaluating injectable carriers for osteogenic factors. J Bone Jt Surg Am 2003; 85A(Suppl 3): 96-108.
190.Zhang Y, Zhang M. Synthesis and characterization ofmacroporous chitosan/calcium phosphate composite scaffolds for tissue engineering. J Biomed Mater Res 2001; 55: 304-312.
191.Zhang Y, Zhang M. Calcium phosphate/chitosan composite scaffolds for controlled in vitro antibiotic drug release. J Biomed Mater Res 2002; 62: 378-386.
192.Hung SC, Chen NJ, Hsieh SL, et al. Isolation and characterization of size--sieved stem cells from hum an bone marrow. Stem Cells, 2002, 20(3): 249-253
193.Zhang Y, Ni M, Zhang M, Rather B. Calcium phosphatechitosan composite scaffolds for bone tissue engineering. Tissue Eng 2003; 9: 337-345.
194.Ge Z, Baguenard S, Lim LY, Wee A, Khor E. Hydroxyapatitechitin materials as potential tissue engineered bone substitutes.Biomaterials 2004; 25: 1049-1058.
195.Kawakami T, Antoh M, Hasegawa H, Yamagishi T, Ito M, Eda S. Experimental study on osteoconductive properties ofa chitosan-bonded hydroxyapatite self-hardening paste. Biomaterials 1992; 13: 759-763.
196.Yin Y, Ye F, Cui J, Zhang F, Li X, Yao K. Preparation and characterization ofmacroporous chitosan-gelatin/beta-tricalcium phosphate composite scaffolds for bone tissue engineering.J Biomed Mater Res 2003; 67A: 844-855.
197.Fleming JE Jr, Cornell CN, Muschler GF. Bone cells and matrices in orthopedic tissue engineering. Orthop Clin North Am, 2000, 3t(3): 357-362
198.Zuk PA, Zhu M, Mizuno H. Muhilineage cells from humall adipose tisue: implications for cell--based therapies. Tisue Eng. 2001, 7(2):211-213.
199.Leroux L, Hatim Z, Freche M, Lacout JL. Effects of various adjuvants (lactic acid, glycerol, and chitosan) on the injectability of a calcium phosphate cement. Bone 1999; 25(Suppl 2): 31-41.
200.Gutowska A, Jeong B, Jasionowski M. Injectable gels for tissue engineering. Anat Rec 2001; 263: 342-349.
201.Wang X, Ma J, Wang Y, He B. Bone repair in radii and tibias of rabbits with phosphorylated chitosan reinforced calcium phosphate cements. Biomaterials 2002; 23: 4167-4176.
202.Xu HH, Quinn JB, Takagi S, Chow LC. Synergistic reinforcement of in situ hardening calcium phosphate composite scaffold for bone tissue engineering. Biomaterials 2004; 25: 1029-1037.
203.Kim SB, Kim YJ, Yoon TL, Park SA, Cho IH, Kim EJ, Kim IA, Shin JW. The characteristics of a hydroxyapatite-chitosan-PMMA bone cement. Biomaterials 2004; 25: 5715-5723.
204.Buttery LD, Bourne S, Xynos JD, et al. Differentiation of 08-teoblast and in vitro bone formation from murine embryonic stem cells. Tissue Eng, 2001, 7(1): 89-93
205.Park Y J, Lee YM, Park SN, Sheen SY, Chung CP, Lee SJ. Platelet derived growth factor releasing chitosan sponge for periodontal bone regeneration. Biomaterials 2000; 21: 153-159.
206.Lee JY, Nam SH, Im SY, Park YJ, Lee YM, Seol YJ, Chung CP,Lee SJ. Enhanced bone formation by controlled growth factor delivery from chitosan-based biomaterials. J Control Release 2002; 78: 187-197
207.Petite MF, Mackay AM, Beck SC. Multilineage Ix)tential of adult human mesenchymal stem ceils. Science, 1999, 284(5411): 143-147
208.Lee YM, Park Y J, Lee S J, Ku Y, Han SB, Choi SM, Klokkevold PR, Chung CP. Tissue engineered bone formation using chitosan/tricalcium phosphate sponges. J Periodontol 2000; 71: 410--417.
209.Muzzarelli RA, Mattioli-Belmonte M, Tietz C, Biagini R, Ferioli G, Brunelli MA, Fini M, Giardino R, Ilari P, Biagini G. Stimulatory effect on bone formation exerted by a modified chitosan. Biomaterials1994;15:1075-1081.
210. Lee YM, Park YJ, Lee SJ, Ku Y, Han SB, Klokkevold PR, Chung CP. The bone regenerative effect of platelet-dedved growth factor-BB delivered with a chitosan/tdcalcium phosphate sponge carder. J Periodontol 2000;71:418-424.
211. Kim IS, Park JW, Kwon IC, Baik BS, Cho BC. Role ofBMP,betaig-h3, and chitosan in early bony consolidation in distraction osteogenesis in a dog model. Plast Reconstr Surg 2002; 109:1966-1977.
212. Cho BC, Park JW, Balk BS, Kwon IC, Kim IS. The role of hyaluronic acid, chitosan, and calcium sulfate and their combined effect on early bony consolidation in distraction osteogenesis of a canine model. J Craniofac Surg 2002; 13:783-793.
213. Bumgardner JD, Wiser R, Gerard PD, Bergin P, Chestnutt B,Marin M, Ramsey V, Elder SH, Gilbert JA. Chitosan: potential use as a bioactive coating for orthopaedic and craniofacial/dental implants. J Biomater Sci Polym Ed 2003;14:423-438.
214. Bumgardner JD, Wiser R, Elder SH, Jouett R, Yang Y, Ong JL.Contact angle, protein adsorption and osteoblast precursor cell attachment to chitosan coatings bonded to titanium. J Biomater Sci Polym Ed 2003;14:1401-1409.
215．郭刚，王臻．彭磊，等．成骨细胞异体移植的免疫学初探．第四军医大学学报，2002，23(11)：976-980
216. Anselme K, Noel B, Flautre B, et al. Association of porous hydroxyapatite and bone marrow cells for bone regeneration. Bone, 1999, 25(2Suppl 1): 51-53
217. Kadiyala S, Young RG, Thiede MA, et al. Culture expanded can ine mesenchymal stem ceUs po ses osteoe hondrog enic potential in vivo and invitro. CeU Transplant, 1997, 6(2): 125-131
218. Chen D, Ji X, Harris MA, et al. Differential roles for bone morphogenetic protein(BMP)receptor type IB and IA in differentiation an dspecification of mesenchymal precursor cells to osteoblast and adipoeytelineages. J CellBiol, 1998, 142(1): 295-301219.He TC, Zhou S, da Costa LT, Yu J, Kinzler KW, Vogelstein B. A simplified system for generating recombinant adenovirus.Proc Natl Acad Sci USA,1998; 95(5): 2509-2514.
220.220.Jadlowiec JA, Celil AB, Hollinger JO. Bone tissue engineering: recent advances and promising therapeutic agents. Expert Opin Biol Ther, 2003, 3(3): 409-423.
221.Freed L, Novakovic, Biron RJ. Biodegradable polymer scaffolds fortissue engineering. Biotechnics, 1994, 12(7): 689-676.
222.Sun JS, Liu HC, Chang WH, Li J, Lin FH, Tai HC. Influence of hydroxyapatite particle size on bone cell activities: an in vitro study. J Biomed Mater Res, 1998, 39(3): 390-398.
223.Goshima J, Goldberg VM, Caplan AI. The osteogenic potential of culture expanded rat marrow mesenchymal cells assayed in vivo in calcium phosphate ceramic blocks. Clin Orthop, 1991, (262): 298-305.
