摘要
正畸牙移动过程中，在牙槽骨发生活跃骨吸收和骨形成的同时，也存在牙骨质的吸收和修复，成牙骨质细胞在这个过程中具有重要意义。正常情况下，位于牙骨质表面的成牙骨质细胞可以抵抗破骨细胞的粘附，从而避免牙根吸收；在已吸收的牙根表面，成牙骨质细胞分泌形成新的牙骨质样组织对其进行修复。当牙根吸收与修复的平衡被打破后，牙根吸收发生。正畸牙移动过程中的牙根吸收，由于其存在的广泛性，不可预见性等特点，成为目前研究的热点。许多研究试图寻找正畸力作用下牙根吸收和修复的发生机制，已取得一定的成果，但仍未清楚。
已往对牙移动过程中牙根吸收和修复的研究多集中在组织形态学上的观察，而对成牙骨质细胞的研究又多集中在牙根发育过程和牙周组织的再生工程中，关于机械应力作用下成牙骨质细胞功能变化的研究还未见报道。本课题通过研究机械应力刺激对成牙骨质细胞重要的矿化相关基因-骨涎蛋白(Bone sialoprotein BSP)和骨桥蛋白(Osteopontin OPN)表达的影响，有助于从细胞力学和分子生物学水平上进一步了解正畸牙移动过程中牙根吸收和修复的发生机制。
本课题利用四点弯曲细胞力学加载装置，以体外培养的成牙骨质样细胞OCCM-30为研究对象，运用实时荧光定量PCR(real-time fluorescentquantitativePCR，FQ-PCR)技术，研究了不同性质、不同强度、不同作用时间的应力应变对成牙骨质细胞矿化相关基因BSP／OPN mRNA表达的影响。从细胞力学和分子生物学水平对正畸力作用下成牙骨质细胞的功能变化与牙根吸收和修复的关系进行了初步揭示，为进一步阐明牙根吸收和修复的机制提供实验基础。
根据实验结果得出以下结论：
1．2000、4000μstrain的张应力和压应力在四个时间点都抑制成牙骨质细胞BSP mRNA的表达，4000μstrain力值比2000μstrain力值对成牙骨质细胞BSP mRNA表达的抑制作用更强。表明机械张、压应力刺激均抑制成牙骨质细胞矿化功能，抑制的强弱与应力的大小和加载时间有关。
2．2000、4000μstrain的压应力在作用初期促进成牙骨质细胞OPN mRNA的表达，12h后表现为抑制作用。2000μstrain和4000μstrain的张应力抑制OPN mRNA的表达，抑制作用随着加载时间的延长而增强。表明机械应力刺激可以调节成牙骨质细胞OPN mRNA的表达，进而可能影响成牙骨质细胞的矿化功能和破牙骨质细胞的粘附能力。
综上所述，成牙骨质细胞是力学敏感性细胞，机械应力刺激可以调节成牙骨质细胞BSP／OPN mRNA的表达，进而可能影响成牙骨质细胞的矿化功能和破牙骨质细胞的粘附能力。机械应力刺激对成牙骨质细胞BSP／OPN mRNA表达的影响与应力的性质，大小和加载时间有关。
Background and objective: During orthodontic tooth movement ,The remodeling of alveolar bone is active, Meanwile ,cementum also undergoes both resorption and deposition. Cementoblast plays an important role in the progress of root resorption and repair. Cementoblast is located on the surface of root cementum to inhibit the attachment of the osteoclast and form the new cementum-like tissue in the lacuna on the resorpted root surface. The pathologic resorption is formed when the balance of resorption and repair is disturbed. Orthodontically induced root resorption is the most frequent unwanted side effect of orthodontic tooth movement that is unavoidable. Although many researchers focus on it ,the pathogenesis of root resorption is still not clear.It is not known how orthodontic force influences the function of cementoblast and what is the relationship between the function change of cementoblast with root resorption and repair.The aim of present research is to determine the effects of the mechanical strain on the express of the mineral associated gene BSP and OPN in cementoblast in vitro.
Methods: Cementoblasts OCCM30 were cultured in DMEM with 10% FBS for 48. hours, then subjected to mechanical strain by four-point bending system with tension and compression stress at 2000μstrain and 4000μstrain at 0.5Hz frequency for3h, 6h, 12h, 24h. BSPand OPN mRNA were analysed with real time quantitive RT-PCR after mechanical loading. Statistical significance was determined for each comparison using the one-way ANOVA, and p<0.05 was statistically considered significant.
Results:
1. Cementoblasts are sensitive cells to mechanical strain.
2. 2000/4000μstrain tension and compression both suppressed BSP mRNA expression of Cementoblasts. 4000μstrain suppressed stronger than 2000μstrain.
3. OPN mRNA expression of Cementoblasts was increased in response to compression stress after 31k 6h mechanical loading,then suppressed after 12h、24h loading. while Tension stress almost suppressed OPN mRNA expression all times.
Conclusion:
1. Mechanical strain suppressed BSP mRNA expression of Cementoblasts and is believed inhibit the capability of cementoblast to synthesize cementum matrix.
2. It is suggest that cementoblast regulates the function of osteoclast by the different expression of OPNmRNA after mechanical loading .
引文
[1] Brudvik P, Rygh P. The repair of orthodontic root resorption: an ultrastructural study[J]. Eur J Orthod, 1995, 17(3):189-198.
[2] Harris DA, Jones AS, Darendeliler MA. Physical properties of root cementum: part 8. Volumetric analysis of root resorption craters after application of controlled intrusive light and heavy orthodontic forces: a microcomputed tomography scan study [J]. Am J Orthod Dentofacial Orthop, 2006,130(5):639-647.
[3] Chan E, Darendeliler MA. Physical properties of root cementum: Part 5. Volumetric analysis of root resorption craters after application of light and heavy orthodontic forces [J]. Am J Orthod Dentofacial Orthop, 2005, 127(2):186-195.
[4] Weiland F. External root resorptions and orthodontic forces: correlations and clinical consequences[J]. Prog Orthod , 2006, 7(2): 156-163.
[5] Kurol J. Time-related root resorption after application of a controlled continuos orthodontic force[J]. Am J Orthod Dentofacial Orthop, 1996,110(3):303-310.
[6] El-Bialy T, El-Shamy I, Graber TM. Repair of orthodontically induced rootresorption by ultrasound in humans[J].Am J Orthod Dentofacial Orthop ,2004, 126(2):186-193
[7] Chrissavgi S, Marita H, Sven L. Healing of the root surface-associated periodontium: an immunohistochemical study of orthodontic root resorption in man [J]. Eur J Orthod, 1996, 18(1):435-444.
[8] Fisher LW, Whitson SW, Avioli LV, et al. Matrix sialoprotein of developing bone[J]. J Biol Chem, 1983, 258(20):12723-12727.
[9] Ganss B, Kim RH, Sodek J. Bone sialoprotein[J]. Crit Rev Oral Biol Med , 1999,10(1): 79-98.
[10] Nanci A.Content and distribution of noncollagenous matrix proteins in bone and cementum: relationship to speed of formation and collagen packing density[J]. J Struct Biol, 1999, 126 (3): 256-269.
[11] Saygin NE, Tokiyasu Y, Giannobile WV. Growth factors regulate expression of mineral associated genes in cementoblasts[J]. J Periodontol, 2000 ,71(10):1591-1600.
[12] Fernanda B, Janice EB, Amy JK et al.The Role of Parathyroid Hormone-Related Protein in the Regulation of Osteoclastogenesis by Cementoblasts[J]. J Periodontol, 2004, 75(9):1247-1254.
[13] Hakki SS. Nohutcu RM. Hakki EE. Dexamethasone and basic - fibroblast growth factor regulate markers of mineralization in cementoblasts in vitro[J]. J Periodontol, 2005 ,76(9):1550-1558.
[14] Yong-Hee PC, Brian L, Patricia AL et al. Bisphosphonate Modulates Cementoblast Behavior In Vitro[J]. J Periodont, 2005, 76(11): 1890-1900.
[15] Foster BL, Nociti FH , Swanson E C. Regulation of cementoblast gene expression by inorganic phosphate in vitro[J].Calcified Tissue International, 2006, 78 (2):103-112.
[16] Arzate H, Olson SW, Page RC. Isolation of human tumor cells that produce cementum protein in culture[J]. Bone Miner , 1992, 18(1) : 15-30 [17] Grzesik WJ , Kuzentsov SA, Uzawa K. Normal Human Cementum-Derived Cells: Isolation, Clonal Expansion, and In Vitro and In Vivo Characterization[J]. J Bone Miner Res, 1998,13(10):1547 - 1554.
[18] Kitagawa M, Kitagawa S, Kudo Y et al. Establishment of cementoblast cell lines from rat cementum lining cells by transfection with temperature-sensitive simian virus-40 T-antigen gene[J].Bone, 2005, 37(2): 220-226.
[19] D'Errico JA, MacNeil RL, Takata T et al. Expression of bone associated markers by tooth lining cells, in situ and in vitro[J]. Bone 1997,20 (2):117-126.
[20] D' Errico JA, Ouyang H, Berry J et al. Immortalized cementoblasts and periodontal ligament cells in culture [J]. Bone, 1999, 25(1):39-47.
[21] D'Errico JA, Berry JE, Ouyang H et al. Employing a transgenic animal model to obtain cementoblasts in vitro[J]. J Periodontol, 2000, 71(1): 63-72.
[22] Wang N, Butler JP, Ingber DE. Mechanotransduction across the cell surface and through the cytoskeleton[J]. Science, 1993,260(5111): 1124-1127.
[23] Nguyen KT, Eskin SG, Patterson C et al. Shear stress reduces protease activated receptor-1 expression in human endothelial cells[J]. Ann Biomed Eng ,2001, 29(2): 145-152
[24] Yamamoto T, Kita M, Kimura I, et al. Mechanical stress induces expression of cytokines in human periodontal ligament cells[J]. Oral Diseases ,2006, 12 (2) : 171-175.
[25] Owan I, Burr DB, Turner CH, et al. Mechanotransduction in bone: osteoblasts are more responsive to fluid forces than mechanical strain[J]. Am J Physiol,1997,273(3ptl): C810-815.
[26] Rubin J, Murphy T, Nanes MS, et al. Mechanical strain inhibits expression of osteoclast differentiation factor by murine stromal cells[J]. Am J Physiol Cell Physiol, 2000, 278(6): C1126 - C1132
[27] Burr DB, Milgrom C, Fyhrie D et al .In vivo measurement of human tibial strains during vigorous activity[J]. Bone , 1996,18(5): 405- 410.
[28] Frost HM. Perspectives: bone' s mechanical usage windows [J]. Bone Miner , 1992,19(3) :257-271.
[29] Fermor B, Gundle R, Evans M et al. Primary human osteoblast proliferation and prostaglandin E2 release in response to mechanical strain in vitro[J]. Bone, 1998, 22(6):637-43.
[30] Willard MF, Stephen W, Kent EV. Quantitative RT-PCR: Pitfalls and Potential [J]. Biotechniques, 1999,26(1):112-125.
[31] Fisher LW, Whitson SW, Avioli LV et al. Matrix sialoprotein of developing bone[J]. J Biol Chem , 1998, 258(20):12723-12727.
[32] Fujisawa R, Butler WT, Brunn JC et al. Differences in composition . of cell-attachment sialoproteins between dentin and bone[J]. J Dent Res , 1993, 72(8):1222-1226.
[33] Hunter GK, Goldberg HA . Nucleation of hydroxyapatite by bone sialoprotein[J]. Proc Natl Acad Sci USA ,1993,90(18):8562-8565.
[34] Hunter GK, Goldberg HA . Modulation of crystal formation by bone phosphoproteins: role of glutamic acid-rich sequences in the nucleation of hydroxyapatite by bone sialoprotein[J]. Biochem . J ,1994, 302(1): 175-179.
[35] Hunter GK, Kyle CL, Goldberg HA . Modulation of crystal formation by bone phosphoproteins: structural specificity of the osteopontin- mediated inhibition of hydroxyapatite formation[J]. Biochem J, 1994, 300(3):723-728.
[36] Alford AI, Hankenson KD.Matricellular proteins: Extracellular modulators of bone development, remodeling, and regeneration [J] Bone, 2006, 38(6) :749-57.
[37] MacNeil RL, Sheng N, Strayhorn C et al. Bone sialoprotein is localized to the root surface during cementogenesis[J]. J Bone Min Res, 1994, 9(10):1597-1606.
