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Effects of several storage media on viability and proliferation capacity of periodontal ligament cells

  • Original Scientific Article
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European Archives of Paediatric Dentistry Aims and scope Submit manuscript

Abstract

Aim

The purpose of this study was to investigate the effect of different storage media on viability and proliferation capacity of periodontal ligament cells.

Methods

Plates with periodontal ligament fibroblasts (PDLF) cells were incubated in skimmed and whole milk, recently prepared Hank’s balanced salt solution (HBSS), Save-A-Tooth system’s, coconut water, propylene glycol with 20% propolis, egg white, tap water (negative control) at 5 °C and 20 °C, for 24 h. In one of the plates of each temperature, cell viability was determined by MTT assay. In the remaining plates, the wells were filled and incubated with Minimum Essential Medium (MEM) at 37 °C for 24, 48, 72, 96, and 120 h. The proliferation capacity of PDFL cells was also evaluated by MTT assay. Data were statistically analyzed by Kruskal–Wallis, Scheffé and Mann–Whitney tests (α = 5%).

Results

At 5 °C, milk maintained more viable cells than other storage media immediately after exposure (0 h) and allowed greater proliferation capacity. At 20 °C, milk and HBSS had similar and allowed similar proliferation ability at 24 and 48 h. From 72 h onwards, capacity to maintain cell viability the proliferation rate of cells incubated in HBSS was superior than milk. At both temperature and experiments, Save-A-Tooth system was similar to tap water.

Conclusion

Milk and HBSS were more effective in maintaining cellular viability and proliferation capacity than any other storage media. At 5 °C, the most viable alternative was milk. At 20 °C, HBSS had better results.

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References

  • Altschuler SJ, Wu LF. Cellular heterogeneity: do differences make a difference? Cell. 2010;141:559–63.

    Article  Google Scholar 

  • Ashkenazi M, Sarnat H, Keila S. In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in six different media. End Dent Traumatol. 1999;15:149–56.

    Article  Google Scholar 

  • Barile F. Introduction to in vitro cytotoxicology: mechanisms and methods. Boca Raton: CRC Press; 1994. p. 12–4.

    Google Scholar 

  • Belford DA, Rogers ML, Regester GO, et al. Milk-derived growth factors as serum supplements for the growth of fibroblast and epithelial cells. Vitro Cell Dev Biol Anim. 1995;31:752–60.

    Article  Google Scholar 

  • Blomlof L. Milk and saliva as possible storage media for traumatically exarticulated teeth prior to replantation. Swed Dent J. 1981;8:1–26.

    Google Scholar 

  • Blomlof L, Otteskog P. Viability of human periodontal ligament cells after storage in milk or saliva. Scand J Dent Res. 1980;88:436–40.

    PubMed  Google Scholar 

  • Casaroto AR, Hidalgo MM, Sell AM, et al. Study of the effectiveness of propolis extract as a storage medium for avulsed teeth. Dent Traumatol. 2010;26:323–31.

    Article  Google Scholar 

  • Chen F, Qi S, Lu L, Xu Y. Effect of storage temperature on the viability of human periodontal ligament fibroblasts. Dent Traumatol. 2015;31:24–8.

    Article  Google Scholar 

  • Choi SC, Seo YH, Bae WJ, et al. Milk activates the expression of cytokines via nrf2/ho-1 pathway in human periodontal ligament cells. Dent Traumatol. 2015;31:457–64.

    Article  Google Scholar 

  • Gauthier SF, Pouliot Y, Maubois JL. Growth factors from bovine milk and colostrum: composition, extraction and biological activities. Le Lait. 2006;86:90–125.

    Article  Google Scholar 

  • Gulinelli JL, Panzarini SR, Fattah CM, et al. Effect of root surface treatment with propolis and fluoride in delayed tooth replantation in rats. Dent Traumatol. 2008;24:651–7.

    Article  Google Scholar 

  • Harkacz OM Sr, Carnes DL Jr, Walker WA III. Determination of periodontal ligament cell viability in the oral rehydration fluid gatorade and milks of varying fat content. J Endod. 1997;23:687–90.

    Article  Google Scholar 

  • Hwang JY, Choi SC, Park JH, Kang SW. The use of green tea extract as a storage medium for the avulsed tooth. J Endod. 2011;37:962–7.

    Article  Google Scholar 

  • Khademi AA, Saei S, Mohajeri MR, et al. A new storage medium for an avulsed tooth. J Contemp Dent Pract. 2008;9:25–32.

    Article  Google Scholar 

  • Kokkali VV, Bendgude V, Sharangpani G. Comparative evaluation of post-traumatic periodontal ligament cell viability using three storage media. Eur Arch Paediatr Dent. 2017;18:209–14.

