Cytocompatibility and antibacterial properties of capping materials

Claudio Poggio, Carla Renata Arciola, Riccardo Beltrami, Annachiara Monaco, Alberto Dagna, Marco Lombardini, Livia Visai, Claudio Poggio, Carla Renata Arciola, Riccardo Beltrami, Annachiara Monaco, Alberto Dagna, Marco Lombardini, Livia Visai

Abstract

The aim of this study was to evaluate and compare the antimicrobial activity and cytocompatibility of six different pulp-capping materials: Dycal (Dentsply), Calcicur (Voco), Calcimol LC (Voco), TheraCal LC (Bisco), MTA Angelus (Angelus), and Biodentine (Septodont). To evaluate antimicrobial activity, materials were challenged in vitro with Streptococcus mutans, Streptococcus salivarius, and Streptococcus sanguis in the agar disc diffusion test. Cytocompatibility of the assayed materials towards rat MDPC-23 cells was evaluated at different times by both MTT and apoptosis assays. Results significantly differed among the different materials tested. Both bacterial growth inhibition halos and cytocompatibility performances were significantly different among materials with different composition. MTA-based products showed lower cytotoxicity and valuable antibacterial activity, different from calcium hydroxide-based materials, which exhibited not only higher antibacterial activity but also higher cytotoxicity.

Figures

Figure 1
Figure 1
Antibacterial activity of the different pulp-capping materials evaluated by agar diffusion test. Each paper disk impregnated with the different pulp-capping materials was placed on agar plates previously incubated with the indicated streptococcal strains and incubated at 37°C for 24 h. The positive control was represented by a 10% dilution of 30% H2O2. All the assays were conducted in triplicate and the results were recorded in terms of the average diameter of inhibition zone (mm). Error bars indicate standard errors of the means.
Figure 2
Figure 2
MDPC-23 cells cytocompatibility of the different pulp-capping materials using the Transwell method. MDPC-23 cells were incubated with the different pulp-capping materials at 37°C for 24 h, 48 h, and 72 h in a Transwell culture plate as reported in Materials and Methods Section. The cell viability was assessed with MMT test. The data are presented as percent of the control incubated in absence of any materials and set as 100%. Five replicates for each pulp-capping material were used for each experiment performed in duplicate. Error bars indicate standard errors of the means.
Figure 3
Figure 3
CLSM images of apoptosis assay. MDPC-23 cells were cultured in a 24-Transwell-tissue culture plate for 24 h at 37°C in the absence of any material (a) or in the presence of H2O2 (b), MTA Angelus (c), Biodentine (d), Calcicur (e), Calcimol LC (f), Dycal (g), and TheraCal LC (h). PSVue480 reagent was used for staining apoptotic cells. CLSM images were taken at 20x magnification.

