Effect of storage medium of extracted human teeth on the result of in vitro studies – a scoping review
DOI:
https://doi.org/10.2340/biid.v12.44183Keywords:
Storage media, composite, resins, microleakage, bond strength, micromechanical propertiesAbstract
Purpose: This scoping review aimed to investigate how different storage media of extracted human teeth before and after preparation of specimens influence the outcomes of various dental materials intended for clinical application.
Materials and methods: Literature search in four databases and by manual searching was carried out on a predefined PIO as follows: Population – extracted human teeth; Intervention – different storage media; Outcome(s) – material properties.
Results: After screening 110 studies, 11 were included in the scoping review. Eight studies examined storage media effects before specimen preparation and three after specimen preparation. Four studies focused on enamel bonding, six on dentine, and one both on enamel and dentine. Concerning storage of extracted teeth prior to specimen preparation, cryopreservation, Chloramine T and thymol had no negative effect on bond strength to enamel. One study found Chloramine T to increase microleakage at enamel margins whereas another study found no negative effect. Storage in formalin, ethanol and thymol also had no negative effect on microleakage at enamel margins. Cryopreservation decreased bond strength to coronal dentine, but not to mid-coronal or deep dentine. Formalin, Chloramine T, ethanol and thymol had no negative effect on bond strength to dentine nor on microleakage at dentine margins. However, one study found long-term storage in Chloramine T to increase microleakage at dentine margins. Concerning storage of extracted teeth after specimen preparation, thymol significantly lowered bond strength to enamel, whereas formalin had no negative effect. Sodium hypochlorite had no negative effect on bond strength to dentine, and finally, formalin, Chloramine T, ethanol, and thymol had no negative effect on microleakage at dentine margins. Risk of bias assessment showed one high quality study, while the remaining 10 had questionable quality.
Conclusions: The effects of storage media on in-vitro dental material testing are variable and substrate-dependent, with dentine showing greater susceptibility than enamel. While distilled water and cryopreservation showed relative stability, other media like thymol and Chloramine T produced inconsistent results. Standardized protocols and further research are needed to ensure reliable and comparable outcomes.
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Nawrocka A, Łukomska-Szymańska M. Extracted human teeth and their utility in dental research. Recommendations on proper preservation: a literature review. Dent Med Probl. 2019;56(2):185–90. https://doi.org/10.17219/dmp/105252 DOI: https://doi.org/10.17219/dmp/105252
Boruziniat A, Danaifard N, Gifani M, Hakimi N. Effect of storage media and sterilization method on shear bond strength of composite to enamel of extracted teeth. J Dent Mater Tech. 2017;6(3):96–102.
Meza MS, Michel IM, Rivas RA, Benítez GB, Solaeche SM, Salas-de la Cruz D, et al. Obtaining human teeth for dental education: a cross-sectional study to create ethical and transparent processes. J Dent Educ. 2023;1(87):50–9. https://doi.org/10.1002/jdd.13094 DOI: https://doi.org/10.1002/jdd.13094
Aydın B, Pamir T, Baltaci A, Orman MN, Turk T. Effect of storage solutions on microhardness of crown enamel and dentin. Eur J Dent. 2015;9(2):262–6. https://doi.org/10.4103/1305-7456.156848 DOI: https://doi.org/10.4103/1305-7456.156848
Takagi S, Liao H, Chow LC. Effect of tooth-bound fluoride on enamel demineralization/remineralization in vitro. Caries Res. 2000;34(4):281–8. https://doi.org/10.1159/000016603 DOI: https://doi.org/10.1159/000016603
Yaguchi T. Layering mechanism of MDP-Ca salt produced in demineralization of enamel and dentin apatite. Dent Mater. 2017;33(1):23–32. https://doi.org/10.1016/j.dental.2016.09.037 DOI: https://doi.org/10.1016/j.dental.2016.09.037
Soares FZ, Follak A, Da Rosa LS, Montagner AF, Lenzi TL, Rocha RO. Bovine tooth is a substitute for human tooth on bond strength studies: A systematic review and meta-analysis of in vitro studies. Dent Mater. 2016;32(11):1385–93. https://doi.org/10.1016/j.dental.2016.09.019 DOI: https://doi.org/10.1016/j.dental.2016.09.019