    Article  Google Scholar 

  • Lee W, Stover S, Rasoulianboroujeni M, et al. The efficacy of commercial tooth storage media for maintaining the viability of human periodontal ligament fibroblasts. Int Endod J. 2018;51:58–68.

    Article  Google Scholar 

  • Macway-Gomez S, Lallier TE. Pedialyte promotes periodontal ligament cell survival and motility. J Endod. 2013;39:202–7.

    Article  Google Scholar 

  • Mahal NK, Singh N, Thomas AM, Kakkar N. Effect of three different storage media on survival of periodontal ligament cells using collagenase-dispase assay. Int Endod J. 2013;46:365–70.

    Article  Google Scholar 

  • Marino TG, West LA, Liewehr FR, et al. Determination of periodontal ligament cell viability in long shelf-life milk. J Endod. 2000;26:699–702.

    Article  Google Scholar 

  • Moreira-Neto JJ, Gondim JO, Raddi MS, Pansani CA. Viability of human fibroblasts in coconut water as a storage medium. Int Endod J. 2009;42:827–30.

    Article  Google Scholar 

  • Moura CC, Soares PB, de Paula Reis MV, et al. Potential of coconut water and soy milk for use as storage media to preserve the viability of periodontal ligament cells: an in vitro study. Dent Traumatol. 2014;30:22–6.

    Article  Google Scholar 

  • Olson BD, Mailhot JM, Anderson RW, Schuster GS, Weller RN. Comparison of various transport media on human periodontal ligament cell viability. J Endod. 1997;23:676–9.

    Article  Google Scholar 

  • Ozan F, Polat ZA, Er K, Ozan U, Deger O. Effect of propolis on survival of periodontal ligament cells: new storage media for avulsed teeth. J Endod. 2007;33:570–3.

    Article  Google Scholar 

  • Riss TL, Moravec RA, Niles AL, et al. Cell viability assays. Assay Guidance Manual 2013 (Updated 2016):1–31.

  • Sigalas E, Regan JD, Kramer PR, Witherspoon DE, Opperman LA. Survival of human periodontal ligament cells in media proposed for transport of avulsed teeth. Dent Traumatol. 2004;20:21–8.

    Article  Google Scholar 

  • Silva EJ, Rollemberg CB, de Souza Coutinho-Filho T, Zaia AA. A multiparametric assay to compare the cytotoxicity of soy milk with different storage media. Dent Traumatol. 2013;29:319–22.

    Article  Google Scholar 

  • Souza BD, Bortoluzzi EA, da Silveira Teixeira C, et al. Effect of HBSS storage time on human periodontal ligament fibroblast viability. Dent Traumatol. 2010;26:481–3.

    Article  Google Scholar 

  • Souza BDM, Luckemeyer DD, Reyes-Carmona JF, et al. Viability of human periodontal ligament fibroblasts in milk, hank’s balanced salt solution and coconut water as storage media. Int Endod J. 2011;44:111–5.

    Article  Google Scholar 

  • Souza BD, Alves AM, Santos LG, et al. Fibroblast viability after storage at 20 degrees c in milk, hank’s balanced salt solution and coconut water. Braz Dent J. 2016;27:404–7.

    Article  Google Scholar 

  • Souza BDM, Bortoluzzi EA, Reyes-Carmona J, et al. Effect of temperature and seven storage media on human periodontal ligament fibroblast viability. Dent Traumatol. 2017;33:100–5.

    Article  Google Scholar 

  • Trope M. Avulsion of permanent teeth: theory to practice. Dent Traumatol. 2011;27:281–94.

    Article  Google Scholar 

  • Udoye CI, Jafarzadeh H, Abbott PV. Transport media for avulsed teeth: a review. Aust Endod J. 2012;38:129–36.

    Article  Google Scholar 

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Acknowledgements

The authors would like to thank Morgane Marion Kuntze, Ana Maria Hecke Alves and Jessie Reyes-Carmona for their precious contribution to this study.

Funding

The authors would like to thank the National Council for Scientific and Technological Development (CNPq) (Process No. 432572/2016-3) for the financial support.

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Correspondence to L. F. R. Garcia.

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The Ethics Committee for Research with Humans has approved this study (Protocol #073/08).

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Souza, B.D.M., Garcia, L.F.R., Bortoluzzi, E.A. et al. Effects of several storage media on viability and proliferation capacity of periodontal ligament cells. Eur Arch Paediatr Dent 21, 53–59 (2020). https://doi.org/10.1007/s40368-019-00450-8

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  • DOI: https://doi.org/10.1007/s40368-019-00450-8

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