BMP-4/ALK-3在小鼠胫骨牵引成骨过程中的表达
|
摘要
目的：建立小鼠胫骨牵引成骨模型,分析骨形态形成蛋白4 (Bone Morphogenetic Protein 4,BMP-4)及其受体(activin-like kinase3,ALK-3)在牵引成骨过程中的表达,探讨机械牵张力转化为生物信号从而调节骨再生过程的机制。
方法：选取36只健康雄性8周龄CD-1小鼠,腹腔内注射苯巴比妥钠(30mg/kg)麻醉下,所有小鼠均接受左胫骨中上段骨干横行截骨,安置特制延长外固定架。方法为常规消毒皮肤后,于左胫骨中上段截骨处,用27号注射针头钻孔数个后,无菌剪刀剪断胫骨干。然后于截骨处远近端分别经皮穿交叉27号注射针头,并安置延长外固定架。胫骨牵引过程中包括5天静止期,12天牵引期和14天固塑期。牵引速率为0.1mm／次,2次／天,共0.2mm／天,术后于第5,9,13,17,24,31天分组杀取动物,采集左胫骨标本,分别作组织学检查、RT-PCR和原位杂交试验分析,每个时间点6只小鼠。
结果：组织学检查显示静止期其修复过程基本与骨折愈合过程相似。牵引期,被牵引骨痂显示出三个典型的生物学功能区：纤维间区,初始骨基质前沿和微骨柱形成区。在固塑期早期,牵引骨痂骨性愈合。RT-PCR和原位杂交分析显示机械牵张力能提高BMP-4、ALK-3的表达。
结论：1、牵引成骨是一种持续的骨再生过程,机械张力通过刺激局部生长因子及其受体(如BMP-4和ALK-3)以及细胞外基质(如骨钙素)的持续表达,维持骨痂的不断形成和再塑,从而填充被逐渐延长的骨间隙。2、牵引期新生骨的主要形成方式是膜内成骨。
Objective:Establish model of mouse tibia for distraction osteogenesis,To understand how the mechanical forces are translated into biological signals that regulate bone regeneration and repair, we investigated the expression of Bone Morphogenetic protein 4(BMP-4) and its receptor activin -like kinase3(ALK-3) using a mouse tibia for distraction osteogenesis.
Methods:Thirty - six male CD - 1 mice,8 weeks old, were used in the study. All animals received placement of the external fixators and osteotomy on the left tibia. Distraction protocol included 5 days latency, 12 days distraction, and 14 days consolidation. Distraction rate was 0.2 mm/day. The animals were sacrificed at 5,9,13,17,24, and 31days after the operation. The lengthened tibiae were harvested and distraction gaps were analyzed by histology, RT - PCR, and in situ hybridization.
Results:Histological analysis demonstrated that the healing process of osteotomic gap was similar to fracture repair in the latency. The distraction callus consisted of fibrous interzone, primary matrix front, and microcolumn formation during the distraction. In the early stage of consolidation the distraction gap was bridged by bony callus. RT - PCR and in situ hybridization analysis showed that mechanical forces enhanced expression of BMP - 4 and ALK-3.
Conclusions:1.Our data suggested that distraction osteogenesis is a unique form of bone regeneration. The mechanic forces induce expression of local growth factors and receptors (such as BMP-4 and ALK-3) that maintain callus formation and remodelling which fill osteotomic gap gradually enlarged by mechanical forces.2.The primary mode of bone formation in the new bone is intramembranous ossification.
方法：选取36只健康雄性8周龄CD-1小鼠,腹腔内注射苯巴比妥钠(30mg/kg)麻醉下,所有小鼠均接受左胫骨中上段骨干横行截骨,安置特制延长外固定架。方法为常规消毒皮肤后,于左胫骨中上段截骨处,用27号注射针头钻孔数个后,无菌剪刀剪断胫骨干。然后于截骨处远近端分别经皮穿交叉27号注射针头,并安置延长外固定架。胫骨牵引过程中包括5天静止期,12天牵引期和14天固塑期。牵引速率为0.1mm／次,2次／天,共0.2mm／天,术后于第5,9,13,17,24,31天分组杀取动物,采集左胫骨标本,分别作组织学检查、RT-PCR和原位杂交试验分析,每个时间点6只小鼠。
结果：组织学检查显示静止期其修复过程基本与骨折愈合过程相似。牵引期,被牵引骨痂显示出三个典型的生物学功能区：纤维间区,初始骨基质前沿和微骨柱形成区。在固塑期早期,牵引骨痂骨性愈合。RT-PCR和原位杂交分析显示机械牵张力能提高BMP-4、ALK-3的表达。
结论：1、牵引成骨是一种持续的骨再生过程,机械张力通过刺激局部生长因子及其受体(如BMP-4和ALK-3)以及细胞外基质(如骨钙素)的持续表达,维持骨痂的不断形成和再塑,从而填充被逐渐延长的骨间隙。2、牵引期新生骨的主要形成方式是膜内成骨。
Objective:Establish model of mouse tibia for distraction osteogenesis,To understand how the mechanical forces are translated into biological signals that regulate bone regeneration and repair, we investigated the expression of Bone Morphogenetic protein 4(BMP-4) and its receptor activin -like kinase3(ALK-3) using a mouse tibia for distraction osteogenesis.
Methods:Thirty - six male CD - 1 mice,8 weeks old, were used in the study. All animals received placement of the external fixators and osteotomy on the left tibia. Distraction protocol included 5 days latency, 12 days distraction, and 14 days consolidation. Distraction rate was 0.2 mm/day. The animals were sacrificed at 5,9,13,17,24, and 31days after the operation. The lengthened tibiae were harvested and distraction gaps were analyzed by histology, RT - PCR, and in situ hybridization.
