Sepehr Talebian

Nanoreinforced Hydrogels for Tissue Engineering: Biomaterials that are Compatible with Load-Bearing and Electroactive Tissues

Given their highly porous nature and excellent water retention, hydrogel-based biomaterials can mimic critical properties of the native cellular environment. However, their potential to emulate the electromechanical milieu of native tissues or conform well with the curved topology of human organs needs to be further explored to address a broad range of physiological demands of the body. In this regard, the incorporation of nanomaterials within hydrogels has shown great promise, as a simple one-step approach, to generate multifunctional scaffolds with previously unattainable biological, mechanical, and electrical properties. Here, recent advances in the fabrication and application of nanocomposite hydrogels in tissue engineering applications are described, with specific attention toward skeletal and electroactive tissues, such as cardiac, nerve, bone, cartilage, and skeletal muscle. Additionally, some potential uses of nanoreinforced hydrogels within the emerging disciplines of cyborganics, bionics, and soft biorobotics are highlighted.

A Comparative Study on In Vitro Osteogenic Priming Potential of Electron Spun Scaffold PLLA/HA/Col, PLLA/HA, and PLLA/Col for Tissue Engineering Application

A comparative study on the in vitro osteogenic potential of electrospun poly-L-lactide/hydroxyapatite/collagen (PLLA/HA/Col, PLLA/HA, and PLLA/Col) scaffolds was conducted. The morphology, chemical composition, and surface roughness of the fibrous scaffolds were examined. Furthermore, cell attachment, distribution, morphology, mineralization, extracellular matrix protein localization, and gene expression of human mesenchymal stromal cells (hMSCs) differentiated on the fibrous scaffolds PLLA/Col/HA, PLLA/Col, and PLLA/HA were also analyzed. The electrospun scaffolds with a diameter of 200–950 nm demonstrated well-formed interconnected fibrous network structure, which supported the growth of hMSCs. When compared with PLLA/H%A and PLLA/Col scaffolds, PLLA/Col/HA scaffolds presented a higher density of viable cells and significant upregulation of genes associated with osteogenic lineage, which were achieved without the use of specific medium or growth factors. These results were supported by the elevated levels of calcium, osteocalcin, and mineralization (P<0.05) observed at different time points (0, 7, 14, and 21 days). Furthermore, electron microscopic observations and fibronectin localization revealed that PLLA/Col/HA scaffolds exhibited superior osteoinductivity, when compared with PLLA/Col or PLLA/HA scaffolds. These findings indicated that the fibrous structure and synergistic action of Col and nano-HA with high-molecular-weight PLLA played a vital role in inducing osteogenic differentiation of hMSCs. The data obtained in this study demonstrated that the developed fibrous PLLA/Col/HA biocomposite scaffold may be supportive for stem cell based therapies for bone repair, when compared with the other two scaffolds.

Chitosan (PEO)/bioactive glass hybrid nanofibers for bone tissue engineering

A novel hybrid nanofibrous scaffold prepared with chitosan [containing 1.2 wt% polyethylene oxide (PEO)] and bioactive glass (BG) was fabricated by an electrospinning technique. The morphological and physicochemical properties of scaffolds were studied by scanning electron microscopy (SEM) and spectroscopy. The measurements of tensile strength and water-contact angles suggested that the incorporation of BG into the nanofibers improves the mechanical properties and hydrophilicity of the scaffolds. Biomineralization of the nanofibers was evaluated by soaking them in simulated body fluid (SBF), and the formation of hydroxycarbonate apatite (HCA) layer was determined by EDX and FE-SEM. The results showed that BG-containing nanofibers could induce the formation of HCA on the surface of the composite after 14 days of immersion in SBF. In vitro-cell viability of human mesenchymal stromal cells (hMSCs) on nanofibers was assessed by using the MTT assay. The cell-adhesion results showed that hMSCs were viable at variable time points on the chitosan/PEO/BG nanofiber scaffolds. In addition, the presence of BG enhanced the alkaline phosphatase (ALP) activity of hMSCs cultured on composite scaffolds at day 14 compared to that on pure chitosan/PEO scaffolds. Our results suggest that a chitosan/PEO/BG nanofibrous composite could be a potential candidate for application in tissue engineering.

