Background The wound healing process can be optimized through the addition of a biomaterial such as recombinant secretory leukocyte protease inhibitor (rSLPI). The SLPI is a non-glycosylated proteomic material that inhibits protease enzymes and has anti-inflammatory properties, thus accelerating wound healing. This study analyzed the administration of rSLPI doses 0.04 cc and 0.06 cc in skin wound healing on the CD163 expression of macrophages and cytokines such as interleukin 1 (IL-1), interleukin 6 (IL-6) and fibroblast growth factor 2 (FGF-2). Materials and Methods rSLPI produced from Escherichia coli TOP10 as the cloning host, BL21 (DE3) strains as the expression host and pET30a plasmids were used for the expression system construction. The wound was created on Wistar rat dorsal skin, then rSLPI 0.04 cc and 0.06 cc was administered. In the next four days, the back skin was biopsied and stained by immunohistochemistry to analyze the CD163, FGF-2, IL-1 and IL-6 expression. Results The administration of rSLPI increased CD163 and FGF-2 expression dependent on dose (p<0.05). On the other hand, administration of rSLPI decreased IL-1 and IL-6 expression depending on dose (p <0.05). Conclusion The administration of rSLPI is able to accelerate the wound healing process by increasing the CD163 and FGF-2 expression. The cytokines such as IL-1 and IL-6 decreased depending on rSLPI doses.