Results:Histological analysis demonstrated that the healing process of osteotomic gap was similar to fracture repair in the latency. The distraction callus consisted of fibrous interzone, primary matrix front, and microcolumn formation during the distraction. In the early stage of consolidation the distraction gap was bridged by bony callus. RT - PCR and in situ hybridization analysis showed that mechanical forces enhanced expression of BMP - 4 and ALK-3.
Conclusions:1.Our data suggested that distraction osteogenesis is a unique form of bone regeneration. The mechanic forces induce expression of local growth factors and receptors (such as BMP-4 and ALK-3) that maintain callus formation and remodelling which fill osteotomic gap gradually enlarged by mechanical forces.2.The primary mode of bone formation in the new bone is intramembranous ossification.
引文
[1]G.Shen and M,Ali Darendeliler. The Adaptive Remodeling of Condylar Cartilage-A Transition from Chondrogenesis to Osteogenesis. [J] Dent Res.2005, 84(8):691-699
[2]Neil Ghodadra,Kern Singh.Recombinant human bone morphogenetic protein-2 in the treatment of bone fractures. Biologics.2008,2(3):345-54
[3]李长有,梁德勇,宋今丹.鼠骨折愈合过程中膜内化骨和软骨内化骨的细胞组织学变化[J].中国医科大学学报.2005,2(1)：43-44
[4]Michael J. Zuscik, Matthew J. Hilton, Xinping Zhang. Regulation of chondrogenesis and chondrocyte differentiation by stress. [J] Clin Invest.2008, 118(2):429-38
[5]刘振东,钟杰林,高敏伟,等.牵引成骨与小鼠早期骨折愈合[J].中国组织工程研究与临床康复.2009,13(2)：225-228
[6]王守丰,邱勇.软骨内成骨的调控[J].中华外科杂志.2006,44(16)：1147-49
[7]万贤凤,兰泽栋,闫福华.骨形成蛋白-4的作用机制及功能研究现状[J].广东牙病防治.2007,15(4)：184-185
[8]Chen D,Zhao M,Mundy GR.Bone morphogenetic proteins.Growth Factors.2004,22(4):233-41
[9]马真胜,胡蕴玉,王臻,等.骨形态发生蛋白4(BMP-4)基因表达在骨折愈合过程中的定位研究.中华骨科杂志,1997,8(17)：517-519
[10]Jeanette C,Perron,Jane Dodd.ActRIIA and BMPRII Type II BMP Receptor Subunits Selectively Required for Smad4-Independent BMP7-Evoked Chemotaxis.PLoS ONE.2009,4(12):1-14
[11]Yeh LC,ZavalaMC,Lee JC.Osteogenic protein-1 and interleukin-6 with its soluble receptor synergistically stimulate rat osteoblastic cell differentiation[J].J Cell Physiol,2002,190(3):322-331
[12]吴在德.外科学.北京：人民卫生出版社,2004.746-748
[13]Liu ZD,James Aronson,Elizabeth C. Wahl,et al. A novel rat model for the study of deficits in bone formation in type-2 diabetes. Acta Orthop.2007, 78(1):46-55
[14]Reddi AH,Cunningham NS.Initiation and promotion of bone differentiation by bone morphogenetic proteins. J Bone Miner Res.1993,8 (2):499-502
[15]Hernandez CJ,Tang SY, Baumbach BM, et al. Trabecular micro-fracture and the influence of pyridinium and non-enzymatic glycation-mediated collagen cross-links[J].Bone,2005,37(6):825-832
[16]Viguet Carrin S, Roux JP, ArlotME, et al. Contribution of the advanced glycation end product pentosidine and ofinaturation of type I collagen to compressive biomechanical properties of human lumbar vertebrae [J]Bone,2006,39(5):1073-1079
[17]Lammens J, Liu Z, Aerssens J,et al. Distraction bone healing versus osteotomy healing:a comparative biochemical analysis[J].J Bone Miner Res,1998,13(2):279-86
[18]Liu Z,Luyten FP,Lammens J,et al. Molecular signaling in bone fracture healing and distraction osteogenesis[J].Histol Histopathol,1999,14 (2):587-95
[19]Schep NW,van Lieshout EM, Patka P,et al. Long-term functional and quality of live assessment following post-traumatic distraction osteogenesis of the lower limb. Strategies Trauma Limb Reconstr.2009,24(4):107-112
[20]Kimberly A Jacobsen, Zainab S Al-Aql,Chao Wan,et al. Bone Formation During Distraction Osteogenesis Is Dependent on Both VEGFR1 and VEGFR2 Signaling. J Bone Miner Res.2008,23(5):596-609
[21]倪滨,李明.BMP生物活性的研究现状.重庆医学,2007,36(10)：970-973
[22]Jane JA J r,Dunford BA, Kron A, et al.Ectopic osteogenesis using adenoviral bone morphogenetic protein(BMP)-4 and BMP-6 gene transfer[J]. Mol Ther,2002,6(4):464-70
[23]Yaoita H,Orimo H,Shirai Y,et al.Expression of bone morphogenetic proteins and rat distal-less homolog genes following rat femoral fracture[J].J Bone Miner Metab,2000,18(2):63-70
[24]刘振东,张朝跃,詹瑞森,等.BMP-4和BMP-6在小鼠胫骨牵引成骨过程中的表达.实用预防医学.2004,11(4)：743-746
[25]Sato M,Ochi T,Nakase T,et al.Mechanical sension-stress induces expression of bone morphngenetic protein(BMP)-2 and BMP-4,but not BMP-6,BMP-7,and GDF-5mRNA,during distraction osteogenesis[J].J BoneMinerRes,1999,14(7):1084-1095
[26]武玉刚,王自立.BMP-2／4在脊柱结核病灶骨中的表达.宁夏医学杂志.2005,27(2)：75-77
[27]房可一,王稚英.骨形态发生蛋白-4的研究进展.社区医学杂志.2007,5(9)：42-44
[28]Kohei Miyazono,Yuto Kamiya,Masato Morikawa. Bone morphogenetic protein receptors and signal transduction.J. Biochem.2010,147(1):35-51
[29]Nam Y. Lee, Kellye C. Kirkbride, Richard D. Sheu,et al. The Transforming Growth Factor-PType III Receptor Mediates Distinct Subcellular Trafficking and Downstream Signaling of Activin-like Kinase (ALK)3 and ALK6 Receptors. Molecular Biology of the Cell.2009,15(20):4362-4370
[30]Noel D, Gazit D, Bouquet C. Short-Term BMP-2 Expression Is Sufficient for In Vivo Osteochondral Differentiation of Mesenchymal Stem Cells. Stem Cells.2004,22(1):74-85
[31]Chang J, Sonoyama W, Wang Z. Noncanonical Wnt-4 Signaling Enhances Bone Regeneration of Mesenchymal Stem Cells in Craniofacial Defects through Activation of p38 MAPK. J Biol Chem.2007,282(42):30938-48
[1]Bjoern Rath,Jin Nam,Thomas J Knobloch.Compressive forces induce osteogenic gene expression in calvarial osteoblasts.J Biomech.2008; 41(5):1095-1103
[2]Sato M,Ochi T,Nakase T,et al.Mechanical sension-stress induces expression of bone morphngenetic protein(BMP)-2 and BMP-4,but not BMP-6,BMP-7,and GDF-5mRNA,during distraction osteogenesis[J].J BoneMinerRes,1999,14(7):1084-1095
[3]Palomares KT, Gleason RE, Mason ZD,et al. Mechanical stimulation alters tissue differentiation and molecular expression during bone healing.J Orthop Res. 2009 September;27(9):1123-1132
[4]吴在德主编.外科学[M].第6版.人民卫生出版社.2004,746-748
[5]苏佳灿,许硕贵,张春才.骨重建中细胞因子的作用厦机制[J].中国矫形外科杂志,2001,8(6)：593-595
[6]苏佳灿.张春才.骨折愈合中基因直接转入的作用及机制[J].现代康复.2001,5(1)：62-63
[7]Kloen P,Dory SB,Gordon E,et al.Expression and activatioft of the BMP-signaling components in human fracture nonunions.J Bone Joint Surg Am,2002,84:1909-1918
[8]Ling Yu, Manjong Han, Mingquan Yan,et al. BMP signaling induces digit regeneration in neonatal mice. Development.2010 February 15;137(4):551-559
[9]Chen D, Zhao M, Mundy GR.Bone morphogenetic proteins. Growth Factors.2004,22(4):233-41
[10]Kishimoto KN,W atan abe Y,Nakamura H,et al.Ectopic bone formalion by electroporatic transfer of bone morphogenetie protein-4 gene.Bone,2002,31:340-347
[11]Namkung MH,Appleyard R,Jansen J.Osteoporosis influences the early period of fracture healing in a rat osteo-pomtic model.Bone,2001,28:80-86
[12]Ten Dijke P,Yamashita H, Sampath,et al. Identification of type I receptors for osteogenic protein-1 and bone morphogenetic protein-4. J. Biol. Chem.1994, 26(9):16985-16988
[13]Kohei Miyazono,Yuto Kamiya,Masato Morikawa. Bone morphogenetic protein receptors and signal transduction.J. Biochem.2010;147(1):35-51
[14]Jean-Charles Guimond, Mathieu Levesque, Pierre-Luc Michaud,et al. BMP-2 functions independently of SHH signaling and triggers cell condensation and apoptosis in regenerating axolotl limbs. BMC Dev Biol.2010 Feb 12;10:15
[15]Murtangh LC,Chyung JH.Lassar AB.Sonic hedgehog promotes somatic chondrogenesis by altering the cellular response to BMP signaling.Genes Dev,1999,13:225-237
[16]孙洋,赵宇,汪春兰,等.骨形态发生蛋白4转基因治疗大鼠的胫骨缺损.中国组织工程研究与临床康复.2007Mat；11(19)：3778-3780
[17]Bonadio J, Smiley E, Patil P, et al. Localized, direct plasmid gene delivery in vivo:prolonged therapy results in reproducible tissue regeneration. Na t Med 1999; 5(7):753-759
[18]Liu R, Ginn SL, Lek M. Myoblast sensitivity and fibroblast insensitivity to osteogenic conversion by BMP-2 correlates with the expression of Bmpr-la. BMC Musculoskelet Disord.2009 May 15; 10:51
