Changes in the Microbiome of the Inner Surface of Clear Aligners After Different Usage Periods.

Clear aligners are removable orthodontic appliances that cover the tooth surface. The microbial composition and pH of the inner surface of aligners directly affect the enamel health. In this study, eight subjects who used the same type of clear aligners were instructed to brush their teeth normally and to not clean their aligners until sampling. Saliva and the contents of the inner surface of the aligners (liquid and plaque) were collected at 0 h (T0), 4 h (T4), 8 h (T8), 12 h (T12), and 24 h (T24) after usage, and pH values and microbial compositions were measured. The microbial composition was analyzed with 16S rRNA gene sequencing, and changes were assessed based on operational taxonomic unit abundance. The pH, alpha diversity values, and abundance of specific microbes on the inner surface of the aligners gradually decreased from T0 to T24 (P < 0.05). An insignificant increase in microbial community beta diversity was observed from T0 to T24. Principal component analysis revealed that the microbial composition at T0 was different from at T12 and T24. The relative abundances of phylum Firmicutes (P < 0.01), orders Lactobacillales and Bacteroidales (P < 0.05), and genus Streptococcus and species Streptococcus infantis increased significantly, while those of genera Actinomyces and Rothia and species Rothia aeria decreased significantly at T24 (P < 0.05). These findings reveal that uncleaned aligners might lead to enamel damage, especially after continuous usage for 12 h. Thus, clear aligners should be cleaned after 12 h of usage or at least within 24 h of usage.