Introducing biological factors into dental implant by phase-transited lysozyme priming and layer-by-layer self-assembly system

概要

Purpose: Icariin (ICA) is one of the main active constituents of Herba Epimedii for improving osteogenesis. It is necessary to create a simple and efficient method to load ICA onto the surface of titanium (Ti) implant. The purpose of this study was to establish a local ICA delivery system via a layer-by-layer self-assembly (LbL) system on phase-transited lysozyme (PTL) primed Ti surface.

Materials and methods: A PTL nanofilm was first firmly coated on the pristine Ti. Then, the ICA loaded hyaluronic acid/chitosan (HA/CS) multilayer was applied via the LbL system to form the HA/CS-ICA surface. This established HA/CS-ICA surface was characterized by scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and contact angle measurement. The ICA release pattern of the HA/CS-ICA surface was also examined. MC3T3-E1 osteoblasts and RAW264.7 macrophages culture test and a rat model were used to evaluate the effects of the HA/CS-ICA surface in vitro and in vivo

Results: SEM, XPS and contact angle measurement demonstrated successful fabrication of the HA/CS-ICA surface. The HA/CS-ICA surfaces with different ICA concentrations all revealed a controlled release profile of ICA during a 2-week monitoring span. Osteoblasts grown on the coated substrates displayed higher adhesion, viability, proliferation and alkaline phosphatase activity than those on the polished Ti surface. Macrophages grown on the coated substrates showed higher adhesion, proliferation and M2 polarization than those on the polished Ti surface. Furthermore, in vivo histological evaluation revealed much obvious bone formation in the ICA coated group by histological staining and double fluorescent labeling at 2 weeks after implantation.

Conclusion: The present study demonstrated that ICA immobilized HA/CS multilayer on the PTL primed Ti surface had a sustained release pattern of ICA which could promote the osteogenesis of osteoblasts, modify the macrophage polarization state in vitro and improve early osseointegration in vivo. This study provides a novel method for creating a sustained ICA delivery system to improve osteoblasts and macrophages response and osseointegration.