[19]Mukherjee A, Rotwein P. Akt promotes BMP2-mediated osteoblast differentiation and bone development. J Cell Sci.2009 Mar 1;122(Pt 5):716-26
[20]王霖霞,李玉坤.BMP-2信号通路与成骨细胞分化.国际骨科学杂志.2009,30(2)：132-136
[21]Neil Ghodadra,Kern Singh. Recombinant human bone morphogenetic protein-2 in the treatment of bone fractures. Biologics.2008 September; 2(3): 345-354
[22]Chhabra A,Zijerdi D,Zhang J,et al.BMP-4 deciency inhibits longbone fracture healing:a biochemical,histologic,and radiographic assessment.Orthop Trauma,2005,19:629-634
[23]Shida JI,Jingushi S,Izumi T,et al.Basic fihroblast growth factor regulates expression of growth factors in rat epiphyseal chondrocytes.J Orthop Res,2001,19:259-264
[24]王立平,党耕町.碱性成纤维细胞生长因子在骨愈合过程中的表达与合成.中华骨科杂志,1999,19：488-490
[25]张永刚,卢世璧.骨形态发生蛋白和转化生长因子.B及碱性成纤维细胞生长因子在骨折修复中的表达.中华创伤杂志,2000,16：170-172
[26]Tanaka H,Wakisaka A,Ogasa H,et al.Effects of basic fibroblast growth factor on osteoblast-related gene expression in the process of medullary bone formation induced in rat femur.J Bone Miner Metab.2003,21:74-79
[27]Tardif G, Pelletier JP, Hum D, et al. Differential regulation of the bone morphogenic protein antagonist chordin in human normal andosteoarthritic chondrocytes. Ann Rheum Dis.2006; 65(2):261-264
[28]Gong YK, Peng WX, Li XH, et al. Revascularizing effect of basicfibroblast growth factor and partially deproteinised bone on femoral head defects in rabbits. Zhongguo Linchuang Kangfu.2005; 9(42):137-139
[29]Nie SP, Cao LS, Liu XQ, et al. Enhanced expression of vascular growth factors and cell proliferation by controlled-releasing basic fibroblast growth factors in canine myocardium. Zhongguo Linchuang Kangfu.2005; 9(46):153-156
[30]陈建庭,杨德鸿,景宗森,等.bFGF调节PDGF对人成骨细胞的促增殖作用.中华外科杂志.2000Sep；38(9)：707-710
[31]Tatsuyama K,Maezawa Y,Baloe H,et al.Expression of various growth factors for cell proliferatior and cytocifferentiation during fracture repairof bone.Eur J Histooem 2000;4(33):269-278
[32]Ekam EC,Ravarti L,Kaharl V,et al.Expression of extraceluler matrix genes transforming growth factor(TGF)-beta and ras in tibial fracture healing of athynitic rat.Bone 2000;27(4):551-557
[33]陈永龙,雷明全,兰英,等.高原环境对大鼠桡骨骨折愈合过程中TGF-β1表达的影响.解放军医学杂志.2003Jan；28(1)：88-89
[34]杨柳,李起鸿,谭祖键,等.实验性骨延长区TGF-β的基因表达和细胞定位.第三军医大学学报.2001Jan；23(1)：66-68
[35]卢卫忠,唐康来,杨柳,等.不同时间给予转化生长因子p对兔骨折愈合作用的研究.2003 May；25(9)：753-757
[36]马维,牛彦平,张华.合并脑损伤大鼠骨折愈合过程中转化生长因子β1血清含量及在骨折位点的表达.2006Nov；10(42)：88-91
[37]王守君,张柳,孙良智,等.大鼠股骨骨折合并脑外伤时骨愈合过程中TGF-β1的作用.第四军医大学学报.2006；27(15)：1407-1410
[38]Dominguez LJ,Muratore M,Quarta E,et al.Osteoporosis and diabetes. Euatismo 2004;56(4):235-241
[39]Wengreen H J, Munger R G, Wset N A, et al. Dietary protein intake and risk of osteoporotic hip fracture in elderly residents of Utah. J Bone Miner Res.2004; 19(4):537-543
[40]王敏,辛海霞,黄象艳.胰岛素样生长因子-Ⅰ对成骨细胞生长影响的实验研究.中国矫形外科杂志.2004,2(12)：236-238
[41]Spencer CP,Codsland IF,Cooper AJ,et al. Effects of oral for IGF-1 with oral 17 beta-estradiol on insulin sensitivity,Secretion,and elimintion in postmenopausal women. J Metablism,2000,49(6):742
[42]Kurland ES,Rosen CJ,Cosman F.Insulin-like growth factor-I in men with idiopathic osteoporosis[J].J Clin Endocrinol Metab,1997,82:279
[43]Johnsson AG,Lander JM.Human insulin-like growth factor-I binding protein-I(rhIGFBP-I) in transgenic mice:characterization and insights into the regulation of IGFBP-I expression[J].J Clin Endocrinol Metab,1996,81:44
[44]李伟明,王伟,吕建国,等.胰岛素样生长因子-1对骨质疏松大鼠骨形成作用.骨与关节损伤杂志.2004,19(4)：252-254
[45]朱国强,吴织芬,王勤涛,等.IGF-1对骨髓基质细胞分泌BMP-2和VEGF的影响.临床口腔医学杂志.2004,20(4)：215-217
[46]Sakata T, Halloran BP, Elalieh HZ, et al. Skeletal unloading induces resistance to insulin-like growth factor I on bone formation[J]. Bone.2003, 32(6):669-680
[47]徐萍,张克勤,刘超,等.重组人胰岛素样生长因子-Ⅰ(rhIGF-I)对大鼠成骨细胞增殖和功能的影响.南京医科大学学报(自然科学版).2005 Jul,25(7)：472-475
[48]Jehle P,Schulten K,Schulz W,et al.Serum level of insulin-like growth factor(IGF)-I and IGF binding protein(IGFBP)-I to-6 and their relationship to bone metabolism in osteoporosis patients [J]. Eur J Intern Med.2003,14(1):32-38
[49]初同伟,王正国,朱佩芳,等.骨折愈合过程中VEGF及其受体的表达.中华创伤杂志,2001,17：344-346
[50]武永刚,陈君长,王坤正.血管内皮细胞生长因子在骨折愈合过程中的表达.西安医科大学学报.2001,22(1)：51-53
[51]初同伟,王正国,朱佩芳,等.骨折愈合过程中血管内皮生长因子基因扩增及蛋白表达研究.中华实验外科杂志.2002Jul；19(4)：367-368
[52]Streeten EA, Brandi ML. Biology of bone endothelial cells [J]. Bone Miner, 1990,10:85-94
[53]马维,张英泽,宋永周,等.血小板浓缩液对兔尺骨骨折影响的探讨.江苏医药,2003,8：592-59
[54]崔猛,陈建庭.血小板衍生生长因子促进鼠骨早期修复中增殖细胞核抗原与c-Jun蛋白的表达.中国临床康复,2006,10：53-55
[55]Lee JY. Nam SH, Im SY, et al. Enhanced bone form ation by controlledgrowth factor deliyery from chitosan-based biomaterials. J Control Release,2002,78:187-197
[56]Zhang L, Leeman E, Carnes DC, et al. Human osteoblast synthesize and respond to platelet-derived growth factor. Am J Physiol,1991,261:348-354
[57]Horner A, Bord S, Kemp P, et al. Distribution of platelet-derived growth factor (PDGF):a chain mRNA, protein and PDGF-areceptor in rapid ly forming human bone. Bone,1996,19:353-362
[58]张冬根,谭颖徽,裘松波,等.血小板衍生生长因子、骨保护素对体外培养成骨细胞的功能影响.激光杂志.2005；26(3)：94-95
[59]张忠民,陈建庭,金大地,等.血小板衍生生长因子2BB在成骨细胞-破骨细胞培养体系中的生物学作用.中华医学杂志.2001Apr；81(7)：425-428
[60]Alini M,Matsui Y,Dodge GR,et al.The extracellular matrix of cartilage in the growth plate before and during calcification:changes in composition and degradation of type II collagen.Calcif Tissue Int.1992;50(4):327-335.
[61]Grant WT,Wang GJ,Balian G.Type X collagen synthesis during endochondral ossification in fracture repair.J Biol Chem.l987;262(20):9844-9849
[62]Warman ML, Abbott M, Apte SS, et al.A type X collagen mutation causes Schmid metaphyseal chondrodysplasia. Nat Genet.1993;5(l):79-82
[63]郭嫒,黄健,许善锦,等.Ⅱ、Ⅹ型胶原对钙磷酸盐生物矿化生物动力学的影响[J].自然科学进展,2001,11(6)：573-575
[64]Hiltulen A,Aro HT,Vuotle E.Regulation of extracelular matrix genes during fracture healing in mice.Clip Orthop,1993,297:23-27
[65]王立平,党耕町.骨连接素在骨愈合中作用的探讨[J].中华骨科杂志.1998,18(8)：497
[66]Hirakawa K,Hirota S,Ikeda T,et al.Localization of the mRNA for bone matrix proteins during fracture healing as determined by in site hybridization.J Bone Mine Res 1994;9:1551
[67]麻松,阎景龙,王新涛,等.微小颗粒骨复合细胞移植治疗大鼠颅骨.缺损中华实验外科杂志.2005,22：154-155
[68]Xiao G.Fibroblast growth factor 2 induction of the osteocalingene requires MARK activity and phosphorylation of the osteoblast trscription factor,Cbfl/Runx2.J Biol Chem,2002,227:36181-36187
[2]Neil Ghodadra,Kern Singh.Recombinant human bone morphogenetic protein-2 in the treatment of bone fractures. Biologics.2008,2(3):345-54
[3]李长有,梁德勇,宋今丹.鼠骨折愈合过程中膜内化骨和软骨内化骨的细胞组织学变化[J].中国医科大学学报.2005,2(1)：43-44
[4]Michael J. Zuscik, Matthew J. Hilton, Xinping Zhang. Regulation of chondrogenesis and chondrocyte differentiation by stress. [J] Clin Invest.2008, 118(2):429-38
[5]刘振东,钟杰林,高敏伟,等.牵引成骨与小鼠早期骨折愈合[J].中国组织工程研究与临床康复.2009,13(2)：225-228
[6]王守丰,邱勇.软骨内成骨的调控[J].中华外科杂志.2006,44(16)：1147-49
[7]万贤凤,兰泽栋,闫福华.骨形成蛋白-4的作用机制及功能研究现状[J].广东牙病防治.2007,15(4)：184-185
[8]Chen D,Zhao M,Mundy GR.Bone morphogenetic proteins.Growth Factors.2004,22(4):233-41
[9]马真胜,胡蕴玉,王臻,等.骨形态发生蛋白4(BMP-4)基因表达在骨折愈合过程中的定位研究.中华骨科杂志,1997,8(17)：517-519
[10]Jeanette C,Perron,Jane Dodd.ActRIIA and BMPRII Type II BMP Receptor Subunits Selectively Required for Smad4-Independent BMP7-Evoked Chemotaxis.PLoS ONE.2009,4(12):1-14
[11]Yeh LC,ZavalaMC,Lee JC.Osteogenic protein-1 and interleukin-6 with its soluble receptor synergistically stimulate rat osteoblastic cell differentiation[J].J Cell Physiol,2002,190(3):322-331
[12]吴在德.外科学.北京：人民卫生出版社,2004.746-748
[13]Liu ZD,James Aronson,Elizabeth C. Wahl,et al. A novel rat model for the study of deficits in bone formation in type-2 diabetes. Acta Orthop.2007, 78(1):46-55
[14]Reddi AH,Cunningham NS.Initiation and promotion of bone differentiation by bone morphogenetic proteins. J Bone Miner Res.1993,8 (2):499-502
[15]Hernandez CJ,Tang SY, Baumbach BM, et al. Trabecular micro-fracture and the influence of pyridinium and non-enzymatic glycation-mediated collagen cross-links[J].Bone,2005,37(6):825-832
[16]Viguet Carrin S, Roux JP, ArlotME, et al. Contribution of the advanced glycation end product pentosidine and ofinaturation of type I collagen to compressive biomechanical properties of human lumbar vertebrae [J]Bone,2006,39(5):1073-1079
[17]Lammens J, Liu Z, Aerssens J,et al. Distraction bone healing versus osteotomy healing:a comparative biochemical analysis[J].J Bone Miner Res,1998,13(2):279-86
[18]Liu Z,Luyten FP,Lammens J,et al. Molecular signaling in bone fracture healing and distraction osteogenesis[J].Histol Histopathol,1999,14 (2):587-95
[19]Schep NW,van Lieshout EM, Patka P,et al. Long-term functional and quality of live assessment following post-traumatic distraction osteogenesis of the lower limb. Strategies Trauma Limb Reconstr.2009,24(4):107-112
[20]Kimberly A Jacobsen, Zainab S Al-Aql,Chao Wan,et al. Bone Formation During Distraction Osteogenesis Is Dependent on Both VEGFR1 and VEGFR2 Signaling. J Bone Miner Res.2008,23(5):596-609
[21]倪滨,李明.BMP生物活性的研究现状.重庆医学,2007,36(10)：970-973
[22]Jane JA J r,Dunford BA, Kron A, et al.Ectopic osteogenesis using adenoviral bone morphogenetic protein(BMP)-4 and BMP-6 gene transfer[J]. Mol Ther,2002,6(4):464-70
[23]Yaoita H,Orimo H,Shirai Y,et al.Expression of bone morphogenetic proteins and rat distal-less homolog genes following rat femoral fracture[J].J Bone Miner Metab,2000,18(2):63-70
[24]刘振东,张朝跃,詹瑞森,等.BMP-4和BMP-6在小鼠胫骨牵引成骨过程中的表达.实用预防医学.2004,11(4)：743-746
[25]Sato M,Ochi T,Nakase T,et al.Mechanical sension-stress induces expression of bone morphngenetic protein(BMP)-2 and BMP-4,but not BMP-6,BMP-7,and GDF-5mRNA,during distraction osteogenesis[J].J BoneMinerRes,1999,14(7):1084-1095
[26]武玉刚,王自立.BMP-2／4在脊柱结核病灶骨中的表达.宁夏医学杂志.2005,27(2)：75-77
[27]房可一,王稚英.骨形态发生蛋白-4的研究进展.社区医学杂志.2007,5(9)：42-44
[28]Kohei Miyazono,Yuto Kamiya,Masato Morikawa. Bone morphogenetic protein receptors and signal transduction.J. Biochem.2010,147(1):35-51
[29]Nam Y. Lee, Kellye C. Kirkbride, Richard D. Sheu,et al. The Transforming Growth Factor-PType III Receptor Mediates Distinct Subcellular Trafficking and Downstream Signaling of Activin-like Kinase (ALK)3 and ALK6 Receptors. Molecular Biology of the Cell.2009,15(20):4362-4370
[30]Noel D, Gazit D, Bouquet C. Short-Term BMP-2 Expression Is Sufficient for In Vivo Osteochondral Differentiation of Mesenchymal Stem Cells. Stem Cells.2004,22(1):74-85
[31]Chang J, Sonoyama W, Wang Z. Noncanonical Wnt-4 Signaling Enhances Bone Regeneration of Mesenchymal Stem Cells in Craniofacial Defects through Activation of p38 MAPK. J Biol Chem.2007,282(42):30938-48
[1]Bjoern Rath,Jin Nam,Thomas J Knobloch.Compressive forces induce osteogenic gene expression in calvarial osteoblasts.J Biomech.2008; 41(5):1095-1103
[2]Sato M,Ochi T,Nakase T,et al.Mechanical sension-stress induces expression of bone morphngenetic protein(BMP)-2 and BMP-4,but not BMP-6,BMP-7,and GDF-5mRNA,during distraction osteogenesis[J].J BoneMinerRes,1999,14(7):1084-1095
[3]Palomares KT, Gleason RE, Mason ZD,et al. Mechanical stimulation alters tissue differentiation and molecular expression during bone healing.J Orthop Res. 2009 September;27(9):1123-1132
[4]吴在德主编.外科学[M].第6版.人民卫生出版社.2004,746-748
[5]苏佳灿,许硕贵,张春才.骨重建中细胞因子的作用厦机制[J].中国矫形外科杂志,2001,8(6)：593-595
[6]苏佳灿.张春才.骨折愈合中基因直接转入的作用及机制[J].现代康复.2001,5(1)：62-63
[7]Kloen P,Dory SB,Gordon E,et al.Expression and activatioft of the BMP-signaling components in human fracture nonunions.J Bone Joint Surg Am,2002,84:1909-1918
[8]Ling Yu, Manjong Han, Mingquan Yan,et al. BMP signaling induces digit regeneration in neonatal mice. Development.2010 February 15;137(4):551-559
[9]Chen D, Zhao M, Mundy GR.Bone morphogenetic proteins. Growth Factors.2004,22(4):233-41
