In vitro cytotoxic evaluation of conventional denture base material and soft lining material using colorimetric MTT assay

Abstract

Abstract

Purpose: In this study, it was aimed to evaluate the time course of cytotoxic effects of the conventional base material and soft lining material on the mouse fibroblast cells.

Material and Method:

Disc-shaped test samples of denture base material (rodex) and soft lining material( dentusil) were fabricated according to manufacturers' instructions under aseptic conditions. The samples were subjected to 5,000 thermal cycling to mimic the oral environment. Following aging procedures, the cytotoxic effect of the materials was assessed by [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide]  assay using L929 mouse fibroblast cells after 24 h, 48 h and 72 h cell incubation period. Cell viability values were calculated for each group. Statistical analysis of the data was performed using a two-way repeated measurement method. (p<0,001)

Results: For 24 hours and 48 hours incubation period soft lining material, and  for 72 hour incubation period the base material showed more cell viability. Statistically, there was a significant difference between the two materials. During the incubation period, the group incubated 24 hours is statistically different from 48 hours and 72 hours. No significant difference was found between 72 hours and 48 hours.

Conclusion: The soft lining material we used under the base materials is more biocompatible than the base material.

Keywords

• Conventional denture base material,soft liners,cytotoxicity,thermal cycles

Kolorimetrik MTT testi kullanarak geleneksel protez kaide materyali ile yumuşak astar materyalinin in vitro sitotoksik özelliklerinin değerlendirilmesi

Abstract

Özet

Amaç: Bu çalışmada protez yapımında kullanılan geleneksel kaide materyali ile yumuşak astar materyalinin fare fibroblast hücreleri üzerinde zamanla meydana gelen sitotoksik etkilerinin değerlendirilmesi amaçlandı.

Materyal ve Metod: Protez kaide materyali (rodeks) ve yumuşak astar materyalinin (dentusil) disk şekilli test numuneleri üreticinin talimatlarına göre aseptik şartlar altında hazırlandı.. Örnekler, ağız ortamını taklit etmek için 5.000 termal döngüye tabi tutuldu. Yaşlanma prosedürlerini takiben, materyallerin sitotoksik etkisi, 24 saat, 48 saat ve 72 saatlik hücre inkubasyon döneminden sonra L929 fare fibroblast hücreleri kullanılarak [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] testi ile değerlendirildi. Her grup için hücre canlılığı değerleri hesaplandı. Verilerin istatistiksel analizi, iki yönlü tekrarlanan bir ölçüm yöntemi kullanılarak gerçekleştirildi. (P <0.001)

Bulgular: 24 saat ve 48 saat inkubasyon periyodunda yumuşak astar materyali, 72 saat inkübasyon periyodunda ise kaide materyali daha fazla hücre canlılığı göstermiştir. İstatistiksel olarak iki materyal arasında anlamlı fark bulunmuştur. İnkübasyon periyotları arasında ise 24 saat inkübe edilen grup 48 saat ve 72 saatden istatistiksel olarak farklıdır. 72 saat ve 48 saat arasında anlamlı fark bulunamamıştır.

Sonuç: Kaide materyallerinin altına kullanmış olduğumuz yumuşak astar materyali kaide materyaline göre daha biyouyumludur.

Keywords

• Geleneksel kaide materyali,yumuşak astar,sitotoksisite,termal siklus

References

1. 1. Song YH, Song H J, Han MK, Yang HS, Park YJ. Cytotoxicity of soft denture lining materials depending on their component types. Int J Prosthodont, 2014;27(3):229-35
2. 2. Gonçalves TS, Morganti MA, Campos LC, Rizzatto SM, Menezes LM. Allergy to autopolymerized acrylic resin in an orthodontic patient. Am J Orthod Dentofacial Orthop 2006;129:431-5
3. 3. Chaves CDAL, Machado AL, Vergani CE, de Souza RF, Giampaolo ET. Cytotoxicity of denture base and hard chairside reline materials: a systematic review. J Prosthet Dent, 2012:107(2); 114-127
4. 4. Karakış D, Akay C, Erdönmez D, Doğan A. Farklı yumuşak astar materyallerinin Candida albicans biyofilm formasyonu açısından değerlendirilmesi. Acta Odontol Turc 2015;32(1), 19-25
5. 5. Akay C, Tanış MÇ, Sevim H. Effect of artificial saliva with different pH levels on the cytotoxicity of soft denture lining materials. Int J Artif Organs 2017 Jun 27:0. doi: 10.5301/ijao.5000614. [Epub ahead of print]
6. 6. Silva CRC, Pellissari C VG, Sanitá PV, Arbeláez M IA, Massetti P, Jorge JH. Metabolism of L929 cells after contact with acrylic resins. Part 1: Acrylic denture base resins. International Journal of Dentistry and Oral Science, 2015; 1-8
7. 7. Jorge JH, Silva CRC, Pavarina AC, Amaya MI, Masetti P, Pellissari CV. Metabolism of L929 Cells After Contact With Acrylic Resins. Part 2: Soft Relines. Int J Dentistry Oral Sci. S,2015; 1, 6-11
8. 8. Straioto FG, Ricomini Filho AP, Fernandes Neto AJ, et al: Polytetrafluorethylene added to acrylic resins: mechanical properties. Braz Dent J 2010;21:55-59

