Cytotoxicity and effect on protease activity of copolymer extracts containing catechin

Author(s)

Zarella, Bruno Lara ^[1] ; Rabelo Buzalaf, Marilia Afonso ^[1] ; Kato, Melissa Thiemi ^[2] ; Hannas, Angelica Reis ^[1] ; Salo, Tuula ^{[3, 4, 5, 6, 7]} ; Tjaderhane, Leo ^{[3, 4, 5, 6, 7]} ; Prakki, Anuradha ^[8]

Affiliation

^[1] Univ Sao Paulo, Bauru Sch Dent, Dept Biol Sci, BR-17012901 Bauru, SP - Brazil

^[2] Univ Sagrado Coracao, Dept Dent, BR-17011160 Bauru, SP - Brazil

^[3] Univ Oulu, Inst Dent, FI-90014 Oulu - Finland

^[4] Univ Oulu, FI-90014 Oulu - Finland

^[5] Oulu Univ Hosp, Med Res Ctr Oulu, FI-90014 Oulu - Finland

^[6] Univ Helsinki, Dept Oral & Maxillofacial Dis, FI-00014 Helsinki - Finland

^[7] Helsinki Univ Hosp, FI-00014 Helsinki - Finland

^[8] Univ Toronto, Fac Dent, Dept Clin Sci Restorat, Toronto, ON M5G 1G6 - Canada

• Document type: Journal article
• Source: ARCHIVES OF ORAL BIOLOGY; v. 65, p. 66-71, MAY 2016.
• Web of Science Citations: 3

Abstract

Objective: To evaluate cytotoxicity and effect on protease activity of epigallocatechin-gallate extracted from experimental restorative dental copolymers in comparison to the control compound chlorhexidine. Methods: Copolymer disks were prepared from bis-GMA/TEGDMA (70/30 mol%) containing no compound (control) or 1% w/w of either epigallocatechin-gallate or chlorhexidine. MDPC-23 odontoblast-like cells were seeded with the copolymer extracts leached out into deionized water. Cell metabolic activity was quantified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MIT) assay at 24, 48, 72 h. Inhibition of protease activity by resin extracts was measured by a collagenolytic/genatinolytic enzyme activity assay and gelatin zymography. Data for MIT and protease inhibition were analyzed using two-way ANOVA followed by Tukey or Bonferroni post hoc tests (alpha = 0.05). Results: The MIT revealed that at 72 h, extracts from control (16.7%) and chlorhexidine (22.3%) copolymers induced significant reduction in cell metabolism (p < 0.05). All copolymer extracts caused enzymatic inhibition in a dose dependent manner (p < 0.01). Even when highly diluted, epigallocatechingallate extract had a significant antiproteolytic activity (p < 0.05). Zymograms showed that all extracts reduced activity of MMP-2 and MMP-9 (pro- and active forms), with MMP-9 exhibiting the highest percentage inhibition revealed by densitometry. Conclusions: Epigallocatechin-gallate and chlorhexidine extracts did not exert cytotoxicity on evaluated cells when compared to control extracts. Both compounds retained antiproteolytic activity after extraction from a dental copolymer. Clinical significance: Once extracted from a dental copolymer, epigallocatechin-gallate is not cytotoxic and retains antiproteolytic activity. These results may allow incorporation of epigallocatechin-gallate as a natural-safe alternative to chlorhexidine in functionalized restorative materials. (C) 2016 Elsevier Ltd. All rights reserved. (AU)

• FAPESP's process: 11/03618-0 - Effects of experimental resins containing inhibitor of matrix metaloproteinases (MMPs) on gelatinases
• Grantee: Bruno Lara Zarella
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 12/03033-4 - Effects of experimental resins containing inhibitor of matrix metaloproteinases (MMPs) on gelatinases
• Grantee: Bruno Lara Zarella
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree