Effective healing of skin injury involves complex simultaneous processes. Extracts of meniran (Phyllanthus niruri L.), a plant medicine found in almost all parts of Indonesia, have been essential in traditional wound healing treatments. Phyllanthin is the major therapeutically active lignan present in Phyllanthus species. This study investigated the wound-healing activity. of ointments prepared from a phyllanthin-rich subfraction of meniran leaf in an animal model. Two parameters were measured, namely, hydroxyproline concentration and wound healing percentage. Hydroxyproline is a biochemical marker representing collagen production in wound healing. Seventy-two rats were divided into four groups of 18, including a control group and three groups administered 5, 10, and 15% phyllanthin-rich fraction ointment. Each of the four groups was subdivided according to the treatment duration of 5, 10, and 15 days. The hydroxyproline concentration and wound healing percentage were analyzed by one-way ANOVA followed by Duncan’s post-hoc test. The increase in hydroxyproline concentration and wound healing percentage showed that the effect of each ointment concentration was significantly different (p <0.05) compared to the control group. The optimal ointment concentration was 10%, with optimal results after 15 days. The study revealed that the phyllanthin-rich sub-fraction isolated from meniran (Phyllanthus niruri L.) had a wound healing activity on Adult male Wistar rats (Rattus novergicus). © 2024 Tobat et al.