Synthesis, Characterization and Cytotoxic Evaluation of Insoles Impregnated with Copper Nanoparticles

Tay, Ying Shan (2019) Synthesis, Characterization and Cytotoxic Evaluation of Insoles Impregnated with Copper Nanoparticles. Final Year Project (Bachelor), Tunku Abdul Rahman University College.

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Abstract

Copper nanoparticles (Cu NPs) has been extensively synthesized in various industrial and commercial applications to perform its multifunctional properties such as antifungal and antibacterial. In this study, copper nanoparticle synthesis, characterization using scanning electron microscope (SEM), X-ray diffraction (XRD) and cytotoxic effect of the impregnated copper nanoparticles insoles towards mammalian cell lines were performed. This study aims to synthesize and characterize the copper particles and to evaluate the of cytotoxicity effect of the insole impregnated with Cu NPs towards mammalian cells via an in vitro model. Cu NPs were synthesized by using chemical reduction method and the sizes were confirmed by SEM and XRD analysis. The SEM results showed low distribution of Cu NPs and size in the range of 89.80 nm to 393.9 nm. XRD results showed the crystalline nature with peaks corresponding to Cu NPs and silica particles, the average size of Cu NPs was between 28 to 57 nm. The insoles impregnated with Cu NPs were then assessed for toxicity effect in human dermal fibroblast (HDF) cells and human hepatocellular carcinoma cells (HepG2) by using microculture tetrazolium (MTT) assay for cell viability. This biological evaluation was performed via indirect contact method. The leachate growth medium were prepared for cytotoxicity analysis instead of exposing cell lines to impregnated insoles directly. The six treatments include insole coated with Cu particles, Cu with ethylene glycol (EG), copper precursor (Cu(NH3)4(OH)2), Cu(NH3)4(OH)2 with EG, EG and insole without coating. The positive control was 30% v/v DMSO whereas negative control was medium without insole. The insoles were immersed into Dulbecco modified Eagle medium (DMEM) for 4 hours. The leachates medium solution from 4 hours were incubated with the cell lines for 24 hours prior for cytotoxicity testing. The comparative effects on the types of cell lines on cell viability were studied in MTT assay, but in HDF cells showed negative result with no significant data between positive and negative control. Morphologically, the HDF passage 17 showed healthy after treatments but unable to uptake MTT reagent. HepG2 cells showed significant difference (P<0.05) between treated and untreated cells. In conclusion, Cu NPs synthesis method is environmental friendly and cost effective. Cu NPs coated insoles have the potential for practical application based on its targeted antimicrobial effect reported previously. The toxicity of Cu NPs, copper precursor and ethylene glycol was observed in HepG2 and caused the cell viability to decrease. However, cytotoxicity effect on HDF cells was not conclusive in the current investigation due to cell condition was not in ideal condition.