Gene Expression during Osteo/Odontogenic Differentiation of Mesenchymal Stem Cells with Platelet Rich Plasma and Mineral Trioxide Aggregate


Affiliations

  • Pacific Academy of Higher Education and Research University, Udaipur, Rajasthan, 313003, India
  • Ibn Sina National College for Medical Studies, Department of Preventive Dental Sciences, Jeddah, 22421, Saudi Arabia
  • Deccan College of Medical Sciences, Central Laboratory for Stem Cell Research and Translational Medicine, CLRD, Hyderabad, Telangana, 500 058, India
  • Jaipur Dental College, Department of Pedodontics and Preventive Dentistry, Jaipur, Rajasthan, 302015, India
  • Pacific Dental College and Hospital, Department of Pedodontics and Preventive Dentistry, Udaipur, Rajasthan, 313011, India
  • Ibn Sina National College for Medical Studies, Jeddah, 22421, Saudi Arabia

Abstract

Platelet Rich Plasma (PRP) has the potential to regenerate pulp in immature pulpless teeth. Mineral trioxide is commonly used to seal the PRP into the pulp canal space. We investigated the effect of PRP and MTA individually and combined on osteo/odontogenic differentiation potential and the phenotype of tissue formed. MSCs were cultured in vitro with MTA, 5% PRP, 10% PRP, MTA with 5% PRP and MTA with 10% PRP. Osteo/odontogenic differentiation was assessed and quantified with alizarin red staining. Relative expression of Alkaline Phosphatase (ALP) activity, type 1 collagen (COL1A1), Dental Sialo-Phospho Protein (DSPP), Dentin Matrix Protein (DMP-1), Bone Gamma-carboxyglutamate Protein (BGLAP), Runt-related transcription factor 2 (Runx2), Osterix (Osx) and TGF-β1 was identified by RT-q PCR. 10% PRP with MTA displayed significantly higher calcium deposition during differentiation and high ALP levels. Significantly enhanced levels of DSPP, DMP1, COL1A1, BGLAP, Runx2, and Osx and TGF-β1 transcripts were observed. Within limitations of the in vitro environment, results imply enhanced osteodentin formation, on combining PRP with MTA.

Keywords

Platelet rich plasma, Mineral trioxide aggregate, Mesenchymal stem cells, odontogenic, differentiation

Subject Discipline

Dentistry

Full Text:

References

Thomson A, Kahler B. Regenerative endodonticsbiologicallybased treatment for immature permanent teeth: a case report and review of the literature. Aust Dent J. 2010; 55(4):446-52. https://doi.org/10.1111/j.18347819.2010.01268.x. PMid: 21133946.

da Silva L, Nelson-Filho P, da Silva R, et al. Revascularization and periapical repair after endodontic treatment using apical negative pressure irrigation versus conventional irrigation plus triantibiotic intracanal dressing in dogs’ teeth with apical periodontitis. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2010; 109:779-87.

Wang X, Thibodeau B, Trope M, Lin L, Huang G. Histologic characterization of regenerated tissues in canal space after the revitalization/revascularization procedure of immature dog teeth with apical periodontitis. J Endod. 2010; 36:5663.https://doi.org/10.1016/j.joen.2009.09.039. PMid: 20003936.

Yamauchi N, Yamauchi S, Nagaoka H, et al. Tissue engineering strategies for immature teeth with apical periodontitis. J Endod. 2011; 37:390-97. https://doi.org/10.1016/j.joen.2010.11.010. PMid: 21329828.

Mao JJ, Kim SG, Zhou J, Ye L, Cho S, Suzuki T, Fu SY, Yang R, Zhou X. Regenerative endodontics: Barriers and strategies for clinical translation. Dental Clinics. 2012 Jul 1; 56(3):639-49. https://doi.org/10.1016/j.cden.2012.05.005. PMid:22835543. PMCid: PMC4093795.

Hwang YJ, Choi JY. Addition of mesenchymal stem cells to the scaffold of platelet-rich plasma is beneficial for the reduction of the consolidation period in mandibular distraction osteogenesis. J Oral Maxillofac Surg. 2010; 68:1112-24. https://doi.org/10.1016/j.joms.2008.08.038. PMid: 20223574.

D’antò V, Di Caprio MP, Ametrano G, Simeone M, Rengo S, Spagnuolo G. Effect of mineral trioxide aggregate on mesenchymal stem cells. J Endod. 2010; 36:1839-43. https://doi.org/10.1016/j.joen.2010.08.010. PMid: 20951297.

Vanka A, Sandeep KV, Shanthi G, Manohar KB, Wali O, Khan AA. Osteo/odontogenic differentiation of human mesenchymal stem cells with Platelet Rich Plasma (PRP) and Mineral Trioxide Aggregate (MTA). J Contemp Dent Pract (accepted for publication).

Bausset O, Giraudo L, Veran J. Formulation and storage of platelet-rich plasma homemade product. Biores Open Access. 2012; 1:115-23. https://doi.org/10.1089/ biores.2012.0225. PMid: 23516671, PMCid: PMC3559222.

Vanka A, Sandeep KV, Shanthi G, Manohar KB, Khan AA. In vitro proliferation of MSCs using mineral trioxide aggregate: A most recent material for in situ stem cells mobilization. Int J of Adv Res. 2014; 2:561-67.

Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenolchloroform extraction. Anal Biochem. 1987; 162:156-59. https://doi.org/10.1016/0003-2697(87)90021-2, https://doi.org/10.1006/abio.1987.9999. PMid: 2440339.

Marx RE. Platelet-rich plasma: evidence to support its use. J Oral Maxillofac Surg. 2004; 62:489-96. https://doi.org/10.1016/j.joms.2003.12.003. PMid: 15085519.

Dohan Ehrenfest DM, Rasmusson L, Albrektsson T. Classification of platelet concentrates: From Pure PlateletRich Plasma (P-PRP) to Leukocyte- and Platelet-Rich Fibrin (L-PRF). Trends Biotechnol. 2009; 27:158-67. https://doi.org/10.1016/j.tibtech.2008.11.009. PMid: 19187989.

Paranjpe A, Smoot T, Zhang H, Johnson JD. Direct contact with mineral trioxide aggregate activates and differentiates human dental pulp cells. J Endod. 2011 Dec; 37(12):169195. https://doi.org/10.1016/j.joen.2011.09.012. PMid: 22099907, PMCid: PMC3223385.

Wang Y, Li J, Song W, Yu J. Mineral trioxide aggregate upregulates odonto/osteogenic capacity of bone marrow stromal cells from craniofacial bones via JNK and ERK MAPK signalling pathways. Cell proliferation. 2014 Jun; 47(3):241-48. https://doi.org/10.1111/cpr.12099. PMid: 24635197.

Perut F, Filardo G, Mariani E, Cenacchi A, Pratelli L, Devescovi V, Kon E, Marcacci M, Facchini A, Baldini N, Granchi D. Preparation method and growth factor content of platelet concentrate influence the osteogenic differentiation of bone marrow stromal cells. Cytotherapy. 2013 Jul 1; 15(7):830-39. https://doi.org/10.1016/j.jcyt.2013.01.220. PMid: 23731763.

Smith AJ, Scheven BA, Takahashi Y, Ferracane JL, Shelton RM, Cooper PR. Dentine as a bioactive extracellular matrix. Arch Oral Biol. 2012; 57:109-21. https://doi.org/10.1016/j.archoralbio.2011.07.008. PMid:21855856.

Huang W, Yang S, Shao J, Li YP. Signaling and transcriptional regulation in osteoblast commitment and differentiation. Frontiers in Bioscience: A Journal and Virtual Library. 2007 May 1; 12:30-68. https://doi.org/10.2741/2296. PMid: 17485283, PMCid: PMC3571113.

Linde A, Granstro¨ MG. Odontoblast alkaline phosphatases and Ca2þ transport. J Biol Buccale. 1978; 6:293-308.

Couchourel D, Aubry I, Delalandre A. Altered mineralization of human osteoarthritic osteoblasts is attributable to abnormal type I collagen production. Arthritis and Rheumatology. 2009; 60:1438-50. https:// doi.org/10.1002/art.24489. PMid: 19404930, PMCid: PMC5250342.

Miron RJ, Zhang YF. Osteoinduction: a review of old concepts with new standards. Journal of Dental Research. 2012 Aug; 91(8):736-44. https://doi.org/10.1177/0022034511435260. PMid:22318372.

Nakashima K, Zhou X, Kunkel G, Zhang Z, Deng JM, Behringer RR, De Crombrugghe B. The novel zinc fingercontaining transcription factor osterix is required for osteoblast differentiation and bone formation. Cell. 2002 Jan 11; 108(1):17-29. https://doi.org/10.1016/S00928674(01)00622-5.

Nakamura A, Dohi Y, Akahane M, Ohgushi H, Nakajima H, Funaoka H, Takakura Y. Osteocalcin secretion as an early marker of in vitro osteogenic differentiation of rat mesenchymal stem cells. Tissue Eng Part C Methods. 2009; 15:169-80. https://doi.org/10.1089/ten.tec.2007.0334. PMid: 19191495.

Qin C, Brunn JC, Cadena E, Ridall A, Tsujigiwa H, Nagatsuka H, Nagai N, Butler WT. The expression of dentin sialophosphoprotein gene in bone. J Dent Res. 2002; 81:392-94. https://doi.org/10.1177/154405910208100607. PMid: 12097430.

Massa LF, Ramachandran A, George A, Arana-Chavez VE. Developmental appearance of dentin matrix protein 1 during the early dentinogenesis in rat molars as identified by high-resolution immunocytochemistry. Histochem Cell Biol. 2005; 124:197-205. https://doi.org/10.1007/s00418005-0009-9. PMid: 16049693.

Teti G, Salvatore V, Ruggeri A, Manzoli L, GESI M, Orsini G, Falconi M. In vitro reparative dentin: A biochemical and morphological study. Eur J Histochem. 2013; 57:e23.

https://doi.org/10.4081/ejh.2013.e23. PMid: 24085272, PMCid: PMC3794354.

Bakopoulou A, Leyhausen G, Volk J, Tsiftsoglou A, Garefis P, Koidis P, Geurtsen W. Comparative analysis of in vitro osteo/odontogenic differentiation potential of human Dental Pulp Stem Cells (DPSCs) and Stem Cells from the Apical Papilla (SCAP). Arch Oral Biol. 2011; 56:709-21. https://doi.org/10.1016/j.archoralbio.2010.12.008. PMid: 21227403.

Zhao L, Hantash BM. TGF-β1 regulates differentiation of bone marrow mesenchymal stem cells. Vitam Horm. 2011; 87:12741. https://doi.org/10.1016/B978-0-12-386015-6.00042-1. PMid: 22127241.

Jadlowiec JA, Zhang X, Li J, Campbell PG and Sfeir C. Extracellular matrix-mediated signaling by dentin phosphophoryn involves activation of the Smad pathway independent of bone morphogenetic protein. The Journal of Biological Chemistry. 2006; 281(9):5341-47. https://doi.org/10.1074/jbc.M506158200. PMid: 16326713.


Refbacks

  • There are currently no refbacks.