Previous studies on honey have shown its potential as an alternative approach in optimizing medical resources for periodontal treatment. However, to date, there has been no study published on the effects of honey on human periodontal ligament fibroblasts (HPDLFs). The aimed of this study was to investigate the effect of Tualang honey on HPDLF proliferation and alkaline phosphatase (ALP) level with an intention to establish honey as one of the natural products which may promote healing and regeneration of periodontium. The HPDLFs were cultured and treated with honey at four different concentrations (0.02, 0.3, 1 and 5%) in a 96-well plate and incubated at 37°C with 5% CO2. Proliferation test (MTT test) was assessed up to day 3 whereas ALP level (ALP assay) was assessed up to day 7. The αMEM supplemented with 10% FBS and 1% penicillin streptomycin was used as control. The results for MTT showed that the cell proliferation for 0.02% honey concentration was significantly higher (p<0.05) on day 3 whereas the proliferation were decreased for 1% honey and 5% honey concentration (p<0.05) than control on all the 3 days. There was increased in ALP level in a similar pattern for all concentration groups from day 0 to day 7 without any significant difference compared with control. Therefore, the present data suggests that Tualang honey stimulated HPDLF proliferation at low concentrations and has an inhibitory effect at high concentrations. However, its role in osteoblastic differentiation requires further investigation.