It is crucial to prevent microbial and viral infections as a results of microbial contamination, epidemiological factors, and infection risks in many areas, especially in clinics, hospital, food packaging sector, and dental equipments. Therefore, attention of many researchers was focused on the fabrication and application of a new generation of functional polymer nanostructures, and polymer-biopolymer/inorganic nanofiller hybrid composites with desired properties such as antimicrobial and superhydrophobicity [1][2][3][4][5][6].Among them, silver nanomaterials include silver nanoparticles, stabilized silver salts, silver-dendrimers, silver-polymer and metal oxide composites, and activated carbon materials demonstrated antibacterial effect against high toxic organisms [7]. In addition, silver-enhanced zeolite, antibacterial polymer thin films containing silver cations-zeolite additives can be also included into this classification [8]. The effect of synthetic and biopolymer based surfactants on the stability and bioactivity of silver nan particles were investigated. Additionally, the antibacterial activity Abstract. This work presents a new approach to synthesize the colloidal ODA-MMT-poly(maleic anhydride-alt-1-dodecene)-g-α,ω-methoxyhydroxyl-PEO/silver nanoparticles (AgNPs) nanohybrid composites (NHC) using the following synthetic pathways: (1) complex-radical alternating copolymerization of maleic anhydride with 1-dodecene α-olefin comonomer, (2) grafting of PEO onto alternating copolymer through esterification, (3) intercalating a copolymer-g-PEO between organoclay layers via complex formation of maleate carboxyl with octadecyl amine, and (4) in situ generation of AgNPs in polymer nanocomposite by annealing method under vacuum. The obtained multifunctional NHCs with different contents of AgNPs were characterized by UV spectroscopy, ζ-potential and size analysis methods. It was demonstrated that annealing of the colloidal NHC is accompanied with in situ generation of stable and partially protonated AgNPs due to specific reducing and stabilizing effects of multifunctional matrix polymer contained positively charged reactive and bioactive sites. Antibacterial and antifungal activities against Gram-negative and Gram-positive bacteria and fungal microorganism were investigated. The cytotoxic, apoptotic and necrotic effects in NHC/L929 fibroblast cells systems were evaluated. The synthesized watersoluble, biocompatible, and bioactive colloidal NHCs are promising candidate for a wide-range of applications in air filtration, food packaging systems, bioengineering, especially in tissue regeneration and nanomedicine.