Abstract
Multifunctional nanofibrous scaffolds for effective bone tissue engineering (BTE) application must incorporate factors to promote neovascularization and tissue regeneration. In this study, silica-coated gold nanoparticles Au(SiO2) were tested for their ability to promote differentiation of human mesenchymal stem cells (hMSCs) into osteoblasts. Biocompatible poly-ε-caprolactone (PCL), PCL/silk fibroin (SF) and PCL/SF/Au(SiO2) loaded nanofibrous scaffolds were first fabricated by an electrospinning method. Electrospun nanofibrous scaffolds were characterized for fiber architecture, porosity, pore size distribution, fiber wettability and the relevant mechanical properties using field emission scanning electron microscopy (FESEM), porosimetry, determination of water contact angle, measurements by a surface analyzer and tabletop tensile-tester measurements. FESEM images of the scaffolds revealed beadless, porous, uniform fibers with diameters in the range of 164 ± 18.65 nm to 215 ± 32.12 nm and porosity of around 88–92% and pore size distribution around 1.45–2.35 µm. Following hMSCs were cultured on the composite scaffolds. Cell-scaffold interaction, morphology and proliferation of were analyzed by FESEM analysis, MTS (3-(4,5-dimethyl thiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt) and CMFDA (5-choromethyl fluorescein acetate) dye assays. Osteogenic differentiation of MSCs into osteogenic cells were determined by alkaline phosphatase (ALP) activity, mineralization by alizarin red S (ARS) staining and osteocalcin expression by immunofluorescence staining. The results revealed that the addition of SF and Au(SiO2) to PCL scaffolds enhanced the mechanical strength, interconnecting porous structure and surface roughness of the scaffolds. This, in turn, led to successful osteogenic differentiation of hMSCs with improved cell adhesion, proliferation, differentiation, mineralization and expression of pro-osteogenic cellular proteins. This provides huge support for Au(SiO2) as a suitable material in BTE.
Highlights
Bone, which is an integral part of the skeleton, is known to be active throughout its lifetime as it undertakes structural remodeling in order to provide mechanical protection for internal organs
Silica is a vital substance for the formation of bone as it helps to improve the function of osteoblasts, inhibits the function of osteoclasts and promote mineralization by precipitating calcium phosphate in its early stages [14]
The surface morphology of electrospun nanofibrous scaffolds were analyzed with field emission scanning electron microscopy (FESEM) at an accelerating voltage of 10kV by sputter coating
Summary
Bone, which is an integral part of the skeleton, is known to be active throughout its lifetime as it undertakes structural remodeling in order to provide mechanical protection for internal organs. Micro and nano structural scaffolds with a high surface area to volume ratios, an interconnected porous structure and suitable mechanical properties can be fabricated using synthetic and natural polymer, thereby creating a fibrous matrix that closely mimics the ECM and provides support for the healing process [10]. AuNPs are appropriate agents for functionalization of electrospun scaffolds for bone regeneration, due to their properties of encouraging osteogenic differentiation in stem cells and osteoclast reticence [12,22,23,24] They are water-soluble and hold desirable features, such as biocompatibility and ease of synthesis [25]. PCL, PCL/SF and PCL/SF/Au(SiO2) composite nanofibrous scaffolds were fabricated by electrospinning
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