Effect of Butyrate on Collagen Expression, Cell Viability, Cell Cycle Progression and Related Proteins Expression of MG-63 Osteoblastic Cells.

Mei-Chi Chang,Yi-Ling Tsai,Hsin-Cheng Liu,Ming-Wei Liao,Sin-Yuet Yeung,Eric Jein-Wein Liou,Chia-Mei Tang,Chiu-Po Chan,Jiiang-Huei Jeng,Tong-Mei Wang,Yuan-Soon Ho

doi:10.1371/journal.pone.0165438

Mei-Chi Chang, Yi-Ling Tsai + Show 9 more

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https://doi.org/10.1371/journal.pone.0165438

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Journal: PloS one	Publication Date: Nov 28, 2016
Citations: 14	License type: CC BY 4.0

Affiliation: Chang Gung Memorial Hospital

Abstract

AimsButyric acid is one major metabolic product generated by anaerobic Gram-negative bacteria of periodontal and root canal infection. Butyric acid affects the activity of periodontal cells such as osteoblasts. The purposes of this study were to investigate the effects of butyrate on MG-63 osteoblasts.MethodsMG-63 cells were exposed to butyrate and cell viability was estimated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The mRNA and protein expression of type I collagen and cell cycle-related proteins were measured by reverse-transcriptase polymerase chain reaction (RT-PCR), western blotting or immunofluorescent staining. Cellular production of reactive oxygen species (ROS) was analyzed by 2',7'-dichlorofluorescein (DCF) fluorescence flow cytometry.ResultsExposure to butyrate suppressed cell proliferation, and induced G2/M (8 and 16 mM) cell cycle arrest of MG-63 cells. Some cell apoptosis was noted. The mRNA expression of cdc2 and cyclin-B1 decreased after exposure to butyrate. The protein expression of type I collagen, cdc2 and cyclin B1 were decreased, whereas the expression of p21, p27 and p57 was stimulated. Under the treatment of butyrate, ROS production in MG-63 cells markedly increased.ConclusionsThe secretion of butyric acid by periodontal and root canal microorganisms may inhibit bone cell growth and matrix turnover. This is possibly due to induction of cell cycle arrest and ROS generation and inhibition of collagen expression. These results suggest the involvement of butyric acid in the pathogenesis of periodontal and periapical tissue destruction by impairing bone healing responses.

Highlights

Microorganisms are shown to play important roles in the diseased processes of periodontal and pulpal/periapical lesions
During the metabolism of amino acids, hexose or pentose by microorganisms, significant amounts of butyric acid are produced in the periodontal pockets and root canals [2,3,4,5], and affect the biological activities of adjacent periodontal cells
MG-63 osteoblastic cells were from American Type Culture Collection (ATCC, USA)

Summary

Introduction

Microorganisms are shown to play important roles in the diseased processes of periodontal and pulpal/periapical lesions. May be involved in the initiation and propagation of these diseased processes by generation a number of toxic products such as lipopolysaccharide, short chain fatty acids (SCFA), proteases etc. During the metabolism of amino acids, hexose or pentose by microorganisms, significant amounts of butyric acid are produced in the periodontal pockets and root canals [2,3,4,5], and affect the biological activities of adjacent periodontal cells (e.g., gingival fibroblasts, bone cells, periodontal ligament cells). The concentration of SCFAs (e.g., acetic acid, propionic acid and butyric acid) in gingival crevicular fluid (GCF) from diseased periodontal pocket is generally at mM concentration, and associated with the severity of periodontal diseases. The other paper shows the level of butyric acid to be 0.5–16 mM in GCF from sites with different diseased status [7]

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