Poole DF, Shellis RP, Tyler JE. Rates of formation in vitro of dental caries-like enamel lesions in man and some non-human primates. Arch Oral Biol. 1981;26(5):413–7. https://doi.org/10.1016/0003-9969(81)90039-X DOI: https://doi.org/10.1016/0003-9969(81)90039-X
Fricke HC,O’Neil JR. Inter-and intra-tooth variation in the oxygen isotope composition of mammalian tooth enamel phosphate: implications for palaeoclimatological and palaeobiological research. Palaeogeogr Palaeoclimatol Palaeoecol. 1996;126(1–2):91–9. https://doi.org/10.1016/S0031-0182(96)00072-7 DOI: https://doi.org/10.1016/S0031-0182(96)00072-7
Nakajima Y, Shimada Y, Miyashin M, Takagi Y, Tagami J, Sumi Y. Noninvasive cross-sectional imaging of incomplete crown fractures (cracks) using swept-source optical coherence tomography. Int Endod J. 2012;45(10):933–41. https://doi.org/10.1111/j.1365-2591.2012.02052.x DOI: https://doi.org/10.1111/j.1365-2591.2012.02052.x
Freitas AR, Aznar FD, Silva AL, Sales-Peres A, Sales-Peres SHC. Assessment of the effects of decontamination and storage methods on the structural integrity of human enamel. Rev Odontol UNESP. 2016;45(19):59–64. https://doi.org/10.1590/1807-2577.05915 DOI: https://doi.org/10.1590/1807-2577.05915
Oesterle LJ, Shellhart WC,Belanger GK. The use of bovine enamel in bonding studies. Am J Orthod Dentofacial Orthop. 1998;114(5):514–9. https://doi.org/10.1016/S0889-5406(98)70171-4 DOI: https://doi.org/10.1016/S0889-5406(98)70171-4
Attin T, Wegehaupt F, Gries D, Wiegand A. The potential of deciduous and permanent bovine enamel as substitute for deciduous and permanent human enamel: Erosion-abrasion experiments. J Dent. 2007;35(10):773–7. https://doi.org/10.1016/j.jdent.2007.07.007 DOI: https://doi.org/10.1016/j.jdent.2007.07.007
Camps J, Baudry X, Bordes V, Dejou J, Pignoly C, Ladeque P. Influence of tooth cryopreservation and storage time on microleakage. Dent Mater. 1996;12(2):121–6. https://doi.org/10.1016/S0109-5641(96)80079-0 DOI: https://doi.org/10.1016/S0109-5641(96)80079-0
Jaffer S, Oesterle LJ, Newman SM. Storage media effect on bond strength of orthodontic brackets. Am J Orthod Dentofacial Orthop. 2009;136(1):83–6. https://doi.org/10.1016/j.ajodo.2007.07.028 DOI: https://doi.org/10.1016/j.ajodo.2007.07.028
Sultana S, Nikaido T, Asafujjoha M, Tagami J, Matin K. Storage media to preserve dentin and their effects on surface properties. Int Chin J Dent. 2006;6:123–9.
Oilo G. Bond strength testing – what does it mean? Int Dent J. 1993;43(5):492–8.
Dominici JT, Eleazer PD, Clark SJ, Staat RH, Scheetz JP. Disinfection/sterilization of extracted teeth for dental student use. J Dent Educ 2001;65(11):1278–81. https://doi.org/10.1002/j.0022-0337.2001.65.11.tb03488.x DOI: https://doi.org/10.1002/j.0022-0337.2001.65.11.tb03488.x
White J, Goodis H, Marshall S, Marshall GW. Sterilization of teeth by gamma radiation. J Dent Res. 1994;73(9):1560–7. https://doi.org/10.1177/00220345940730091201 DOI: https://doi.org/10.1177/00220345940730091201
Tijare M, Smitha D, Kasetty S, Kallianpur S, Gupta S, Amith HV. Vinegar as a disinfectant of extracted human teeth for dental educational use. J Oral Maxillofac Pathol. 2014;18(1):14. https://doi.org/10.4103/0973-029X.131883 DOI: https://doi.org/10.4103/0973-029X.131883
Skene L. Ownership of human tissue and the law. Nat Rev Genet. 2002;3(2):145–8. https://doi.org/10.1038/nrg725 DOI: https://doi.org/10.1038/nrg725
Malek S, Darendeliler MA, Rex T, Kharbanda OP, Srivicharnkul P, Swain MV, et al. Physical properties of root cementum: part 2. Effect of different storage methods. Am J Orthod Dentofacial Orthop. 2003;124(5):561–70. https://doi.org/10.1016/S0889-5406(03)00398-6 DOI: https://doi.org/10.1016/S0889-5406(03)00398-6
Tosun G, Sener Y, Sengun A. Effect of storage duration/solution on microshear bond strength of composite to enamel. Dent Mater J. 2007;26(1):116–21. https://doi.org/10.4012/dmj.26.116 DOI: https://doi.org/10.4012/dmj.26.116
Toledano M, Osorio R, Osorio E, Aguilera FS, Yamauti M, Pashley DH, et al. Durability of resin–dentin bonds: effects of direct/indirect exposure and storage media. Dent Mater. 2007;23(7):885–92. https://doi.org/10.1016/j.dental.2006.06.030 DOI: https://doi.org/10.1016/j.dental.2006.06.030
