Neuronal differentiation of mesenchymal stem cells by polyvinyl alcohol/Gelatin/crocin and beta-carotene.

BACKGROUND: Nerve tissues are important in coordinating the motions and movements of the body. Nerve tissue repair and regeneration is a slow process that might take a long time and cost a lot of money. As a result, tissue engineering was employed to treat nerve tissue lesions. The aim of this study was to investigate the proliferation of C6 cells and human mesenchymal stem cells derived bone marrow (hBMMSCs) differentiate into neuronal-like cells on the polyvinyl alcohol/gelatin/crocin (PVA/Gel/Cro) nanofiber scaffolds in vitro. METHODS: PVA/Gel scaffolds containing crocin in three concentrations (1%, 3%, and 5%) were prepared by the electrospinning method. The human bone marrow-derived mesenchymal stem cells (hBMSCs) differentiation on the PVA/Gel/Cro 5% that induced by beta-carotene (ßC), was analyzed during 10 days. Morphology of differentiated cells on the scaffolds was taken by scanning electron microscope (SEM). The expression of the neural cell markers was studied by quantitative reverse transcription- polymerase chain reaction (qRT-PCR) and immunocytochemistry (ICC). RESULTS: MTT results of C6 cells culture on the scaffolds showed that proliferation and metabolic activity on PVA/Gel scaffold containing crocin 5% (PVA/Gel/Cro 5%) are significantly more than the other concentrations (P = 0.01). MSC differentiation to nerve-like cells was approved by MAP-2 expression at the mRNA level and NESTIN and MAP-2 at the protein level. CONCLUSIONS: These results suggested that PVA/Gel/Cro 5% and ßC could lead to hBMSCs differentiation to neural cells.

Polysaccharide-based (kappa carrageenan/carboxymethyl chitosan) nanofibrous membrane loaded with antifibrinolytic drug for rapid hemostasis- in vitro and in vivo evaluation.

This work aimed to establish a novel membrane consisting of hemostatic polysaccharides, kappa-carrageenan (KC), and carboxymethyl chitosan (CMC) in tandem with polyvinyl alcohol that spun together as a matrix and loaded with tranexamic acid (TXA) as antifibrinolytic agent for further coagulation effect during and after oral surgeries. The electrospinning of KC was done for the first time and in comparison of CMC has better hemostatic efficacy. The effect of the hemostat was investigated by its surface morphology (SEM), FTIR/ATR analysis, swelling behavior in both PBS and blood, hydrophilicity, porosity, mechanical properties, and cumulative release rate. The effect of materials and the drug concentration ratio were considered. The effect of acetic acid percent in aqueous solutions of CMC/PVA and KC/PVA on morphology was investigated. The cell culture assay showed that all membranes interacted well (98 %) with fibroblast cells attached and grown on the fabricated substrate. Furthermore, the membranes are evaluated by clotting time, whole blood clotting, hemocompatibility, and platelet and RBC adhesion tests. Also, the hemostatic performance of the membrane was analyzed in vivo, using the tail and liver bleeding model in rats. Therefore, TXA loading into CMC and KC dressing could be an attractive hemostatic system for various clinical applications.

Multi-antibacterial agent-based electrospun polycaprolactone for active wound dressing.

Today, due to the greater knowledge of the side effects of chemical drugs and the favorable pharmacological properties of herbal compounds, the use of these compounds is increasing. Since wounds need fast and efficient healing, wound dressing fabrication methods play an important role in wound healing. In this research, electrospinning process was used to prepare samples. Natural antibacterial compounds, such as curcumin, piperine, eugenol, and rutin were loaded in electrospun nano-fibrous based on polycaprolactone. Three-component novel systems of curcumin-piperine-eugenol (PCPiEu), and curcumin-piperine-rutin (PCPiR) were designed and prepared. Their synergistic effect was investigated and also compared with one- and two-component systems. The results showed that medium diameter nanofibers of PCPiEu and PCPiR samples was 198.38 and 142.60, respectively, and they were obtained in smooth, uniform and bead free morphology using optimization of process parameters. The amount of water absorption and water vapor permeability of the three-component samples were in the appropriate range (8.33-10.42 mg cm2 h-1) for wound dressings. The mechanical properties of samples were reduced compared to the control sample, which required further investigation. Antibacterial tests showed good results for partial toxicity of PCPiEu and PCPiR samples. Antibacterial tests showed minor toxicity in PCPiR samples and good results were obtained for PCPiEu samples. In addition, the results showed that PCPiEu and PCPiR samples exhibited antibacterial activity against Gram-positive bacterium Staphylococcus aureus and Gram-negative Enterococcus faecalis bacterium, so that killing ability of 74% and 75% against Gram-positive bacterium and 99.47% and 96.88% against Gram-negative bacterium were obtained for these three systems, respectively.

Multilayered 3-D nanofibrous scaffold with chondroitin sulfate sustained release as dermal substitute.

Electrospun nanofibers for skin tissue engineering applications face two main challenges. The low thickness of electrospun mats is the main reason for their weak load-bearing performance at clinical applications and limited cell penetration due to their small pore sizes. We have developed multi-layered nanofibrous 3D (M3DN) scaffolds comprising gelatin, polyvinyl alcohol, and chondroitin sulfate (CS) by an electrospinning method and attaching three electrospun layers via ethanol to cause interface fibers to come in contact with each other. Prepared M3DN scaffolds revealed a sustained CS release profile. The improved mechanical performance, stable release of CS, and penetration capability of the cells and blood vessels through the spaces between layers in the prepared multi-layered nanofibrous scaffolds demonstrate their potential applications in response to the increasing demand for replacement of damaged dermis. The results of animal studies on the dorsal skin of Rat with full-thickness wounds have shown that the reconstruction of full-thickness skin lesions is significantly higher for M3DN scaffolds than a control group (treated with sterile gauze). The amount of epithelization, collagen arrangement, and inflammatory cells (acute and chronic) has been investigated, and their associated results demonstrated that M3DN scaffolds have great potential for full-thickness wound restoration.

Fabrication and In Vitro Evaluation of A Chondroitin Sulphate-Polycaprolactone Composite Nanofibrous Scaffold for Potential Use in Dermal Tissue Engineering.

OBJECTIVE: Poly(ε-caprolactone) (PCL) has considerable mechanical and biological properties that make it a good candidate for tissue engineering applications. PCL alongside proteins and polysaccharides, like gelatin (GEL) and chondroitin sulphate (CS), can be used to fabricate composite scaffolds that provide mechanical and biological requirements for skin tissue engineering scaffolds. The aim of this study was fabricating novel composite nanofibrous scaffold containing various ratios of GEL/CS and PCL using co-electrospinning process. MATERIALS AND METHODS: In this experimental study, PCL mixed with a GEL/CS blend has limitations in miscibility and the lack of a common solvent. Here, we electrospun PCL and GEL/CS coincide separately on the same drum by using different nozzles to create composite nanofibrous scaffolds with different ratios (2:1, 1:1 and 1:2) of GEL to CSPCL, and we mixed them at the micro/nanoscale. Morphology, porosity, phosphate-buffered saline (PBS) absorption, chemical structure, mechanical property and in vitro bioactivity of the prepared composite scaffolds were analysed. RESULTS: Scanning electron microscopy (SEM) images showed beadless nanofibres at all ratios of GEL to CS-PCL. The composite scaffolds (2:1, 1:1 and 1:2) had increased porosity compared to the PCL nanofibrous scaffolds, in addition to a significant increase in PBS absorption. The mechanical properties of the composite scaffolds were investigated under different conditions. The results demonstrated that all of the composite specimens had better strength when compared with the GEL/CS nanofibres. The increase in PCL ratio led to an increase in tensile strength of the nanofibres. Human dermal fibroblasts (HDF) were cultured on the fabricated composite scaffolds and evaluated by 3-(4,5-dimethylthiazol- 2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) analysis and SEM. The results showed the bioactivity of these nanofibres and the potential for these scaffolds to be used for skin tissue engineering applications. CONCLUSION: The fabricated co-electrospun composite scaffolds had higher porosity and PBS absorption in comparison with the PCL nanofibrous scaffolds, in addition to significant improvements in mechanical properties under wet and dry conditions compared to the GEL/CS scaffold.

Stimuli-responsive electrospun nanofibers based on PNVCL-PVAc copolymer in biomedical applications.

