Introduction: The aim of this study was to synthesize an optimized chitosan/carbon dot hydrogel and measure its effects on the proliferation of dental pulp stem cells. Methods: 1.5% w/v chitosan hydrogel samples were fabricated with 300 mg/ml aqueous β-glycerol phosphate (β-GP) to create an optimized chitosan with a gelation onset temperature below 37°C. Spermidine-functionalized carbon dots (CDs) were added to test groups in concentrations of 25 µg/ml and 50 µg/ml. Results: The optimized chitosan hydrogel samples had a gelation onset temperature of 34.6°C, which was not appreciably increased with the addition of CDs (34.7°C). All experimental chitosan hydrogels demonstrated superior cellular proliferation when compared to calcium hydroxide paste. Conclusions: Chitosan/CD hydrogels may be a promising medicament for endodontic use and a viable alternative to either Ca(OH)2 or TAP in regenerative endodontic cases. The CD structure contains surface characteristics that are open to customization for individual therapeutic needs. This hydrogel is thermally responsive and becomes a solid gel when administered at human body temperature.