224.Musgrave DS, Fu FH, Huard J.Gene therapy and tissue engineering in orthopaedic surgery. J Am Acad Orthop surg, 2002, 10(1): 6-15.
2. Hogan BLM: Bone morphogenetic proteins: multifunctionalregulators of vertebrate development. Genes Dev 1996, 10: 1580-1594.
3. Tabin C J: Retinoids, homeoboxes, and growth factors: towardmolecular models for limb development. Cell 1991, 66: 199-217.
4. Hall BK, Miyake T: The membranous skeleton: the role of cell condensations in vertebrate skeletogenesis. Anat Embryo/1992, 186: 107-124.
5. Horton WA: Morphology of connective tissue: cartilage.In Connective Tissue and Its Heritable Disorders. New York: Wiley-Liss, Inc; 1993: 73-84.
6. Lui VC, Ng L J, Nicholls J, Tam PP, Cheah KS: Tissue-specific and differential expression of alternatively spliced alpha 1 (11) collagenmRNAs in early human embryos. Dev Dyn 1995, 203: 198-211.
7. Ryan MC, Sandell L J: Differential expression of a cystein-dchdomain in the amino-terminal propeptide of type II (cartilage)procollagen by alternative splicing of mRNA, J Bio/Chem 1990, 265: 10334-10339.
8. Sandell U, Morris N, Robbins JR, Goldring MB: Alternatively splicedtype II procollagen mRNAs define distinct populations of cellsduring vertebral development: differential expression of theamino-propeptide. J Ce//Bio/1991, 114: 1307-1319.
9. Luo G, D'Souza R, Hogue D, Karsenty G: The matrix Gla proteingene is a marker of the chondrogenesis cell lineage during mouse development. J Bone Miner Res 1995, 10: 325-334.
10. Thorogood P: Differentiation and morphogenesis of cranial skeletal tissues. In The Skull. Edited by J Hanken, BK Hall. Chicago:University of Chicago Press; 1993.
11. Grigodadis AE, Heersche JNM, Aubin JE: Analysis of chondroprogenitor frequency and cartilage differentiation in anovelfamily of clonal chondrogenic rat cell lines. Differentiation 1996, 60: 299-307.
12. Suda T, Takaharashi N, Martin TJ: Modulation of osteoclast differentiation. Endocr Rev 1992, 13: 66-80.
13. Ng L J, Wheatley S, Muscat GE, Conway-Campbell J, Bowles J, o Wright E, Bell DM, Tam PP, Cheah KS, Koopman P: SOX9 binds DNA, activates transcription, and coexpresses with type II collagen during chondrogenesis in the mouse. Dev Biol 1997, 183: 108-121.
14. Zhao Q, Eberspaecher H, Lefebvre V, De Crombrugghe B: Parallelexpression of Sox9 and Col2al in cells undergoingchondrogenesis. Dev Oyn 1997, 209: 377-386.
15. Wagner T, Wirth J, Meyer J, Zabel B, Held M, Zimmer J, Pasantes J,Bricarelli FD, Keutel J, Hustert E: Autosomal sex reversal andcampomelic dysplasia are caused by mutations in and around theSRY-related gene SOX9. Cell 1994, 79: 1111-1120.
16. Lefebvre V, Huang W, Harley VR, Goodfellow PN, De Crombrugghe B: SOX9 is a potent activator of the chondroeyte-specific enhancerof the pro alphal (11) collagen gene. Mol Cell Biol 1997, 17: 2336-2346.
17. Bell DM, Leung KK, Wheatley SC, Ng L J, Zhou S, Ling KW,Do Sham MH, Koopman P, Tam PP, Cheah KS: SOX9 directly regulatesthe type-II collagen gene. Nat Genet 1997, 16: 174-178.
18. Suva U, Winslow GA, Wettenhall RE, Hammonds RG, Moseley JM,Diefenbach-Jagger H, Rodda CP, Kemp BE, Rodriguez H, Chen EYet aL: A parathyroid hormone-related protein implicated inmalignant hypercalcemia: cloning and expression. Science 1987, 237: 893-896.
19. Karaplis AC, Luz A, Glowacki J, Bronson RT, Tybulewicz VL,Kronenberg HM, Mulligan RC: Lethal skeletal dysplasia fromtargeted disruption of the parathyroid hormone-related peptidegene. Genes Dev 1994, 8: 277-289.
20. Lanske B, Karaplis AC, Lee K, Luz A, Vortkamp A, Pirro A,Karperien M, Defize LHK, Ho C, Mulligan RC et al.: PTH/PTHrP receptor in early development and Indian hedgehog-regulated bonegrowth. Science 1996, 273: 663-666.
21. Schipani E, Kruse K, Juppner H: A constitutively active mutant PTHPTHrP receptor in Jansen-type metaphyseal chondrodysplasia.Science 1995, 268: 98-100.
22. Bitgood M J, McMahon AP: Hedgehog and Bmp genes arecoexpressed at many diverse sites of cell-cell interaction in themouse embryo. Dev Biol 1993, 172: 126-138.
23. Vortkamp A, Lee K, Lanske B, Segre GV, Kronenberg HM, Tabin CJ:Regulation of rate of cartilage differentiation by Indian hedgehogand PTH-related protein. Science 1996, 273: 613-622.
24. Chiang C, Litingtung Y, Lee E, Young KE, Corden JL, Westphal H,Beachy PA: Cyclopia and defective axial patterning in mice lackingSonic hedgehog gene function. Nature 1998, 383: 407-413.
25. Naski MC, Wang Q, Xu J, Omitz DM: Graded activation of fibroblast growth factor receptor 3 by mutations causing achondroplasiaand thanatophoric dysplasia. Nat Genet 1996, 13: 233-237.
26. Colvin JS, Bohne BA, Harding GW, McEwen DG, Ornitz DM:Skeletal overgrowth and deafness in mice lacking fibroblastgrowth factor receptor 3. Nat Genet 1996, 12: 390-397.
27. Vu TH, Shipley JM, Bergers G, Berger JE, Helms JA, Hanahan D,te Shapiro SD, Senior RM, Werb Z: M MP-9/Gelatinase B is a keyregulator of growth plate angiogenesis and apoptosis ofhypertrophic chomdrocytes. Cell 1998, 93: 411-422.
28. Ducy P, Karsenty G: Two distinct osteoblast-specific cis-actingelements control expression of a mouse osteocalcin gene.Mo/Cell Biol 1995, 15: 1858-1869.
29. Geoffroy V, Duty P, Karsenty G: A PEBP2/AML-I-related factorincreases osteocalcin promoter activity through its binding to anosteoblast-specific cis-acUng elemenL J BiD/Chem 1995, 270: 30973-30979.
30. Merriman HL, van Wijnen A J, Hiebert S, Bidwell JP, Fey E, Lian J,Stein J, Stein GS: The tissue-specific nuclear matrix protein,NMP-2, is a member of the AM L/CBF/PEBP2/Runt domaintranscription factorfamily: interactions with the osteocalcin gene promoter. Biochemistry 1995, 34: 13125-13132.
31. Gergen JP, Wieschaus E: The localized requirements for a gene affecting segmentation in Drosophila: analysis of larvae mosaic for runL Dev Bio11985, 109: 321-335.
32. Kagoshima H, Shigesada K, Satake M, Ito Y, Miyoshi H, Ohki M,Pepling M, Gergen P: The Runt domain identifies a new family ofheteromeric transcriptional regulators. Trends Genet 1993, 9: 338-341.
33. Ito Y, Bae S-C: The runt domain transcription factor PEBP2/CBF,and its involvment in human leukemia. In Oncogenes asTranscriptional Regulators, vol 2. Edited by Yaniv M, Ghysdael J.Basel: Birkhauser Verlag; 1997: 107-132.