Poly(N-vinylcaprolactam) (PNVCL) is a suitable alternative for biomedical applications due to its biocompatibility, biodegradability, non-toxicity, and showing phase transition at the human body temperature range. The purpose of this study was to synthesize a high molecular weight PNVCL-PVAc thermo-responsive copolymer with broad mass distribution suitable for electrospun nanofiber fabrication. The chemical structure of the synthesized materials was detected by FTIR and 1HNMR spectroscopies. N-Vinyl caprolactam/vinyl acetate copolymers (159,680 molecular weight (g/mol) and 2.51 PDI) were synthesized by radical polymerization. The phase transition temperature of N-vinyl caprolactam/vinyl acetate copolymer was determined by conducting a contact angle test at various temperatures (25, 26, 28, and 30 [Formula: see text]). The biocompatibility of the nanofibers was also evaluated, and both qualitative and quantitative results showed that the growth and proliferation of 929L mouse fibroblast cells increased to 80% within 48 h. These results revealed that the synthesized nanofibers were biocompatible and not cytotoxic. The results confirmed that the synthesized copolymers have good characteristics for biomedical applications.

Fish cartilage: A promising source of biomaterial for biological scaffold fabrication in cartilage tissue engineering.

Here, engineered cartilage-like scaffold using an extracellular matrix (ECM) from sturgeon fish cartilage provided a chondroinductive environment to stimulate cartilaginous matrix synthesis in human adipose stem cells (hASCs). Three dimensional porous and degradable fish cartilage ECM-derived scaffold (FCS) was produced using a protocol containing chemical decellularization, enzymatic solubilization, freeze-drying and EDC-crosslinking treatments and the effect of different ECM concentrations (10, 20, 30, and 40 mg/ml) on prepared scaffolds was investigated through physical, mechanical and biological analysis. The histological and scanning electron microscopy analysis revealed the elimination of the cell fragments and a 3-D interconnected porous structure, respectively. Cell viability assay displayed no cytotoxic effects. The prepared porous constructs of fish cartilage ECM were seeded with hASCs for 21 days and compared to collagen (Col) and collagen-10% hyaluronic acid (Col-HA) scaffolds. Cell culture results evidenced that the fabricated scaffolds could provide a proper 3-D structure to support the adhesion, proliferation and chondrogenic differentiation of hASCs considering the synthesis of specific proteins of cartilage, collagen type II (Col II) and aggrecan (ACAN). Based on the results of the present study, it can be concluded that the porous scaffold derived from fish cartilage ECM possesses an excellent potential for cartilage tissue engineering.

PEGylated curcumin-loaded nanofibrous mats with controlled burst release through bead knot-on-spring design.

APEGylatedcurcumin (PCU) loaded electrospuns based on poly(ε-caprolactone) (PCL) andpolyvinyl alcohol (PVA) were fabricated for wound dressing applications. The main reason for this wound dressing design is antibacterialactivity enhancement, and wound exudates management. PEGylation increases curcuminsantibacterial properties and PVA can help exudates management. For optimal wound dressing, first, response surface methodology (RSM) was applied to optimize the electrospinning parameters to achieve appropriate nanofibrous mats. Then a three-layer electrospun was designed by considering the water absorbability, PCU release profile as well as antibacterial and biocompatibility of the final wound dressing. The burst release in controlled release systems could be evaluated for prevention of the higher initial drug release and control the effective life time. The PCU release results illustrated that the bead knot plays a positive role in controlling the release profile andby increase in the number of beads per unit area from 3000 to 9000 mm-2,the PCU burst release will be reduced; Also in vitro studies show that optimized three-layer dressing based on PCL/PVA/PCU can support water vapour transmission rate in optimal range and also absorb more than three times exudates in comparison with mono-layerdressing. Antibacterial tests show that the electrospun wound dressing containing 5% PCU exhibits100% antibacterial activityas well as cell viability level within an acceptable range.

Polyvinyl alcohol/sulfated alginate nanofibers induced the neuronal differentiation of human bone marrow stem cells.

