Decellularization and Delipidation Protocols of Bovine Bone and Pericardium for Bone Grafting and Guided Bone Regeneration Procedures.

Chiara Gardin,Riccardo Guazzo,Letizia Ferroni,Maurizio Isola,Luca Finotti,Sara Ricci,Giulia De Benedictis,Barbara Zavan,Luca Sbricoli,Eriberto Bressan

doi:10.1371/journal.pone.0132344

Chiara Gardin, Riccardo Guazzo + Show 8 more

Open Access

https://doi.org/10.1371/journal.pone.0132344

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Journal: PloS one	Publication Date: Jul 20, 2015
Citations: 84	License type: CC BY 4.0

Affiliation: University of Padua

Abstract

The combination of bone grafting materials with guided bone regeneration (GBR) membranes seems to provide promising results to restore bone defects in dental clinical practice. In the first part of this work, a novel protocol for decellularization and delipidation of bovine bone, based on multiple steps of thermal shock, washes with detergent and dehydration with alcohol, is described. This protocol is more effective in removal of cellular materials, and shows superior biocompatibility compared to other three methods tested in this study. Furthermore, histological and morphological analyses confirm the maintenance of an intact bone extracellular matrix (ECM). In vitro and in vivo experiments evidence osteoinductive and osteoconductive properties of the produced scaffold, respectively. In the second part of this study, two methods of bovine pericardium decellularization are compared. The osmotic shock-based protocol gives better results in terms of removal of cell components, biocompatibility, maintenance of native ECM structure, and host tissue reaction, in respect to the freeze/thaw method. Overall, the results of this study demonstrate the characterization of a novel protocol for the decellularization of bovine bone to be used as bone graft, and the acquisition of a method to produce a pericardium membrane suitable for GBR applications.

Highlights

In the field of oral surgery and dental implantology, bone deficiency represents the main problem that clinicians have to overcome in order to ensure the implant stability and the complete functional restoration
The use of bone grafting materials associated with the guided bone regeneration (GBR) technique seems to provide the most promising result to restore bone defects in dental clinical practice
In order to generate a resorbable membrane to be used in the GBR technique, in the second part of this study we developed a method of bovine pericardium decellularization

Summary

Introduction

In the field of oral surgery and dental implantology, bone deficiency represents the main problem that clinicians have to overcome in order to ensure the implant stability and the complete functional restoration. Osteoinduction refers to the ability of the scaffold to recruit multipotent mesenchymal stem cells (MSCs) from the surrounding tissue, and to induce their differentiation into bone-forming osteoblasts. An ideal bone graft should function through all three mechanisms by providing a substrate that directs three-dimensional (3D) bone growth, recruits and induces differentiation of resident bone-forming cells, and supplies more bone-forming cells to the recipient site. Another fundamental property of an ideal bone graft is osseointegration, which is the ability to bind to the surrounding bone without an intervening layer of fibrous tissue, allowing incorporation of the graft at the host site. Osseointegration is not an isolated phenomenon but depends on previous osteoinduction and osteoconduction [3]

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