References

    1. Bachoo IK, Seymour D, Brunton PA. Biocompatible and bioactive replacement for dentine: is this a reality? The properties and uses of a novel calcium-based cement. British Dental Journal. 2013;214(2):p. E5.
    1. Dominguez MS, Witherspoon DE, Gutmann JL, Opperman LA. Histological and scanning electron microscopy assessment of various vital pulp-therapy materials. Journal of Endodontics. 2003;29(5):324–333.
    1. Camilleri J, Pitt Ford TR. Mineral trioxide aggregate: a review of the constituents and biological properties of the material. International Endodontic Journal. 2006;39(10):747–754.
    1. Prati C, Siboni F, Polimeni A, Bossu M, Gandolfi MG. Use of calcium-containing endodontic sealers as apical barrier in fluid-contaminated wide-open apices. Journal of Applied Biomaterials & Functional Materials. 2014
    1. Hasheminia SM, Nejad SL, Dianat O, Modaresi J, Mahjour F. Comparing the sealing properties of mineral trioxide aggregate and an experimental ceramic based root end filling material in different environments. Indian Journal of Dental Research. 2013;24(4):474–477.
    1. Malik G, Bogra P, Singh S, Samra RK. Comparative evaluation of intracanal sealing ability of mineral trioxide aggregate and glass ionomer cement: an in vitro study. Journal of Conservative Dentistry. 2013;16(6):540–545.
    1. Desai S, Chandler N. The restoration of permanent immature anterior teeth, root filled using MTA: a review. Journal of Dentistry. 2009;37(9):652–657.
    1. Mohammadi Z, Dummer PMH. Properties and applications of calcium hydroxide in endodontics and dental traumatology. International Endodontic Journal. 2011;44(8):697–730.
    1. Fulzele P, Baliga S, Thosar N, Pradhan D. Evaluation of calcium ion, hydroxyl ion release and pH levels in various calcium hydroxide based intracanal medicaments: an in vitro study. Contemporary Clinical Dentistry. 2011;2(4):291–295.
    1. Laurent P, Camps J, About I. Biodentine induces TGF-β1 release from human pulp cells and early dental pulp mineralization. International Endodontic Journal. 2012;45(5):439–448.
    1. Téclès O, Laurent P, Aubut V, About I. Human tooth culture: a study model for reparative dentinogenesis and direct pulp capping materials biocompatibility. Journal of Biomedical Materials Research B: Applied Biomaterials. 2008;85(1):180–187.
    1. Goldberg M, Six N, Decup F, et al. Bioactive molecules and the future of pulp therapy. The American Journal of Dentistry. 2003;16(1):66–76.
    1. Almushayt A, Narayanan K, Zaki AE, George A. Dentin matrix protein 1 induces cytodifferentiation of dental pulp stem cells into odontoblasts. Gene Therapy. 2006;13(7):611–620.
    1. Pinho Veloso HH, do Santos RA, de Araújo TP, Leonardi DP, Baratto Filho F. Histological analysis of the biocompatibility of three different calcium hydroxide-based root canal sealers. Journal of Applied Oral Science. 2006;14(5):376–381.
    1. Nosrat A, Peimani A, Asgary S. A preliminary report on histological outcome of pulpotomy with endodontic biomaterials vs calcium hydroxide. Restorative Dentistry & Endodontics. 2013;38(4):227–233.
    1. Furey A, Hjelmhaug J, Lobner D. Toxicity of flow line, durafill VS, and Dycal to dental pulp cells: effects of growth factors. Journal of Endodontics. 2010;36(7):1149–1153.
    1. Torabinejad M, Parirokh M. Mineral trioxide aggregate: a comprehensive literature review-part II: leakage and biocompatibility investigations. Journal of Endodontics. 2010;36(2):190–202.
    1. Torabinejad M, Watson TF, Pitt Ford TR. Sealing ability of a mineral trioxide aggregate when used as a root end filling material. Journal of Endodontics. 1993;19(12):591–595.
    1. Parirokh M, Torabinejad M. Mineral trioxide aggregate: a comprehensive literature review-part I: chemical, physical, and antibacterial properties. Journal of Endodontics. 2010;36(1):16–27.
    1. Asgary S, Eghbal MJ, Parirokh M, Ghoddusi J. Effect of two storage solutions on surface topography of two root-end fillings. Australian Endodontic Journal. 2009;35(3):147–152.
    1. Parirokh M, Torabinejad M. Mineral trioxide aggregate: a comprehensive literature review-part III: clinical applications, drawbacks, and mechanism of action. Journal of Endodontics. 2010;36(3):400–413.
    1. Gandolfi MG, Shah SN, Feng R, Prati C, Akintoye SO. Biomimetic calcium-silicate cements support differentiation of human orofacial mesenchymal stem cells. Journal of Endodontics. 2011;37(8):1102–1108.
