Epithelial cell adhesion efficacy of a novel peptide identified by panning on a smooth titanium surface

Hidemichi Kihara,Hidemichi Kihara,Masazumi Nagai,Chia-Yu Chen,Toshiki Nojiri,Shigemi Nagai,Wataru Hatakeyama,Wataru Hatakeyama,Hisatomo Kondo,David M Kim,John Da Silva,Cliff Lee

doi:10.1038/s41368-018-0022-1

Abstract

Epithelial attachment via the basal lamina on the tooth surface provides an important structural defence mechanism against bacterial invasion in combating periodontal disease. However, when considering dental implants, strong epithelial attachment does not exist throughout the titanium-soft tissue interface, making soft tissues more susceptible to peri-implant disease. This study introduced a novel synthetic peptide (A10) to enhance epithelial attachment. A10 was identified from a bacterial peptide display library and synthesized. A10 and protease-activated receptor 4-activating peptide (PAR4-AP, positive control) were immobilized on commercially pure titanium. The peptide-treated titanium showed high epithelial cell migration ability during incubation in platelet-rich plasma. We confirmed the development of dense and expanded BL (stained by Ln5) with pericellular junctions (stained by ZO1) on the peptide-treated titanium surface. In an adhesion assay of epithelial cells on A10-treated titanium, PAR4-AP-treated titanium, bovine root and non-treated titanium, A10-treated titanium and PAR4-AP-treated titanium showed significantly stronger adhesion than non-treated titanium. PAR4-AP-treated titanium showed significantly higher inflammatory cytokine release than non-treated titanium. There was no significant difference in inflammatory cytokine release between A10-treated and non-treated titanium. These results indicated that A10 could induce the adhesion and migration of epithelial cells with low inflammatory cytokine release. This novel peptide has a potentially useful application that could improve clinical outcomes with titanium implants and abutments by reducing or preventing peri-implant disease.

Highlights

The structure and mechanism of epithelial attachment around dental implants and abutments are different from those around natural teeth.[1]
Peri-implantitis is a significant complication of dental implants that is caused by bacterial infection; it is difficult to predictably treat and can lead to implant failure.[11,12]
The five novel peptides identified in the present study showed comparable or better effects than protease-activated receptor-4 activating peptide (PAR4-AP) in terms of the induction of epithelial chemokines and growth factors

Summary

INTRODUCTION

The structure and mechanism of epithelial attachment around dental implants and abutments are different from those around natural teeth.[1]. Sugawara et al.[16] reported the establishment of BL-mediated epithelial cell attachment to the treated smooth titanium surface through the use of platelets. Our previous study indicated that the platelet-induced epithelial sheet on PAR4-AP-treated titanium blocked bacterial invasion.[17]. This result indicated the establishment of strong epithelial adhesion on the smooth titanium surface, but the adhesion strength could not be quantified. The induction chemokines and growth factors and establish strong epithelial of EGF (Fig. 1a) and IGF-1 (Fig. 1b) by 96 peptides identified in the attachment. We introduced novel peptides selected from a peptides for EGF and IGF-1 induction included A10, A1, B11, D2, bacterial peptide library that promoted increased chemokine and and A6 (Fig. 1c). The top 15 peptides that stimulated release are shown in c

A1: CGPAEKAYPNNSPLFGPC TGCGGCCCAGCGGAAAAGGCTTATCCAAATAATTCCCCGCTTTTTGGCCCCTGT A10

DISCUSSION

MATERIALS AND METHODS