Document Type : Original Article


1 Postgraduate, Dept. of Periodontics, School of Dentistry, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran.

2 Dept. of Periodontics, School of Dentistry, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran.


Statement of the Problem: Osteoblastic differentiation of periodontal ligament stem cells (PLSCs) is essential for alveolar bone regeneration.
Purpose: The purpose of this study was to compare the potential of two β-tricalcium phosphate (βTCP) products to induce osteoblastic differentiation of human PLSCs.
Materials and Method: In this in vitro study, human PLSCs were cultured in mediums supplemented with Guidor Easy-Graft [βTCP+polylactide-co-glycolide(PLCG)+n-methyl-2-pyrrolidone(NMP)] [Sunstar Company, Swiss] or Sorbone [βTCP] [Meta Company, South Korea] as two alloplasts experimental groups, mesenchymal cells differentiated into osteoblasts without alloplast as positive control group, and mesenchymal cells without osteoblastic induction as negative control group. Osteoblastic differentiation was evaluated using Alizarine Red staining and spectrophotometry to assay calcium deposits and real-time polymerase chain reaction to examine expression of alkaline phosphatase (ALP) and osteopontin (OPN) antigens on day 21. Data were analyzed by using SPSS 22 software and one-way ANOVA and Bonferoni tests (p< 0.05).
Results: Spectrophotometry confirmed that calcium deposits were higher in Guidor Easy-Graft group compared to Sorbone group (p< 0.001) and higher in two experimental groups than controls (p< 0.05). According to real-time polymerase chain reaction, level of ALP expression was higher in Sorbone than Guidor and the levels of Guidor, positive control and negative control were equal; OPN levels of the positive control were more than the other groups. OPN levels of Sobone, Guidor and negative control were the same.
Conclusion: These findings indicated that Guidor Easy-Graft and Sorbone enhanced differentiation of human PLSCs to osteoblasts, and could be employed as appropriate bone-graft materials.


  • Pilipchuk SP, Plonka AB, Monje A, Taut AD, Lanis A, Kang B, et al. Tissue engineering for bone regeneration and osseointegration in the oral cavity. Dent Mater. 2015; 31: 317-338.
  • Johansson B, Grepe A, Wannfors K, Hirsch J. A clinical study of changes in the volume of bone grafts in the atrophic maxilla. Dentomaxillofac Radiol. 2001; 30: 57-61.
  • Lee MJ, Kim BO, Yu SJ. Clinical evaluation of a biphasic calcium phosphate grafting material in the treatment of human periodontal intrabony defects. J Periodontal Implant Sci. 2012; 42: 127-135.
  • Sukumar S, Drízhal I, Paulusová V, Bukac J. Surgical treatment of periodontal intrabony defects with calcium sulphate in combination with beta-tricalcium phosphate: clinical observations two years post-surgery. Acta Medica (Hradec Kralove). 2011; 54: 13-20.
  • Neamat A, Gawish A, Gamal-Eldeen AM. Beta-Tricalcium phosphate promotes cell proliferation, osteogenesis and bone regeneration in intrabony defects in dogs. Arch Oral Biol. 2009; 54: 1083-1090.
  • Shim JH, Won JY, Park JH, Bae JH, Ahn G, Kim CH, et al. Effects of 3D-Printed Polycaprolactone/β-Tricalcium Phosphate Membranes on Guided Bone Regeneration. Int J Mol Sci. 2017; 18: pii: E899.
  • Piccinini M, Prosperi S, Preve E, Rebaudi A, Bucciotti F. In vitro Biocompatibility Assessment and in vivo Behavior of a New Osteoconductive βTCP Bone Substitute. Implant Dent. 2016; 25: 456-463.
  • Leventis MD, Fairbairn P, Kakar A, Leventis AD, Margaritis V, Lückerath W, et al. Minimally invasive alveolar ridge preservation utilizing an in situ hardening β-tricalcium phosphate bone substitute: a multicenter case series. Int J Dent. 2016; 2016: 5406736.
  • Chen S, Ye X, Yu X, Xu Q, Pan K, Lu S, et al. Co-culture with periodontal ligament stem cells enhanced osteoblastic differentiation of MC3T3-E1 cells and osteoclastic differentiation of RAW264.7 cells. Int J Clin Exp Pathol. 2015; 8: 14596-14607.
  • Li G, Hu J, Chen H, Chen L, Zhang N, Zhao L, et al. Enamel matrix derivative enhances the proliferation and osteogenic differentiation of human periodontal ligament stem cells on the titanium implant surface. Organogenesis. 2017; 13: 103-113.
  • Ge S, Zhao N, Wang L, Yu M, Liu H, Song A, et al. Bone repair by periodontal ligament stem cellseeded nanohydroxyapatite-chitosan scaffold. Int J Nanomedicine. 2012; 7: 5405-5414.
  • Houshmand B, Behnia H, Khoshzaban A, Morad G, Behrouzi G, Dashti SG, et al. Osteoblastic differentiation of human stem cells derived from bone marrow and periodontal ligament under the effect of enamel matrix derivative and transforming growth factor-beta. Int J Oral Maxillofac Implants. 2013; 28: e440-e450.
  • Zhang QB, Cao W, Liu YR, Cui SM, Yan YY. Effects of Sirtuin 1 on the proliferation and osteoblastic differentiation of periodontal ligament stem cells and stem cells from apical papilla. Genet Mol Res. 2016; 15: 5234.
  • Chandrasekaran S, Ramachandran A, Eapen A, George A. Stimulation of periodontal ligament stem cells by dentin matrix protein 1 activates mitogen-activated protein kinase and osteoblast differentiation. J Periodontol. 2013; 84: 389-395.
  • Yang S, Guo L, Su Y, Wen J, Du J, Li X, et al. Nitric oxide balances osteoblast and adipocyte lineage differentiation via the JNK/MAPK signaling pathway in periodontal ligament stem cells. Stem Cell Res Ther. 2018; 9: 118.
  • Choi MH, Noh WC, Park JW, Lee JM, Suh JY. Gene expression pattern during osteogenic differentiation of human periodontal ligament cells in vitro. J Periodontal Implant Sci. 2011; 41: 167-175.
  • Miron RJ, Bosshardt DD, Gemperli AC, Dard M, Buser D, Gruber R, et al. In vitro characterization of a synthetic calcium phosphate bone graft on periodontal ligament cell and osteoblast behavior and its combination with an enamel matrix derivative. Clinical Oral Investigations. 2014; 18: 443-451.
  • Saini N, Sikri P, Gupta H. Evaluation of the relative efficacy of autologous platelet-rich plasma in combination with β-tricalcium phosphate alloplast versus an alloplast alone in the treatment of human periodontal infrabony defects: a clinical and radiological study. Indian J Dent Res. 2011; 22: 107-115.
  • An S, Gao Y, Huang X, Ling J, Liu Z, Xiao Y. A comparative study of the proliferation and osteogenic differentiation of human periodontal ligament cells cultured on β-TCP ceramics and demineralized bone matrix with or without osteogenic inducers in vitro. Int J Mol Med. 2015; 35: 1341-1346.
  • Xia L, Zhang Z, Chen L, Zhang W, Zeng D, Zhang X, et al. Proliferation and osteogenic differentiation of human periodontal ligament cells on akermanite and β-TCP bioceramics. Eur Cell Mater. 2011; 22: 68-82.
  • Yao S, Zhao W, Ou Q, Liang L, Lin X, Wang Y. MicroRNA-214 Suppresses Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Targeting ATF4. Stem Cells Int. 2017; 2017: 3028647.
  • Alves LB, Mariguela VC, Grisi MF, Souza SL, Novaes*Junior AB, Taba*Junior M, et al. Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel. J Appl Oral Sci. 2015; 23: 206-214.
  • Miron RJ, Sculean A, Shuang Y, Bosshardt DD, Gruber R, Buser D, et al. Osteoinductive potential of a novel biphasic calcium phosphate bone graft in comparison with autographs, xenografts, and DFDBA. Clin Oral Implants Res. 2016; 27: 668-675.
  • Winning L, Robinson L, Boyd AR, El Karim IA, Lundy FT, Meenan BJ. Osteoblastic differentiation of periodontal ligament stem cells on non-stoichiometric calcium phosphate and titanium surfaces. J Biomed Mater Res A. 2017; 105: 1692-1702.
  • Naineni R, Ravi V, Subbaraya DK, Prasanna JS, Panthula VR, Koduganti RR. Effect of alendronate with β-TCP bone substitute in surgical therapy of periodontal intra-osseous defects: a randomized controlled clinical trial. J Clin Diagn Res. 2016; 10: ZC113-ZC117.
  • Amalakara J, Reddy K, Avula H, Mishra A, Kalakonda B, Pandey R. Evaluation of cyclosporine a with β-TCP in the treatment of human infra bony defects- a randomized controlled pilot study. J Clin Diagn Res. 2017; 11: ZC66-ZC70.
  • Cochran DL, Oh TJ, Mills MP, Clem DS, McClain PK, Schallhorn RA, et al. A Randomized Clinical Trial Evaluating rh-FGF-2/β-TCP in Periodontal Defects. J Dent Res. 2016; 95: 523-530.
  • Maroo S, Murthy KR. Treatment of periodontal intrabony defects using β-TCP alone or in combination with rhPDGF-BB: a randomized controlled clinical and radiographic study. Int J Periodontics Restorative Dent. 2014; 34: 841-847.
  • McCafferty MM, Burke GA, Meenan BJ. Mesenchymal stem cell response to conformal sputter deposited calcium phosphate thin films on nanostructured titanium surfaces. J Biomed Mater Res A. 2014; 102: 3585-3597.
  • Kouhestani F, Dehabadi F, Hasan*Shahriari M, Motamedian SR. Allogenic vs. synthetic granules for bone tissue engineering: an in vitro study. Prog Biomater. 2018; 7: 133-141.
  • Beck GR*Jr, Sullivan EC, Moran E, Zerler B. Relationship between alkaline phosphatase levels, osteopontin expression, and mineralization in differentiating MC3T3-E1 osteoblasts. J Cell Biochem. 1998; 68: 269-280.