Autologous platelet-rich plasma (PRP) (also called PRP) has been widely used in medicine, but the mechanism by which PRP heals the wound remains unknown. This study is aimed to analyze the effect of PRP on the production of pro- and anti-inflammatory mediators from macrophage culture because macrophages have an important role in wound healing process. This experimental research used macrophage culture and PRP which was taken from peripheral blood of healthy subjects. Macrophages were obtained from monocyte isolation which was cultured for 1 week, using Roswell Park Memorial Institute (RPMI) medium and granulocyte macrophage-colony stimulating factor (GM-CSF). The cultured macrophages were divided into 5 for different treatments to determine PRP’s effect on pro- and anti-inflammatory mediator production. Pro- and anti-inflammatory mediators were measured using enzyme-linked immunosorbent assay (ELISA) technique. TNF-α levels on day 10 were high and decreased on day 14. Treatment groups receiving PRP experienced significant decrease in TNF-α production. IL-10 levels on day 10 were still low and began to rise on day 14. There was no significant difference between the treatment groups in terms of pro- and anti-inflammatory mediator level changes. PRP increased the production level of TNF-α when the macrophage was activated by LPS (Lipopolysaccharide) and after the macrophage activation began to decrease, the TNF-α production also decreased. PRP did not affect the production of IL-10 at the onset of macrophage activation, but may increase the production of IL-10 at the end of macrophage activation.