Topical application of bFGF on acid-conditioned and non-conditioned dentin: effect on cell proliferation and gene expression in cells relevant for periodontal regeneration.

Fernanda Regina Godoy Rocha,João Antônio Chaves De Souza,Carlos Rossa Junior,José Eduardo Cezar Sampaio,Morgana Rodrigues Guimarães-Stabili

doi:10.1590/1678-7757-2017-0051

Abstract

Periodontal regeneration is still a challenge in terms of predictability and magnitude of effect. In this study we assess the biological effects of combining chemical root conditioning and biological mediators on three relevant cell types for periodontal regeneration.Material and Methods:Bovine dentin slices were conditioned with 25% citric acid followed by topical application of basic fibroblast growth factor (bFGF, 10 and 50 ng). We used ELISA to assess the dynamics of bFGF release from the dentin surface and RT-qPCR to study the expression of Runx2, Col1a1, Bglap and fibronectin by periodontal ligament (PDL) fibroblasts, cementoblasts and bone marrow stromal cells (BMSC) grown onto these dentin slices. We also assessed the effects of topical application of bFGF on cell proliferation by quantification of genomic DNA.Results:Acid conditioning significantly increased the release of bFGF from dentin slices. Overall, bFGF application significantly (p<0.05) increased cell proliferation, except for BMSC grown on non-conditioned dentin slices. Dentin substrate discretely increased expression of Col1a1 in all cell types. Expression of Runx2, Col1a1 and Fn was either unaffected or inhibited by bFGF application in all cell types. We could not detect expression of the target genes on BMSC grown onto conditioned dentin.Conclusion:Acid conditioning of dentin improves the release of topically-applied bFGF. Topical application of bFGF had a stimulatory effect on proliferation of PDL fibroblasts, cementoblasts and BMSC, but did not affect expression of Runx2, Col1a1, Bglap and fibronectin by these cells.

Highlights

Evidence indicates that current cause-related periodontal therapy is effective in stopping disease progression and reducing local inflammation, but regeneration of periodontal tissues lost as a consequence of disease still poses a challenge both in terms of predictability and magnitude of effect5
One such product includes in its surgical procedure protocol acid conditioning of the root surface to remove smear layer from the dentin4 and there is evidence indicating that chemical conditioning with EDTA improved PDGF-induced adhesion of periodontal OLJDPHQW ¿EUREODVWV LQ FRPSDULVRQ ZLWK WKH XVH RI PDGF without previous EDTA conditioning3
We determined the effect of the culture substrate by comparing the quantity of genomic DNA of cells cultured on cell culturetreated plastic or onto dentin slices. bFGF treatment RI SHULRGRQWDO OLJDPHQW ¿EUREODVWV JURZQ RQ SODVWLF VXEVWUDWH VLJQL¿FDQWO\ LQFUHDVHG FHOO SUROLIHUDWLRQ DW K:KHQWKHVH¿EUREODVWVZHUHJURZQRQWRGHQWLQ slices, topical application of bFGF increased cell

Summary

Introduction

Evidence indicates that current cause-related periodontal therapy is effective in stopping disease progression and reducing local inflammation, but regeneration of periodontal tissues lost as a consequence of disease still poses a challenge both in terms of predictability and magnitude of effect. Acid root conditioning is one of WKH¿UVWWKHUDSHXWLFDSSURDFKHVGHYHORSHGZLWKWKLV intent. There are products currently available commercially for applications in periodontal regeneration based on the biological properties of growth factors. One such product includes in its surgical procedure protocol acid conditioning of the root surface to remove smear layer from the dentin and there is evidence indicating that chemical conditioning with EDTA improved PDGF-induced adhesion of periodontal OLJDPHQW ¿EUREODVWV LQ FRPSDULVRQ ZLWK WKH XVH RI PDGF without previous EDTA conditioning

Methods

Results

Conclusion