34. Satake M, Nomura S, Yamagushi-lwai Y, Takahama Y, Hashimoto Y,Niki M, Kitamura Y, Ito Y: Expression of the runt domain-encodingPEBP'2a genes in T cells during thymic developmenL Mo/Cell BiD/1995, 15: 1662-1670.
35. D,,cy P, Zhang R, Geoffroy V, Ridall A, Karsenty G: Osf2/Cbfal: a transcriptional activator of ostaoblast differentiation. Cell 1997, 89: 747-754.
36. Otto F, Thornell AP, Crompton T, Denzel A, Gilmour KC, Rosewell IR, Stamp GWH, Beddington RSP, Mundlos S, Olsen B: Cbfal, a candidate gene for cleidocranial dysplasia syndrome, is essentialfor osteoblast differentiation and bone development. Cell 1997, 89: 765-771.
37. Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K,Shimizu Y, Bronson RT, GaD Y-H, Inada Met a/.: Targeted disruptionof Cbfal results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell 1997, 89: 755-764.
38. Mundlos S, Otto F, Mundlos C, Mulliken JB, Aylsworth AS, Albright S, Lindhout D, Cole WG, Henri W, Knoll JHM et aL: Mutations involvingthe transcription factor CBFA1 cause cleidocranial dysplasia. Ce111997, 89: 773-779.
39. Lee B, Thirunavukkarasu K, Zhou L, Pastore L, Baldini A, Geoffroy V, Duty P, Karsenty G: Missense mutations abolishing DNA bindingOSF2/CBFA1 in patients affected with cleidocranial dysplasia. Nat Genet 199'7, 16: 307-311.
40. Tondravi MM, McKercher SR, Anderson K, Erdmann JM, Ouiroz M,Maki R, Teitelbaum SL: Osteopetrosis in mice lackinghaematopoietic transcription factor PU.1. Nature 1997, 386: 81-84.
41. Yoshida H, Hayashi S, Kunisada T, Ogawa M, Nishikawa S,Okamura H, Sudo T, Shultz LD, Nishikawa S: The murine mutationosteopetrosis is in the coding region of the macrophage colonystimulating factor gene. Nature 1990, 345: 442-444.
42. Simonet WS, Lacey DL, Dunstan CR, Kelley M, Chang MS, Luthy R,Lee Nguyen HQ, Wooden S, Bennett L, Boone T et al.: Osteoprotegerin:a novel secreted protein involved in the regulation of bone density. Cell 1997, 89: 309-319.
43. Bucay N, Sarosi I, Dunstan CR, Morony S, Tarpley J, Capparelli C,Scully S, Tan HL, Xu W, Lacey DL eta/.: Osteoprotegedn-deflcient mice develop early onset osteoporosis and arterial calcification.Genes Dev 1998, 12: 1260-1268.
44. Lacey DL, Timms E, Tan HL, Kelley M J, Dunstan CR, Brugess T, Elliot R, Colombero A, Elliot G, Scully S, Hsu Her.:Osteoprotegerin ligand is a cytokine that regulates osteoclastdifferentiation and activation. Cell 1998, 93: 165-168.
45. Yasuda H, Shima N, Nakagawa N, Yamaguchi K, Kinosaki M, Mochizuki S, Tomoyasu A, Yano K, Goto M, Murakami A:Osteoclast differentiation factor is a ligand for osteoprotegerin/osteclastogenesis-inhibitory factor and isidentical to TRANCE/RANKL Proc Nat/Acad Sci USA 1998, 95: 3597-35602.
46. Wang ZQ, Ovitt C, Grigoriadis AE, Mohle-Steinlein U, Ruther U,Wagner EF: Bone and haeroatopoiatic defects in mice lackingc-los. Nature 1999, 360: 741-745.
47. Grigoriadis AE, Wang ZCI, Cecchini MG, Hofstetter W, FelixR,Fleisch HA, Wagner EF: c-Fos: a key regulator of osteoclastroacrephagelineage determination and bone remodeling.Science 1994, 266: 443-448.
48. Johnson RS, Spiegelman BM, Papaioannou V: Pleiotropic effects of a null mutation in the c-los proto-oncogene. Cell 1992, 71: 577-586.
49. Franzoso G, Carlson L, Xing L, Poljak L, Shores EW, Brown KD, Leonardi A, Tran T, Boyce BF, Siebenlist U: Requirement forNF-kappa B in osteoclast and B-cell development. Genes Dev1997, 11: 3482-3496.
50. Hodgkinson CA, Moore KJ, Nakayama A, Steingrimsson E,Copeland NG, Jenkins NA, Arnheiter H: Mutations at the mouseroicrophthalroia locus are associated with defects in a geneencoding a novel basic-helix-loop-helix-zipper protein. Cell 1993, 74: 395-404.
51. Satake M, Nomura S, Yamaguchi-lwai Y, Brown KD. Expression of the Runt domain-encoding PEBP2 alpha genes in T cells during thymic development. Mol Cell Biol. 1995; 15(3): 1662-70.
52. Mundlos S, Otto F, Mundlos C, Xing L. Mutations involving the transcription factor CBFA1 cause cleidocranial dysplasia. Cell. 1997; 89(5): 773-9.
53. Xiao ZS,Thomas R.,Hinson TK.. Genomic structure and isoform expression of the mouse, rat and human Cbfal/Osf2 transcription factor. Gene. 1998; 214(1-2): 187-97.
54. Harada H, Tagashira S, Fujiwara M, McLarren KW. Cbfal isoforms exert functional differences in osteoblast differentiation. J Biol Chem. 1999; 274(11): 6972-8.
55. Marks SC, Hermey DC: The structure and development of bone. In Principles of Bone Biology. Edtted by Bllezikian JP, Ralsz LG, RodanGA. London: Academic Press; 1996: 3-14.
56. Suds T, Udagawa N, Takahashl N: Cells of bone: osteoclastgeneration. in Prinuples of Bone Biology. Edited by Bilezikian JP,Ralsz LG, Rodan GA. London: Academic Press; 1996: 87-102.
57. Aubln JE, Turksen K, Heersche JNM: Osteoblastic lineage. In Cellularand Molecular Bology of Bone. Edited by Noda M. London:Academic Press; 1993: 1-45.
58. Welntraub H: The MyoD family and myogenesis: redundancy,networks, and thresholds. Cell 1993, 75: 1241-I 244.
59. Tontonoz P, Hu E, SpIegelman BM: Stimulation of adipogenesis infibroblasts by PPARr2, a lipid-activated transcription factor. Cell 1994, 79: 1147-1156.
60. Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K,Shimlzu Y, Bronson RT, Gao YH, lnada Met a/.: Targeted disruption of Cbfal results in a complete lack of bone formation owi& tomaturational arrest of osteoblasts. Cell 1997, 89: 755-764.
61. Otto F, Thornell AP, Crompton T, Denzel A, Gllmour KC, Rosewell IR, Stamo GWH. Beddlnaton RSP. Mundlos S. Olsen BR et al.: Cbfal. acandidate g&e for cieidocranial dysplasia syndrome, is essentialfor osteoblast differentiation and bone development. Cell 1997, 89: 765-771.
62. Duty P, Zhang R, Geoffroy V, Rldall AL, Karsenty G: Osf2/Cbfal: a transcriptional activator of osteoblast differentiation. Cell 1997, 89: 747-754.
63. Ogawa E, Maruyama M, Kagoshlma H, lnuzuka M, Lu J, Satake M,Shiaesada K. Ito Y: PEBP2IPEA2 represents a new familv of traiscription factor homologous toihe products of the drosophila runt and the human AMLf gene. Proc Nat/ Acad So USA 1993, 90: 6859-6863.
