层状纳米颗粒Laponite对成牙本质细胞矿化促进作用的研究
|
摘要
目的:研究层状纳米粒颗粒Laponite对成牙本质细胞(OLCs)的矿化促进作用。方法:采用透射电镜(TEM)和动态光散射法(DLS)对Laponite进行形态表征观察;通过CCK-8和活/死细胞染色法检测不同浓度的Laponite对OLCs增殖和活力的影响,并分别通过划痕实验比较各组细胞的迁移能力,q-PCR检测各组细胞成牙本质相关基因的表达水平,茜素红染色和半定量分析比较各组细胞的钙化结节形成情况。结果:Laponite在水溶液中可分散形成纳米圆盘状颗粒,其平均粒径为(30. 9±7. 4) nm;当Laponite浓度小于100μg/mL时,对OLCs的增殖和活力无显著影响(P> 0. 05);而10μg/mL和100μg/mL的Laponite均能明显增强OLCs的迁移能力,并显著提高其成牙本质相关基因的表达水平以及钙化结节的形成量(P <0. 05)。结论:10μg/mL和100μg/mL的Laponite能够促进OLCs的生物矿化。
AIM: To investigate the biomineralization of the nanoplatelet Laponite on odontoblast-lineage cells in vitro. METHODS: The morphology of Laponite was characterized by TEM and DLS. The effects of Laponite on cell proliferation and viability were examined by CCK-8 and LIVE/DEAD cell staining. The cell migration rate was observed by wound healing assay. q-PCR was used to analyze the expression of the dentinogenesis-related genes( COL-1,DSPP,DMP1 and BMP2). Calcium accumulation was detected by using alizarin red staining and cetylpyridinium chloride assay. RESULTS: Laponite dispersed in aqueous solution and formed nanoplatelets of which particle size was( 30. 9 ± 7. 4) nm. When the concentration of Laponite was less than 100 μg/mL,cell proliferation and viability were not affected( P > 0. 05). The cell migration,expression of the dentinogenesis-related genes and calcium accumulation were promoted by Laponite at 10 μg/mL and 100 μg/mL( P < 0. 05). CONCLUSION: Laponite promotes the biomineralization of the OLCs.
AIM: To investigate the biomineralization of the nanoplatelet Laponite on odontoblast-lineage cells in vitro. METHODS: The morphology of Laponite was characterized by TEM and DLS. The effects of Laponite on cell proliferation and viability were examined by CCK-8 and LIVE/DEAD cell staining. The cell migration rate was observed by wound healing assay. q-PCR was used to analyze the expression of the dentinogenesis-related genes( COL-1,DSPP,DMP1 and BMP2). Calcium accumulation was detected by using alizarin red staining and cetylpyridinium chloride assay. RESULTS: Laponite dispersed in aqueous solution and formed nanoplatelets of which particle size was( 30. 9 ± 7. 4) nm. When the concentration of Laponite was less than 100 μg/mL,cell proliferation and viability were not affected( P > 0. 05). The cell migration,expression of the dentinogenesis-related genes and calcium accumulation were promoted by Laponite at 10 μg/mL and 100 μg/mL( P < 0. 05). CONCLUSION: Laponite promotes the biomineralization of the OLCs.
引文
[1]丛明宇,赵亮,常蓓,等.修复性牙本质形成的机理与调控[J].牙体牙髓牙周病学杂志,2014,24(10):615-619.
[2] Galler KM. Clinical procedures for revitalization:current knowledge and considerations[J]. Int Endod J,2016,49(10):926-936.
[3] Albuquerque MTP,Valera MC,Nakashima M,et al. Tissue-engineering-based Strategies for Regenerative Endodontics[J]. J Dent Res,2014,93(12):1222-1231.
[4] Dawson JI,Oreffo RO. Clay:new opportunities for tissue regeneration and biomaterial design[J]. Adv Mater,2013,25(30):4069-4086.
[5] Mihaila SM,Gaharwar AK,Reis RL,et al.The osteogenic differentiation of SSEA-4sub-population of human adipose derived stem cells using silicate nanoplatelets[J]. Biomaterials,2014,35(33):9087-9099.
[6] Zhang F,Phiel CJ,Spece L,et al. Inhibitory phosphorylation of glycogen synthase kinase-3(GSK-3)in response to lithium. evidence for autoregulation of GSK-3[J]. J Biol Chem,2003,278(35):33067-33077.
[7] Tamura M,Nemoto E. Role of the wnt signaling molecules in the tooth[J]. Jpn Dent Sci Rev,2016,52(4):75-83.
[8] Qu T,Jing J,Jiang Y,et al. Magnesiumcontaining nanostructured hybrid scaffolds for enhanced dentin regeneration[J]. Tissue Eng Pt A,2014,20(17-18):2422-2433.
