硅酸二钙离子浸提液对人牙周膜细胞增殖和矿化能力的影响
|
摘要
目的:探讨硅酸二钙离子浸提液对人牙周膜细胞增殖与矿化能力的影响。方法:组织块消化法分离培养人牙周膜细胞,制备硅酸二钙离子浸提液,并配置为条件培养基诱导培养细胞,采用细胞增殖-毒性检测试剂盒检测细胞增殖情况,检测碱性磷酸酶活性及矿化结节形成情况。结果:硅酸二钙离子浸提液对人牙周膜细胞无细胞毒性,促进细胞增殖,可有效提高细胞的碱性磷酸酶活性,与对照组比较差异具有统计学意义(P<0.05)。该条件培养基可显著促进牙周膜细胞形成矿化颗粒,与对照组比较差异具有统计学意义(P<0.05)。结论:硅酸二钙离子浸提液对人牙周膜细胞无细胞毒性,促进了人牙周膜细胞的增殖和矿化能力。
Objective:To investigated the effect of ionic products of dicalcium silicate on human periodontal ligament cells proliferation and mineralization. Methods: Human periodontal ligament cells were isolated and cultured by enzymolytic method. Ionic products of dicalcium silicate was prepared as previously reported. The cells were co-cluture with ionic products of dicalcium silicate or with normal DMEM. Then CCK-8,Alkaline Phosphatase(ALP) analyses and mineralization assay were used to identify the cell proliferation and mineralization effects. Results: The induced cells showed high cell proliferation rate, higher ALP activity(P<0.05) and increased calcified nodules formation(P<0.05).Conclusion:Ionic products of dicalcium silicate promote human periodontal ligament cell proliferation and mineralization capacity.
Objective:To investigated the effect of ionic products of dicalcium silicate on human periodontal ligament cells proliferation and mineralization. Methods: Human periodontal ligament cells were isolated and cultured by enzymolytic method. Ionic products of dicalcium silicate was prepared as previously reported. The cells were co-cluture with ionic products of dicalcium silicate or with normal DMEM. Then CCK-8,Alkaline Phosphatase(ALP) analyses and mineralization assay were used to identify the cell proliferation and mineralization effects. Results: The induced cells showed high cell proliferation rate, higher ALP activity(P<0.05) and increased calcified nodules formation(P<0.05).Conclusion:Ionic products of dicalcium silicate promote human periodontal ligament cell proliferation and mineralization capacity.
引文
[1]Liu X,Morra M,Carpi A,et al.Bioactive calcium silicate ceramics and coatings[J].Biomed Pharmacother,2008,62(8):526-529
[2]Gandolfi MG,Ciapetti G,Taddei P,et al.Apatite formation on bioactive calcium-silicate cements for dentistry affects surface topography and human marrow stromal cells proliferation[J].Dent Mater,2010,26(10):974-992
[3]Chen CC,Shie MY,Ding SJ.Human dental pulp cell responses to new calcium silicate-based endodontic materials[J].Int Endod J,2011,44(9):836-842
[4]Wu BC,Wei CK,Hsueh NS,et al.Comparative cell attachment,cytotoxicity and antibacterial activity of radiopaque dicalcium silicate cement and white-coloured mineral trioxide aggregate[J].Int Endod J,2015,48(3):268-276
[5]Chiang TY,Ding SJ.Physicochemical properties of radiopaque dicalcium silicate cement as a root-end filling material in an acidic environment[J].Int Endod J,2013,46(3):234-241
[6]Chiang TY,Ding SJ.Comparative physicochemical and biocompatible properties of radiopaque dicalcium silicate cement and mineral trioxide aggregate[J].J Endod,2010,36(10):1683-1687
[7]Sun J,Li J,Liu X,et al.Proliferation and gene expression of osteoblasts cultured in DMEM containing the ionic products of dicalcium silicate coating[J].Biomedicine&pharmacotherapy,2009,63(9):650-657
[8]Chen LJ,Zhang YL,Liu J,et al.Exposure of the Murine RAW 264.7 Macrophage Cell Line to Dicalcium Silicate Coating Surface:Assessment of Cytotoxicity and Pro-inflammatory effects[J].J Mater Sci-Mater M,2016,27(3):59
[9]Wang Y,Medvid R,Melton C,et al.DGCR8 is essential for microRNA biogenesis and silencing of embryonic stem cell self-renewal[J].Nat Genet,2007,39(3):380-385
[10]Liu X,Tao S,Ding C.Bioactivity of plasma sprayed dicalcium silicate coatings[J].Biomaterials,2002,23(3):963-968
