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Silkworm silk - CNT composite fibers: fabrication, characterization and application in cell stimulation for tissue regeneration
Title
Silkworm silk - CNT composite fibers: fabrication, characterization and application in cell stimulation for tissue regeneration
Creator
Zheng, Shuyao
Date
2019
Description
Silk fibroin, a structural protein of the Bombyx mori cocoons, is widely studied for biomedical applications because of its strong mechanical...
Show moreSilk fibroin, a structural protein of the Bombyx mori cocoons, is widely studied for biomedical applications because of its strong mechanical property, biocompatibility, low degradation rate and ease of processing. In this study, we incorporated functionalized CNT in silkworm silk proteins to generate biocomposite fibers by electrospinning. The electro-spun (E-spun) fibers were unidirectionally aligned with a diameter at the sub-micrometer scale, mimicking the locally oriented ECM proteins in native tissues. The addition of a minute amount of CNT not only reinforced the stability and the mechanical strength of the flexible scaffolds, but also rendered the fibers electrical conductivity to not only facilitate the E-spun fiber formation but also grant the fibers an additional functionality that can be utilized for cell stimulation. It is known that endogenous electric field is present in the human body at various locations and plays critical physiological roles through its effect on cell proliferation, migration and activation mediated by its interference with growth factor secretion and intracellular signaling pathways. Electrical stimulation (ES) has been widely investigated as a potential method to regulate cell functions for the treatment of a number of diseases. It has been reported that in dermal fibroblasts, ES induces the secretion of FGF-1 and FGF-2 which promotes cell proliferation, contraction, migration, and activation/differentiation into myofibroblasts for increased collagen synthesis in favor of tissue repair. In this work, the modulation effect of the aligned E-spun SWS-CNT fibers was examined by their application in inducing polarization and activation of fibroblasts with cellular deficit, like those from pelvic organ prolapse (POP) patients. These cells are characterized by low productivity of collagen with abnormally high COLI/COLIII ratio, leading to a loose and fragile collagen network, decreased integration among the tissue constituents and reduced tissue strength. Electrical stimulation boosted the collagen productivity by 20 folds in cells on silk-CNT than on pure silk due to silk-CNT’s high electrical conductivity. The developed approach can be potentially utilized to remedy the dysfunctional fibroblasts for therapeutic treatment of diseases and health conditions associated with collagen disorder.
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ELECTROSPUN SILKWORM SILK FIBROIN - INDOCYANINE GREEN BIOCOMPOSITE FIBERS: FABRICATION, CHARACTERIZATION AND APPLICATION TOWARDS HEMORRHAGE CONTROL
Title
ELECTROSPUN SILKWORM SILK FIBROIN - INDOCYANINE GREEN BIOCOMPOSITE FIBERS: FABRICATION, CHARACTERIZATION AND APPLICATION TOWARDS HEMORRHAGE CONTROL
Creator
Siddiqua, Ayesha
Date
2022
Description
Silk fibroin (SF), a structural protein found in the Bombyx mori cocoons has gained attention in several biomedical applications as tissue...
Show moreSilk fibroin (SF), a structural protein found in the Bombyx mori cocoons has gained attention in several biomedical applications as tissue engineering scaffolds and wound dressings owing to its properties such as biocompatibility, water vapor permeability and biodegradability. Indocyanine Green (ICG) is an FDA approved tricarbocyanine dye used in medical diagnostics due to its unique photothermal and fluorescent properties. Electrospinning is a highly efficient, easy, and inexpensive technique used to generate nanometer to micrometer thick fibers. In this study, SF and ICG were co-spun to generate flexible microfibers with high surface area to volume ratios. Pure silk, SF-ICG (0.1%) and SF-ICG (0.4%) were chosen for the purpose of this study. Since, as-spun fibers are unstable in aqueous solutions, post treatment methods were explored to enhance the durability of the fibers and to minimize ICG leaching. It was found that ethanol vapor treatment (EVT) not only induced β-sheet formation in SF but also improved the SF-ICG interaction thereby reducing ICG leaching from the composite fibers. Ethanol vapor treated SF-ICG fibers showed less ICG leaching than liquid ethanol treated (LET) SF-ICG fibers indicating the efficacy of the EVT. The increase in SF solution viscosity with ICG concentration suggested a strong silk-ICG interaction which was further confirmed by DSC. The 1h water uptake and the three-day mass loss experiments indicated that the fibers are stable and highly absorbent material. Heat evolution was evaluated by measuring the temperature change in water of a fixed volume after irradiation with a 500 mW, 808 nm diode laser. The heat evolved by the flat fiber scaffolds was higher than the 3D fiber balls, indicating improved light penetration in the former. Pure silk produced negligible heat and it was used as a control. With 14.9 W/cm2 irradiation, the post-treated SF-ICG (0.4%) 3D fibrous ball of 2-3 mg dry weight, solidified a drop of bovine blood in 40 s. In contrast, a single layer fiber matrix required 3 min. to achieve the same clotting effect. Fibers folded into flat scaffolds were able to solidify a blood drop in 25 s. Pure silk fibers in all the cases showed negligible change after irradiation. The results suggest that a larger contact area of fibers is desirable for faster blood clotting, and EVT prompted better ICG retention in SF fibers. Based on the above results, SF-ICG (0.4%) fibers were utilized in a device developed to mimic blood flowing at a rate of 0.5 mL/h through a damaged blood vessel. It was found that irradiation of SF-ICG locally placed at the “damage” region effectively stopped “bleeding” whereas irradiated pure silk was unable to control the blood flow, which demonstrated the success of our SF-ICG fibers towards hemorrhage control.
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