64. Bae SC, Yamaguchl-lwai Y, Ogawa E, Maruyama M, lnuzuka M,Kaaoshima H. Shiaesada K. Satake M. Ito Y: Isolation of PEBP2aB cDiA repre&ntiig the m&se homolog of human acute myeloidleukemia gene, AMLI. Oncogene 1993, 8: 809-814.
65. Bae SC, Takahashi E, Zhang VW, Ogawa E, Shigesada K, Namba Y,Satake M, Ito Y: Cloning, mapping and expression of PEBP2c&, athird gene encoding the mammalian Runt domain. Gene 1995, 159: 245-248.
66. Kanla MA, Banner AS, Duffy JB, Gergen JP: The Drosophilasegmentation gene runt encodes a novel nuclear regulatoryprotein that is also expressed in the developing nervous system.GenesDev1990, 4: 1701-1713.
67. Ogawa E, lnuzaka M, Maruyama M, Satake M, Naito-Fqmoto M, Ito Y,ShIResada K: Molecular cloning and characterization of PEBPSP,theheterodimeric partner of a novel Drosophila runt-related DNA binding protein PEBP2a V/rology 1993, 194: 314-331.
68. Vaillant F, Blyth K, Terry A. A full-length Cbfal gene product perturbs T-cell development and promotes lymphomagenesis in synergy with myc. Oncogene. 199925; 18(50): 7124-34.
69. Wang S, Wang a, Crute BE, Melnlkova IN, Keller SR, Speck NA:Cloning and characterization of subunits of the T-cell receptorand murine leukemia virus enhancer core-binding factor. MO/Cell Biol 1993, 13: 3324-3339.
70. Zeng C, Wtjnen AJV, Stein JL, Meyers S, Sun W, Shopland L,Lawrence JB, Penman S, Llan JB, Stein GS, Hiebert SW:Identification of a nuclear matrix targeting signal in the leukemiaand bone-related AML/CBF-a transcription factors. Proc Nat/ Acad SC; USA 1997, 94: 6746-6751
71. Kamachl Y, Ogawa E, Asano M, lshlda S, Murakami Y, Satake M, Ito Y,Shigesada K: Purification of a mouse nuclear factor that binds to both the A and B cores of the polyomavirus enhancer. J Viral 1990, 64: 4808-4819.
72. Drissi H,Lue Q, Shakoori R, Gibert L. Transcriptional autoregulation of the bone related CBFA 1/RUNX2gene.J Cell Physiol,2000, 1284(3): 341-350.
73. Wang S, Speck NA: Purification of core-binding factor, a protein that binds the conserved core site in murine leukemia virus enhancers. MO/ Cell B/o/1992, 12: 89-102.
74. Okuda T, van Deursen J, Hlebert SW, Grosveld G, Downing JR:AMLI, the target of multiple chromosomal translocation in humanleukemia, is essential for normal fetal liver hematopoiesis. Cell 1996, 84: 321-330.
75. Wang a, Stancy T, Binder M, Marin-PadlIla M, Sharpe AH, SpeckNA:Disruption of the Cbfa2 gene causes necrosis and hemorrh agingin the central nervous system and blocks definitive hematopoiesis. Proc Nat/Acad Sci USA 1996, 93:3444-3449.
76. Sasakl K, Yagl H, Bronson RT, Tomlnaga K, Matsunashl T, Deguchl K,Tani Y, Klshlmoto T, Komorl T: Absence of fetal liver hematopoiesis in mice deficient in transcriptional coactivator core binding factor. Proc Nat/AcadSo USA 1996, 93:i 2359-Ⅰ 2363.
77. Marzia M,Sims NA, Voit A. Decteaed c-Src expression enhances osteoblast differentiation and bone formation.J Cell Biol,2000,151 (2):311-320.
78. Wang Q, Stacy T, Miller JD, Lewis AF, Gu TL, Huang X, Bushweller JH, Bodes JC, Alt FW, Ryan Get al.: The CBFb subunit is essential for CBFa2 (AMLI) function in viva. Cell 1996, 87:697-708
79. Satake M, Nomura S, Yamaguchl-lwal Y, Takahama Y, Hashlmoto Y.Nikl M, Kltamura Y, Ito Y: Expression of the runt domain-encoding PEBP2a genes in T cells during thymic development. MO/Cell Bfol 1995, 15:1662-1670.
80. Wargnler A, Legros-Malda S, Bosselut R, Bourge JF, Lafaurle C, Ghysdael J, Sasportes M, Paul P: Identification of human granzyme B promoter regulatory elements interacting with activated T-cellspecific proteins: implication of Ikaros and CBF binding sites in promoter activation, froc Nat/AcadSo USA 1995, 92:6930-6934.
81. Geoffroy V, Corral DA, Zhou L, Lee B, Karsenty G: Genomic organization, expression of the human CBFAI gene, and evidence for an alternative splicing event affecting protein function. Mamm Genome 1998, 9:54-57.
82. Ahn MY, Bae SC, Maruyama M, Ito Y: Comparison of the humangenomic structure of the Runt domain-encoding PEBP2/CBFagene family. Gene 1996, 168:279-280.
83. Mundlos S. Otto F. Mundlos C. Mulliken JB. Avlsworth AS. Albrlaht S. Lindhout D, Cole WG, Henri W, Knoll JI-IM'etal.: Mutations invilvingthe transcription factor CBFAI cause eleidoeranial dysplasia. Cell 1997, 89:?73-779.84. Banerjee C, McCabe LR, Chow J, Hiebert SW, Stetn JL, Stein GS, Llan JB: Runt homology domain proteins in osteoblast differentiation: AMLI/CBFA1 is a maior component of a bonespecific complex. J Cell Biochem 1967, 66: 1-8.
85. Jarvls JL, Keats TE: Cleidocranial dysostosis, a review of 40 new cases. Am J Radio/1974, 121: 5-16.
86. Mundlos S, Mulllken JB, Abramson DL, Warman ML, Knoll JHM, Olsen BR: Genetic mapping of cleidocranial dysplasia and evidence of a microdeletion in one family. Hum MO/Genet 1995, 4: 71-75.
87. Mundlos S, Huang LF, Selby P, Olsen BR: Cleidocranial dysplasia in mice. Ann NY Acad So 1996, 785: 301-302.
88. Kvrishman V, Ma YL,Moseley JM. Bone anabolic effects of sonic/Indian hedgehog are mediated by BMP-2/4-dependent pathways in the neonatal rat metatarsal model. Endocrinology, 2001, 142(2): 940-947.
89. Fagenholz PJ,Warren SM,Greenwald JA, Llan JB. Osteoblast gene expression is differentially regulated by TGF-β isoforms.J Craniofac Surg,2001, 12(2): 183-190.
90. Gallea S,Lallemand F, Atfi A. Activiation of mitogen-activated protein kinase cascades is involved in regulation of bone morphogenetic protein 2-induced osteoblast differentiation in pluripotent C2C 12 cells.Bone,2001, 28(5): 491498.
91. Gibert L, He X,Farmer P. Expression of the osteoblast differentiation factor RUNX2(Cbfal/AML3/Pebp2alphaA) is inhibited by tumor necrosis factor-alpha.J Biol Chem,2002, 277: 2695-2701.
92. Levanon D, Negreanu V, Bernstein Y, Bar-Am I, AVIVI L, Groner Y:AMLI, AML2, and AML3, the human members of the runt domain gene-family: cDNA structure, expression, and chromosomal localization. Genomlcs 1994, 23: 425-432.
