Restor Dent Endod.  2021 Feb;46(1):e1. 10.5395/rde.2021.46.e1.

Biomineralization of three calcium silicate-based cements after implantation in rat subcutaneous tissue

• Talabani RM
• Garib BT
• Masaeli R
• Zandsalimi K
• Ketabat F

• KMID: 2548051
• DOI: http://doi.org/10.5395/rde.2021.46.e1

Abstract

Objectives
The aim of this study was to evaluate the dystrophic mineralization deposits from 3 calcium silicate-based cements (Micro-Mega mineral trioxide aggregate [MM-MTA], Biodentine [BD], and EndoSequence Root Repair Material [ESRRM] putty) over time after subcutaneous implantation into rats.
Materials and Methods
Forty-five silicon tubes containing the tested materials and 15 empty tubes (serving as a control group) were subcutaneously implanted into the backs of 15 Wistar rats. At 1, 4, and 8 weeks after implantation, the animals were euthanized (n = 5 animals/ group), and the silicon tubes were removed with the surrounding tissues. Histopathological tissue sections were stained with von Kossa stain to assess mineralization. Scanning electron microscopy and energy-dispersive X-ray spectroscopy (SEM/EDX) were also used to assess the chemical components of the surface precipitates deposited on the implant and the pattern of calcium and phosphorus distribution at the material-tissue interface. The calciumto-phosphorus ratios were compared using the non-parametric Kruskal-Wallis test at a significance level of 5%.
Results
The von Kossa staining showed that both BD and ESRRM putty induced mineralization starting at week 1; this mineralization increased further until the end of the study. In contrast, MM-MTA induced dystrophic calcification later, from 4 weeks onward. SEM/EDX showed no statistically significant differences in the calcium- and phosphorus-rich areas among the 3 materials at any time point (p > 0.05).
Conclusions
After subcutaneous implantation, biomineralization of the 3-calcium silicatebased cements started early and increased over time, and all 3 tested cements generated calcium- and phosphorus-containing surface precipitates.

Keyword

Biodentine; Biomineralization; EndoSequence

Figure

• 

  Figure 1 Histologic tissue sections demonstrating the biomineralization (calcium deposition) in the experimental groups at the open ends of the subcutaneously-implanted silicon tubes at the 3 time points (weeks 1, 4, and 8). (A-C) Empty tubes: no dystrophic calcification was seen at any time point (×10). (D) MM-MTA containing tubes: no calcification were seen at week 1 (×10). (E and F) Black dystrophic calcifications were seen at weeks 4 and 8 (arrows, ×10). (G) BD-containing tubes: discrete deposits were seen at week 1 (arrow, ×10), and (H and I) more obvious dystrophic calcifications were seen at weeks 4 and 8 (arrows, ×10). (J-L) ESRRM putty-containing tubes: noticeable black dystrophic calcifications were obvious at all periods (arrows, ×10) (von Kossa stain, scale bar = 200 μm).MM-MTA, MicroMega mineral trioxide aggregate; BD, Biodentine; ESRRM, EndoSequence Root Repair Material; M, material; I, interface.

• 

  Figure 2 Scanning electron microscopy observations and elemental composition obtained using energy-dispersive X-ray analysis of the surface precipitates produced by (A-C) MM-MTA, (D-F) BD, and (G-I) ESRRM putty at 7 days after the subcutaneous implantation of the materials. Mineralization is indicated by the red arrows.MM-MTA, MicroMega mineral trioxide aggregate; BD, Biodentine; ESRRM, EndoSequence Root Repair Material; M, material.

• 

  Figure 3 Scanning electron microscopy observations and elemental composition obtained using energy-dispersive X-ray analysis of the surface precipitates produced by (A-C) MM-MTA, (D-F) BD, and (G-I) ESRRM putty at 4 weeks after the subcutaneous implantation of the materials. Mineralization is indicated by the red arrows.MTA, MicroMega mineral trioxide aggregate; BD, Biodentine; ESRRM, EndoSequence Root Repair Material; M, material.

