In this study, pure silica nanofibers (SNFs) were fabricated by the electrospinning technique. Subsequently, the as-prepared SNFs were modified with (3-aminopropyl) trimethoxysilane (APTS) for applications in neural tissue engineering. The structure and properties of the as-prepared SNFs and the modified SNFs (SNFAPxM, x ¼ 1-3) were evaluated with FTIR, TGA, nitrogen adsorption/desorption isotherms, and SEM. It was found that the surface hydrophilicity of SNF-APxM was lowered upon increasing the amino alkyl group content. The SEM and confocal images revealed that neural stem cells (NSCs) cultured on the electrospun SNFs and SNF-APxM substrates were elongated along the fibers in comparison to poly-Dlysine-coated (PDL-coated) substrate. In addition, a higher degree of proliferation and more responsive cells were observed for the NSCs cultured on the SNF-AP3M substrate than those on the SNFs and the PDL-coated substrates. The results indicated that the APTS-modified silica nanofibers can be potential substrates for regulating growth and differentiation of NSCs in culture.
In this study, we first synthesized a slow-degrading silica nanofiber (SNF2) through an electrospun solution with an optimized tetraethyl orthosilicate (TEOS) to polyvinyl pyrrolidone (PVP) ratio. Then, laminin-modified SNF2, namely SNF2-AP-S-L, was obtained through a series of chemical reactions to attach the extracellular matrix protein, laminin, to its surface. The SNF2-AP-S-L substrate was characterized by a combination of scanning electron microscopy (SEM), Fourier transform–infrared (FTIR) spectroscopy, nitrogen adsorption/desorption isotherms, and contact angle measurements. The results of further functional assays show that this substrate is a biocompatible, bioactive and biodegradable scaffold with good structural integrity that persisted beyond 18 days. Moreover, a synergistic effect of sustained structure support and prolonged biochemical stimulation for cell differentiation on SNF2-AP-S-L was found when neuron-like PC12 cells were seeded onto its surface. Specifically, neurite extensions on the covalently modified SNF2-AP-S-L were significantly longer than those observed on unmodified SNF and SNF subjected to physical adsorption of laminin. Together, these results indicate that the SNF2-AP-S-L substrate prepared in this study is a promising 3D biocompatible substrate capable of sustaining longer neuronal growth for tissue-engineering applications.
In this study, a previously known high-affinity silica binding protein (SB) was genetically engineered to fuse with an integrin-binding peptide (RGD) to create a recombinant protein (SB-RGD). SB-RGD was successfully expressed in Escherichia coli and purified using silica beads through a simple and fast centrifugation method. A further functionality assay showed that SB-RGD bound to the silica surface with an extremely high affinity that required 2 M MgCl2 for elution. Through a single-step incubation, the purified SB-RGD proteins were noncovalently coated onto an electrospun silica nanofiber (SNF) substrate to fabricate the SNF-SB-RGD substrate. SNF-SB-RGD was characterized by a combination of scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, and immunostaining fluorescence microscopy. As PC12 cells were seeded onto the SNF-SB-RGD surface, significantly higher cell viability and longer neurite extensions were observed when compared to those on the control surfaces. These results indicated that SB-RGD could serve as a noncovalent coating biologic to support and promote neuron growth and differentiation on silica-based substrates for neuronal tissue engineering. It also provides proof of concept for the possibility to genetically engineer protein-based signaling molecules to noncovalently modify silica-based substrates as bioinspired material.
A detailed genomic and epigenomic analyses of neural stem cells (NSCs) differentiation in synthetic microenvironments is essential for the advancement of regenerative medicine and therapeutic treatment of diseases. This study identified the changes in mRNA and miRNA expression profile during NSC differentiation on an artificial matrix. NSCs were grown on a surface-modified, electrospun tetraethyl-orthosilicate nanofiber (designated as SNF-AP) by providing a 3D-environment for cell growth and differentiation. Differentially expressed mRNAs and miRNAs of NSC differentiated in this microenvironment were identified through microarray analysis. The genes and miRNA targets responsible for the differentiation fate of NSCs and neuron development process were determined using Ingenuity Pathway Analysis (IPA). SNF-AP enhanced the expression of genes that activates the proliferation, development, and outgrowth of neurons, differentiation and generation of cells, neuritogenesis, outgrowth of neurites, microtubule dynamics, formation of cellular protrusions, and long-term potentiation during NSC differentiation. On the other hand, PDL inhibited neuritogenesis, microtubule dynamics, and proliferation and differentiation of cells and activated the apoptosis function. Moreover, the nanomaterial promoted the expression of more let-7 miRNAs, which have vital roles in NSC differentiation. Overall, SNF-AP is biocompatible and applicable scaffold for NSC differentiation in the development of neural tissue engineering. These findings are useful in enhancing in vitro NSC differentiation potential for preclinical studies and future clinical applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2730-2743, 2016.
In this work, silica nanofibers (SNFs) were prepared by an electrospinning method and modified with poly- d -lysine (PDL) or (3-aminopropyl) trimethoxysilane (APTS) making biocompatible and degradable substrates for neuronal growth. The as-prepared SNF, modified SNF-PDL, and SNF-APTS were evaluated using scanning electron microscopy, nitrogen adsorption/desorption isotherms, contact angle measurements, and inductively coupled plasma atomic emission spectroscopy. Herein, the scanning electron microscopic images revealed that dissolution occurred in a corrosion-like manner by enlarging porous structures, which led to loss of structural integrity. In addition, covalently modified SNF-APTS with more hydrophobic surfaces and smaller surface areas resulted in significantly slower dissolution compared to SNF and physically modified SNF-PDL, revealing that different surface modifications can be used to tune the dissolution rate. Growth of primary hippocampal neuron on all substrates led to a slower dissolution rate. The three-dimensional SNF with larger surface area and higher surface density of the amino group promoted better cell attachment and resulted in an increased neurite density. This is the first known work addressing the degradability of SNF substrate in physiological conditions with neuron growth in vitro, suggesting a strong potential for the applications of the material in controlled drug release.
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