[11]Li J,Wei L,Sun J.Effect of ionic products of dicalcium silicate coating on osteoblast differentiation and collagen production via TGF-β1 pathway[J].J Biomater Appl,2011,9(7):98-103
[12]Liu X,Xie Y,Ding C,et al.Early apatite deposition and osteoblast growth on plasma-sprayed dicalcium silicate coating[J].J Biomed Mater Res,2005,74(3):356-365
[13]Velasquez P,Luklinska ZB,Meseguer-Olmo L,et al.Α-TCP ceramic doped with dicalcium silicate for bone regeneration applications prepared by powder metallurgy method:in vitro and in vivo studies[J].J Biomed Mater Res A,2013,101(7):1943-1954
[14]Meseguer-Olmo L,Aznar-Cervantes S,Mazón P,et al.“In vitro”behavior of adult mesenchymal stem cells of human bone marrow origin seeded on a novel bioactive ceramics in the Ca2SiO4-Ca3(PO4)2 system[J].J Mater Sci Mater Med,2012,23(12):3003-3014
[15]De Aza PN,Zuleta F,Velasquez P,et al.(α'(H))-Dicalcium silicate bone cement doped with tricalcium phosphate:characterization,bioactivity and biocompatibility[J].J Mater Sci Mater Med,2014,25(2):445-452
[16]Correa D,Almirall A,Carrodeguas RG,et al.α-Tricalcium phosphate cements modified withβ-dicalcium silicate and tricalcium aluminate:Physicochemical characterization,in vitro bioactivity and cytotoxicity[J].J Biomed Mater Res B Appl Bio-mater,2015,103(1):72-83
[17]de Aza PN,Luklinska ZB,Mate Sanchez,et al.Biodegradation process ofα-tricalcium phosphate andα-tricalcium phosphate solid solution bioceramics in vivo:a comparative study[J].Microsc Microanal,2013,19(5):1350-1357
[18]Dong S,Sun J,Li Y,et al.Electrospun nanofibrous scaffolds of poly(L-lactic acid)-dicalcium silicate composite via ultrasonic-aging technique for bone regeneration[J].Mater Sci Eng C Mater Biol Appl,2014,35:426-433
[19]Wang N,Zhou Z,Xia L,et al.Fabrication and characterization of bioactiveβ-Ca2SiO4/PHBV composite scaffolds[J].Mater Sci Eng C Mater Biol Appl,2013,33(4):2294-2301
[20]朱文俊,谭媛元,梁敏.人CD146+牙周膜干细胞的分选及其干细胞特性的鉴定[J].中华老年口腔医学杂志,2013,11(2):74-78
[21]Li HW,Sun JY.Effects of dicalcium silicate coating ionic dissolution products on human mesenchymal stem-cell proliferation and osteogenic differentiation[J].J Int Med Res,2011,39(1):112-128
[22]Sun J,Wei L,Liu X,et al.Influences of ionic dissolution products of dicalcium silicate coating on osteoblastic proliferation,differentiation and gene expression[J].Acta Biomater,2009,5(4):1284-1293
[23]Kim HK,Kim JH,Abbas AA,et al.Alendronate enhances osteogenic differentiation of bone marrow stromal cells:a preliminary study[J].Clin Orthop Relat Res,2009,467(12):3121-3128
[24]Klar RM,Duarte R,Dix-Peek T,et al.Calcium ions and osteoclastogenesis initiate the induction of bone formation by coral-derived macroporous constructs[J].J Cell Mol Med,2013,17(11):1444-1457
[25]Wu C,Chen Z,Yi D,et al.Multidirectional effects of Sr-,Mg-,and Si-containing bioceramic coatings with high bonding strength on inflammation,osteoclastogenesis and osteogenesis[J].ACS Appl Mater Interfaces,2014,6(6):4264-4276
[2]Gandolfi MG,Ciapetti G,Taddei P,et al.Apatite formation on bioactive calcium-silicate cements for dentistry affects surface topography and human marrow stromal cells proliferation[J].Dent Mater,2010,26(10):974-992
[3]Chen CC,Shie MY,Ding SJ.Human dental pulp cell responses to new calcium silicate-based endodontic materials[J].Int Endod J,2011,44(9):836-842
[4]Wu BC,Wei CK,Hsueh NS,et al.Comparative cell attachment,cytotoxicity and antibacterial activity of radiopaque dicalcium silicate cement and white-coloured mineral trioxide aggregate[J].Int Endod J,2015,48(3):268-276
[5]Chiang TY,Ding SJ.Physicochemical properties of radiopaque dicalcium silicate cement as a root-end filling material in an acidic environment[J].Int Endod J,2013,46(3):234-241