• 

  Figure 4 Scanning electron microscopy observations and elemental composition obtained using energy-dispersive X-ray analysis of the surface precipitates produced by (A-C) MM-MTA, (D-F) BD, and (G-I) EndoSequence Root Repair Material putty at 8 weeks after the subcutaneous implantation of the materials. Mineralization is indicated by the red arrows.MTA, MicroMega mineral trioxide aggregate; BD, Biodentine; ESRRM, EndoSequence Root Repair Material; M, material.

Reference

1. Wang Z. Bioceramic materials in endodontics. Endod Topics. 2015; 32:3–30.

2. Koch KA, Brave DG. Bioceramics, part I: the clinician's viewpoint. Dent Today. 2012; 31:130–135.

3. Bueno CR, Valentim D, Marques VA, Gomes-Filho JE, Cintra LT, Jacinto RC, Dezan-Junior E. Biocompatibility and biomineralization assessment of bioceramic-, epoxy-, and calcium hydroxide-based sealers. Braz Oral Res. 2016; 30:S1806-83242016000100267.

4. Jitaru S, Hodisan I, Timis L, Lucian A, Bud M. The use of bioceramics in endodontics - literature review. Clujul Med. 2016; 89:470–473. PMID: 27857514.

5. Prati C, Gandolfi MG. Calcium silicate bioactive cements: biological perspectives and clinical applications. Dent Mater. 2015; 31:351–370. PMID: 25662204.

6. Gandolfi MG, Siboni F, Botero T, Bossù M, Riccitiello F, Prati C. Calcium silicate and calcium hydroxide materials for pulp capping: biointeractivity, porosity, solubility and bioactivity of current formulations. J Appl Biomater Funct Mater. 2015; 13:43–60. PMID: 25199071.

7. Reyes-Carmona JF, Felippe MS, Felippe WT. Biomineralization ability and interaction of mineral trioxide aggregate and white Portland cement with dentin in a phosphate-containing fluid. J Endod. 2009; 35:731–736. PMID: 19410094.

8. Torabinejad M, Parirokh M, Dummer PM. Mineral trioxide aggregate and other bioactive endodontic cements: an updated overview - part II: other clinical applications and complications. Int Endod J. 2018; 51:284–317. PMID: 28846134.

9. Köseoğlu S, Pekbağr Yan K T, Kucukyilmaz E, Sağlam M, Enhos S, Akgün A. Biological response of commercially available different tricalcium silicate-based cements and pozzolan cement. Microsc Res Tech. 2017; 80:994–999. PMID: 28504319.

10. Raghavendra SS, Jadhav GR, Gathani KM, Kotadia P. Bioceramics in endodontics - a review. J Istanb Univ Fac Dent. 2017; 51(Supplement 1):S128–S137. PMID: 29354316.

11. Köseoğlu S, Pekbağr Yan K T, Kucukyilmaz E, Sağlam M, Enhos S, Akgün A. Biological response of commercially available different tricalcium silicate-based cements and pozzolan cement. Microsc Res Tech. 2017; 80:994–999. PMID: 28504319.

12. Escobar-García DM, Aguirre-López E, Méndez-González V, Pozos-Guillén A. Cytotoxicity and initial biocompatibility of endodontic biomaterials (MTA and Biodentine) used as root-end filling materials. BioMed Res Int. 2016; 2016:7926961. PMID: 27595108.

13. Han L, Okiji T. Uptake of calcium and silicon released from calcium silicate-based endodontic materials into root canal dentine. Int Endod J. 2011; 44:1081–1087. PMID: 21777256.

14. Atmeh AR, Chong EZ, Richard G, Festy F, Watson TF. Dentin-cement interfacial interaction: calcium silicates and polyalkenoates. J Dent Res. 2012; 91:454–459. PMID: 22436906.

15. Charland T, Hartwell GR, Hirschberg C, Patel R. An evaluation of setting time of mineral trioxide aggregate and EndoSequence Root Repair Material in the presence of human blood and minimal essential media. J Endod. 2013; 39:1071–1072. PMID: 23880280.