93. Wotton D, Ghysdael J, Wang S, Speck NA, Owen M J: Cooperative binding of ets-1 and core binding factor to DNA. MO/Cell Biol 1994, 14: 840-850.
94. Zhang Y, Bae S, Takahashi E, Ito Y: The cDNA cloning of thetranscripts of human PEBP2aA/CBFAI mapped to 6p12.3--21.I,the locus for cleidocranial dysplasia. Oncogene 1997, 15: 367-371.
95. Lomri A,Lemonnier J,Delannoy P. Increased expression of protein kinase Calpha, interleukin-Ialpha, and RhoA guanosine 5'-triphosphatase in osteoblasts expressing the Ser252Trp fibroblast growth factor 2 Apert mutation:identifieation by analysis of complementary DNA microarray.J Bone Miner Res,2001, 16(4): 705-712
96. Abe, E, Yamamoto M., Taguchi Y, Lecka-Czernik B, O'Brien CA.,Economides A. N, Stahl N, Jilka R L, and Manolagas SC. Essential requirement of BMPs-2/4 for both osteoblast and osteoclast formation in murine bone marrow cultures from adult mice: Antagonism by noggin. J Bone Miner Res 15: 663-673; 2000.
97. Anderson D M., Maraskovsky E, Billingsley WL, Dougall WC, Tometsko ME, Roux ER., Teepe MC, DuBose RF, Cosman D, and Galibert L.A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function. Nature 390: 175-179; 1997.
98. Ashton BA, Allen TD, Howlett CR, Eaglesom CC, Hattori A,and Owen M. Formation of bone and cartilage by marrow stromal cells in diffusion chambers in vivo. Clin Orthopaed Rel Res 151: 294-307; 1980.
99. Benson MD, Aubin JE, Xiao G, Thomas PE, and Franceschi RT.Cloning of a 2.5 kb murine bone sialoprotein promoter fragment and functional analysis of putative Osf2 binding sites. J Bone Miner Res 14: 396-405; 1999.
100. Corral DA., Amling M., Priemel M, Loyer E, Fuchs S, Duty P, Baron R, and Karsenty G. Dissociation between bone resorption and bone formation in osteopenic transgenic mice. Proc Natl Acad Sci USA 95: 13835-13840; 1998.
101. Ducy P, Amling M., Takeda S, Priemel M., Schilling A F, Beil FT, Shen J, Vinson C, Rueger J M., and Karsenty G. Leptin inhibits bone formation through a hypothalamic relay: A central control of bone mass. Cell 100: 197-207; 2000.
102.Zhou YX,Xu X,Ch L1. A Pro250 Arg substitution in mouse Fgfrl causes increased expression of cbfal and premature fusion of calvarial sutures.Hum Mol Genet, 2000, 9(13): 2001-2008.
103.Kannan S, Lasmoles F, Oyajobi BO, Chen H. Reciproal control of osteoblast/chondroblast and osteoblast/adipocyte differentiation of multipotential clonal human marrow stromal F/STRO-1 +cells.J Cell Biochem,2001, 81(1): 23-38.
104.Mernman HL, van Wljnen A J, Hiebert S, BIdwell JP, Fey E, Llan J,Stein J, Stein GS: The tissue-specific nuclear matrix protein. NMP-2. is a member of the AML/CBF/PEBP2/Runt domain transcription factor family: interaction with the osteocalcin gene promoter. Bochemistry 1995, 34: 13125-13132.
105.Geoffroy V, Duty P, Karsenty G: A PEBP2cL/AML-I -related factor increases osteocalcin promoter activity through its binding to an osteoblast-specific cis-acting element. J Viol Chem 1995, 270: 30973-30979.
106.Banerlee C, Hlebert SW, Stein JL, Liar- JB, Stein GS: An AML-1 consensus sequence binds an osteoblast-specific complex and transcriptionally activates the osteocalcin gene. Proc Nail Acad SC/ USA 1996, 93: 4968-4973.
107.Mlyazakl Y. Setoguchl M, Yoshlda S, Hlguchl Y, Aklzukl S, Yamamoto S:The mouse osteopontin gene. J B/o/ Chem 1990, 265: 14432-14438.
108.Yang R, Gerstenfeld LC: Structural analysis and characterization of tissue and hormonal responsive expression of the avian bone sialoprotein (BSP) gene. J Cell Bwchem 1996, 64: 128-139.
109.Rossert J, Eberspaecher H, Crombrugghe BD: Separate cis-acting DNA elements of the mouse pro-al (I) collagen promoter direct expression of reporter genes to different type I collagen-producingcells in transgenic mice. J Cell.&o/1995, 129: 142 1-1432.
110.Bedalov A, Salvatorl R, Dodlg M, Kronenberg MS, Kapural B,Bogdanovlc Z, Kream BE, Woody CO, Clark SH, Mack K:Regulation of collal expression in type I collagen producing tissues:identification of a 49 base pair region which is required for transgene expression in bone of transgenic mice. J Bone Miner Res 1995. 10: 1443-1451.
111.TSUJI K, Ito Y, Noda M: Expression of the PEBP2aA/AML3/CBFAl gene is regulated by BMP4/7 heterodimer and its overexpressionsuppresses type I collagen and osteocalcin gene expression inosteoblastic and nonosteoblastic mesenchymal cells. Bone 1998, 22. 87-92.
112.Chang DJ, JI C, Kim KK, Casmghlno S, McCarthy TL, Centrella M:Reduction in transforming growth factor p receptor I expression and transcription factor CBFal on bone cells by glucocorticoid. J 6101 Chem 1998, 273: 4892-4896.
113.Lee B, Thirunavukkarasu K, Zhou L, Pastore L, BaldIn A, Hecht J, Geoffroy V, Duty P, Karsenty G: Missense mutations abolishing DNA binding of the osteoblast-specific transcription factor OSF2/CBFAi in cleidocranial dysplasia. Nat Genet 1997, 16: 307-309
114.Stewart M, Terry A, Hu M, O'hara M, Blyth K, Baxter E, Cameron E,Onions DE, Neil JC: Proviral insertions induce the expression of bone-specific isoforms of PEBP2aA (CBFAI): evidence for a newmyc collaborating oncogene. Proc Nat/Acad So USA 1997, 94: 8646-8651.
115.Pend& AM, Balbin M, Llano E, Jimenez MG, Lbpez-Ottn C: Structural analysis and promoter characterization of the human collagenase-3 (MMPI3). Genomtcs 1997, 40: 222-233.
116.Mc Cauley LK.Kolr Paige AJ.Chen H. Parath.yroid hormone stimulates fra-2 expression in osteoblastic cells in vitro and in vivo.Endocrinology, 2001, 142(5): 1975-1981.
117.Naot D,Callon KE,Grey A. A potential role for adrenomedullin as a local regulator of bone growth[J ]. Endocrinology,2001, 142(5): 1849-1857.
118.Hay E.Lemonnier J,Modrowski D. N- and E-cadherin mediated early human calvaria osteoblast differentiation promoted by bone morphgenetie protein-2.J Cell Physiol,2000, 183(1): 117-128.
119.Zalman AL, Lenz J: Transcriptional activation of a retrovirus enhancerby CBF (AMLI) requires a second factor: evidence for cooperativity with c-Myb. J I/iro/1996, 70: 5618-5629.
120.Schorpp M, Mattel M-G, Herr I, Gack S, Schaper J, Angel P:Structural organization and chromosomal localization of the mouse collagenase type I gene. B--ochem J 1995, 308: 211-217.
