Cytotoxicity assessment of two biogenic silver nanoparticles and their effects when incorporated into an experimental resin-based adhesive system

Abstract

Objectives: To evaluate and compare the in vitro cytotoxicity of two biogenic silver nanoparticles (AgNPs) synthesized using Phanerochaete chrysosporium and Penicillium expansum, as well as their effects when incorporated into an experimental resin-based adhesive system.

Methods: Murine NIH/3T3 fibroblasts will be used as the cellular model. Experimental groups will include: both biogenic AgNPs alone, the unmodified adhesive system, and the adhesive system modified with different concentrations of each AgNP type. Concentrations will be established through pilot studies. Cell viability and half-maximal inhibitory concentration (IC50) will be assessed using MTT assays at 24 and 48 hours. Briefly, experimental materials will be incubated in DMEM culture medium to obtain eluates, which will subsequently be applied to cells seeded in 96-well plates. After the corresponding incubation periods, absorbance will be measured at 570 nm, and IC50 values will be calculated using GraphPad Prism 8 software. In addition, cellular morphological changes will be monitored in real time using high-resolution microscopy (Nanolive 3D Cell Explorer), acquiring images every 15 minutes over a 48-hour period. Morphometric and cell viability parameters will be quantified using Steve and ImageJ/FIJI software.

Expected Results: The study is expected to determine the cytotoxic profile of both biogenic AgNPs and the modified adhesive systems. Likewise, the nAg from *P. chrysosporium* are expected to exhibit better biocompatibility than those from *P. expansum*, contributing to the development of safer dental biomaterials.