    1. Tuna D, Ölmez A. Clinical long-term evaluation of MTA as a direct pulp capping material in primary teeth. International Endodontic Journal. 2008;41(4):273–278.
    1. Takita T, Hayashi M, Takeichi O, et al. Effect of mineral trioxide aggregate on proliferation of cultured human dental pulp cells. International Endodontic Journal. 2006;39(5):415–422.
    1. Moghaddame-Jafari S, Mantellini MG, Botero TM, McDonald NJ, Nör JE. Effect of ProRoot MTA on pulp cell apoptosis and proliferation in vitro. Journal of Endodontics. 2005;31(5):387–391.
    1. Okiji T, Yoshiba K. Reparative dentinogenesis induced by mineral trioxide aggregate: a review from the biological and physicochemical points of view. International Journal of Dentistry. 2009;2009:12 pages.464280
    1. Bogen G, Kim JS, Bakland LK. Direct pulp capping with mineral trioxide aggregate: an observational study. Journal of the American Dental Association. 2008;139(3):305–315.
    1. Arciola CR, Montanaro L, Costerton JW. New trends in diagnosis and control strategies for implant infections. International Journal of Artificial Organs. 2011;34(9):727–736.
    1. Hebling J, Lessa FCR, Nogueira I, de Carvalho RM, de Costa CAS. Cytotoxicity of resin-based light-cured liners. The American Journal of Dentistry. 2009;22(3):137–142.
    1. Zanini M, Sautier JM, Berdal A, Simon S. Biodentine induces immortalized murine pulp cell differentiation into odontoblast-like cells and stimulates biomineralization. Journal of Endodontics. 2012;38(9):1220–1226.
    1. Accorinte MDLR, Holland R, Reis A, et al. Evaluation of mineral trioxide aggregate and calcium hydroxide cement as pulp-capping agents in human teeth. Journal of Endodontics. 2008;34(1):1–6.
    1. Carvalho TL, Teófilo JM, Araújo CA, Brentegani LG. Chronology of alveolar healing following immediate implantation of Ricinus communis polyurethane resin: histometric analysis in rats. Journal of Biomedical Materials Research. 1997;37(4):449–452.
    1. Mantellini MG, Botero TM, Yaman P, Dennison JB, Hanks CT, Nör JE. Adhesive resin induces apoptosis and cell-cycle arrest of pulp cells. Journal of Dental Research. 2003;82(8):592–596.
    1. Schmalz G, Schweikl H. Characterization of an in vitro dentin barrier test using a standard toxicant. Journal of Endodontics. 1994;20(12):592–594.
    1. de Souza CG, Girardo NS, Costa MA, Peralta RM. Influence of growth conditions on the production of xylanolytic enzymes by Aspergillus flavus. Journal of Basic Microbiology. 1999;39(3):155–160.
    1. Babich H, Sinensky MC. Indirect cytotoxicity of dental materials: a study with Transwell inserts and the neutral red uptake assay. Alternatives to Laboratory Animals. 2001;29(1):9–13.
    1. Saino E, Grandi S, Quartarone E, et al. In vitro calcified matrix deposition by human osteoblasts onto a zinc-containing bioactive glass. European Cells & Materials. 2011;21:59–72.
    1. van Engeland M, Nieland LJW, Ramaekers FCS, Schutte B, Reutelingsperger CPM. Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure. Cytometry. 1998;31(1):1–9.
    1. Al-Khatib ZZ, Baum RH, Morse DR, Yesilsoy C, Bhambhani S, Furst ML. The antimicrobial effects of various endodontic sealers. Oral Surgery Oral Medicine and Oral Pathology. 1990;70(6):784–790.
    1. Cohen S, Burns RC. Pathways of the Pulp. 8th edition. St. Louis, Mo, USA: Mosby; 2002.
    1. Çobankara FK, Altinöz HC, Erganiş O, Kav K, Belli S. In vitro antibacterial activities of root-canal sealers by using two different methods. Journal of Endodontics. 2004;30(1):57–60.
    1. Tobias RS. Antibacterial properties of dental restorative materials: a review. International Endodontic Journal. 1988;21(2):155–160.
    1. Lai CC, Huang FM, Yang HW, et al. Antimicrobial activity of four root canal sealers against endodontic pathogens. Clinical oral investigations. 2001;5(4):236–239.
    1. Lu Y, Liu T, Li H, Pi G. Histological evaluation of direct pulp capping with a self-etching adhesive and calcium hydroxide on human pulp tissue. International Endodontic Journal. 2008;41(8):643–650.
    1. Siqueira JF., Jr. Strategies to treat infected root canals. Journal of the California Dental Association. 2001;29(12):825–837.
    1. Roberts HW, Toth JM, Berzins DW, Charlton DG. Mineral trioxide aggregate material use in endodontic treatment: a review of the literature. Dental Materials. 2008;24(2):149–164.
