![]() ![]() After solubilization, each well was diluted 10-fold, and aliquoted into one column of 96-well plate. An extraction buffer (250 μL, 20% SDS/50% DMF, pH 7.4) was added to each well, and the gels were kept at 37 ☌, overnight. After the MTT treatment, the plates were centrifuged at 800 × g for 5 min, the supernatant was removed, and the pellet was washed once with PBS. In order to quantify L-929 cells viability, MTT (50 μL, 5 mg/mL PBS) was added to each well and the plate was incubated for 3 h at 37 ☌. Thus, the number of viable cells can be correlated with the absorbance at 540 nm (λ max). The MTT chromophore represents the number of viable cells, by measuring the amount of formazan generated by the mitochondrial enzymes of metabolically active cells. The number of viable cells was assayed over a period of 14 days, with the assumption that any toxic effects would be manifested within that period of time. The toxicity of PAMAM-grafted SF nanofibers was measured using the MTT assay. The cells were suspended in McCoy's 5 A medium and they were supplemented with 10% Fetal Bovine Serum (FBS) and 1% Penicillin-Streptomycin, with the exception of transfection experiments, where cells were grown in complete media without antibiotics. L-929 cells were cultured in an incubator (5% CO 2, 37 ☌). In order to evaluate the properties of SF nanofibers scaffold prepared by this methods, the dissolve process, morphology, structure, biocompatibility and cytotoxicity were studied. SF nanofibers were modified by crosslinking PAMAM dendrimers to carboxylic-terminated Fibroin, via EDC and NHS. The electrospinning of SF sponge was performed with formic acid as a spinning solvent. In the present study, silk was dissolved in CaCl 2-C 2H 5OH-H 2O in order to obtain silk fibroin solution and it later was lyophilized in a freeze-dryer. A cylinder collector rotating at a high speed can obtain aligned fiber a simple and common way, compared to other methods, to obtain aligned fiber. Electrospinning is a unique method, capable of producing nanoscale fibers from both synthetic as well as natural polymers for biomedical applications. Silk fibroin has been widely explored in many biomedical applications, due to its impressive biocompatibility, biodegradability and minimal inflammatory reactions. It has been reported that the biological features of silk fibroin make it a very good choice for use for regeneration. Silk fibroin is a natural protein and is known to promote cell attachment, proliferation and tissue regeneration. For biomedical applications, only silk fibroin is useful, and sericin, due to its immunogenicity, needs to be removed. Sericin is the gummy substance surrounding fibroin. Silk fibers are built by two proteins, namely, fibroin and sericin. Natural silk from the silkworm cocoons and Bombyx Mori has been used for centuries as biomedical suture material. Silk proteins are produced by various species of silkworms, spiders, scorpions and bees. thin films, gels, 3 D sponges as well as micro- and nano-particles. There are many structures that can be used as biomaterials, e.g. Tissue engineering tries to resolve these problems by using natural and synthetic polymeric matrices to produce appropriate biomaterial shapes. Some of the main problems of human health are related to accidents and illnesses that cause a loss of organ or tissue damages. Therefore, fibroin nanofibers scaffolds appear to be a probable choice for potential use in tissue engineering. Adhesion and proliferation of fibroblasts were investigated by MTT assay, which showed no cytotoxicity. Biocompatibility tests were carried out through seeding fibroblasts cell line L929 on nanofibers. The modification results of analysis demonstrated that the amino groups were more established on the surface of the SF nanofiber scaffold. FTIR spectrum showed amide peaks, which confirmed the existence of fibroin. Through treatment, conformational transitions of the SF nanofibers from random coil to β-sheet occurred rapidly, confirmed by FTIR and X-Ray diffraction. SEM micrograph showed that the electrospun containing 7% (w/w) fibroin had continuous fibers and an average diameter of 80 nm. The modified SF nanofibers scaffold was evaluated by SEM, X-Ray diffraction, FTIR and MTT assay. SF nanofibers were modified by crosslinking PAMAM dendrimers to carboxylic-terminated fibroin, via EDC and NHS. The electrospinning of SF sponge was performed at different fibroin concentrations. In the present study, silk was dissolved in CaCl 2-C 2H 5OH-H 2O in order to obtain silk fibroin (SF) solution, and it was then lyophilized in freeze-dryer. Fibroin is a protein that has been extensively studied for biomedical application.
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