Abstract:-Enzyme engineering via immobilization techniques is a suitable approach for improving enzyme function and stability and is superior to the other chemical or biological methods. In this study chitosan nanoparticles were synthesized using the Ionic Gelation method and were characterized by Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). Alkaline phosphatase was successfully immobilized on the chitosan nanoparticles in optimum conditi… Show more
“…In the FTIR spectrum of chitosan nanoparticles, a broad peak at 3436 cm −1 corresponds to the -NH and -OH stretching vibrations. The weak band at 2930 cm −1 corresponds to –CH stretching, whereas vibrational bands at 1640 cm −1 , 1560 cm −1 , and 1320 cm −1 may be attributed to the amide carbonyl stretch and -NH bend of the amine groups in the chitosan nanoparticles [32]. The band at 1099 cm −1 represents C–O bond stretching.…”
The aim of the present investigation was to study the anti-oxidant effect of chitosan nanoparticles on a human SH-SY5Y neuroblastoma cell line using a rotenone model to generate reactive oxygen species. Chitosan nanoparticles were synthesized using an ionotropic gelation method. The obtained nanoparticles were characterized using various analytical techniques such as Dynamic Light Scattering, Scanning Electron Microscopy, Transmission Electron Microscopy, Fourier Transmission Infrared spectroscopy and Atomic Force Microscopy. Incubation of SH-SY5Y cells with 50 µM rotenone resulted in 35–50% cell death within 24 h of incubation time. Annexin V/Propidium iodide dual staining verified that the majority of neuronal cell death occurred via the apoptotic pathway. The incubation of cells with chitosan nanoparticles reduced rotenone-initiated cytotoxicity and apoptotic cell death. Given that rotenone insult to cells causes oxidative stress, our results suggest that Chitosan nanoparticles have antioxidant and anti-apoptotic properties. Chitosan can not only serve as a novel therapeutic drug in the near future but also as a carrier for combo-therapy.
“…In the FTIR spectrum of chitosan nanoparticles, a broad peak at 3436 cm −1 corresponds to the -NH and -OH stretching vibrations. The weak band at 2930 cm −1 corresponds to –CH stretching, whereas vibrational bands at 1640 cm −1 , 1560 cm −1 , and 1320 cm −1 may be attributed to the amide carbonyl stretch and -NH bend of the amine groups in the chitosan nanoparticles [32]. The band at 1099 cm −1 represents C–O bond stretching.…”
The aim of the present investigation was to study the anti-oxidant effect of chitosan nanoparticles on a human SH-SY5Y neuroblastoma cell line using a rotenone model to generate reactive oxygen species. Chitosan nanoparticles were synthesized using an ionotropic gelation method. The obtained nanoparticles were characterized using various analytical techniques such as Dynamic Light Scattering, Scanning Electron Microscopy, Transmission Electron Microscopy, Fourier Transmission Infrared spectroscopy and Atomic Force Microscopy. Incubation of SH-SY5Y cells with 50 µM rotenone resulted in 35–50% cell death within 24 h of incubation time. Annexin V/Propidium iodide dual staining verified that the majority of neuronal cell death occurred via the apoptotic pathway. The incubation of cells with chitosan nanoparticles reduced rotenone-initiated cytotoxicity and apoptotic cell death. Given that rotenone insult to cells causes oxidative stress, our results suggest that Chitosan nanoparticles have antioxidant and anti-apoptotic properties. Chitosan can not only serve as a novel therapeutic drug in the near future but also as a carrier for combo-therapy.
“…The optimum temperature for immobilized enzyme was specified 45 °C, which is 5 °C higher than that of free inulinase. The increase in the structural rigidity and decrease in the flexibility of the immobilized inulinase via covalent bonding can justify the higher optimum temperature of immobilized enzyme compared with free enzyme [ [23] , [24] ]. Fig.…”
“…A similar appearance of CNPs was also observed in previous studies. 40,41 The different surface characteristics and morphology of CNPs from CS and TPP molecules suggested that the nano-sized CNP system was formed through association between these molecules and constituted a population of spherical particles. The differences in surface characteristics of CNPs from CS and TPP molecules were significant, as smaller-sized CNPs can only be formed through cross-linking between CS and TPP molecules at a specific volume ratio as exhibited by DLS data.…”
Section: Morphology and Size Of Cnp Ga-cnp And Fga-cnpmentioning
BackgroundInefficient cellular delivery and poor intracellular accumulation are major drawbacks towards achieving favorable therapeutic responses from many therapeutic drugs and biomolecules. To tackle this issue, nanoparticle-mediated delivery vectors have been aptly explored as a promising delivery strategy capable of enhancing the cellular localization of biomolecules and improve their therapeutic efficacies. However, the dynamics of intracellular biomolecule release and accumulation from such nanoparticle systems has currently remained scarcely studied.ObjectivesThe objective of this study was to utilize a chitosan-based nanoparticle system as the delivery carrier for glutamic acid, a model for encapsulated biomolecules to visualize the in vitro release and accumulation of the encapsulated glutamic acid from chitosan nanoparticle (CNP) systems.MethodsCNP was synthesized via ionic gelation routes utilizing tripolyphosphate (TPP) as a cross-linker. In order to track glutamic acid release, the glutamic acid was fluorescently-labeled with fluorescein isothiocyanate prior encapsulation into CNP.ResultsLight Scattering data concluded the successful formation of small-sized and mono-dispersed CNP at a specific volume ratio of chitosan to TPP. Encapsulation of glutamic acid as a model cargo into CNP led to an increase in particle size to >100 nm. The synthesized CNP exhibited spherical shape under Electron Microscopy. The formation of CNP was reflected by the reduction in free amine groups of chitosan following ionic crosslinking reactions. The encapsulation of glutamic acid was further confirmed by Fourier Transform Infrared (FTIR) analysis. Cell viability assay showed 70% cell viability at the maximum concentration of 0.5 mg/mL CS and 0.7 mg/mL TPP used, indicating the low inherent toxicity property of this system. In vitro release study using fluorescently-tagged glutamic acids demonstrated the release and accumulation of the encapsulated glutamic acids at 6 hours post treatment. A significant accumulation was observed at 24 hours and 48 hours later. Flow cytometry data demonstrated a gradual increase in intracellular fluorescence signal from 30 minutes to 48 hours post treatment with fluorescently-labeled glutamic acids encapsulated CNP.ConclusionThese results therefore suggested the potential of CNP system towards enhancing the intracellular delivery and release of the encapsulated glutamic acids. This CNP system thus may serves as a potential candidate vector capable to improve the therapeutic efficacy for drugs and biomolecules in medical as well as pharmaceutical applications through the enhanced intracellular release and accumulation of the encapsulated cargo.
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