Pengaruh reuterin isolat lactobacillus reuteri terhadap ekspresi gen asa1, gelE, salB, dan atlA biofilm enterococcus faecalis dalam saluran akar gigi
L Latar Belakang: Enterococcus faecalis berperan penting pada kegagalan perawatan saluran akar. Atas kemampuannya membentuk biofilm, E. faecalis menjadi resisten terhadap beberapa larutan irigasi dan medikamen intrakanal. Sebagian gen telah diidentifikasi terkait dengan pembentukan biofilm E. faecalis. Penggunaan probiotik Lactobacillus reuteri dikenal dapat menghambat mikroorganisme dengan spektrum luas termasuk bakteri Gram-positif, bakteri Gram-negatif, jamur, serta protozoa. L. reuteri juga mampu menurunkan gangguan kesehatan rongga mulut, seperti gingivitis dan indeks plak. Tujuan: Untuk mengevaluasi pengaruh reuterin isolat L. reuteri terhadap ekspresi gen asa1, gelE, salB, dan atlA dalam pembentukan biofilm E. faecalis dengan model saluran akar secara ex vivo. Metode: Dua puluh empat gigi manusia yang telah dicabut dipilih. Setelah preparasi biomekanis, saluran akar diinokulasi dengan E. faecalis yang telah dikultur selama 24 jam. Kelompok penelitian dibuat menurut jenis bahan irigasi yaitu salin (kontrol negatif), NaOCl 2.5% (kontrol positif), reuterin isolat 25 μg/mL, and reuterin prodentis 25 μg/mL. Sampel irigasi diambil dalam waktu 5 menit dan 30 menit pasca irigasi untuk menentukan unit pembentuk koloni yang tersisa. RT-qPCR digunakan untuk mendeteksi ekspresi gen dalam pembentukan biofilm isolat E. faecalis dari saluran akar. Hasil: Reuterin mampu menurunkan regulasi transkripsi ekspresi gen gelE dan atlA terkait pembentukan biofilm terutama saat proses maturasi biofilm secara signifikan (p<0.05). Hasil juga menunjukkan adanya pengaruh reuterin terhadap ekspresi gen asa1 dan salB walaupun tidak secara signifikan. Kesimpulan: Penelitian ini menunjukkan bahwa reuterin isolat L. reuteri 25 μg/mL memiliki potensi sebagai bahan irigasi dalam perawatan saluran akar gigi melalui kemampuan antibiofilm yang dimilikinya. Reuterin juga dapat mempengaruhi ekspresi gen dalam pembentukan biofilm E. faecalis secara ex vivo. Penelitian lebih lanjut secara in vivo, ex vivo, maupun studi klinis untuk mengetahui komponen reuterin isolat L. reuteri dan mekanismenya pada ekspresi gen lain terkait biofilm E. faecalis masih perlu dilakukan.
B Background: Enterococcus faecalis plays an important role mainly in root canal failures. Due to its ability to form biofilm, E. faecalis become resistant to some of irrigating solutions and intracanal medicaments. Multiple genes have been identified that are associated to biofilm formation of E. faecalis. The use of probiotic Lactobacillus reuteri is well-known for inhibiting a wide spectrum of microorganism including Gram positive bacteria, Gram-negative bacteria, fungi, and protozoa. L. reuteri also capable to reduce oral health problems, such as gingivitis and plaque index. Objective: To evaluate the effect of reuterin isolate L. reuteri on asa1, gelE, salB, and atlA genes expression in biofilm formation of E. faecalis using ex vivo root canal model. Methods: Twenty four extracted human teeth were selected. After biomechanical preparation, root canals were inoculated with E. faecalis that has been cultured for 24 hours. Research groups samples were made according to the type of irrigation solution namely saline (negative control), NaOCl 2.5% (positive control), reuterin isolate 25 μg/mL, and reuterin prodentis 25 μg/mL. The irrigation samples were obtained in 5 minutes and 30 minutes postirrigation to determine the remaining colony forming units. RT-qPCR were used for detecting the genes expression in biofilm formation of E. faecalis isolates from root canal. Results: Reuterin was able to significantly (p<0,05) down-regulated transcription of gelE and atlA genes expression related to biofilm formation, mainly during the biofilm maturation process. The results also showed an effect of reuterin on the expression of asa1 and salB genes, although not significant. Conclusion: This study shows that reuterin isolate L. reuteri 25 μg/mL has potential as an irrigation agent in root canal treatment through its antibiofilm ability. Reuterin can also affects the gene expression in E. faecalis biofilm formation ex vivo. Further in vivo, ex vivo, and clinical studies to determine the component of reuterin isolate L. reuteri and their mechanisms on other gene expression related to E. faecalis biofilm is still needed.