Background: Toothbrushing is the most widespread method of oral hygiene maintenance at a personal level. With the variety of toothbrushes available on the market and increasing awareness about making sustainable choices, bamboo toothbrushes are gaining popularity. Since worn-out toothbrushes lose their effectiveness, this study aims to compare the wear between nylon-bristled toothbrushes and bamboo toothbrushes with plant-based bio-bristles. Materials and methods: Toothbrush bristle wear was checked between bamboo toothbrushes with plant-based bio-bristles and nylon-bristled toothbrushes after 7 days of use by 20 participants aged 8–11 years, using the wear index (WI) and acceptable percentage of bristle end morphology. The Mann–Whitney U test and Fisher's exact test were used to compare measurements of WI and acceptable percentages of bristle end morphology. The difference was considered significant if p < 0.05. Results: The WI of nylon bristles was −0.0120 ± 0.0986, with a standard error of the mean of 0.0311. For bamboo bristles, the mean was 0.1001 ± 0.0920, with a standard error of the mean of 0.0291. In the nylon bristles group, nine (90%) of bristles had acceptable morphology, whereas one (10%) had nonacceptable morphology. In the bamboo bristles group, three (30%) of bristles had acceptable morphology, whereas seven (70%) had nonacceptable morphology. Conclusion: This study compares the wear between nylon-bristled toothbrushes and bamboo toothbrushes with plant-based bio-bristles and indicates that nylon-bristled toothbrushes exhibit lower wear compared to bamboo toothbrushes with plant-based bio-bristles.