[9] Jurki'c LM,Cepanec I,Paveli'c SK,et al. Biological and therapeutic effects of ortho-silicic acid and some ortho-silicic acid-releasing compounds:New perspectives for therapy[J].Nut Metab,2013,10(1):1-12.
[10] Opal S,Garg S,Dhindsa A,et al. Minimally invasive clinical approach in indirect pulp therapy and healing of deep carious lesions[J]. J Clin Pediatr Dent,2014,38(3):185-192.
[11] Dawson JI,Kanczler JM,Yang XB,et al. Clay gels for the delivery of regenerative microenvironments[J]. Adv Mater,2011,23(29):3304-3308.
[12] Arany S,Nakata A,Kameda T,et al. Phenotype properties of a novel spontaneously immortalized odontoblast-lineage cell line[J]. Biochem Biophys Res Commun,2006,342(3):718-724.
[13] Li M,Sun X,Liang M,et al. SDF-1/CXCR4 axis induces human dental pulp stem cell migration through FAK/PI3K/Akt and GSK3β/β-catenin pathways[J]. Sci Rep,2017,7:40161.
[14] Liu Q,Ma Y,Wang J,et al. Demineralized bone matrix used for direct pulp capping in rats[J]. Plos One,2017,12(3):e0172693.
[15] Smith AJ,Scheven BA,Takahashi Y,et al.Dentine as a bioactive extracellular matrix[J].Arch Oral Biol,2012,57(2):109-121.
[16] Ili'c J,Radovi K,Roganovi J,et al. The levels of vascular endothelial growth factor and bone morphogenetic protein 2 in dental pulp tissue of healthy and diabetic patients[J]. J Endod,2012,38(6):764-768.
[2] Galler KM. Clinical procedures for revitalization:current knowledge and considerations[J]. Int Endod J,2016,49(10):926-936.
[3] Albuquerque MTP,Valera MC,Nakashima M,et al. Tissue-engineering-based Strategies for Regenerative Endodontics[J]. J Dent Res,2014,93(12):1222-1231.
[4] Dawson JI,Oreffo RO. Clay:new opportunities for tissue regeneration and biomaterial design[J]. Adv Mater,2013,25(30):4069-4086.
[5] Mihaila SM,Gaharwar AK,Reis RL,et al.The osteogenic differentiation of SSEA-4sub-population of human adipose derived stem cells using silicate nanoplatelets[J]. Biomaterials,2014,35(33):9087-9099.
[6] Zhang F,Phiel CJ,Spece L,et al. Inhibitory phosphorylation of glycogen synthase kinase-3(GSK-3)in response to lithium. evidence for autoregulation of GSK-3[J]. J Biol Chem,2003,278(35):33067-33077.
[7] Tamura M,Nemoto E. Role of the wnt signaling molecules in the tooth[J]. Jpn Dent Sci Rev,2016,52(4):75-83.
[8] Qu T,Jing J,Jiang Y,et al. Magnesiumcontaining nanostructured hybrid scaffolds for enhanced dentin regeneration[J]. Tissue Eng Pt A,2014,20(17-18):2422-2433.
[9] Jurki'c LM,Cepanec I,Paveli'c SK,et al. Biological and therapeutic effects of ortho-silicic acid and some ortho-silicic acid-releasing compounds:New perspectives for therapy[J].Nut Metab,2013,10(1):1-12.
[10] Opal S,Garg S,Dhindsa A,et al. Minimally invasive clinical approach in indirect pulp therapy and healing of deep carious lesions[J]. J Clin Pediatr Dent,2014,38(3):185-192.
[11] Dawson JI,Kanczler JM,Yang XB,et al. Clay gels for the delivery of regenerative microenvironments[J]. Adv Mater,2011,23(29):3304-3308.
[12] Arany S,Nakata A,Kameda T,et al. Phenotype properties of a novel spontaneously immortalized odontoblast-lineage cell line[J]. Biochem Biophys Res Commun,2006,342(3):718-724.
[13] Li M,Sun X,Liang M,et al. SDF-1/CXCR4 axis induces human dental pulp stem cell migration through FAK/PI3K/Akt and GSK3β/β-catenin pathways[J]. Sci Rep,2017,7:40161.
[14] Liu Q,Ma Y,Wang J,et al. Demineralized bone matrix used for direct pulp capping in rats[J]. Plos One,2017,12(3):e0172693.
[15] Smith AJ,Scheven BA,Takahashi Y,et al.Dentine as a bioactive extracellular matrix[J].Arch Oral Biol,2012,57(2):109-121.
[16] Ili'c J,Radovi K,Roganovi J,et al. The levels of vascular endothelial growth factor and bone morphogenetic protein 2 in dental pulp tissue of healthy and diabetic patients[J]. J Endod,2012,38(6):764-768.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |