Effect of iRoot BP Plus on biological behavior of deciduous tooth pulp stem cells and human pulp stem cells

doi:10.19439/j.sjos.2019.03.006

Abstract

Abstract: PURPOSE: To evaluate the effect of iRoot BP Plus as pulp-capping agents on the biological behaviors of stem cells from human exfoliated deciduous teeth (SHED) and human dental pulp stem cells (DPSC). Methods: iRoot BP Plus and ProRoot MTA sample disks were prepared and the extractive solution was extracted from iRoot BP Plus and ProRoot MTA sample disks. The influence of iRoot BP Plus and MTA extracts on the SHED and DPSC proliferation capacity was detected by CCK-8 method at 1, 3, 5 and 7 days.The influence of iRoot BP Plus and MTA extract on the SHED and DPSC migration capacity was observed by Transwell chamber and scratch repair experiments. SHED and DPSC were respectively inoculated on the sample disks of iRoot BP Plus and MTA for culture. Phalloidin and DAPI (4',6-diamidino-2-phenylindole) were used for immunofluorescence staining on 1, 3 and 5 days respectively to observe cytoskeleton changes. SHED and DPSC underwent mineralization induction respectively in osteoblastic induction medium, the osteoblastic induction medium containing MTA extract and the osteoblastic induction medium containing iRoot BP Plus extract. Alkaline phosphatase (ALP) staining and quantitative analysis of ALP were performed at 7 and 14 days, Alizarin Red staining and semi-quantitative analysis of calcium salt deposition were performed at 21 days. SPSS 19.0 software package was used for statistical analysis of the data. Results: iRoot BP Plus and MTA extracts could promote cell proliferation of SHED and DPSC. In cell migration and adhesion experiments, iRoot BP Plus and MTA both promoted migration and adhesion of SHED and DPSC, and iRoot BP Plus played a more significant role (P=0.000). After mineralization induction, the ALP activity of SHED and DPSC in iRoot BP Plus group was significantly greater than that of MTA. Alizarin red staining and semi-quantitative analysis of calcium salt deposition showed that both iRoot BP Plus and MTA could promote cell mineralization. Moreover, the ability of iRoot BP Plus to promote cell mineralization was significantly stronger than that of MTA (P=0.000). Conclusions: iRoot BP Plus and MTA has good biocompatibility and good osteogenetic differentiation ability, it can promote SHED and DPSC cell proliferation, adhesion, migration, and BP Plus has better affect of promoting iRoot SHED and DPSC adhesion, migration and distribution of differentiation than MTA, therefore iRoot BP Plus and MTA may be used as pulp capping agent both for deciduous and permanent teeth.

Key words: SHED, DPSC, iRoot BP Plus, MTA, Proliferation, Migration, Adhesion, Osteoblast induction

CLC Number:

R788.2

WANG Jing, FANGTENG Jiao-zi, LIU He. Effect of iRoot BP Plus on biological behavior of deciduous tooth pulp stem cells and human pulp stem cells[J]. Shanghai Journal of Stomatology, 2019, 28(3): 251-258.

References

[1] Dalli M, Colak H, Mustafa Hamidi M.Minimal intervention concept: a new paradigm for operative dentistry[J]. J Investig Clin Dent, 2012, 3(3): 167-175.
[2] Seale NS, Glickman GN.Contemporary perspectives on vital pulp therapy: views from the endodontists and pediatric dentists[J]. J Endod, 2008, 34(7 Suppl): S57-S61.
[3] Nair PN, Duncan HF, Pitt Ford TR, et al.Histological, ultrastructural and quantitative investigations on the response of healthy human pulps to experimental capping with mineral trioxide aggregate: a randomized controlled trial[J]. Int Endod J, 2009, 42(5):422-444.
[4] Schröder U.Effects of calcium hydroxide-containing pulp-capping agents on pulp cell migration, proliferation, and differentiation[J]. J Dent Res, 1985, 64(Spec No): 541-548.
[5] Al-Hezaimi K, Al-Tayar BA, Bajuaifer YS, et al.A hybrid approach to direct pulp capping by using emdogain with a capping material[J]. J Endod, 2011, 37(5): 667-672.
[6] Eid AA, Hussein KA, Niu LN, et al.Effects of tricalcium silicate cements on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells in vitro[J]. Acta Biomaterialia, 2014, 10(7): 3327-3334.
[7] Kang JY, Lee BN, Son HJ, et al.Biocompatibility of mineral trioxide aggregate mixed with hydration accelerators[J]. J Endod, 2013, 39(4): 497-500.
[8] Hirschman WR, Wheater MA, Bringas JS, et al.Cytotoxicity comparison of three current direct pulp-capping agents with a new bioceramic root repair putty[J]. J Endod, 2012, 38(3): 385-388.
[9] Nair U, Ghattas S, Saber M, et al.A comparative evaluation of the sealing ability of 2 root-end filling materials: an in vitro leakage study using Enterococcus faecalis[J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2011, 112(2): e74-e77.
[10] Lovato KF, Sedgley CM.Antibacterial activity of EndoSequence root repair material and ProRoot MTA against clinical isolates of Enterococcus faecalis[J]. J Endod, 2011, 37(11): 1542-1546.
[11] Zhang S, Yang X, Fan M.BioAggregate and iRoot BP Plus optimize the proliferation and mineralization ability of human dental pulp cells[J]. Int Endod J, 2013, 46(10): 923-929.
[12] 路博闻, 刘娜, 徐璐璐, 等. 人脱落乳牙牙髓干细胞与人恒牙牙髓干细胞成骨分化及破骨能力的差异[J]. 南方医科大学学报, 2016, 36(2): 180-185.
[13] Jiang Y, Zheng Q, Zhou X, et al.A comparative study on root canal repair materials: a cytocompatibility assessment in L929 and MG63 cells[J]. Sci World J, 2014, 2014: 463826.
[14] Chang SW, Lee SY, Kang SK, et al.In vitro biocompatibility, inflammatory response,and osteogenic potential of 4 root canal sealers: Sealapex, Sankin Apatite Root Sealer, MTA Fillapex, and iRoot SP root canal sealer[J]. J Endod, 2014, 40(10): 1642-1648.
[15] Liu S, Wang S, Dong Y.Evaluation of a bioceramic as a pulp capping agent in vitro and in vivo[J]. J Endod, 2015, 41(5): 652-657.
[16] Zhu L, Yang J, Zhang J, et al.In vitro and in vivo evaluation of a nanoparticulate bioceramic paste for dental pulp repair[J]. Acta Biomater, 2014, 10(12): 5156-5168.
[17] Zhang H, Shen Y, Ruse ND, et al.Antibacterial activity of endodontic sealers by modified direct contact test against Enterococcus faecalis[J]. J Endod, 2009, 35(7): 1051-1055.
[18] Park JW, Hong SH, Kim JH, et al.X-Ray diffraction analysis of white ProRoot MTA and Diadent BioAggregate[J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2010, 109(1): 155-158.
[19] Camilleri J, Montesin FE, Papaioannou S, et al.Biocompatibility of two commercial forms of mineral trioxide aggregate[J]. Int Endod J, 2004, 37(10): 699-704.
[20] Ahmed HM, Luddin N, Kannan TP, et al.Cell attachment properties of Portland cementbased endodontic materials: biological and methodological considerations[J]. J Endod, 2014, 40(10): 1517-1523.
[21] Zhu L, Yang J, Zhang J, et al.A comparative study of BioAggregate and ProRoot MTA on adhesion, migration, and attachment of human dental pulp cells[J]. J Endod, 2014, 40(8): 1118-1123.
[22] Lv F, Zhu L, Zhang J, et al.Evaluation of the in vitro biocompatibility of a new fast-setting ready-to-use root filling and repair material[J]. Int Endod J, 2017, 50(6): 540-548.
[23] Balcerzak M, Hamade E, Zhang L, et al.The roles of annexins and alkaline phosphatase in mineralization process[J]. Acta Biochim Pol, 2003, 50(4): 1019-1038.
[24] Vishwanath VR, Nadig RR, Nadig R, et al.Differentiation of isolated and characterized human dental pulp stem cells and stem cells from human exfoliated deciduous teeth: An in vitro study[J]. J Conserv Dent, 2013, 16(5): 423-428.