[38] MacNeil RL, Berry I, D' Errico I et al. Role of two mineral-associated adhesion molecules, osteopontin and bone sialoprotein, during cementogenesis[J]. Connect Tissue Res , 1995, 33(1-3):1-7.
[39] MacNeil RL, Berry J, Strayhorn C et al. Expression of bone sialoprotein mRNA by cells lining the mouse tooth root during cementogenesis[J]. Archives of Oral Biology, 1996, 41(8-9): 827-35.
[40] Matias MA, Li H, Young WG et al. Immunohistochemical localisation of extracellular matrix proteins in the periodontium during cementogenesis in the rat molar[J]. Archives of Oral Biology, 2003, 48(10): 709-16.
[41] McKee MD, Zalzal S, Nanci A . Extracellular matrix in tooth cementum and mantle dentin localization of osteopontin and other noncollagenous proteins, plasma proteins and glycoconjugates by electronmicroscopy[J]. Anat Rec , 1996,245(2):293- 312.
[43] Shigeyama Y, Grove TK, Strayhorn C et al. Expression of adhesion molecules during tooth resorption in feline teeth: a model system for aggressive osteoclastic activity[J]. Journal of Dental Research, 75(9): 1650-1657.
[44] Carvalho RS, Schaffer JL, Gerstenfeld LC. Osteoblasts induce osteopontin expression in response to attachment on fibronectin: demonstration of a common role for integrin receptors in the signal transduction processes of cell attachment and mechanical stimulation[J]. Journal of Cellular Biochemistry, 1998,70(3): 376-390.
[45] Pavlin D, Gluhak-Heinrich J. Effect of mechanical loading on periodontal cells [J]. Crit Rev Oral Biol Med , 2001,12 (5) : 414-424.
[46] Meikle MC. The tissue, cellular, and molecular regulation of orthodontic tooth movement: 100 years after Carl Sandstedt [J]. Eur J Orthod ,2006, 28(3):221-240.
[47] Brudvik P, Rygh P. Transition and determinants of orthodontic root resorption-repair sequence [J]. Eur J Orthodo, 1995,17(3):177-188.
[48] Casa MA, Faltin RM, Faltin K et al. Root resorption on torqued human premolars shown by tartrate-resistant acid phosphatase histochemistry and transmission electron microscopy[J]. Angle Orthod, 2006, 76(6) :1015-1021.
[49] Ramanathan C,Hofman Z. Root resorption in relation to orthodontic tooth movement[J]. Acta Medica, 2006, 49(2):91-95.
[50] Sismanidou C, Lindskog S. Spatial and temporal repair patterns of orthodontically induced surface resorption patches [J]. Eur J Oral Sciences, 1995,103(5):292-298.
[51] Carvalho RS, Bumann A, Schaffer JL. Predominant Integrin Ligands Expressed by Osteoblasts Show Preferential Regulation in Response to Both Cell Adhesion and Mechanical Perturbation [J]. Journal of Cellular Biochemistry ,2004, 84(3):497-508.
[52] MacNeil RL, Berry I, D' Errico I et al. Role of two mineral- a ssociated adhesion molecules, osteopontin and bone sialoprotein, during cementogenesis[J]. Connect Tissue Res , 1995, 33(1-3):l-7.
[53] Hasegawa N, Kawaguchi H, Ogawa T et al. Immunohistochemical characteristics of epithelial cell rests of Malassez during cementum repair[J]. J Periodont Res, 2003, 38(1):51 - 56.
[54] Miyauchi A, Alvarez J, Greenfield EM et al. Recognition of osteopontin and related peptides by an alpha v beta 3 integrin stimulates immediate cell signals in osteoclasts. [J]. J Biol Chem , 1991, 266(30):20369-20374.
[55] Weber GF, Ashkar S, Glimcher MJ et al. Receptor-ligand interaction between CD44 and osteopontin (Eta-1) [J]. Science, 1996, 271(5248): 509-512.
[56] Asou Y, Rittling SR, Yoshitake H et al. Osteopontin facilitates angiogenesis, accumulation of osteoclasts, and resorption in ectopic bone[J]. Endocrinology, 2001,142(3):1325-1332.
[57] Dolce C, Vakani A, Archer L et al.Effects of echistatin and an RGD peptide on orthodontic tooth movement[J]. J Dent Res, 2003, 82(9) : 682-686
[58] Talic NF, Evans C, Zaki AM. Inhibition of orthodontically induced root resorption with mechistatin, an RGD-containing peptide[J]. Am J Orthod Dentofacial Orthop, 2006,129(2):252-60.
[59] J. Sodek B, Ganss MD,McKee. Osteopontin[J]. Crit Rev Oral Biol Med,2000, 11(3): 279-303 .
[60] McKee MD, Nanci A. Osteopontin at mineralized tissue interfaces in bone, teeth, and osseointegrated implants: ultrastructural distribution and implications for mineralized tissue formation, turnover and repair [J]. Microsc Res Tech, 1996, 33(2):141-164.
[61] McKee MD, Nanci A . Secretion of osteopontin by macrophages and its accumulation at tissue surfaces during wound healing in mineralized tissues: a potential requirement for macrophage adhesion and phagocytosis[J]. Anat Rec , 1996, 245(2):394-409.
[62] . McKee MD, Nanci A. Osteopontin: an interfacial extracellular matrix protein in mineralized tissues [J]. Connect Tissue Res, 1996, 35(1-4): 197-205.
[63] McKee MD, Nanci A . Osteopontin deposition in remodeling bone: an osteoblast mediated event [J], Bone Miner Res ,1996, 11(6): 873-874.
[64] McKee MD, Nanci A. Osteopontin deposition in remodeling bone: an osteoblast mediated event[J].J Bone Miner Res, 1996, 11(6) :873-875.
[65] Noda M, Rittling S, Muguruma Y et al. Osteopontin-deficient mice (0D-)cells exhibit enhanced actin fiber formation and spreading while OD-osteoclasts are less efficient in removing hydroxyapatite coated on glass and in recruitment to ectopically implanted bone matrix (abstract) [J]. J Bone Miner Res, 1998, 23:S220.
[66] YoshitakeH, Rittling SR, Denhardt DT er al. Osteopontin-deficient mice are resistant to ovariectomy-induced bone resorption [J]. Proc NatI Acad Sci USA , 1999,96(14):8156-8160.
[67] Rittling SR, Denhardt DT . Osteopontin function in pat:hology lessons from osteopontin-deficient mice[J].ExL Nephrol ,1999,7 (2): 103-113.
[68] Ishijima M, Rittling SR, Yamashita T et al. Enhancement of osteoclastic bone resorption and suppression of osteoblastic bone formation in response to reduced mechanical stress do not occur in the absence of osteopontin [J]. J Exp Med, 2001,193(3):399-404.
[69] Talic NF, Evans C, Zaki AM. Inhibition of orthodontically induced root resorption with mechistatin,. an RGD-containing peptide[J]. Am J Orthod Dentofacial Orthop, 2006,129(2):252-60.
[70] Adam Lee. Root resorption: The possible role of extracellular matrix proteins [J]. Am J Orthod Dentofacial Orthop , 2004,126(8): 173-177.
[71] Klein NJ, Roelofsen J, Semeins CM et al. Mechanical stimulation of osteopontin mRNA expression and synthesis in bone cell cultures [J]. Journal of Cellular Physiology, 1997, 170(2):174-181.
[72] Carvalho RS. Schaffer JL. Gerstenfeld LC. Osteoblasts induce osteopontin expression in response to attachment on fibronectin: demonstration of a common role for integrin receptors in the signal transduction processes of cell attachment and mechanical stimulation [J]. Journal of Cellular Biochemistry , 1998 , 70(3): 376-90.
[73] Kubota T, Yaraauchi M, Onozaki J et al. Influence of an intermittent compressive force on matrix protein expression by ROS 17/2. 8 cells, with selective stimulation of osteopontin[J]. Archives of Oral Biology, 1993, 38(1): 23-30.
[1] Roach HI. Why does bone matrix contain non-collagenous proteins? The possible roles of osteocalcin, osteonectin, osteopontin and bone sialoprotein in bone mineralisation and resorption[J]. Cell Biol Int, 1994,18(6): 617-628.
[2] Sako D, Comess KM, Barone Km et al. A sulfated peptide segment at the amino terminus of PSGL-1 is critical for P-selectin binding[J]. Cell, 1995, 83 (2) :323-331.