Scaffolds that are used for neural tissue engineering are fabricated to mimic the extracellular matrix. In this paper, we have fabricated polyvinyl alcohol/sulfated alginate (PVA/SA) nanofibers with different concentrations (10, 20 and 30 wt%) of sulfated alginate by electrospinning technique. The average fibers diameters of 169-488 nm were achieved by electrospinning of polymers blend (PVA/SA). The results of the MTT assay and scanning electron microscopy showed that PVA/sulfated alginate nanofibrous scaffold with 30 wt% SA provided more desirable surface attachment of C6, Schwann cells (SCs) and human bone marrow mesenchymal stem cells (hBMSCs). RT-PCR and immunocytochemistry for MAP-2 marker were conducted to confirm the neural-differentiation of hBMSCs. The expression of MAP-2 confirmed neural differentiation for up to 14 days. Our results showed that PVA/SA nanofibrous scaffold with 30 wt% SA is a suitable substrate for mesenchymal stem cells growth and is capable of inducing neuronal differentiation.

Controlled curcumin release from nanofibers based on amphiphilic-block segmented polyurethanes.

A series of biodegradable amphiphilic-block segmented polyurethanes (SPUs) are designed and synthesized based on di-block and tri-block macrodiols of polycaprolactone (PCL) and polyethylene glycol (PEG). Curcumin, as a model herbal antibacterial agent, is used due to its effective inhibitory action against Gram-positive and Gram-negative bacteria. Curcumin-loaded nanofibers, with 400-900 nm diameter range, have been prepared by electrospinning of SPUs. The synthesized SPUs can be used for wound dressing applications due to their excellent mechanical properties and higher hydrophilicity in comparison to PCL-based polyurethane. The elongation-at-break of tri-block SPU with PEG-PCL-PEG soft segments is 350% when produced as an electrospun mat and that for film is 1500%. In vitro release of curcumin, examined by UV-Vis spectroscopy, shows a steady release during 18 days. The inclusion of PEG chains in the soft segment increases the hydrophilicity and biodegradation rate of the electrospun mats compared to a PCL-based polyurethane, which eventually results in a higher curcumin release rate. The antibacterial activity of 50 mg of 10% curcumin-loaded SPU nanofibers is about 100% and 93% against Escherichia coli (E. coli ATCC: 25922) and Staphylococcus aureus (S. aureus ATCC: 6538), respectively. Nontoxic behavior of the scaffolds is evaluated through MTT assay against L929 mouse fibroblast cells. The results show that the synthesized SPUs can be used as a nanoscale sustained release carrier. The SPU with PEG-PCL-PEG soft segments is an excellent candidate for wound dressing in tissues undergoing large deformations during normal activities.

ChABC-loaded PLGA nanoparticles: A comprehensive study on biocompatibility, functional recovery, and axonal regeneration in animal model of spinal cord injury.

Spinal Cord Injury (SCI) is one of the leading causes of physical disability. In this study, spherical PLGA nanoparticles (NPs) containing ChABC enzyme were manufactured and fully characterized for SCI therapy. The NPs were used in the rat's contused spinal cord to assess the functional improvement and scar digestion. Twenty-three adult male Wistar rats (275 ± 25 g) were assigned into four groups of control, sham, blank-treated particle, and ChABC-treated particle. Throughout the survey, the BBB scores were obtained for all the groups. Finally, the injured sections of animals were dissected, and histological studies were conducted using Luxol fast blue and Bielschowsky. The biocompatibility and non-toxicity effects of the NPs on olfactory ensheathing cells (OECs) were confirmed by the MTT test. The flow-cytometry revealed the purity of cultured OECs with p75+/GFAP+ at around 87.9 ± 2.4%. Animals in the control and the blank-treated groups exhibited significantly lower BBB scores compared with the ChABC-treated particle group. Histological results confirmed the induced contusion models in the injured site. Myelin was observed in the treated groups, especially when the ChABC-loaded nanoparticles were utilized. The immunohistochemistry results indicated the scar glial degradation in animals treated by the ChABC-loaded particles. According to this study, the loaded particles can potentially serve as a suitable candidate for spinal cord repair, functional recovery and axonal regeneration.

Tough, hybrid chondroitin sulfate nanofibers as a promising scaffold for skin tissue engineering.

Fabrication of gelatin/polyvinyl alcohol/chondroitin sulfate (GEL/PVA/CS) hybrid nanofibrous scaffolds using acetic acid and water as an environmentally friendly solvent system via electrospinning for skin tissue engineering was investigated. Modeling and optimization of the nanofibers were performed using response surface methodology (RSM). The influence of CS ratio on mechanical, physical and biological properties of the nanofibers was studied. PVA was used as a carrier and enhancer of mechanical properties. The mechanical properties of hybrid nanofibers were investigated in dry and wet states. The results showed that in the cross-linked dry state the tensile strength was up to 4 MPa. In the wet state, nanofibers exhibited 200% elongation at break, indicating a toughness behavior which enhances the flexibility for clinical applications. Scanning electron microscope (SEM) confirmed the stability of nanofibrous morphology during degradation up to 21 days. Human dermal fibroblast-green fluorescent protein-positive (HDF-GFP+) cells were cultured on the scaffolds and results showed the appropriate biocompatibility. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was employed to study cell proliferation, and the results confirmed the positive effect of CS ratio on HDF cells attachment as well as proliferation on the nanofibers. Considering the results of in vitro assay, nanofibers containing 15% CS ratio suggested as an optimum CS ratio.

Chondroitin sulfate immobilized PCL nanofibers enhance chondrogenic differentiation of mesenchymal stem cells.

Cold Atmospheric Plasma (CAP) is used as a promising method in surface modification for immobilization of chondroitin sulfate functional biomacromolecules on PCL nanofibrous substrates for cartilage tissue engineering. The GAG-grafted scaffolds are able to successfully support the attachment and proliferation of mesenchymal stem cells (MSCs). The seeded scaffolds show the chondro-differentiation of MSCs during a 21-days cell culture in a non-differential medium. Expression of SOX9, Collagen10 and Collagen2 proved the chondro-inductive effect of GAG-grafted scaffolds. Besides, no external chondro-genic differential agent was used in the differentiation of MSCs to chondrocyte. The cells passed the last phase of chondrogenesis after 14 days of incubation. Thus, the GAG-fabricated fibrous scaffolds using CAP are potential candidates for cartilage tissue engineering.

Enhanced chondrogenic differentiation of bone marrow mesenchymal stem cells on gelatin/glycosaminoglycan electrospun nanofibers with different amount of glycosaminoglycan.

Tissue engineering is a new technique to help damaged cartilage treatment using cells and scaffolds. In this study we tried to evaluate electrospun scaffolds composed of gelatin/glycosaminoglycan (G/GAG) blend nanofibers in chondrogenesis of bone marrow-derived mesenchymal stem cells (BMMSCs). Scaffolds were fabricated by electrospinning technique with different concentration of glycosaminoglycan (0%, 5%, 10%, and 15%) in gelatin matrix. BMMSCs were cultured on the scaffolds for chondrogenesis process. MTT assay was done for scaffold's biocompatibility and cells viability evaluation. Alcian blue staining was carried out to determine the release of GAG and reverse transcription polymerase chain reaction (RT-PCR) was done for expression of COL2A1 and also immunocytochemistry assay were used to confirm expression of type II collagen. Scaffold with 15% GAG showed better result for biocompatibility (p =0.02). Scanning electron microscopy (SEM) micrographs showed that MSCs have good attachment to the scaffolds. Alcian blue staining result confirmed that cells produce GAG during differentiation time different from GAG in the scaffolds. Also the results for RT-PCR showed the expression of COL2A1 marker. Immunocytochemistry assay for type II collagen confirm that this protein expressed. Scaffold comprising 15% GAG is better results for chondrogenesis and it can be a good applicant for cartilage tissue engineering. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 38-48, 2019.

Reduction in protein absorption on ophthalmic lenses by PEGDA bulk modification of silicone acrylate-based formulation.

The absorption of protein and formation of biofilms on the surface of ophthalmic lenses is one of the factors that destroy their useful performance by causing severe visual impairment, inflammation, dryness and ultimate eye discomfort. Therefore, eye lenses need to be resilient to protein absorption, which is one of the opacity factors in minimizing protein absorption on the lenses. The purpose of this study was to investigate and reduce sediment biotransformation on the surface of the semi-hardened lens based on acrylate by bulk-free radical polymerization method. In this respect, the effect of poly(ethylene glycol) diacrylate (PEGDA) with two different molecular weights of 200 and 600 g/mol on the surface roughness, protein absorption, and hydrophilicity of the lenses were studied. The surface hardness of the lenses, on shore D scale, was measured using a durometer hardness test. The presence of higher molecular weight of PEGDA hydrophilic polymeric monomers reduced the hardness of the lenses. The effect of introducing PEGDA, with two molecular weights, into lens fabrication formulations was studied with respect to their water content parameters and hydrophilicity. The presence of a crosslinker such as poly(ethylene glycol) diacrylates, at two different molecular weights, increased the water content and hydrophilicity of the produced lenses. Surface roughness is associated with the formation of bio-film and accumulation of microorganisms on the surface. Due to the roughness of the lens surface developed in this research, the lenses containing PEGDA 600 exhibited less roughness compared to that of PEGDA 200, which could also affect the absorption of protein. Therefore, according to the results of protein absorption test, the PEGDA 600 lenses showed lower protein absorption, which could be due to their high degree of water absorption and hydrophilicity.

Cold atmospheric plasma as a promising approach for gelatin immobilization on poly(ε-caprolactone) electrospun scaffolds.

Poly(Ɛ-caprolactone) (PCL) is a biocompatible polymer with a high potential to be used in tissue engineering especially in tight tissues. In the current study, cold atmospheric plasma (CAP) is used as a promising method for immobilization of gelatin as a functional biomacromolecule on PCL nanofibrous substrates. The CAP surface modification leads to oxidation of chemical groups existing on the PCL surface without doing any damage to the bulk properties of biomaterials for gelatin biomacromolecule grafting. The water contact angle (WCA) of the CAP-treated surface and gelatin-grafted PCL using CAP indicates an effective increment in the hydrophilicity of the PCL surface. Also to achieve the highest levels of gelatin grafting on the PCL surface, two different grafting methods and gelatin concentration diversity are utilized in the grafting process. The immobilization of gelatin biomacromolecules onto the CAP surface-modified PCL nanofibers is investigated using scanning electron microscope (SEM) and Fourier transform infrared spectroscopy (FTIR). The gelatin-modified PCL substrates revealed uniform nanofibrous morphology with increased average fiber diameter. The results of FTIR spectra, including hydroxyl groups, NH groups, and amide II of gelatin-grafting peaks, confirm the gelatin immobilization on the surface of nanofibers. The metabolic activity of cultured mesenchymal stem cells (MSCs) on the surface-modified scaffolds is evaluated using MTT analysis (P ≤ 0.05). The results of metabolic activity and also SEM and DAPI staining observations indicate proper attachment on the surface and viability for MSCs on the surface-immobilized nanofibrous scaffolds. Therefore, CAP treatment would be an effective method for biomacromolecule immobilization on nanofibers towards the enhancement of cell behavior.

Vitamin C Loaded Poly(urethane-urea)/ZnAl-LDH Aligned Scaffolds Increase Proliferation of Corneal Keratocytes and Up-Regulate Vimentin Secretion.

A novel poly(urethane-urea) (PUU) based on poly(glycolide-co-ε-caprolactone) macro-diol with tunable mechanical properties and biodegradation behavior is reported for corneal stromal tissue regeneration. Zn-Al layered double hydroxide (LDH) nanoparticles were synthesized and loaded with vitamin C (VC, VC-LDH) and dispersed in the PUU to control VC release in the cell culturing medium. To mimic the corneal stromal EC, scaffolds of the PUU and its nanocomposites with VC-LDH (PUU-LDH and PUU-VC-LDH) were fabricated via electrospinning. Average diameters of the aligned nanofibers were recorded as 325 ± 168, 343 ± 171, and 414 ± 275 nm for the PUU, PUU-LDH, and PUU-VC-LDH scaffolds, respectively. Results of hydrophilicity and mechanical properties measurements showed increased hydrophobicity and reduced tensile strength and elongation at break upon addition of nanoparticles to the PUU scaffold. VC release studies represented that intercalation of the drug in Zn-Al-LDH controlled the burst release and extended the release period from a few hours to 5 days. Viability and proliferation of stromal keratocyte cells on the scaffolds were investigated via AlamarBlue assay. After 24 h, the cells showed similar viability on the scaffolds and the control. After 1 week, the cells showed some degree of proliferation on the scaffolds, with the highest value recorded for PUU-VC-LDH. SEM images of the scaffolds after 24 h and 1 week confirmed good penetration and attachment of keratocytes on all the scaffolds and the cells oriented with the direction of nanofibers. After 1 week, the PUU-VC-LDH scaffold was fully covered by the cells. Immunocytochemistry assay (ICC) was performed to investigate secretion of vimentin protein, ALDH3A1, and α-SMA by the cells. After 24h and 1 week, remarkably higher levels of vimentin and ALDH3A1 and lower level of α-SMA were secreted by keratocytes on PUU-VC-LDH compared to those on the PUU and PUU-LDH scaffolds and the control. Our results suggest that the aligned PUU-VC-LDH is a promising candidate for corneal stromal tissue engineering due to the presence of zinc and vitamin C.

Electrospun polyvinyl alcohol/gelatin/chondroitin sulfate nanofibrous scaffold: Fabrication and in vitro evaluation.

Electrospun nanofibers have attracted a lot of attention in recent years in tissue engineering applications. In this research, novel polyvinyl alcohol/gelatin/chondroitin sulfate (PVA/GE/Cs) nanofibrous scaffolds using non-carcinogen solvent system via electrospinning technique was evaluated. A solvent system containing water and acetic acid was used as a safe solvent system to obtain a homogenous mixture with suitable solvent properties and finally non-toxic nanofibrous scaffolds. The effect of water to the acetic acid ratio in the solvent system (7:3, 6:4, 5:5, 4:6, 3:7) and also polymer concentration (8, 9, 10w/v %) on nanofibers morphology was investigated. The appropriate flow rate and voltage ranges to obtain uniform and bead-free electrospun scaffold were investigated. Effect of different Cs ratio (0, 10, 15 and 20wt%) on solution properties was evaluated. Influence of Cs ratio on chemical, physical and thermal properties of the electrospun scaffolds was studied. The results of cell toxicity indicated that prepared PVA/GE/Cs scaffolds have no cell toxicity. SEM results demonstrated that L929 mouse fibroblast cells have suitable interaction with scaffold surface and also attached and proliferated well on the prepared substrate after 24 and 48h and also have a potential for using in tissue engineering.

Tailoring the gelatin/chitosan electrospun scaffold for application in skin tissue engineering: an in vitro study.

The nanofibrous structure containing protein and polysaccharide has good potential in tissue engineering. The present work aims to study the role of chitosan in gelatin/chitosan nanofibrous scaffolds fabricated through electrospinning process under optimized condition. The performance of chitosan in gelatin/chitosan nanofibrous scaffolds was evaluated by mechanical tests, scanning electron microscopy (SEM), Fourier transform infrared (FTIR) and in vitro cell culture on scaffolds with different gelatin/chitosan blend ratios. To assay the influence of chitosan ratio on biocompatibility of the electrospun gelatin/chitosan scaffolds for skin tissue engineering, the culturing of the human dermal fibroblast cells (HDF) on nanofibers in terms of attachment, morphology and proliferation was evaluated. Morphological observation showed that HDF cells were attached and spread well on highly porous gelatin/chitosan nanofibrous scaffolds displaying spindle-like shapes and stretching. The fibrous morphologies of electrospun gelatin/chitosan scaffolds in culture medium were maintained during 7 days. Cell proliferation on electrospun gelatin/chitosan scaffolds was quantified by MTS assay, which revealed the positive effect of chitosan content (around 30%) as well as the nanofibrous structure on the biocompatibility (cell proliferation and attachment) of substrates.

Biodegradable bead-on-spring nanofibers releasing ß-carotene for bone tissue engineering.

Bead-on-string mats based on poly(lactide-co-glycolide) (PLGA) releasing ß-carotene (ßC) as a natural osteogen were fabricated and used for bone tissue engineering. Mesenchymal stem cells (MSCs) seeded on the scaffolds successfully differentiated to osteoblasts without using any a differential medium. The mats showed a small burst of ß-carotene (24-27%) during the first day and a sustained slow release up to 21 days. The MTT and SEM results indicated good attachment and proliferation of MSCs on the scaffolds. Calcination of scaffolds and expression of RUNX2, SOX9, and osteonectin genes approved the differentiation of seeded MSCs to osteoblasts without using any external osteogenic differential agent. The scaffold loaded with 4% ß-carotene not only induced the early phase of osteogenesis but also advanced the differentiation to the osteoblast maturation phase. Thus, these bead-on-string scaffolds can be used as a substrate for direct bone tissue engineering.