[10]Kishimoto KN,W atan abe Y,Nakamura H,et al.Ectopic bone formalion by electroporatic transfer of bone morphogenetie protein-4 gene.Bone,2002,31:340-347
[11]Namkung MH,Appleyard R,Jansen J.Osteoporosis influences the early period of fracture healing in a rat osteo-pomtic model.Bone,2001,28:80-86
[12]Ten Dijke P,Yamashita H, Sampath,et al. Identification of type I receptors for osteogenic protein-1 and bone morphogenetic protein-4. J. Biol. Chem.1994, 26(9):16985-16988
[13]Kohei Miyazono,Yuto Kamiya,Masato Morikawa. Bone morphogenetic protein receptors and signal transduction.J. Biochem.2010;147(1):35-51
[14]Jean-Charles Guimond, Mathieu Levesque, Pierre-Luc Michaud,et al. BMP-2 functions independently of SHH signaling and triggers cell condensation and apoptosis in regenerating axolotl limbs. BMC Dev Biol.2010 Feb 12;10:15
[15]Murtangh LC,Chyung JH.Lassar AB.Sonic hedgehog promotes somatic chondrogenesis by altering the cellular response to BMP signaling.Genes Dev,1999,13:225-237
[16]孙洋,赵宇,汪春兰,等.骨形态发生蛋白4转基因治疗大鼠的胫骨缺损.中国组织工程研究与临床康复.2007Mat；11(19)：3778-3780
[17]Bonadio J, Smiley E, Patil P, et al. Localized, direct plasmid gene delivery in vivo:prolonged therapy results in reproducible tissue regeneration. Na t Med 1999; 5(7):753-759
[18]Liu R, Ginn SL, Lek M. Myoblast sensitivity and fibroblast insensitivity to osteogenic conversion by BMP-2 correlates with the expression of Bmpr-la. BMC Musculoskelet Disord.2009 May 15; 10:51
[19]Mukherjee A, Rotwein P. Akt promotes BMP2-mediated osteoblast differentiation and bone development. J Cell Sci.2009 Mar 1;122(Pt 5):716-26
[20]王霖霞,李玉坤.BMP-2信号通路与成骨细胞分化.国际骨科学杂志.2009,30(2)：132-136
[21]Neil Ghodadra,Kern Singh. Recombinant human bone morphogenetic protein-2 in the treatment of bone fractures. Biologics.2008 September; 2(3): 345-354
[22]Chhabra A,Zijerdi D,Zhang J,et al.BMP-4 deciency inhibits longbone fracture healing:a biochemical,histologic,and radiographic assessment.Orthop Trauma,2005,19:629-634
[23]Shida JI,Jingushi S,Izumi T,et al.Basic fihroblast growth factor regulates expression of growth factors in rat epiphyseal chondrocytes.J Orthop Res,2001,19:259-264
[24]王立平,党耕町.碱性成纤维细胞生长因子在骨愈合过程中的表达与合成.中华骨科杂志,1999,19：488-490
[25]张永刚,卢世璧.骨形态发生蛋白和转化生长因子.B及碱性成纤维细胞生长因子在骨折修复中的表达.中华创伤杂志,2000,16：170-172
[26]Tanaka H,Wakisaka A,Ogasa H,et al.Effects of basic fibroblast growth factor on osteoblast-related gene expression in the process of medullary bone formation induced in rat femur.J Bone Miner Metab.2003,21:74-79
[27]Tardif G, Pelletier JP, Hum D, et al. Differential regulation of the bone morphogenic protein antagonist chordin in human normal andosteoarthritic chondrocytes. Ann Rheum Dis.2006; 65(2):261-264
[28]Gong YK, Peng WX, Li XH, et al. Revascularizing effect of basicfibroblast growth factor and partially deproteinised bone on femoral head defects in rabbits. Zhongguo Linchuang Kangfu.2005; 9(42):137-139
[29]Nie SP, Cao LS, Liu XQ, et al. Enhanced expression of vascular growth factors and cell proliferation by controlled-releasing basic fibroblast growth factors in canine myocardium. Zhongguo Linchuang Kangfu.2005; 9(46):153-156
[30]陈建庭,杨德鸿,景宗森,等.bFGF调节PDGF对人成骨细胞的促增殖作用.中华外科杂志.2000Sep；38(9)：707-710
[31]Tatsuyama K,Maezawa Y,Baloe H,et al.Expression of various growth factors for cell proliferatior and cytocifferentiation during fracture repairof bone.Eur J Histooem 2000;4(33):269-278
[32]Ekam EC,Ravarti L,Kaharl V,et al.Expression of extraceluler matrix genes transforming growth factor(TGF)-beta and ras in tibial fracture healing of athynitic rat.Bone 2000;27(4):551-557
[33]陈永龙,雷明全,兰英,等.高原环境对大鼠桡骨骨折愈合过程中TGF-β1表达的影响.解放军医学杂志.2003Jan；28(1)：88-89
[34]杨柳,李起鸿,谭祖键,等.实验性骨延长区TGF-β的基因表达和细胞定位.第三军医大学学报.2001Jan；23(1)：66-68
[35]卢卫忠,唐康来,杨柳,等.不同时间给予转化生长因子p对兔骨折愈合作用的研究.2003 May；25(9)：753-757
[36]马维,牛彦平,张华.合并脑损伤大鼠骨折愈合过程中转化生长因子β1血清含量及在骨折位点的表达.2006Nov；10(42)：88-91
[37]王守君,张柳,孙良智,等.大鼠股骨骨折合并脑外伤时骨愈合过程中TGF-β1的作用.第四军医大学学报.2006；27(15)：1407-1410
[38]Dominguez LJ,Muratore M,Quarta E,et al.Osteoporosis and diabetes. Euatismo 2004;56(4):235-241
[39]Wengreen H J, Munger R G, Wset N A, et al. Dietary protein intake and risk of osteoporotic hip fracture in elderly residents of Utah. J Bone Miner Res.2004; 19(4):537-543
[40]王敏,辛海霞,黄象艳.胰岛素样生长因子-Ⅰ对成骨细胞生长影响的实验研究.中国矫形外科杂志.2004,2(12)：236-238
[41]Spencer CP,Codsland IF,Cooper AJ,et al. Effects of oral for IGF-1 with oral 17 beta-estradiol on insulin sensitivity,Secretion,and elimintion in postmenopausal women. J Metablism,2000,49(6):742
[42]Kurland ES,Rosen CJ,Cosman F.Insulin-like growth factor-I in men with idiopathic osteoporosis[J].J Clin Endocrinol Metab,1997,82:279
[43]Johnsson AG,Lander JM.Human insulin-like growth factor-I binding protein-I(rhIGFBP-I) in transgenic mice:characterization and insights into the regulation of IGFBP-I expression[J].J Clin Endocrinol Metab,1996,81:44
[44]李伟明,王伟,吕建国,等.胰岛素样生长因子-1对骨质疏松大鼠骨形成作用.骨与关节损伤杂志.2004,19(4)：252-254
[45]朱国强,吴织芬,王勤涛,等.IGF-1对骨髓基质细胞分泌BMP-2和VEGF的影响.临床口腔医学杂志.2004,20(4)：215-217
[46]Sakata T, Halloran BP, Elalieh HZ, et al. Skeletal unloading induces resistance to insulin-like growth factor I on bone formation[J]. Bone.2003, 32(6):669-680
[47]徐萍,张克勤,刘超,等.重组人胰岛素样生长因子-Ⅰ(rhIGF-I)对大鼠成骨细胞增殖和功能的影响.南京医科大学学报(自然科学版).2005 Jul,25(7)：472-475
[48]Jehle P,Schulten K,Schulz W,et al.Serum level of insulin-like growth factor(IGF)-I and IGF binding protein(IGFBP)-I to-6 and their relationship to bone metabolism in osteoporosis patients [J]. Eur J Intern Med.2003,14(1):32-38
[49]初同伟,王正国,朱佩芳,等.骨折愈合过程中VEGF及其受体的表达.中华创伤杂志,2001,17：344-346
[50]武永刚,陈君长,王坤正.血管内皮细胞生长因子在骨折愈合过程中的表达.西安医科大学学报.2001,22(1)：51-53
[51]初同伟,王正国,朱佩芳,等.骨折愈合过程中血管内皮生长因子基因扩增及蛋白表达研究.中华实验外科杂志.2002Jul；19(4)：367-368
[52]Streeten EA, Brandi ML. Biology of bone endothelial cells [J]. Bone Miner, 1990,10:85-94
[53]马维,张英泽,宋永周,等.血小板浓缩液对兔尺骨骨折影响的探讨.江苏医药,2003,8：592-59
[54]崔猛,陈建庭.血小板衍生生长因子促进鼠骨早期修复中增殖细胞核抗原与c-Jun蛋白的表达.中国临床康复,2006,10：53-55
[55]Lee JY. Nam SH, Im SY, et al. Enhanced bone form ation by controlledgrowth factor deliyery from chitosan-based biomaterials. J Control Release,2002,78:187-197
[56]Zhang L, Leeman E, Carnes DC, et al. Human osteoblast synthesize and respond to platelet-derived growth factor. Am J Physiol,1991,261:348-354
[57]Horner A, Bord S, Kemp P, et al. Distribution of platelet-derived growth factor (PDGF):a chain mRNA, protein and PDGF-areceptor in rapid ly forming human bone. Bone,1996,19:353-362
[58]张冬根,谭颖徽,裘松波,等.血小板衍生生长因子、骨保护素对体外培养成骨细胞的功能影响.激光杂志.2005；26(3)：94-95
[59]张忠民,陈建庭,金大地,等.血小板衍生生长因子2BB在成骨细胞-破骨细胞培养体系中的生物学作用.中华医学杂志.2001Apr；81(7)：425-428
[60]Alini M,Matsui Y,Dodge GR,et al.The extracellular matrix of cartilage in the growth plate before and during calcification:changes in composition and degradation of type II collagen.Calcif Tissue Int.1992;50(4):327-335.
[61]Grant WT,Wang GJ,Balian G.Type X collagen synthesis during endochondral ossification in fracture repair.J Biol Chem.l987;262(20):9844-9849
[62]Warman ML, Abbott M, Apte SS, et al.A type X collagen mutation causes Schmid metaphyseal chondrodysplasia. Nat Genet.1993;5(l):79-82
[63]郭嫒,黄健,许善锦,等.Ⅱ、Ⅹ型胶原对钙磷酸盐生物矿化生物动力学的影响[J].自然科学进展,2001,11(6)：573-575
[64]Hiltulen A,Aro HT,Vuotle E.Regulation of extracelular matrix genes during fracture healing in mice.Clip Orthop,1993,297:23-27
[65]王立平,党耕町.骨连接素在骨愈合中作用的探讨[J].中华骨科杂志.1998,18(8)：497
[66]Hirakawa K,Hirota S,Ikeda T,et al.Localization of the mRNA for bone matrix proteins during fracture healing as determined by in site hybridization.J Bone Mine Res 1994;9:1551
[67]麻松,阎景龙,王新涛,等.微小颗粒骨复合细胞移植治疗大鼠颅骨.缺损中华实验外科杂志.2005,22：154-155
[68]Xiao G.Fibroblast growth factor 2 induction of the osteocalingene requires MARK activity and phosphorylation of the osteoblast trscription factor,Cbfl/Runx2.J Biol Chem,2002,227:36181-36187