Fabrication and in vitro biological activity of βTCP-Chitosan-Fucoidan composite for bone tissue engineering

We developed tricalcium phosphate-chitosan-fucoidan biocomposite scaffold (TCP-Ch-Fu) by using the freeze-drying technique. The fabricated biocomposite scaffolds were analyzed by spectroscopy and porosity measurement. The biomechanical properties of scaffolds were assessed by compression test and the results suggested that the incorporation of Fucoidan into the biocomposite improves the compression strength of scaffolds. Biomineralization of scaffolds was evaluated by soaking them in simulated body fluid and the results revealed that the addition of Fucoidan into the scaffolds enhanced the formation of apatite layer on the surface of biocomposite after 7 days of immersion. Alamar Blue assay confirmed that the cell viability of human-derived bone marrow stromal cell was superior in the TCP-Ch-Fuscaffold. The addition of Fucoidan to TCP-Ch increased the release of osteocalcin, confirming that it can support osteogenic differentiation of human mesenchymal stromal cells in in vitro culture. Thus, TCP-Ch-Fu could be a potential candidate for bone-tissue engineering applications.

Electrospinning of polymethyl methacrylate nanofibers: optimization of processing parameters using the Taguchi design of experiments

The effects of polymer concentration and electrospinning parameters on the diameter of electrospun polymethyl methacrylate (PMMA) fibers were experimentally investigated. It was also studied how the controlled factors would affect the output with the intention of finding the optimal electrospinning settings in order to obtain the smallest PMMA fiber diameter. Subsequently the solution feed rate, needle gauge diameter, supply voltage, polymer concentration and tip-to-collector distance were considered as the control factors. To achieve these aims, Taguchi’s mixed-level parameter design (L18) was employed for the experimental design. Optimal electrospinning conditions were determined using the signal-to-noise (S/N) ratio that was calculated from the electrospun PMMA fiber diameter according to “the-smaller-the-better” approach. Accordingly, the smallest fiber diameter observed was 228 (±76) nm and it was yielded at 15 wt% polymer concentration, 20 kV of supply voltage, 1 ml/h feed rate, 15 cm tip-to-distance and 19 needle gauge. Moreover, the S/N ratio response showed that the polymer concentration was the most effective parameter on determination of fiber diameter followed by feed rate, tip-to distance, needle gauge and voltage, respectively. The Taguchi design of experiments method has been found to be an effective approach to statistically optimize the critical parameters used in electrospinning so as to effectively tailor the resulting electrospun fiber diameters and morphology.

Incorporation of Fucoidan in β-Tricalcium phosphate-Chitosan scaffold prompts the differentiation of human bone marrow stromal cells into osteogenic lineage

In our previous study, we reported the fabrication and characterization of a novel tricalcium phosphate-fucoidan-chitosan (TCP-Fu-Ch) biocomposite scaffold. However, the previous report did not show whether the biocomposite scaffold can exhibit osteogenic differentiation of human bone marrow stromal cells in osteogenic media and normal media supplemented with platelet-derived growth factor (PDGF-BB). On day 15, the release of osteocalcin, was significant in the TCP-Fu-Ch scaffold, when compared with that in the TCP-Ch scaffold, and the level of release was approximately 8 and 6 ng/ml in osteogenic and normal media supplemented with PDGF-BB, respectively. Scanning electron microscopy of the TCP-Fu-Ch scaffold demonstrated mineralization and apatite layer formation on day 14, while the addition of PDGF-BB also improved the osteogenic differentiation of the scaffold. An array of gene expression analysis demonstrated that TCP-Fu-Ch scaffold cultured in osteogenic and normal media supplemented with PDGF-BB showed significant improvement in the expression of collagen 1, Runt-related transcription factor 2, osteonectin, bone gamma-carboxyglutamate protein, alkaline phosphatase, and PPA2, but a decline in the expression of integrin. Altogether, the present study demonstrated that fucoidan-incorporated TCP-Ch scaffold could be used in the differentiation of bone marrow stromal cells and can be a potential candidate for the treatment of bone-related ailments through tissue engineering technology.

Synergistic interaction of platelet derived growth factor (PDGF) with the surface of PLLA/Col/HA and PLLA/HA scaffolds produces rapid osteogenic differentiation.