    1. Torabinejad M, Hong CU, Ford TRP, Kettering JD. Antibacterial effects of some root end filling materials. Journal of Endodontics. 1995;21(8):403–406.
    1. Eldeniz AU, Hadimli HH, Ataoglu H, Ørstavik D. Antibacterial effect of selected root-end filling materials. Journal of Endodontics. 2006;32(4):345–349.
    1. Al-Hezaimi K, Al-Shalan TA, Naghshbandi J, Oglesby S, Simon JHS, Rotstein I. Antibacterial effect of two Mineral Trioxide Aggregate (MTA) preparations against Enterococcus faecalis and Streptococcus sanguis in vitro. Journal of Endodontics. 2006;32(11):1053–1056.
    1. Ribeiro CS, Scelza MFZ, Hirata Jünior R, de Oliveira LMB. The antimicrobial activity of gray-colored mineral trioxide aggregate (GMTA) and white-colored MTA (WMTA) under aerobic and anaerobic conditions. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology and Endodontology. 2010;109(6):e109–e112.
    1. Cabiscol E, Tamarit J, Ros J. Oxidative stress in bacteria and protein damage by reactive oxygen species. International Microbiology. 2000;3(1):3–8.
    1. Reston EG, de Souza Costa CA. Scanning electron microscopy evaluation of the hard tissue barrier after pulp capping with calcium hydroxide, mineral trioxide aggregate (MTA) or ProRoot MTA. Australian Endodontic Journal. 2009;35(2):78–84.
    1. Vahey JW, Simonian PT, Conrad EU., III Carcinogenicity and metallic implants. The American Journal of Orthopedics. 1995;24(4):319–324.
    1. Schmalz G. A cell culture method for screening the biocompatibility of dental materials. In: Winter GD, Gibbons DF, Plenk H Jr., editors. Biomaterials. New York, NY, USA: John Wiley & Sons; 1982. pp. 321–326.
    1. Schmalz G, Garhammer P, Schweiki H. A commercially available cell culture device modified for dentin barrier tests. Journal of Endodontics. 1996;22(5):249–252.
    1. Schmalz G. Agar overlay method. International Endodontic Journal. 1988;21(2):59–66.
    1. Wennberg A, Hasselgren G, Tronstad L. A method for toxicity screening of biomaterials using cells cultured on millipore filters. Journal of Biomedical Materials Research. 1979;13(1):109–120.
    1. Holland R, de Souza V, de Mello W, Nery MJ, Bernabé PF, Otoboni Filho JA. Permeability of the hard tissue bridge formed after pulpotomy with calcium hydroxide: a histologic study. The Journal of the American Dental Association. 1979;99(3):472–475.
    1. Goldberg F, Massone EJ, Spielberg C. Evaluation of the dentinal bridge after pulpotomy and calcium hydroxide dressing. Journal of Endodontics. 1984;10(7):318–320.
    1. Pereira JC, Segala AD, Costa CAS. Human pulpal response to direct pulp capping with an adhesive system. The American Journal of Dentistry. 2000;13(3):139–147.
    1. Stanley HR, Pameijer CH. Dentistry’s friend: calcium hydroxide. Operative Dentistry. 1997;22(1):1–3.
    1. Hwang Y-C, Lee S-H, Hwang I-N, et al. Chemical composition, radiopacity, and biocompatibility of Portland cement with bismuth oxide. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology and Endodontology. 2009;107(3):e96–e102.
    1. Coutinho-Filho T, De-Deus G, Klein L, Manera G, Peixoto C, Gurgel-Filho ED. Radiopacity and histological assessment of Portland cement plus bismuth oxide. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology and Endodontology. 2008;106(6):e69–e77.
    1. Arciola CR, Montanaro L, Moroni A, Giordano M, Pizzoferrato A, Donati ME. Hydroxyapatite-coated orthopaedic screws as infection resistant materials: in vitro study. Biomaterials. 1999;20(4):323–327.
    1. Arciola CR, Campoccia D, Speziale P, Montanaro L, Costerton JW. Biofilm formation in Staphylococcus implant infections. A review of molecular mechanisms and implications for biofilm-resistant materials. Biomaterials. 2012;33(26):5967–5982.
    1. Campoccia D, Montanaro L, Arciola CR. A review of the clinical implications of anti-infective biomaterials and infection-resistant surfaces. Biomaterials. 2013;34(33):8018–8029.
    1. Pye AD, Lockhart DEA, Dawson MP, Murray CA, Smith AJ. A review of dental implants and infection. Journal of Hospital Infection. 2009;72(2):104–110.
    1. Bumgardner JD, Adatrow P, Haggard WO, Norowski PA. Emerging antibacterial biomaterial strategies for the prevention of peri-implant inflammatory diseases. International Journal of Oral & Maxillofacial Implants. 2011;26(3):553–560.

Source: PubMed

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