121.Zhang D, Hethermgton C J, Meyers S, Rhoades KL, Larson CJ,Chen H, Hlebert SW, Tenen DG: CCAAT enhancer-binding protein (C/EBP) and AML1 (CBFa2) synergistically activate the macrophage colony-stimulating factor receptor promoter. Mol Cell Biol 1996, 16: 1231-1240.
122.Tardlf G, Pelletier J-P, Dupuis M, Hambor JE, Mar&-Pelletler J:Cloning, sequencing and characterization of the 5' -flankingregion of the human collagenase-3 gene. B/o&em J 1997, 323: 13-16.
123.Otto T, Sugimoto T, Saijoh K, FUJII M. Chihara K: Cloning and characterization of the 5' -flanking region of the mouse diastrophic dysplasia sulphate transpoter gene. Bochem Bmphys Res Commun 1997, 238: 738-743.
124.Bronckers AL,Sasguri K, Engeles MH.Transcription and immunolocalization of Runx2/Cbfal/PebpzαA in developing osteoblasts phagocytose osteocytes.Microsc Res Tech,2003; 61(6): 540-548.
125.Djouad F, Plence P, Bony C, Schaper J. ImmunosuPpressive elect of ruesenchym stem cells favors tumor growth in allogeneic animals. Blood, 2003, 102(10): 3837--3844.
126.Lazarus HM, Koco N, Devine SM, Hart Q. Cotransplantation of HLA. identical sibling culture, expanded mesenchymal stem cells and hemato. poietic stem cells in hematologic malignancy patients. Biol BloodMarrow Transplant. 2005, 11(5): 389--398.
127.Bartholomew A, Sturgeon C, Siatskas M. Mesenchymal stem cells suppress lymphocyte proliferation in vitro and prolong skin survival in vivo. Exp Hematol, 2002, 30(1): 42-48.
128.Deng W, Han Q, Liao L, Le Blanc K. Long--term distribution of adultstem cells in diferent tissues after transplantation. Stem Cell and CellularTherapy, 2003, 1(1): 62--65.
129.Devine S M, Cobbs C, Jennings M, Mevorach D. Mesenchymal stem cells distribute to a wide range of tissues following systemic infusion into nonhuman primates. Blood, 2003, 101(8): 2999--3001.
130.Aggarwal S, Pittenger M F. Human mesenchymal stem cels modulate allogeneic immune cell responses. Blood, 2005, 105(4): 1815--1822.
131.Le Blanc K, Rasmusson I, Gotherstrem C. Mesenchymal stem cells inhibit the expression of CD25 (interleukin-2 receptor) and CD38 on phytohaemagglutinin--activated lymphocytes. Scand JImmunol, 2004, 60(3): 307--315.
132.Beyth S, Borovsky z, Mevorach D. Human mesenchymal stem cells alter antigen presenting cell maturation an d induce (?). Blood, 2005, 105(5): 2214--2219.
133.Arinzeh T L, Peter S J, Arehambault M P, et al. Allogeneic II chymal stem cells regenerate bone in a critical--sized canine segmental defect. J Bone Joint Surg Am, 2003, 85-A(10): 1927--1935.
134.Vacanti CA, Bonassar LJ. An overview of tissue engineered bone. Clin Orthop, 1999, (367Supp 1): 375.
135.Chen JL, Hunt P, McElvain M. Osteoblast precursor cells are found in CD34+ cells from human bone marrow. Stem Cells, 1997, 15(5): 368-375
136.Vogel W, Grunebach F, Messam CA, et al. Heterogeneity among human bone marrow---derived mesenchymal stem cells and neural progenitor cells. Haematologica, 2003, 88(2): 126-131
137.Nuttall ME, Patton A J, Olivera DL, Hart Q. Human trabecar bone cells are able to expres bo th osteoblastic an d adipo cytic phenotype: implications for osteogenic disorders. J Bone Miner Res, 1998, 13(3): 371-376
138.Khor E, Lim LY. Implantable applications ofchitin andchitosan.Biomaterials 2003; 24: 2339-2349.
139.Domish M, Kaplan D, Skaugrud O. Standards and guidelines for biopolymers in tissue-engineered medical products. Ann N Y Acad Sci 2001; 944: 388-397.
140.Athanasiou KA, Shah AR, Hernandez RJ, LeBaron RG. Basic science ofarticular cartilage repair. Clin Sports Med 2001; 20: 223-247.
141.Madihally SV, Matthew HWT. Porous chitosan scaffolds for tissue engineering. Biomaterials 1999; 20: 1133-1142.
142.Mao JS, Cui YL, Wang XH, Sun Y, Yin YJ, Zhao HM, De Yao K. A preliminary study on chitosan and gelatin polyelectrolyte complex cytocompatibility by cell cycle and apoptosis analysis.Biomaterials 2004; 25: 3973-3981.
143.VandeVord PJ, Matthew HW, DeSilva SP, Mayton L, Wu B,Wooley PH. Evaluation ofthe biocompatibility of a chitosanscaffold in mice. J Biomed Mater Res 2002; 59: 585-590.
144.Suh JKF, Matthew HWT. Application ofchitosan-base d polysaccharide biomaterials in cartilage tissue engineering: a review.Biomaterials 2000; 21: 2589-2598.
145.Hu Q, Li B, Wang M, Shen J. Preparation and characterization of biodegradable chitosan/hydroxyapatite nanocomposite rods via in situ hybridization: a potential material as internal fixation of bone fracture. Biomaterials 2004; 25: 779-785.
146.Risbud MV, Bhonde RR. Polyacrylamide-chitosan hydrogels: in vitro biocompatibility and sustained antibiotic release studies.Drug Deliv 2000; 7: 69-75.
147.Chow KS, Khor E. Novel fabrication of open-pore chitin matrixes. Biomacromolecules 2000; 1: 61--67
148.Geng L, Feng W, Hutmacher DW, Wong Y, Loh H, Fuh JYH.Direct writing of chitosan scaffolds using a robotic system. Rapid Prototyping J 2005; 11: 90-97.
149.Aimin C, Chunlin H, Juliang B, Tinyin Z, Zhichao D. Antibiotic loaded chitosan bar. An in vitro, in vivo study of a possible treatment for osteomyelitis. Clin Orthop 1999; 366: 239-247.
150. Hu SG, Jou CH, Yang MC. Protein adsorption, fibroblast activity and antibacterial properties ofpoly(3-hyd roxybutyric acid-co-3-hydroxyvaleric acid) grafted with chitosan and chitooligosaccharide after immobilized with hyaluronic acid. Biomaterials2003 ;24:2685-2693.
151. Buranapanitkit B, Srinilta V, Ingviga N, Oungbho K, Geater A, Ovatlarnpom C. The efficacy of a hydroxyapatite composite as a biodegradable antibiotic delivery system. Clin Orthop 2004;424:244-252.
152. Chenite A, Chaput C, Wang D, Combes C, Buschmann MD,Hoemann CD, Leroux JC, Atkinson BL, Binette F, Selmani A.Novel injectable neutral solutions ofchitosan form biodegradable gels in situ. Biomaterials 2000;21:2155-2161.
153. Tuzlakoglu K, Alves CM, Mano JF, Reis RL. Production and characterization of chitosan fibers and 3-D fiber mesh scaffolds for tissue engineering applications. Macromol Biosci 2004;4:811-819.
154. Xia W, Liu W, Cui L, Liu Y, Zhong W, Liu D, Wu J, Chua K,Cao Y. Tissue engineering ofcartilage with the use ofchitosan-gelatin complex scaffolds. J Biomed Mater Res 2004;71 B:373-380.
155. Risbud M, Ringe J, Bhonde R, Sittinger M. In vitro expression of cartilage-specific markers by chondrocytes on a biocompatible hydrogel: implications for engineering cartilage tissue. Cell Transplant 2001;10:755-763.
