<![CDATA[Abstract: Periodontal disease is caused by the accumulation of plaque biofilm; one of the bacteria involved is Staphylococcus aureus. Red betel leaf (Piper crocatum) has potential as a therapeutic agent for various diseases, including oral cavity diseases. This study aimed to determine the effect of administering red betel leaf extract (EDSM) in inhibiting the formation of Staphylococcus aureus bacterial biofilms. This was a laboratory experimental study. The test group consisted of EDSM with concentrations of 25%, 50%, 75%, and 100%, while the positive control used 0.2% chlorhexidine/ CHX, and 1% DMSO as a negative control. Red betel leaf extract was prepared using the 96% ethanol maceration method. Inhibition of biofilm formation was measured using the microtiter plate biofilm assay method with a wavelength of 595 nm. Data were analyzed using the Kruskal-Wallis test and the Mann-Whitney post hoc test. The results showed significant differences between the six groups. Post hoc test revealed significant differences between DMSO 1% vs CHX, DMSO 1% vs EDSM 50% concentration, DMSO 1% vs EDSM 25% concentration, CHX vs EDSM 100% concentration, CHX vs EDSM 75% concentration, CHX vs EDSM 50% concentration, CHX vs EDSM 25% concentration, and EDSM 75% vs EDSM 25% concentration with p values <0.05. In conclusion, red betel leaf extract is most effective in inhibiting Staphylococcus aureus biofilm formation at a concentration of 25%. Keywords: antibiofilm; red betel leaf extract; Staphylococcus aureus Abstrak: Penyakit periodontal disebabkan oleh akumulasi biofilm plak; salah satu bakteri yang berperan yaitu Staphylococcus aureus. Daun sirih merah (Piper crocatum) berpotensi sebagai agen terapi berbagai penyakit termasuk penyakit rongga mulut. Penelitian ini bertujuan menentukan efek pemberian ekstrak daun sirih merah (EDSM) dalam menghambat pembentukan biofilm bakteri S. aureus. Jenis penelitian ialah eksperimental laboratorium. Kelompok uji terdiri dari EDSM dengan konsentrasi 25%, 50%, 75%, dan 100%, sedangkan kontrol positif menggunakan klorheksidin/CHX 0,2%, dan DMSO 1% sebagai kontrol negatif. Ekstrak daun sirih merah dibuat menggunakan metode maserasi etanol 96%. Penghambatan pembentukan biofilm diukur menggunakan metode microtiter plate biofilm assay dengan panjang gelombang 595 nm. Data dianalisis dengan uji Kruskal-Wallis dan uji post hoc Mann-Whitney. Hasil analisis menunjukkan perbedaan bermakna antar enam kelompok. Uji post hoc mengungkapkan perbedaan bermakna antara DMSO 1% vs CHX, DMSO 1% vs EDSM konsentrasi 50%, DMSO 1% vs EDSM konsentrasi 25%, CHX vs EDSM konsentrasi 100%, CHX vs EDSM konsentrasi 75%, CHX vs EDSM konsentrasi 50%, CHX vs EDSM konsentrasi 25%, serta EDSM 75% vs EDSM 25% dengan nilai p<0,05. Simpulan penelitian ini yaitu EDSM paling efektif dalam menghambat pembentukan biofilm Staphylococcus aureus pada konsentrasi 25%. Kata kunci: antibiofilm; ekstrak daun sirih merah; Staphylococcus aureus]]>
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