In this study, we developed a fast, simple and novel process to fabricate cross-linked electro-spun gelatin with limited amounts of glutaraldehyde (GA) using trifluoroacetic acid (TFA) as the solvent. Using SEM, the uncross-linked gelatin fibers were determined to have diameters between 50-300 nm, while the cross-linked gelatin electro-spun fibers had diameters between 100-500 nm. FT-IR revealed that the un-cross-linked and cross-linked electro-spun gelatin was fabricated successfully by electro-spinning using TFA as a solvent, which has not been reported until now. Stress-strain curves showed that the addition of small amounts of GA increased the strength of the gelatin by two fold and allowed for the creation of a water insoluble gelatin electro-spun membrane
In this study, two distinct systems of biomaterials were fabricated and their potential use as a bilayer scaffold (BS) for skin bioengineering applications was assessed. The initial biomaterial was a polycaprolactone/poly(lacto-co-glycolic acid) (PCL/PLGA) membrane fabricated using the electrospinning method. The PCL/PLGA membrane M-12 (12% PCL/10% PLGA, 80:20) displayed strong mechanical properties (stress/strain values of 3.01 ± 0.23 MPa/225.39 ± 7.63%) and good biocompatibility as demonstrated by adhesion of keratinocyte cells on the surface and ability to support cell proliferation. The second biomaterial was a hydrogel composed of 2% chitosan and 15% gelatin (50:50) crosslinked with 5% glutaraldehyde. The CG-3.5 hydrogel (with 3.5% glutaraldehyde (v/v)) displayed a high porosity, ≥97%, good compressive strength (2.23 ± 0.25 MPa), ability to swell more than 500% of its dry weight and was able to support fibroblast cell proliferation. A BS was fabricated by underlaying the membrane and hydrogel casting method to combine these two materials. The physical properties and biocompatibility were preliminarily investigated and the properties of the two biomaterials were shown to be complementary when combined. The upper layer membrane provided mechanical support in the scaffold and reduced the degradation rate of the hydrogel layer. Cell viability was similar to that in the hydrogel layer which suggests that addition of the membrane layer did not affect the biocompatibility.
Phyllanthus emblica is an edible nutraceutical and functional food in the Asia area with medicinal and nutritive importance. The fruit extract of P. emblica is currently considered to be one of the effective functional foods for flesh maintenance and disease treatments because of its antioxidative and immunomodulatory properties. We examined the antioxidant abilities of the fruit extract powder by carrying out 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging, iron reducing power, and metal chelating activity analysis and showed excellent antioxidative results. In 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the result showed that the samples had no cytotoxic effect on RAW 264.7 cells even at a high concentration of 2 mg/mL. To investigate its immunomodulatory function, our estimation was to treat it with lipopolysaccharide (LPS) in RAW 264.7 cells to present anti-inflammatory capacities. The extract decreased reactive oxygen species (ROS) production levels in a dose-dependent manner measured by flow cytometry. We also examined various inflammatory mRNAs and proteins, including nuclear factor-κB (NF-κB), inducible nitric oxide synthases (iNOS), and cyclooxygenase-2 (COX-2). In quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blotting assay, all three targets were decreased by the extract, also in a dose-dependent manner. In conclusion, P. emblica fruit extract powder not only lessened antioxidative stress damages, but also inhibited inflammatory reactions.
Three dimensional porous scaffolds composed of various ratios of polycaprolactone and poly(L-lactic acid) (PLLA) were prepared using salt leaching method for bone regeneration applications. Surfaces of the scaffolds were visualized using scanning electron microscope (SEM) and the combination of the polymers was confirmed by FT-IR. Addition of PLLA increased the porosity and pore sizes of the scaffolds and also the scaffolds' compressive strength initially. Osteoblast-like cells were used and it was found that the samples' cell biocompatibility was further promoted with the increase in PLLA content as observed via cell proliferation assays using MTT, gene expression with RT-PCR, and micrographs from SEM and confocal microscopy. Samples were then implanted into male rabbits for 2 months, and histological staining and micro-CT histomorphometry show that new bone formations were detected in the site containing the implants of the scaffolds and that bone regeneration was further promoted with the increased concentration of PLLA in the scaffold.
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