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Release of bio‐active dentine extracellular matrix components by histone deacetylase inhibitors ( HDAC i)
Author(s) -
Duncan H. F.,
Smith A. J.,
Fleming G. J. P.,
Reid C.,
Smith G.,
Cooper P. R.
Publication year - 2017
Publication title -
international endodontic journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.988
H-Index - 119
eISSN - 1365-2591
pISSN - 0143-2885
DOI - 10.1111/iej.12588
Subject(s) - chemistry , histone deacetylase , extracellular matrix , microbiology and biotechnology , trichostatin a , biochemistry , histone , biology , gene
Aim To characterize dentine matrix component ( DMC ) release and smear layer removal by histone deacetylase inhibitors ( HDAC is). Methodology DMC s were extracted from powdered human dentine over 14 days using three HDAC is, valproic acid ( VPA ), trichostatin A ( TSA ) and suberoylanilide hydroxamic acid ( SAHA ) and compared with a control extractant, 10% (w/v) EDTA . Protein compositions of the resultant extracts were analysed by 1D‐polyacrylamide gel electrophoresis (1D‐ PAGE ), TGF ‐β‐1 and MMP ‐9 ELISA s and a high‐throughput growth factor antibody array. Dentine discs with a standardized smear layer were prepared from human molars and treated with EDTA (17% w/v), polyacrylic acid ( PA ) (20% v/v) and the experimental HDAC is prior to analysis by scanning electron microscopy. Parametric ELISA data were analysed using one‐way anova and Tukey's post hoc test, whilst nonparametric smear layer data were analysed by Kruskal–Wallis test and Mann–Whitney U ‐test ( P < 0.05). Results HDAC is did not remove smear layer in the presence or absence of PA pre‐treatment ( P ≥ 0.478). 1D‐ PAGE analysis demonstrated different protein profiles for EDTA and HDAC i extracts. All HDAC i solutions released TGF ‐β‐1 although less effectively than EDTA ( P < 0.001), whilst MMP ‐9 was extracted in significantly higher concentration by EDTA and VPA compared with TSA ( P < 0.012). Antibody array analysis demonstrated the ability of HDAC is to extract a complex cocktail of established/novel growth factors from dentine, albeit significantly less efficiently than EDTA for certain cytokines ( TGF ‐β‐1, PDGF ‐ AA , VEGF ‐A) and significantly more effectively for others ( GDF ‐15, IGF ‐1, EGRF ‐1, NGFR , BDNF , SCF ‐R). Conclusions HDAC i release a range of bioactive DMC s that could promote dentine repair processes in vivo ; however, they are ineffective at removing smear layer.