[3] MacNeil RL, Berry I, D'Errico et al. Role of two mineral- associated adhesion molecules, osteopontin and bone sialoprotein, during cementogenesis[J]. Connect Tissue Res,1995,33(1-3):l-7.
[4] Fisher LW,Whitson SW, Avioli LV et al. Matrix sialoprotein of developing bone[J]. J Biol Chem ,1998,258(20):12723-12727
[5] Fujisawa R, Butler WT, Brunn JC et al. Differences in composition of cell-attachment sialoproteins between dentin and bone [J]. J Dent Res,1993,72(8):1222-1226.
[6] Hunter GK, Goldberg HA . Nucleation of hydroxyapatite by bone sialoprotein[J]. Proc Natl Acad Sci USA ,1993,90(18):8562-8565.
[7] Hunter GK, Goldberg HA . Modulation of crystal formation by bone phosphoproteins: role of glutamic acid-rich sequences in the nucleation of hydroxyapatite by bone sialoprotein[J]. Biochem J,1994,302(1):175-179.
[8] Hunter GK, Kyle CL, Goldberg HA . Modulation of crystal formation by bone phosphoproteins: structural specificity of the osteopontin- mediated inhibition of hydroxyapatite formation[J]. Biochem J,1994, 300(3):723-728.
[9] Fujisawa R, Butler WT, Brunn JC et al. Differences in composition of cell-attachment sialoproteins between dentin and bone[J]. J Dent Res, 1993,72(8): 1222-1226.
[10] Bosshardt DD, Schroeder HE.Cementogenesis reviewed: a comparison between human premolars and rodent molars[J].Anat Rec,1996,245 (2):267-92.
[11] Chen IK, Shapiro HS, Wrana JL et al.Localization of bone sialoprotein(BSP) expression to sites of mineralized tissue formation in fetal rat tissues by in situ hybridization[J]. Matrix,1991, 11 (2):133-143.
[12] Kobayashi D,Takita H, Mizuno M et al. Time-dependent expression of bone sialoprotein fragments in osteogenesis induced by bone morphogenetic protein[J]. J Biochem,1996, 119(3):475-481..
[13] 卫书盛，曹采方，孟焕新．大鼠牙齿发育过程中骨涎蛋白和骨桥蛋白的表达[J]．中华口腔医学杂志，2002．37(1)：47-49． WEISs, CAO Cf, MENG Hx. Expression of bone sialoprotein and osteopontin in developing dental tissues of rats[J]. Chin J.Stomatol, 2002, 37(1):47-49.
[14] Chen J, Sasaguri K, Sodek J , et al. Enamel epithelium expresses bone sialoprotein(BSP). Eur J Oral Sci, 1998,106 (Suppl 1):331-361.
[15] Cho P, Schneider GB, Krizan K. Examination of the bone-implant interface in experimentally induced osteoporotic bone [J]. J Implant Dent, 2004,13 (1):79-87.
[16] O'Toole GC. Salih E. Gallagher C. Bone sialoprotein-coated femoral implants are osteoinductive but mechanically compromised [J]. J Orthop Res,2004,22(3):641-646.
[17] McKee MD, Giachelli CM, Nanci A . Matrix-mineral relationships in calcifying human atherosclerotic plaque: ultrastructural immunodetection of osteopontinand bone sialoprotein at calcification sites (abstract). J Bone Miner Res ,1996,1 l:S330.
[18] Franzen A, Heinegard D.Isolation and characterization of two sialoproteins present only in bone calcified matrix[J]. Biochem J.1985,232(3):715-724.
[19] J. Sodek B,Ganss MD,McKee. Osteopontin[J]. Crit Rev Oral Biol Med,2000,ll(3):279-303.
[20] Hunter GK, Kyle CL, Goldberg HA . Modulation of crystal formation by bone phosphoproteins: structural specificity of the osteopontin- mediated inhibition of hydroxyapatite formation[J]. Biochem J, 1994, 300(3):723-728.
[21] Wada T, McKee MD, Steitz S.Calcification of vascular smooth muscle cell cultures:inhibition by osteopontin[J]. Circ Res ,1999, 84(2):166-178.
[22] Hunter GK, Hauschka PV, Poole AR. Nucleation and inhibition of hydroxyapatite formation by mineralized tissue proteins [J]. Biochem J,1996,317 (Ptl):59-64.
