Polymer-clay based nanocomposites are among the attractive materials to be applied for various applications, including biomedical. The incorporation of the nano sized clay (nanoclay) into polymer matrices can result in their remarkable improvement in mechanical, thermal and barrier properties as long as the nanofillers are well exfoliated and dispersed throughout the matrix. In this work, exfoliation strategy through pre-dispersing process of the organically modified montmorillonite (organo-MMT) nanofiller was done to obtain ethyl vinyl acetate (EVA) nanocomposite with improved flexibility, toughness, thermal stability and biostability. Our results indicated that the degree of organo-MMT exfoliation affects its cytotoxicity level and the properties of the resulting EVA nanocomposite. The pre-dispersed organo-MMT by ultrasonication in water possesses higher degree of exfoliation as compared to its origin condition and significantly performed reduced cytotoxicity level. Beneficially, this nanofiller also enhanced the EVA flexibility, thermal stability and biostability upon the in vitro exposure. We postulated that these were due to plasticizing effect and enhanced EVA-nanofiller interactions contributing to more stable chemical bonds in the main copolymer chains. Improvement in copolymer flexibility is beneficial for close contact with human soft tissue, while enhancement in toughness and biostability is crucial to extend its life expectancy as insulation material for implantable device.
The demand for developing bioindicators to assess environmental pollution has increased significantly due to the awareness of potential threats of diseases. Herein, the eukaryotic ubiquitous microorganism Acanthamoeba sp. was used as a bioindicator to explore further the influence of functionalized organic molecules containing -C≡C- and -CH=N- moieties prior application in the potential electronic components. The acetylide and hybrid acetylide-imine derivatives (FYD3A, FYD4B, and FYD4C) were tested for their cytotoxicity potentials based on dose-response analysis, morphological observation, and mode of cell death assessment on Acanthamoeba sp. (environmental-isolate). The biological activities of optimized compounds were evaluated by HOMO-LUMO energy gap and MEP analysis. The determination of the IC50 value through the MTT assay showed functionalized organic molecules of FYD3A, FYD4B, and FYD4C, revealing the inhibition growth of Acanthamoeba sp. with IC50 values in the 3.515 - 3.845 μg/mL range. Morphological observation displayed encystment with cellular agglutination and overall cell shrinkage. AO/PI-stained moieties-treated Acanthamoeba sp. cells appeared with shades of red to orange in necrotic Acanthamoeba cells whilst green to yellow apoptotic Acanthamoeba cells when compared to entirely green fluorescence untreated cells. Moreover, the results of the mitochondrial membrane potential (MMP) assay demonstrate the integrity and functionality potential of the mitochondrial membrane in cells, where a decrease in the MMP assay is linked to apoptosis. This study confirmed that the functionalized organic molecule featuring acetylide and its designated acetylide-imine moieties exhibit cytotoxicity towards the Acanthamoeba sp. by apoptotic and necrotic mode of cell death. This indicates that seeping these derivatives as electronic components can lead to the leaching of hazardous chemicals and contribute to environmental pollution that negatively affects the ecosystem. This study proposes the selection of efficient systems and elements for functionalized organic molecules that are safe to be released into the environment.