9. 9. Iwasaki N, Yamaki C, Takahashi H, Oki M, Suzuki T. Effect of long-time immersion of soft denture liners in water on viscoelastic properties. Dent Mater J 2017. doi:10.4012/dmj.2016-320
10. 10. Zarb G, Hobkirk JA, Eckert SE, Jacob RF. Prosthodontic treatment for edentulous patients. 13th ed. St. Louis: Mosby; 2013. p. 147-312
11. 11. Palla ES, Karaoglani E, Naka O, Anastassiadou V. Soft denture liners’ effect on the masticatory function in patients wearing complete dentures: A systematic review. J Dent 2015; 43: 1403-1410
12. 12. Ogawa A, Kimoto S, Hiroyuki S, Ono M, Furuse N, Kawai Y. The influence of patient characteristics on acrylic-based resilient denture liners embedded in maxillary complete dentures. J Prosthodont Res 2016; 60: 199-205
13. 13. International Organization for Standardization, ISO 10993-5. Biological evaluation of medical devices-part 5. Tests for cytotoxicity: in vitro methods. Geneve: ISO, 1992
14. 14. Landayan, J IA, Manaloto ACF, Lee JY, Shin SW. Effect of aging on tear strength and cytotoxicity of soft denture lining materials; in vitro. J Adv Prosthodont 2014; 6(2): 115-120
15. 15. Akay C, Cevik P, Karakis D, Sevim H. In Vitro Cytotoxicity of Maxillofacial Silicone Elastomers: Effect of Nano‐particles. J Prosthodont. 2016
16. 16. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65:55–63
17. 17. Lü L, Zhang L, Wai MSM, Yew DTW, Xu J. Exocytosis of MTT formazan could exacerbate cell injury. Toxicol in Vitro. 2012;26:636–44
18. 18. Stockert JC, Blázquez-Casto A, Cañete M, Horobin RW, Villanueva A. MTT assay for cell viability: intracellular localisation of the formazan product is in lipid droplets. Acta Histochem. 2012;114:785–96
19. 19. Van Tonder A, Joubert AM, Cromarty AD. Limitations of the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay when compared to three commonly used cell enumeration assays. BMC research notes 2015; 8(1), 47
20. 20. Moharamzadeh K, Van Noort R, Brook IM, Scutt AM.“Cytotoxicity of resin monomers on human gingival fibroblasts and HaCaT keratinocytes,” Dent Mater 2007; 23(1): 40–44
21. 21. Chaves CDAL, de Souza Costa CA, Vergani CE, Chaves de Souza P P, Machado AL . Effects of soft denture liners on L929 fibroblasts, HaCaT keratinocytes, and RAW 264.7 macrophages. BioMed research international, 2014. http://dx.doi.org/10.1155/2014/840613
22. 22. Bailey LO, Weir MD, Washburn NR. Quantification of macrophage viability and inflammatory response to dental bonding resins. J Bioact Compat Pol 2006; 21(3): 185–206
23. 23. Atay A, Cetintas VB, Cal E, Kosova B, Kesercioglu A, Guneri P. Cytotoxicity of hard and soft denture lining materials. Dent Mater J 2012; 31(6), 1082-6
24. 24. Akin H, Tugut F, Polat ZA. In vitro comparison of the cytotoxicity and water sorption of two different denture base systems. J Prosthodont 2015; 24(2):152-155
25. 25. da Silva EVF, Goiato MC, dos Santos DM, da Rocha Bonatto L, Brito VGB, de Oliveira SHP. Effect of different methods of polymerizing ocular prosthesis acrylic resin on a human conjunctival cell line. J Prosthet Dent, 2016; 116(5), 818-23
26. 26. Anusavice KJ, Shen C, Rawls HR. Phillips’ science of dental materials (12th ed.), Elsevier, St. Louis, MO 2012; 474-498
27. 27. Canadas MD, Garcia LF, Consani S, Pires-de-Souza FC. Color stability, surface roughness, and surface porosity of acrylic resins for eye sclera polymerized by different heat sources J Prosthodont 2010; 19: 52-57
28. 28. Bural C, Aktaş E, Deniz G, Ünlüçerçi Y, Kızılcan N, Bayraktar G. Effect of post-polymerization heat-treatments on degree of conversion, leaching residual MMA and in vitro cytotoxicity of autopolymerizing acrylic repair resin. Dent Mater, 2011; 27: 1135-43
29. 29. Anusavice KJ. Phillips’ Science of Dental Materials, ed 11. Amsterdam: Saunders, 2006:174,753–754
30. 30. Neves CB, Lopes LP, Ferrão HF, Miranda JP, Castro MF, Bettencourt AF. Ethanol postpolymerization treatment for improving the biocompatibility of acrylic reline resins. BioMed Res Int 2013; http://dx.doi.org/10.1155/2013/485246
31. 31. Milheiro A, Nozaki K, Kleverlaan CJ, Muris J, Miura H, Feilzer A J. In vitro cytotoxicity of metallic ions released from dental alloys. Odontology 2016; 104(2): 136-142.