[23] Lekic P,Sodek J,McCulloch CA. Relationship of cellular proliferation to expression of osteopontin and bone sialoprotein in regenerating rat periodontiumn[J]. Cel Tissue Res ,1996,285(3):491 -500.
[24] Liaw L, Skinner MP, Raines EW. The adhesive and migratory effects of osteopontin are mediated via distinct cell surface integrins. Role of alpha v beta 3 in smooth muscle cell migration to osteopontin in vitro [J]. J Clin Invest,1995,95(2):713-724.
[25] Ross FP, Chappel J, Alvarez JI et al.Interactions between the bone matrix proteins osteopontin and bone sialoprotein and the osteoclast integrin alpha v beta 3 potentiate bone resorption [J]. Biol Chem,1993,268(13):9901 -9907.
[26] Miyauchi A, Alvarez J, Greenfield EM et al. Recognition of osteopontin and related peptides by an alpha v beta 3 integrin stimulates immediate cell signals in osteoclasts. [J]. J Biol Chem ,1991,266(30):20369-20374.
[27] Weber GF, Ashkar S, Glimcher MJ et al. Receptor-ligand interaction between CD44 and osteopontin (Eta- 1) [J]. Science,1996, 271(5248): 509-512.
[28] McKee MD, Nanci A. Osteopontin at mineralized tissue interfaces in bone, teeth, and osseointegrated implants: ultrastructural distribution and implications for mineralized tissue formation, turnover and repair [J]. Microsc Res Tech ,1996,33(2):141-164.
[29] McKee MD, Nanci A . Secretion of osteopontin by macrophages and its accumulation at tissue surfaces during wound healing in mineralized tissues: a potential requirement for macrophage adhesion and phagocytosis [J]. Anat Rec,1996,245(2):394-409.
[30] McKee MD, Nanci A.Osteopontin: an interfacial extracellular matrix protein in mineralized tissues[J]. Connect Tissue Res,1996, 35(1-4): 197-205.
[31] McKee MD, Nanci A . Osteopontin deposition in remodeling bone: an osteoblast mediated event[J]. Bone Miner Res ,1996, 11(6): 873-874.
[32] McKee MD, Nanci A. Osteopontin deposition in remodeling bone: an osteoblast mediated event[J].J Bone Miner Res,1996,11(6): 873-875.
[33] Zohar R. Cheifetz S. McCulloch CA et al.Analysis of intracellular osteopontin as a marker of osteoblastic cell differentiation and mesenchymal cell migration [J]. Eur J Oral Sci 106 Suppl 1: 401 -407.
[34] Takazawa Y, Rittling S, Furuya K et al. (1998). Interleukin-1- induced bone resorption does not occur in osteopontin-deficient mice (abstract). J Bone Miner Res,1998, 23:S246.
[35] Dolce C, Vakani A,Archer L et al.Effects of echistatin and an RGD peptide on orthodontic tooth movement[J].J Dent Res,2003,82(9): 682-686
[36] Talic NF, Evans C,Zaki AM.Inhibition of orthodontically induced root resorption with mechistatin, an RGD-containing peptide [J], Am J Orthod Dentofacial Orthop, 2006,129(2):252-60.
[37] Asou Y, Rittling SR, Yoshitake H et al. Osteopontin facilitates angiogenesis, accumulation of osteoclasts, and resorption in ectopic bone [J]. Endocrinology,2001,142(3):1325-1332.
[38] Noda M, Rittling S, Muguruma Y et al. Osteopontin-deficient mice (OD-)cells exhibit enhanced actin fiber formation and spreading while OD-osteoclasts are less efficient in removing hydroxyapatite coated on glass and in recruitment to ectopically implanted bone matrix (abstract) [J]. J Bone Miner Res,1998,23:S220.
[39] Ishijima M, Rittling SR, Yamashita T et al. Enhancement of osteoclastic bone resorption and suppression of osteoblastic bone formation in response to reduced mechanical stress do not occur in the absence of osteopontin[J].J Exp Med, 2001,193(3):399-404.
[40] Bosshardt DD, Schroeder HE . Cementogenesis reviewed: A comparison between human premolars and rodent molars[J]. Anat Rec,1996, 245(2):267-292.
[41] Bosshardt DD.Are cementoblast a subpopulation of osteoblasts or a unique phenotype[J].J Dent Res ,2005, 84(5):390-406。
[42] McKee MD, Zalzal S, Nanci A . Extracellular matrix in tooth cementum and mantle dentin localization of osteopontin and other noncollagenous proteins,plasma proteins and glycoconjugates by electronmicroscopy[J]. Anat Rec ,1996,245(2):293- 312.
[43] Bosshardt DD,Zalzal S,McKee . Developmental appearance and distribution of bone sialoprotein and osteopontin in human and rat cementum. [J] Anat Rec,1998,250(1):13-33.
[44] Bosshardt DD,Degen T,Lang NP. Sequence of protein expression of bone sialoprotein and osteopontin at the developing interface between repair cementum and dentin in human deciduous teeth. [J] Cell Tissue Res,2005, 320(3):399-407.
[45] MacNeil RL, Sheng N, Strayhorn C et al. Bone sialoprotein is localized to the root surface during cementogenesis[J]. J Bone Min Res, 1994, 9(10):1597-1606.
[46] D'Errico JA, MacNeil RL,Strayhorn CL et al.Models for the study of cementogenesis[J]. Connective Tissue Research,1995,33(1-3): 9-17.
[47] MacNeil RL, Berry I, D'Errico I et al. Role of two mineral- associated adhesion molecules, osteopontin and bone sialoprotein, during cementogenesis[J]. Connect Tissue Res ,1995,33(1-3):l-7.
[48] MacNeil RL, Berry J, Strayhorn C et al.Expression of bone sialoprotein mRNA by cells lining the mouse tooth root during cementogenesis[J]. Archives of Oral Biology,1996,41(8-9):827-835.
[49] Matias MA, Li H, Young WG et al.Immunohistochemical localisation of extracellular matrix proteins in the periodontium during cementogenesis in the rat molar[J]. Archives of Oral Biology, 2003,48(10):709-16.
[50] Cho MI,Gsrant PR.Development and general structure of the periodontium[J]. Periodontol,2000,24:9-27
[51] Bosshardt DD, Nanci A: Hertwig's epithelial root sheath, enamel matrix proteins, and. initiation of cementogenesis in porcine teeth[J]. J Clin Periodontol, 2004, 31(3): 184-192.
[52] Bosshardt DD,selvig K. Dental cementum :The dynamic tissue covering the root[J]. Periodontol, 1997,13:41-75.
[53] Saygin NE,Tokiyasu Y,Giannobile WV.Growth factors regulate expression of mineral associated genes in cementoblasts[J]. J Periodontol, 2000,71(10):1591-1600.
[54] Hakki SS. Nohutcu RM. Hakki EE. Dexamethasone and basic - fibroblast growth factor regulate markers of mineralization in cementoblasts in vitro [J]. J Periodontol, 2005 ,76(9):1550-1558.
[55] Foster BL, Nociti FH, Swanson E C. Regulation of cementoblast gene expression by inorganic phosphate in vitro [J] .Calcified Tissue International, 2006,78(2):103-112.
[56] Yong-Hee PC,Brian L,Patricia AL et al.Bisphosphonate Modulates Cementoblast Behavior In Vitro[J]. J Periodont,2005, 76(11): 1890-1900.
[57] Zhao M. Berry JE. Somerman MJ. Bone morphogenetic protein-2 inhibits differentiation and mineralization of cementoblasts in vitro [J]. Journal of Dental Research,2003, 82(1):23-7.
[58] Domon S. Shimokawa H. Yamaguchi S. Temporal and spatial mRNA expression of bone sialoprotein and type I collagen during rodent tooth movement[J]. Eur J Orthod,2001,23(4):339-348.
[59] Shigeyama Y,Grove TK,Strayhorn C et al. Expression of adhesion molecules during tooth resorption in feline teeth: a model system for aggressive osteoclastic activity [J]. Journal of Dental Research,75(9):1650-1657.
[60] Adam Lee. Root resorption: The possible role of extracellular matrix proteins[J]. Am J Orthod Dentofacial Orthop, 2004,126(8): 173-177.
[61] Hasegawa N, Kawaguchi H, Ogawa T et al. Immunohistochemical characteristics of epithelial cell rests of Malassez during cementum repair[J].J Periodont Res, 2003,38(1):51-56.