Under challenging oral environments, the overall performance of resin composites is affected by bio-aging. This study investigated the effects of saliva biofilm-induced bio-aging on the mechanical properties and microbial behavior of composites with different filler types. Microhybrid, nanohybrid, nano-filled and nano-filled flowable composites were bio-aged with saliva biofilm for 30 days. Surface morphology, roughness, mechanical and aesthetic properties were determined. A 48 h saliva biofilm model was used to evaluate the microbial behavior of different composites in vitro. Biofilm metabolic activity, lactic acid production and live/dead bacterial staining were tested. Six volunteers were selected to wear intra-oral appliances with composite slabs for 24 h and biofilms were collected and analyzed using 16S rRNA sequencing to assess the biofilm formation over those materials in situ. Although there were increasing trends, surface roughness, water resorption and material solubility had no significant changes for all groups after bio-aging (p > 0.05). There were no significant changes in elastic modulus for all groups after aging (p > 0.05). However, a decrease in flexural strength in all groups was observed (p < 0.05), except for the nanoflow composite group (p > 0.05). The Vickers hardness remained stable in all groups after aging (p > 0.05), except for the nano-filled group (p < 0.05). The nanoflow composite showed distinct color changes compared to the micro-hybrid group after aging (p < 0.05). Biofilm metabolic activity and lactic acid production in vitro increased slightly after bio-aging in all groups, but with no statistical significance (p > 0.05). The Shannon index diversity of biofilms in situ decreased after aging (p < 0.05), while no significant difference was shown in species composition at the genus level in all groups (p > 0.05). Resin composites with different sized fillers displayed a relatively stable mechanical performance and uncompromised microbial behavior both in vitro and in situ after 30 days of bio-aging. Based on the results, composites with different filler types can be selected flexibly according to clinical needs. However, a longer time for bio-aging is still needed to confirm the mechanical properties and microbial behaviors of composites in the long run.
Keywords: bio-aging; biofilm; caries prevention; mechanical properties; microbial community; resin composites.