Abstract

Recently, stem cell-based bone tissue engineering (BTE) has been recognized as a preferable and clinically significant strategy for bone repair. In this study, a pure 3D silk fibroin (SF) scaffold was fabricated as a BTE material using a lyophilization method. We aimed to investigate the efficacy of the SF scaffold with and without seeded human adipose-derived mesenchymal stem cells (hASCs) in facilitating bone regeneration. The effectiveness of the SF-hASCs scaffold was evaluated based on physical characterization, biocompatibility, osteogenic differentiation in vitro, and bone regeneration in critical rat calvarial defects in vivo. The SF scaffold demonstrated superior biocompatibility and significantly promoted osteogenic differentiation of hASCs in vitro. At six and twelve weeks postimplantation, micro-CT showed no statistical difference in new bone formation amongst all groups. However, histological staining results revealed that the SF-hASCs scaffold exhibited a better bone extracellular matrix deposition in the defect regions compared to other groups. Immunohistochemical staining confirmed this result; expression of osteoblast-related genes (BMP-2, COL1a1, and OCN) with the SF-hASCs scaffold treatment was remarkably positive, indicating their ability to achieve effective bone remodeling. Thus, these findings demonstrate that SF can serve as a potential carrier for stem cells, to be used as an osteoconductive bioscaffold for BTE applications.

Highlights

  • Bone defects, caused by diseases or traumas, are still clinically challenging

  • Silk Fibroin (SF) scaffolds were sterilized overnight by UV irradiation, followed by human adipose-derived mesenchymal stem cells (hASCs) seeding onto the scaffolds at a density of 1 × 106 cells cm−2 and incubating under 5% CO2 at 37 ◦ C in keratinocyte serum-free medium (KSFM) supplemented with 10% fetal bovine serum (FBS)

  • Fourier Transform Infra-Red (FTIR) was performed to examine the structural characteristics of the SF scaffold compared to was performed to examine the2A, structural of the SFbands scaffold to the the SFFTIR

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Summary

Introduction

Bone defects, caused by diseases or traumas, are still clinically challenging. With advancing age, they can lead to a life-long disability in many patients [1]. Successful treatment of bone defects depends on the interaction between the injured area and the implant such that any missing piece of bone is replaced with a proper functional substitute. Bone tissue engineering (BTE) offers potential strategies for bone defect therapies and remains to gain clinical significance annually [5]. The strategies of functional temporary matrices including scaffolds, cells, growth factors, and their interrelation in the microenvironment are crucial and major concerns for the reconstruction of the defective bone

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