156. Sittinger M, Hutmacher DW, Risbud MV. Current strategies for cell delivery in cartilage and bone regeneration. Curr Opin Biotechnol 2004;15:411-418
157．秦辉，许建中，周强，胡光运．两种构建方法对组织工程骨种子细胞粘附和增殖的影响．第三军医大学学报，2004，26(11)：990—992．
158．赵敏，许建中，周强，李新民．脉冲电磁场促进人骨髓问充质干细胞成骨的研究．中国矫形外科杂志，2004，12(6)：43-45
159．朱灏，许建中，周强，李新民．不同年龄人骨髓问充质干细胞体??外增殖及成骨分化的研究．中国矫形外科杂志，2004．12(10)：759—761．
160. Abdallah B M, Haack Sorensen M, Bums J S. Maintenance of diferentiation potential of human bo ne manDw mesenchymal stem tens immortalized by human telomerase reverse transcdptase gene despite of extensive proliferation. Biochem Biophys Res Commun, 2005. 326(3): 527—538.
161．裴国献，金丹．骨组织工程学种子细胞研究进展．中华创伤骨科杂志，2003，5(1)：55-57
162. Lahiji A, Sohrabi A, Hungerford DS, Frondoza CG, Lahiji A,Sohrabi A, Hungerford DS, Frondoza CG. J Biomed Mater Res2000;51(4):586-595;
163. Iwasaki N, Yamane ST, Majima T, Kasahara Y, Minami A,Harada K, Nonaka S, Maekawa N, Tamura H, Tokura S, Shiono M, Monde K, Nishimura S. Feasibility ofpolysacc haride hybrid materials for scaffolds in cartilage tissue engineering: evaluation ofchondrocyte adhesion to polyion complex fibers prepared from alginate and chitosan. Biomacromolecules 2004;5:828-833.
164. Cui YL, Qi AD, Liu WG, Wang XH, Wang H, Ma DM, Yao KD. Biomimetic surface modification of poly(L-lactic acid) with chitosan and its effects on articular chondrocytes in vitro.Biomaterials 2003;24:3859-3868.
165. Yamane S, Iwasaki N, Majima T, Funakoshi T, Masuko T,Harada K, Minami A, Monde K, Nishimura S. Feasibility of chitosan-based hyaluronic acid hybrid biomaterial for a novel scaffold in cartilage tissue engineering. Biomaterials 2005;26:611-619.
166. Niemeyer P, Secldnger A, Simank H G, Helm GA. Allogenic transplantation of human mesenchymal stem cells for tissue engineering purposes: fluirtro study. Orthopade, 2004, 33(12): 1346—1353.
167. Lee JE, Kim KE, Kwon IC, Aim HJ, Lee SH, Cho H, Kim HJ,Seong SC, Lee MC. Effects of the controlled-released TGF-beta 1 from??chitosan microspheres on chondrocytes cultured in a collagen/chitosan/glycosaminoglycan scaffold. Biomaterials 2004; 25: 4163--4173. 168.Taravel MN, Domard A. Collagen and its interactions with chitosan, III some biological and mechanical properties. Biomaterials1996; 17: 451-455.
169.Kim SE, Park JH, Cho YW, Chung H, Jeong SY, Lee EB, Kwon IC. Porous chitosan scaffold containing microspheres loaded with transforming growth factor-betal: implications for cartilage tissue engineering. J Control Release 2003; 91: 365-374.
170.Mattioli-Belmonte M, Gigante A, Muzzarelli RA, Politano R, De Benedittis A, Specchia N, Buffa A, Biagini G, Greco F. N,Ndicarboxymethyl chitosan as delivery agent for bone morphogenetic protein in the repair ofarticular cartilage. Med Biol Eng Comput 1999; 37: 130-134.
171.Lu JX, Prudhommeaux F, Meunier A, Sedel L, Guillemin G.Effects of chitosan on rat knee cartilages. Biomaterials 1999; 20: 1937-1944.
172.Boden SD. Overview ofthe biology of lumbar spine fusion and principles for selecting a bone graft substitute. Spine 2002; 27(Suppl 16): S26-31.
173.Hankey DP, McCabeRE, DohertyMJ, etal. Enhancementof human osteoblast proliferation and phenotypic expression when cultured in hurnfln senl(?)1. Acta()nhp Scand, 2001, 72(4): 395-401
174.Cooper LF, Masuda T, Whitson SW. et al. Formation of miner alizing osteoblast cultures on machined, titan ium oxide grit--blasted, and plasma--sprayed titanium surfaces. Int J Oral Maxillo. fac Implants, 1999, 14(1): 37-41
175.Saito N, Takaoka K. New synthetic biodegradable polymers as BMP carders for bone tissue engineering. Biomaterials 2003; 24: 2287-2293.
176.Alden TD, Varady P, Kallmes DF, Jane Jr JA, Helm GA. Bone morphogenetic protein gene therapy. Spine 2002, 27(Suppl 16): S87-93.
177. Yoon ST, Boden SD. Spine fusion by gene therapy. Gene Ther 2004; 11(4):360-367.
178. Wang J, de Boer J, de Groot K. Preparation and characterization ofelectrodepo sited calcium phosphate/chitosan coating onTi6A14V plates. J Dent Res 2004;83:296-301.
179. Bonadio J. Tissue engineering via local gene delivery: update and future prospects for enhancing the technology. Adv Drug Deliv Rev 2000;44:185-194.
180. Cha CW, Boden SD. Gene therapy applications for spine fusion.Spine 2003;28(Suppl 15):S74-84.
181. Lee KY, Kwon IC, Kim YH, Jo WH, Jeong SY. Preparation of chitosan self-aggregates as a gene delivery system. J Control Release 1998 ;51:213-220.
182. Koping-Hoggard M, Varum KM, Issa M, Danielsen S,Christensen BE, Stokke BT, Artursson P. Improved chitosanmediated gene delivery based on easily dissociated chitosan polyplexes ofhighly defined chitosan oligomers. Gene Ther 2004; 11:1441-1452.
183. Gan JC, Ducheyne P, Vresilovic EJ, Swaim W, Shapiro IM.Intervertebral disc tissue engineering I: characterization of the nucleus pulposus. Clin Orthop 2003;411:305-314.
184. Yung Lee J, Hall R, Pelinkovic D, Cassinelli E, Usas A,Gilbertson L, Huard J, Kang J. New use of a three-dimensional pellet culture system for human intervertebral disc cells: initial characterization and potential use for tissue engineering. Spine 2001 ;26:231 6-2322.
185．王常勇，王永红，郭希民，等．应用旋转生物反应器和微载体技术体外大规模扩增骨髓间充质干细胞．中华实验外科杂志，2002，19(4)：378-380
186. An HS, Thonar EJ, Masuda K. Biological repair ofinterverteb ral disc. Spine 2003;28(Suppl 15):S86-92.
187. Alini M, Roughley PJ, Antoniou J, Stoll T, Aebi M. A biological approach to treating disc degeneration: not for today, but maybe for tomorrow. Eur Spine J 2002;11(Suppl 2):S215-220.
188. Seol YJ, Lee JY, Park YJ, Lee YM, Young-Ku, Rhyu IC, Lee SJ,Han??SB, Chung CP. Chitosan sponges as tissue engineering scaffolds for bone formation. Biotechnol Lett 2004; 26: 1037-1041.
189.Seeherman H, Li R, Wozney J. A review ofpreclinic al program development for evaluating injectable carriers for osteogenic factors. J Bone Jt Surg Am 2003; 85A(Suppl 3): 96-108.
190.Zhang Y, Zhang M. Synthesis and characterization ofmacroporous chitosan/calcium phosphate composite scaffolds for tissue engineering. J Biomed Mater Res 2001; 55: 304-312.
191.Zhang Y, Zhang M. Calcium phosphate/chitosan composite scaffolds for controlled in vitro antibiotic drug release. J Biomed Mater Res 2002; 62: 378-386.
192.Hung SC, Chen NJ, Hsieh SL, et al. Isolation and characterization of size--sieved stem cells from hum an bone marrow. Stem Cells, 2002, 20(3): 249-253
193.Zhang Y, Ni M, Zhang M, Rather B. Calcium phosphatechitosan composite scaffolds for bone tissue engineering. Tissue Eng 2003; 9: 337-345.
194.Ge Z, Baguenard S, Lim LY, Wee A, Khor E. Hydroxyapatitechitin materials as potential tissue engineered bone substitutes.Biomaterials 2004; 25: 1049-1058.
195.Kawakami T, Antoh M, Hasegawa H, Yamagishi T, Ito M, Eda S. Experimental study on osteoconductive properties ofa chitosan-bonded hydroxyapatite self-hardening paste. Biomaterials 1992; 13: 759-763.
196.Yin Y, Ye F, Cui J, Zhang F, Li X, Yao K. Preparation and characterization ofmacroporous chitosan-gelatin/beta-tricalcium phosphate composite scaffolds for bone tissue engineering.J Biomed Mater Res 2003; 67A: 844-855.
197.Fleming JE Jr, Cornell CN, Muschler GF. Bone cells and matrices in orthopedic tissue engineering. Orthop Clin North Am, 2000, 3t(3): 357-362
198.Zuk PA, Zhu M, Mizuno H. Muhilineage cells from humall adipose tisue: implications for cell--based therapies. Tisue Eng. 2001, 7(2):211-213.
199.Leroux L, Hatim Z, Freche M, Lacout JL. Effects of various adjuvants (lactic acid, glycerol, and chitosan) on the injectability of a calcium phosphate cement. Bone 1999; 25(Suppl 2): 31-41.
200.Gutowska A, Jeong B, Jasionowski M. Injectable gels for tissue engineering. Anat Rec 2001; 263: 342-349.
201.Wang X, Ma J, Wang Y, He B. Bone repair in radii and tibias of rabbits with phosphorylated chitosan reinforced calcium phosphate cements. Biomaterials 2002; 23: 4167-4176.
202.Xu HH, Quinn JB, Takagi S, Chow LC. Synergistic reinforcement of in situ hardening calcium phosphate composite scaffold for bone tissue engineering. Biomaterials 2004; 25: 1029-1037.
203.Kim SB, Kim YJ, Yoon TL, Park SA, Cho IH, Kim EJ, Kim IA, Shin JW. The characteristics of a hydroxyapatite-chitosan-PMMA bone cement. Biomaterials 2004; 25: 5715-5723.
204.Buttery LD, Bourne S, Xynos JD, et al. Differentiation of 08-teoblast and in vitro bone formation from murine embryonic stem cells. Tissue Eng, 2001, 7(1): 89-93
205.Park Y J, Lee YM, Park SN, Sheen SY, Chung CP, Lee SJ. Platelet derived growth factor releasing chitosan sponge for periodontal bone regeneration. Biomaterials 2000; 21: 153-159.
206.Lee JY, Nam SH, Im SY, Park YJ, Lee YM, Seol YJ, Chung CP,Lee SJ. Enhanced bone formation by controlled growth factor delivery from chitosan-based biomaterials. J Control Release 2002; 78: 187-197
207.Petite MF, Mackay AM, Beck SC. Multilineage Ix)tential of adult human mesenchymal stem ceils. Science, 1999, 284(5411): 143-147
208.Lee YM, Park Y J, Lee S J, Ku Y, Han SB, Choi SM, Klokkevold PR, Chung CP. Tissue engineered bone formation using chitosan/tricalcium phosphate sponges. J Periodontol 2000; 71: 410--417.
209.Muzzarelli RA, Mattioli-Belmonte M, Tietz C, Biagini R, Ferioli G, Brunelli MA, Fini M, Giardino R, Ilari P, Biagini G. Stimulatory effect on bone formation exerted by a modified chitosan. Biomaterials1994;15:1075-1081.
210. Lee YM, Park YJ, Lee SJ, Ku Y, Han SB, Klokkevold PR, Chung CP. The bone regenerative effect of platelet-dedved growth factor-BB delivered with a chitosan/tdcalcium phosphate sponge carder. J Periodontol 2000;71:418-424.
211. Kim IS, Park JW, Kwon IC, Baik BS, Cho BC. Role ofBMP,betaig-h3, and chitosan in early bony consolidation in distraction osteogenesis in a dog model. Plast Reconstr Surg 2002; 109:1966-1977.
212. Cho BC, Park JW, Balk BS, Kwon IC, Kim IS. The role of hyaluronic acid, chitosan, and calcium sulfate and their combined effect on early bony consolidation in distraction osteogenesis of a canine model. J Craniofac Surg 2002; 13:783-793.
213. Bumgardner JD, Wiser R, Gerard PD, Bergin P, Chestnutt B,Marin M, Ramsey V, Elder SH, Gilbert JA. Chitosan: potential use as a bioactive coating for orthopaedic and craniofacial/dental implants. J Biomater Sci Polym Ed 2003;14:423-438.
214. Bumgardner JD, Wiser R, Elder SH, Jouett R, Yang Y, Ong JL.Contact angle, protein adsorption and osteoblast precursor cell attachment to chitosan coatings bonded to titanium. J Biomater Sci Polym Ed 2003;14:1401-1409.
215．郭刚，王臻．彭磊，等．成骨细胞异体移植的免疫学初探．第四军医大学学报，2002，23(11)：976-980
216. Anselme K, Noel B, Flautre B, et al. Association of porous hydroxyapatite and bone marrow cells for bone regeneration. Bone, 1999, 25(2Suppl 1): 51-53
217. Kadiyala S, Young RG, Thiede MA, et al. Culture expanded can ine mesenchymal stem ceUs po ses osteoe hondrog enic potential in vivo and invitro. CeU Transplant, 1997, 6(2): 125-131
218. Chen D, Ji X, Harris MA, et al. Differential roles for bone morphogenetic protein(BMP)receptor type IB and IA in differentiation an dspecification of mesenchymal precursor cells to osteoblast and adipoeytelineages. J CellBiol, 1998, 142(1): 295-301219.He TC, Zhou S, da Costa LT, Yu J, Kinzler KW, Vogelstein B. A simplified system for generating recombinant adenovirus.Proc Natl Acad Sci USA,1998; 95(5): 2509-2514.
220.220.Jadlowiec JA, Celil AB, Hollinger JO. Bone tissue engineering: recent advances and promising therapeutic agents. Expert Opin Biol Ther, 2003, 3(3): 409-423.
221.Freed L, Novakovic, Biron RJ. Biodegradable polymer scaffolds fortissue engineering. Biotechnics, 1994, 12(7): 689-676.
222.Sun JS, Liu HC, Chang WH, Li J, Lin FH, Tai HC. Influence of hydroxyapatite particle size on bone cell activities: an in vitro study. J Biomed Mater Res, 1998, 39(3): 390-398.
223.Goshima J, Goldberg VM, Caplan AI. The osteogenic potential of culture expanded rat marrow mesenchymal cells assayed in vivo in calcium phosphate ceramic blocks. Clin Orthop, 1991, (262): 298-305.
224.Musgrave DS, Fu FH, Huard J.Gene therapy and tissue engineering in orthopaedic surgery. J Am Acad Orthop surg, 2002, 10(1): 6-15.