Da Silva EM, Glir DH, Gill AW, Giovanini AF, Furuse AY, Gonzaga CC. Effect of chlorhexidine on dentin bond strength of two adhesive systems after storage in different media. Braz Dent J. 2015;26(6):642–7. https://doi.org/10.1590/0103-6440201300159 DOI: https://doi.org/10.1590/0103-6440201300159
Brännström M, Coli P, Blixt M. Effect of tooth storage and cavity cleansing on cervical gap formation in Class II glass-ionomer/composite restorations. Dent Mater. 1992;8(5):327–31. https://doi.org/10.1016/0109-5641(92)90109-P DOI: https://doi.org/10.1016/0109-5641(92)90109-P
Haller B, Hofmann N, Klaiber B, Bloching U. Effect of storage media on microleakage of five dentin bonding agents. Dent Mater. 1993;9(3):191–7. https://doi.org/10.1016/0109-5641(93)90119-B DOI: https://doi.org/10.1016/0109-5641(93)90119-B
Raum K, Kempf K, Hein HJ, Schubert J, Maurer P. Preservation of microelastic properties of dentin and tooth enamel in vitro – a scanning acoustic microscopy study. Dent Mater. 2007;23(10):1221–8. https://doi.org/10.1016/j.dental.2006.11.009 DOI: https://doi.org/10.1016/j.dental.2006.11.009
Francescut P, Zimmerli B, Lussi A. Influence of different storage methods on laser fluorescence values: a two-year study. Caries Res. 2006;40(3):181-5. https://doi.org/10.1159/000092223. PMID: 16707864. DOI: https://doi.org/10.1159/000092223
De Roo NMC, Deboosere E, Thierens LAM, Vercruysse C, Temmerman L, Verbeeck RMH, et al. The orthodontic bonding properties of human enamel after cryopreservation. Dent Mater J. 2020;39(6): 950–5. https://doi.org/10.4012/dmj.2019-208 DOI: https://doi.org/10.4012/dmj.2019-208
Panighi MM, Allart D, Jacquot BM, Camps J, G’Sell C. Influence of human tooth cryopreservation on dentin bond strength. Dent Mater. 1997;13(1):56–61. https://doi.org/10.1016/S0109-5641(97)80009-7 DOI: https://doi.org/10.1016/S0109-5641(97)80009-7
Kühl S, Deyhle H, Zimmerli M, Spagnoli G, Beckmann F, Müller B, et al. Cracks in dentin and enamel after cryopreservation. Oral Surg Oral Med Oral Pathol Oral Radiol. 2012;10(2):e5–10. https://doi.org/10.1016/j.tripleo.2011.06.020 DOI: https://doi.org/10.1016/j.tripleo.2011.06.020
Camps J, Martin P, Ladeque P, Rieu R, Fuseri J. Influence of tooth cryopreservation on human dentin permeability, in vitro. Dent Mater. 1994;10(3):210–4. https://doi.org/10.1016/0109-5641(94)90034-5 DOI: https://doi.org/10.1016/0109-5641(94)90034-5
Lee JJ, Nettey-Marbell A, Cook A Jr, Pimenta LA, Leonard R, Ritter AV. Using extracted teeth for research: the effect of storage medium and sterilization on dentin bond strengths. J Am Dent Assoc. 2007;138(12):1599–603. https://doi.org/10.14219/jada.archive.2007.0110 DOI: https://doi.org/10.14219/jada.archive.2007.0110
Hutton B, Salanti G, Caldwell DM, Chaimani A, Schmid CH, Cameron C, et al. The PRISMA extension statement for reporting of systematic reviews incorporating network meta-analyses of health care interventions: checklist and explanations. Ann Intern Med. 2015;162(11):777–84. https://doi.org/10.7326/M14-2385 DOI: https://doi.org/10.7326/M14-2385
Foster ED, Deardorff A. Open science framework (OSF). J Med Libr Assoc. 2017;105(2):203. https://doi.org/10.5195/jmla.2017.88 DOI: https://doi.org/10.5195/jmla.2017.88
Sheth VH, Shah NP, Jain R, Bhanushali N, Bhatnagar V. Development and validation of a risk-of-bias tool for assessing in vitro studies conducted in dentistry: The QUIN. J Prosthet Dent. 2022;131(6):1038–42. https://doi.org/10.1016/j.prosdent.2022.05.019. DOI: https://doi.org/10.1016/j.prosdent.2022.05.019
Coelho-De-Souza FH, Camacho GB, Demarco FF, Powers JM. Fracture resistance and gap formation of MOD restorations: influence of restorative technique, bevel preparation and water storage. Oper Dent. 2008;33(1):37–43. https://doi.org/10.2341/07-27 DOI: https://doi.org/10.2341/07-27
Mobarak EH, El-Badrawy W, Pashley DH, Jamjoom H. Effect of pretest storage conditions of extracted teeth on their dentin bond strengths. J Prosthet Dent. 2010;104(2):92–7. https://doi.org/10.1016/S0022-3913(10)60098-4 DOI: https://doi.org/10.1016/S0022-3913(10)60098-4
Retief DH, Wendt SL, Bradley EL, Denys FR. The effect of storage media and duration of storage of extracted teeth on the shear bond strength of Scotchbond 2/Silux to dentin. Am J Dent. 1989;2(5):269–73.