16. Tran D, He J, Glickman GN, Woodmansey KF. Comparative analysis of calcium silicate-based root filling materials using an open apex model. J Endod. 2016; 42:654–658. PMID: 26925519.

17. Gomes Filho JE, Queiroz ÍO, Watanabe S, Cintra LT, Ervolino E. Influence of diabetes mellitus on the mineralization ability of two endodontic materials. Braz Oral Res. 2016; 30:S1806-83242016000100218.

18. Holland R, de Souza V, Nery MJ, Otoboni Filho JA, Bernabé PF, Dezan Júnior E. Reaction of rat connective tissue to implanted dentin tubes filled with mineral trioxide aggregate or calcium hydroxide. J Endod. 1999; 25:161–166. PMID: 10321179.

19. Cintra LT, Ribeiro TA, Gomes-Filho JE, Bernabé PF, Watanabe S, Facundo AC, Samuel RO, Dezan-Junior E. Biocompatibility and biomineralization assessment of a new root canal sealer and root-end filling material. Dent Traumatol. 2013; 29:145–150. PMID: 22510310.

20. Hench LL. Bioceramics: from concept to clinic. J Am Ceram. 1991; 74:1487–1510.

21. Bonewald LF, Harris SE, Rosser J, Dallas MR, Dallas SL, Camacho NP, Boyan B, Boskey A. von Kossa staining alone is not sufficient to confirm that mineralization in vitro represents bone formation. Calcif Tissue Int. 2003; 72:537–547. PMID: 12724828.

22. Camilleri J. Characterization of hydration products of mineral trioxide aggregate. Int Endod J. 2008; 41:408–417. PMID: 18298574.

23. Yang WK, Ko HJ, Kim MR. Evaluation of the rat tissue reaction to experimental new resin cement and mineral trioxide aggregate cement. Restor Dent Endod. 2012; 37:194–200. PMID: 23429672.

24. Bósio CC, Felippe GS, Bortoluzzi EA, Felippe MC, Felippe WT, Rivero ER. Subcutaneous connective tissue reactions to iRoot SP, mineral trioxide aggregate (MTA) Fillapex, DiaRoot BioAggregate and MTA. Int Endod J. 2014; 47:667–674. PMID: 25057520.

25. Clark G. Staining procedures. Baltimore, MD: Williams and Wilkins;1981. p. 187.

26. Gerstenfeld LC, Chipman SD, Glowacki J, Lian JB. Expression of differentiated function by mineralizing cultures of chicken osteoblasts. Dev Biol. 1987; 122:49–60. PMID: 3496252.

27. Cosme-Silva L, Dal-Fabbro R, Gonçalves LO, Prado AS, Plazza FA, Viola NV, Cintra LT, Gomes Filho JE. Hypertension affects the biocompatibility and biomineralization of MTA, High-plasticity MTA, and Biodentine^® . Braz Oral Res. 2019; 33:e060. PMID: 31365705.

28. Han L, Okiji T. Bioactivity evaluation of three calcium silicate-based endodontic materials. Int Endod J. 2013; 46:808–814. PMID: 23402321.

29. Hinata G, Yoshiba K, Han L, Edanami N, Yoshiba N, Okiji T. Bioactivity and biomineralization ability of calcium silicate-based pulp-capping materials after subcutaneous implantation. Int Endod J. 2017; 50(Supplement 2):e40–e51. PMID: 28649791.

30. Yaltirik M, Ozbas H, Bilgic B, Issever H. Reactions of connective tissue to mineral trioxide aggregate and amalgam. J Endod. 2004; 30:95–99. PMID: 14977305.

31. Ozbas H, Yaltirik M, Bilgic B, Issever H. Reactions of connective tissue to compomers, composite and amalgam root-end filling materials. Int Endod J. 2003; 36:281–287. PMID: 12702123.