Scaffolds with structural features similar to the extracellular matrix stimulate rapid osteogenic differentiation in favorable microenvironment and with growth factor supplementation. In this study, the osteogenic potential of electrospun poly-l-lactide/hydroxyapatite/collagen (PLLA/Col/HA, PLLA/HA and PLLA/Col) scaffolds were tested in vitro with the supplementation of platelet derived growth factor-BB (PDGF-BB). Cell attachment and topography, mineralization, extracellular matrix protein localization, and gene expression of the human mesenchymal stromal cells were compared between the fibrous scaffolds PLLA/Col/HA, PLLA/Col, and PLLA/HA. The levels of osteocalcin, calcium, and mineralization were significantly greater in the PLLA/Col/HA and PLLA/HA compared with PLLA/Col. High expression of fibronectin, intracellular adhesion molecule, cadherin, and collagen 1 (Col1) suggests that PLLA/Col/HA and PLLA/HA scaffolds had superior osteoinductivity than PLLA/Col. Additionally, osteopontin, osteocalcin, osterix, Runt-related transcription factor 2 (Runx2), and bone morphogenic protein (BMP2) expression were higher in PLLA/Col/HA and PLLA/HA compared with PLLA/Col. In comparison with PLLA/Col, the PLLA/Col/HA and PLLA/HA scaffolds presented a significant upregulation of the genes Runx2, Col 1, Integrin, osteonectin (ON), bone gamma-carboxyglutamic acid-containing protein (BGALP), osteopontin (OPN), and BMP2. The upregulation of these genes was further increased with PDGF-BB supplementation. These results show that PDGF-BB acts synergistically with PLLA/Col/HA and PLLA/HA to enhance the osteogenic differentiation potential. Therefore, this combination can be used for the rapid expansion of bone marrow stromal cells into bone-forming cells for tissue engineering.

Effect of deacetylation on property of electrospun chitosan/PVA nanofibrous membrane and removal of methyl orange, Fe(III) and Cr(VI) ions

In this study, effect of degree of deacetylation on property and adsorption capacity of chitosan/polyvinyl Alcohol electrospun membrane has been investigated. Resulting nanofibers were characterized by FESEM, FTIR, XRD, TGA, tensile testing, weight loss test and adsorption test. FESEM result shows, finer nanofiber was fabricated from 42h hydrolyzed chitosan and PVA blend solution. FTIR and XRD result showed a strong interaction between chitosan and polyvinyl alcohol. Higher tensile strength was observed for the nanofiber having 42h hydrolyzed chitosan. Blend solution of chitosan/PVA having low DD chitosan had higher viscosity. The nanofibrous membrane was stable in distilled water, acidic and basic medium. The isotherm study shows that the adsorption capacity (qm) of nanofiber containing higher DD chitosan was higher for Cr(VI). In contrary, the membrane containing chitosan with lower DD showed the higher adsorption capacity for Fe(III) and methyl orange. Moreover, the effect of DD on removal percentage of adsorbate was dependent on the initial concentration of the adsorbate.

Fabrication and characterisation of chitosan/ poly vinyl alcohol nanofibres via electrospinning

Abstract In this study, chitosan nanofibres were fabricated using electrospinning process. Initially, chitosan powders were further deacetylated with sodium hydroxide to deprotonate the polymer chains and decrease the molecular weight. Subsequently, the treated chitosan (5 wt-% in 90:10 acetic acid/water ratio) were mixed with poly vinyl alcohol (8 wt-% in water) at different ratios in order to facilitate the electrospinning through cross-links formation. Field-emission scanning electron microscopic images showed that 60:40 (chitosan/poly vinyl alcohol) ratio was the threshold for formation of beads (chitosan/poly vinyl alcohol >50:50 contains imperfections) and that increasing chitosan/poly vinyl alcohol ratio from 20:80 to 60:40 decreased the average diameter of fibres from 80 to 40 nm. Fourier transform infrared spectra proved the formation of chitosan/poly vinyl alcohol cross-linked nanofibres.

Preparation and characterisation of electrospun silica nanofibres

Abstract In this study, the silica electrospun nanofibres were fabricated using silica sol containing tetraethylorthosilicate, polyvinylpyrrolidone and butanol (solvent). The perfect concentration for silica sol to undergo the electrospinning was found to be 0·095 gr(pvp)/mL (butanol, tetraethyl orthosilicate), where the tetraethyl orthosilicate/butanol volume ratio was 3:2. Later with the intention of achieving the silica nanofibres, the electrospun samples afterwards were put in a furnace at 700°C for 3 hours. After that the morphological studies on the fibres before and after the thermal treatment were done employing the scanning electron microscopy. The results showed that fibre diameter before thermal treatment fits the micrometre scale, while it decreases dramatically to the 260–360 nm range when it goes through the thermal stage. Furthermore, Fourier transform infrared spectroscopy and energy-dispersive X-ray spectroscopy spectrums proved that polyvinylpyrrolidone and butanol were removed from the fibres after calcination and that the silica nanofibres were fabricated.