釉基质蛋白联合骨髓基质细胞膜片促进类牙骨质异位生成的实验研究
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摘要
目的:研究釉基质蛋白(enamel matrix proteins,EMPs)诱导犬骨髓基质细胞(bone marrow stromal cells,BMSCs)在裸鼠体内促进类牙骨质异位生成的作用和意义。方法:体外分离培养犬骨髓基质细胞并制备细胞膜片,包裹于牙本质根片上,与煅烧骨块紧密贴合捆绑后,制成牙根片/细胞膜片/煅烧骨复合物,体外模拟根周系统。据牙根片是否事先涂覆釉基质蛋白(EMPs),分为实验组和对照组。将2组的"牙根片/细胞膜片/煅烧骨复合物"分别植入裸鼠背部皮下,8周后取材,进行组织学观察和免疫组化检测。结果:实验组的根片上新生出了不规则类牙骨质样物质,并有矿化基质沉淀,有较强骨桥素(osteopontin,OPN)表达;对照组根片上未见类牙骨质样物质或矿化基质沉淀,OPN表达较弱。结论:釉基质蛋白在体内能诱导骨髓基质细胞向类成牙骨质细胞方向分化,从而促进牙周再生。
Objective: To study the potentials of enamel matrix proteins( EMPs) in enhancing the differentiation and mineralization of bone marrow stromal cells(BMSCs) in periodontal regeneration. Methods: Beagle dog's BMSCs were isolated and cultured as seed cells to form cell sheets. Preparation of root dentin sections were obtained from dog's anterior teeth. EMPs were coated on the surface of root dentin sections. BMSCs sheets were used to encapsulate the EMPs coated root dentin sections. These constructs were finally combined with calcinated ceramic bovine bone(CBB) as scaffolds. Study was divided into two groups. Root dentin sections of group A coated with EMPs. In group B, no EMPs was covered in root dentin sections as control. The composite constructs were implanted subcutaneously into the dorsal aspect in nude mice. The histological observation and immunohistochemical detection was carried out 8 weeks after implantation. Results: In the test group, there were new irregular cementum like tissues on the surface of the roots, and the deposition of matrix mineralization was observed with the intensive expression of osteopontin(OPN). While in the control group, no similar cementum like structure or deposition of matrix mineralization was observed on the roots surface, and the expression of osteopontin was weak. Conclusion: EMPs promotes the differentiation of BMSCs into cementoblasts on the surface of dentin roots in vivo, and the mechanism is possibly related to the expression of OPN.
Objective: To study the potentials of enamel matrix proteins( EMPs) in enhancing the differentiation and mineralization of bone marrow stromal cells(BMSCs) in periodontal regeneration. Methods: Beagle dog's BMSCs were isolated and cultured as seed cells to form cell sheets. Preparation of root dentin sections were obtained from dog's anterior teeth. EMPs were coated on the surface of root dentin sections. BMSCs sheets were used to encapsulate the EMPs coated root dentin sections. These constructs were finally combined with calcinated ceramic bovine bone(CBB) as scaffolds. Study was divided into two groups. Root dentin sections of group A coated with EMPs. In group B, no EMPs was covered in root dentin sections as control. The composite constructs were implanted subcutaneously into the dorsal aspect in nude mice. The histological observation and immunohistochemical detection was carried out 8 weeks after implantation. Results: In the test group, there were new irregular cementum like tissues on the surface of the roots, and the deposition of matrix mineralization was observed with the intensive expression of osteopontin(OPN). While in the control group, no similar cementum like structure or deposition of matrix mineralization was observed on the roots surface, and the expression of osteopontin was weak. Conclusion: EMPs promotes the differentiation of BMSCs into cementoblasts on the surface of dentin roots in vivo, and the mechanism is possibly related to the expression of OPN.
引文
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[13]MacNeil RL,Berry J,D'Errico J,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.
[14]MacNeil RL,Berry J,D′Errico J,et al.Localization and expression of osteopontin in mineralized and nonmineralized tissues of the periodontium[J].Ann N Y Acad Sci,1995;760:166-176.
[15]D′Errio JA,MacNeil RL,Takata T,et al.Expression of bone associated markers by toothroot lining cells,in situ and in vitro[J].Bone,1997,20(2):117-126.
[2]Mohr S,Pormann-Lanz CB,Schoeberlein A,et al.Generation of an osteogenic graft from human placenta and placenta-derived mesenchymal stem cells[J].Repord Sci,2010,17(11):1006-1015.
[3]Varga F,Luegmayr E,Fratzl-Zelman N,et al.Tri-iodothyronine inhibits multilayer formation of the osteoblastic cell line,MC3T3-E1,by promoting apoptosis[J].J Endocrinol,1999,160(1):57-65.
[4]Yamaguchi T,Chattopadhyay N,Kifor O,et al.Extracellular calcium(Ca2+(o))-sensing receptor in a murine bone marrow-derived stromal cell line(ST2):potential mediator of the actions of Ca2+(o)on the function of ST2 cells[J].Endocrinology,1998,139(8):3561-3568.
[5]Slavkin HC.What is the role of the host in periodontal disesae[J] Periodontal Abstr,1975,23(3):101-103.
[6]Fukae M,Tanabe T,Yamakoshi Y.Immunoblot detection and expression of enamel proteins at the apical portion of the forming root in porcine permanent incisor tooth germs[J].J Bone Miner Metab,2001,19(4):236-243.
[7]Hommarstrm L,Heijl L,Gestrelius S,et al.Periodontal regeneration in a buccal dehiscence model in monkeys after application of enamel matrix proteins[J].J Clin Periodontol,1997,24(9 Pt 2):669-677.
[8]Filippi A,Pohl Y,von Arx T.Treatment of replacement resorption with Emdogain--a prospective clinic study[J].Dent Traumato1,2002,18(3):138-143.
[9]Caglar E,Tanboga I,Süsal S.Treatment of avulsed teeth with Emdogain--a case report[J].Dent Traumatol,2005,21(1):51-53.
[10]钟永荣,轩东英,黄雁红,等.Emdogain诱导人牙周膜细胞群向成牙骨质细胞分化的研究[J].广东牙病防治,2010,18(3):138-142.
[11]GestreliusS,Andersson C,Johansson AC,et al.Formulation of enamel matrix derivative for surface coating.Kinetics and cell colonization[J].J Clin Periodontol,1997,24(9):678-684.
[12]束蓉,李超伦,刘正,等.猪釉基质蛋白的提取及N末端氨基酸序列分析[J].口腔医学纵横,1999,15(2):67-68.
[13]MacNeil RL,Berry J,D'Errico J,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.
[14]MacNeil RL,Berry J,D′Errico J,et al.Localization and expression of osteopontin in mineralized and nonmineralized tissues of the periodontium[J].Ann N Y Acad Sci,1995;760:166-176.
[15]D′Errio JA,MacNeil RL,Takata T,et al.Expression of bone associated markers by toothroot lining cells,in situ and in vitro[J].Bone,1997,20(2):117-126.