Evaluation of a Decellularisation Process for Developing an Osteogenic Periosteum Membrane In Vitro

Abstract

Introduction

The study addresses the need for a biocompatible, resorbable membrane with osteogenic properties, proposing a periosteum-based scaffold because of its inherent growth proteins and pliability.

Methods

The skin from the skull of a young sheep was removed to expose the connective tissue. A 2 cm x 2 cm periosteum sample was harvested and divided into 4 groups for different decellularisation treatments: sample A (control) was left untreated, sample B was treated with 100% alcohol, sample C was treated with 100% alcohol and 1% sodium hypochlorite, and sample D was treated with 100% alcohol, 1% sodium hypochlorite and 0.1% SDS. The processed periosteum was lyophillised and subjected to histological evaluation, scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDS), RAMAN spectroscopy, cell attachment and viability studies. Statistical analysis was performed using GraphPad Prism.

Results

Group IV scaffold, treated with 100% alcohol, 1% sodium hypochlorite and 0.1% SDS, exhibited the most desirable characteristics for tissue engineering. SEM-EDS analysis revealed a clean, residue-free, well-structured collagen fibril network with significant porosity, while RAMAN spectroscopy confirmed preservation of collagen's triple-helical structure. Histological evaluation showed thorough decellularisation and the absence of cellular remnants. DAPI staining and SEM imaging demonstrated robust osteoblast attachment and proliferation over 24, 48 and 72 hours, suggesting its biomimetic property.

Conclusion

Decellularised ovine periosteum membrane from Group IV demonstrated biocompatibility and supported osteoblast growth, indicating its potential as a soft, osteogenic scaffold for repairing periodontal bone lesions. Further in vivo studies are recommended to confirm its effectiveness in treating periodontal defects for guided bone regeneration.

Clinical relevance

The optimised ovine periosteum scaffold demonstrates promising osteogenic properties and biocompatibility, making it a potential candidate for periodontal bone defect repair. Its resorbable nature and structural integrity offer advantages over synthetic membranes. This novel membrane aligns with the global goals of sustainable health care and waste reduction. Further in vivo validation is warranted before clinical application.