[6]Chiang TY,Ding SJ.Comparative physicochemical and biocompatible properties of radiopaque dicalcium silicate cement and mineral trioxide aggregate[J].J Endod,2010,36(10):1683-1687
[7]Sun J,Li J,Liu X,et al.Proliferation and gene expression of osteoblasts cultured in DMEM containing the ionic products of dicalcium silicate coating[J].Biomedicine&pharmacotherapy,2009,63(9):650-657
[8]Chen LJ,Zhang YL,Liu J,et al.Exposure of the Murine RAW 264.7 Macrophage Cell Line to Dicalcium Silicate Coating Surface:Assessment of Cytotoxicity and Pro-inflammatory effects[J].J Mater Sci-Mater M,2016,27(3):59
[9]Wang Y,Medvid R,Melton C,et al.DGCR8 is essential for microRNA biogenesis and silencing of embryonic stem cell self-renewal[J].Nat Genet,2007,39(3):380-385
[10]Liu X,Tao S,Ding C.Bioactivity of plasma sprayed dicalcium silicate coatings[J].Biomaterials,2002,23(3):963-968
[11]Li J,Wei L,Sun J.Effect of ionic products of dicalcium silicate coating on osteoblast differentiation and collagen production via TGF-β1 pathway[J].J Biomater Appl,2011,9(7):98-103
[12]Liu X,Xie Y,Ding C,et al.Early apatite deposition and osteoblast growth on plasma-sprayed dicalcium silicate coating[J].J Biomed Mater Res,2005,74(3):356-365
[13]Velasquez P,Luklinska ZB,Meseguer-Olmo L,et al.Α-TCP ceramic doped with dicalcium silicate for bone regeneration applications prepared by powder metallurgy method:in vitro and in vivo studies[J].J Biomed Mater Res A,2013,101(7):1943-1954
[14]Meseguer-Olmo L,Aznar-Cervantes S,Mazón P,et al.“In vitro”behavior of adult mesenchymal stem cells of human bone marrow origin seeded on a novel bioactive ceramics in the Ca2SiO4-Ca3(PO4)2 system[J].J Mater Sci Mater Med,2012,23(12):3003-3014
[15]De Aza PN,Zuleta F,Velasquez P,et al.(α'(H))-Dicalcium silicate bone cement doped with tricalcium phosphate:characterization,bioactivity and biocompatibility[J].J Mater Sci Mater Med,2014,25(2):445-452
[16]Correa D,Almirall A,Carrodeguas RG,et al.α-Tricalcium phosphate cements modified withβ-dicalcium silicate and tricalcium aluminate:Physicochemical characterization,in vitro bioactivity and cytotoxicity[J].J Biomed Mater Res B Appl Bio-mater,2015,103(1):72-83
[17]de Aza PN,Luklinska ZB,Mate Sanchez,et al.Biodegradation process ofα-tricalcium phosphate andα-tricalcium phosphate solid solution bioceramics in vivo:a comparative study[J].Microsc Microanal,2013,19(5):1350-1357
[18]Dong S,Sun J,Li Y,et al.Electrospun nanofibrous scaffolds of poly(L-lactic acid)-dicalcium silicate composite via ultrasonic-aging technique for bone regeneration[J].Mater Sci Eng C Mater Biol Appl,2014,35:426-433
[19]Wang N,Zhou Z,Xia L,et al.Fabrication and characterization of bioactiveβ-Ca2SiO4/PHBV composite scaffolds[J].Mater Sci Eng C Mater Biol Appl,2013,33(4):2294-2301
[20]朱文俊,谭媛元,梁敏.人CD146+牙周膜干细胞的分选及其干细胞特性的鉴定[J].中华老年口腔医学杂志,2013,11(2):74-78
[21]Li HW,Sun JY.Effects of dicalcium silicate coating ionic dissolution products on human mesenchymal stem-cell proliferation and osteogenic differentiation[J].J Int Med Res,2011,39(1):112-128
[22]Sun J,Wei L,Liu X,et al.Influences of ionic dissolution products of dicalcium silicate coating on osteoblastic proliferation,differentiation and gene expression[J].Acta Biomater,2009,5(4):1284-1293
[23]Kim HK,Kim JH,Abbas AA,et al.Alendronate enhances osteogenic differentiation of bone marrow stromal cells:a preliminary study[J].Clin Orthop Relat Res,2009,467(12):3121-3128
[24]Klar RM,Duarte R,Dix-Peek T,et al.Calcium ions and osteoclastogenesis initiate the induction of bone formation by coral-derived macroporous constructs[J].J Cell Mol Med,2013,17(11):1444-1457
[25]Wu C,Chen Z,Yi D,et al.Multidirectional effects of Sr-,Mg-,and Si-containing bioceramic coatings with high bonding strength on inflammation,osteoclastogenesis and osteogenesis[J].ACS Appl Mater Interfaces,2014,6(6):4264-4276