32. Bueno CR, Vasques AM, Cury MT, Sivieri-Araújo G, Jacinto RC, Gomes-Filho JE, Cintra LT, Dezan-Júnior E. Biocompatibility and biomineralization assessment of mineral trioxide aggregate flow. Clin Oral Investig. 2019; 23:169–177.

33. Cintra LT, Benetti F, de Azevedo Queiroz ÍO, de Araújo Lopes JM, Penha de Oliveira SH, Sivieri Araújo G, Gomes-Filho JE. Cytotoxicity, biocompatibility, and biomineralization of the new high-plasticity MTA material. J Endod. 2017; 43:774–778. PMID: 28320539.

34. Tziafas D, Pantelidou O, Alvanou A, Belibasakis G, Papadimitriou S. The dentinogenic effect of mineral trioxide aggregate (MTA) in short-term capping experiments. Int Endod J. 2002; 35:245–254. PMID: 11985676.

35. Dominguez MS, Witherspoon DE, Gutmann JL, Opperman LA. Histological and scanning electron microscopy assessment of various vital pulp-therapy materials. J Endod. 2003; 29:324–333. PMID: 12775004.

36. Sarkar NK, Caicedo R, Ritwik P, Moiseyeva R, Kawashima I. Physicochemical basis of the biologic properties of mineral trioxide aggregate. J Endod. 2005; 31:97–100. PMID: 15671817.

37. Seux D, Couble ML, Hartmann DJ, Gauthier JP, Magloire H. Odontoblast-like cytodifferentiation of human dental pulp cells in vitro in the presence of a calcium hydroxide-containing cement. Arch Oral Biol. 1991; 36:117–128. PMID: 2059161.

38. Schröder U. Effects of calcium hydroxide-containing pulp-capping agents on pulp cell migration, proliferation, and differentiation. J Dent Res. 1985; 64:541–548. PMID: 3857254.

39. Camilleri J, Sorrentino F, Damidot D. Investigation of the hydration and bioactivity of radiopacified tricalcium silicate cement, Biodentine and MTA Angelus. Dent Mater. 2013; 29:580–593. PMID: 23537569.

40. Camilleri J. Characterization and hydration kinetics of tricalcium silicate cement for use as a dental biomaterial. Dent Mater. 2011; 27:836–844. PMID: 21600643.

41. Setbon HM, Devaux J, Iserentant A, Leloup G, Leprince JG. Influence of composition on setting kinetics of new injectable and/or fast setting tricalcium silicate cements. Dent Mater. 2014; 30:1291–1303. PMID: 25443159.

42. Murphy S, Wren AW, Towler MR, Boyd D. The effect of ionic dissolution products of Ca-Sr-Na-Zn-Si bioactive glass on in vitro cytocompatibility. J Mater Sci Mater Med. 2010; 21:2827–2834. PMID: 20711638.

43. Kim EJ, Bu SY, Sung MK, Choi MK. Effects of silicon on osteoblast activity and bone mineralization of MC3T3-E1 cells. Biol Trace Elem Res. 2013; 152:105–112. PMID: 23306944.

44. Shokouhinejad N, Nekoofar MH, Razmi H, Sajadi S, Davies TE, Saghiri MA, Gorjestani H, Dummer PM. Bioactivity of EndoSequence root repair material and bioaggregate. Int Endod J. 2012; 45:1127–1134. PMID: 22803801.

45. Ma J, Shen Y, Stojicic S, Haapasalo M. Biocompatibility of two novel root repair materials. J Endod. 2011; 37:793–798. PMID: 21787491.

46. Dawood AE, Parashos P, Wong RH, Reynolds EC, Manton DJ. Calcium silicate-based cements: composition, properties, and clinical applications. J Investig Clin Dent. 2017; 8:e12195.

47. Hansen SW, Marshall JG, Sedgley CM. Comparison of intracanal EndoSequence Root Repair Material and ProRoot MTA to induce pH changes in simulated root resorption defects over 4 weeks in matched pairs of human teeth. J Endod. 2011; 37:502–506. PMID: 21419298.