Rolland SL, Carrick TE, Walls AW, McCabe JF. Dentin decontamination using chloramine T prior to experiments involving bacteria. Dent Mater. 2007 Dec;23(12):1468-72. https://doi.org/10.1016/j.dental.2007.01.001. Epub 2007 Mar 8. PMID: 17349688. DOI: https://doi.org/10.1016/j.dental.2007.01.001
Browne RM. Animal tests for biocompatibility of dental materials--relevance, advantages and limitations. J Dent. 1994;22 Suppl 2:S21-4. https://doi.org/10.1016/0300-5712(94)90035-3. PMID: 7844271. DOI: https://doi.org/10.1016/0300-5712(94)90035-3
Secilmis A, Dilber E, Ozturk N, Yilmaz FG. The effect of storage solutions on mineral content of enamel. Mater Sci Appl. 2013 Jun 28;4(7):439-45. DOI: https://doi.org/10.4236/msa.2013.47053
Fujisawa S, Kadoma Y. Effect of phenolic compounds on the polymerization of methyl methacrylate. Dent Mater. 1992 Sep;8(5):324-6. https://doi.org/10.1016/0109-5641(92)90108-o. PMID: 1303375. DOI: https://doi.org/10.1016/0109-5641(92)90108-O
Pashley DH, Sano H, Ciucchi B, Yoshiyama M, Carvalho RM. Adhesion testing of dentin bonding agents: a review. Dent Mater. 1995;11(2):117–25. https://doi.org/10.1016/0109-5641(95)80046-8 DOI: https://doi.org/10.1016/0109-5641(95)80046-8
Beghetto V, Gatto V, Conca S, Bardella N, Scrivanti A. Polyamidoamide dendrimers and cross-linking agents for stabilized bioenzymatic resistant metal-free bovine collagen. Molecules. 2019;24(19):3611. https://doi.org/10.3390/molecules24193611 DOI: https://doi.org/10.3390/molecules24193611
Oliver RF, Grant RA, Cox RW, Cooke A. Effect of aldehyde cross-linking on human dermal collagen implants in the rat. Br J Exp Pathol. 1980;61(5):544–9.
Sano H, Shono T, Sonoda H, Takatsu T, Ciucchi B, Carvalho R, et al. Relationship between surface area for adhesion and tensile bond strength – evaluation of a micro-tensile bond test. Dent Mater. 1994;10(4):236–40. https://doi.org/10.1016/0109-5641(94)90067-1 DOI: https://doi.org/10.1016/0109-5641(94)90067-1
Carrilho MR, Tay FR, Pashley DH, Tjäderhane L, Carvalho RM. Mechanical stability of resin-dentin bond components. Dent Mater. 2005;21(3):232–41. https://doi.org/10.1016/j.dental.2004.06.001 DOI: https://doi.org/10.1016/j.dental.2004.06.001
Ito S, Hashimoto M, Wadgaonkar B, Svizero N, Carvalho RM, Yiu C, et al. Effects of resin hydrophilicity on water sorption and changes in modulus of elasticity. Biomaterials. 2005;26(33):6449–59. https://doi.org/10.1016/j.biomaterials.2005.04.052 DOI: https://doi.org/10.1016/j.biomaterials.2005.04.052
11405: IT. Dentistry–testing of adhesion to tooth structure. Geneva: International Standards Organization; 2015.
Schwartz O. Cryopreservation as long-term storage of teeth for transplantation or replantation. Int J Oral Maxillofac Surg. 1986;15(1): 30–2. https://doi.org/10.1016/S0300-9785(86)80008-4 DOI: https://doi.org/10.1016/S0300-9785(86)80008-4
Delgado AH, Sauro S, Lima AF, Loguercio AD, Della Bona A, Mazzoni A, et al. RoBDEMAT: A risk of bias tool and guideline to support reporting of pre-clinical dental materials research and assessment of systematic reviews. J Dent 2022;127:104350. https://doi.org/10.1016/j.jdent.2022.104350 DOI: https://doi.org/10.1016/j.jdent.2022.104350
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