RESPON IMUN SELULER PADA PERIODONTITIS DIABETIKA

Ahmad Syaify, drg.Sp.Perio., Prof. dr. Marsetyawan HNE Soesatyo, MSc, PhD,

2012 | Disertasi | S3 Kedokteran Gigi

Keparahan periodotitis diabetika pada pengidap DM tipe 2 diduga kuat disebabkan oleh gangguan respon imun seluler yang diperankan oleh monosit dan neutrofil. Calprotectin merupakan protein sitosol yang hanya dihasilkan oleh neutrofil dan monosit, dan ditemukan di dalam cairan serum serta cairan sulkus gingiva. Tujuan penelitian ini untuk mengkaji peran respon imun seluler pada penderita periodontitis diabetika dilihat dari level calprotetcin dan eskpresi MRP8/14 mRNA serta aktivitas fagositosis. Penelitian observasional secara cross sectional dilakukan untuk mengkaji level calprotectin dalam cairan sulkus gingiva serum, monosit dan neutrofil penderita periodontitis diabetika menggunakan teknik ELISA. Penelitian eksperimental semu untuk mengetahui respon monosit dan neutrofil dalam mengekspresikan MRP8/MRP14 mRNA setelah kedua sel fagositik distimulasi dengan lipopolisakarida (LPS) bakteri E.coli. Deteksi ekspresi kode genetik calprotectin ini menggunakan teknik RT-PCR, dilanjutkan dengan elektroforesis, dan pembacaan densitas pita hasil PCR dengan thin layer chromatography (TLC). Uji aktivitas fagositosis monosit dan neutrofil menggunakan partikel lateks, dan dilanjutkan penghitungan indeks fagositosis. Hasil penelitian memperlihatkan adanya peningkatan kadar calprotetcin pada penderita periodontitis diabetika tidak terkontrol, baik di dalam cairan sulkus gingiva, serum, maupun monosit serta neutrofil darah tepi. Analisis statistik Oneway ANOVA menunjukkan perbedaan yang signifikan (p<0.05) antara kadar calprotectin penderita periodontitis disertai DM tipe 2 tidak terkontrol, dengan penderita periodontitis DM tipe 2 terkontrol, dan periodontitis tanpa DM. Ekspresi MRP8/MRP14 mRNA pada monosit maupun neutrofil tertinggi pada periodontitis dengan DM tipe 2 tidak terkontrol dan terendah pada periodontitis non DM, secara statistik perbedaannya signifikan (p<0.05). Adapun indeks fagositosis, terendah pada penderita periodontitis dengan DM tipe 2 tidak terkontrol Secara statistik dengan uji Oneway ANOVA menunjukkan bahwa perbedaan indeks fagositosis ini signifikan (p<0.05) antara penderita periodontitis diabetika tidak terkontrol dengan periodontitis non DM. Kesimpulan dari penelitian ialah terjadi peningkatan kadar calprotetcin maupun ekspresi MRP8/14 mRNA pada penderita periodontitis diabetika, serta penurunan aktifitas fagositosis baik oleh monosit maupun neutrofil. Kadar calprotectin dan ekspresi MRP8/14 mRNA tertinggi yaitu pada penderita periodontitis dengan DM tipe 2 tidak terkontrol, sehingga diduga kuat bahwa perubahan kadar calprotectin dan ekspresi MRP8/14 terkait dengan regulasi keparahan dari periodontitis diabetika.

The severity of diaberic periodontitis was strongly thought caused by immune response disorder, especially by decreasing functions of monocytes and neutrophiles. Calprotectin is cytosolic protein that was only produced by neutrophiles and monoctyes. It was also existed in serum and gingival crevicular fluid. The aims of this study was to determine the role of innate immune response in diabetic periodontitis on calprotectin level in terms of gingival crevicular fluid, serum, monocytes and neutrophiles and the expression MRP8/14 on both these phagocytic cells, and their phagocytosis function. Observational research with cross sectional approach was taken to assess levels of calprotectin in gingival crevicular fluid serum, monocytes and neutrophils of diabetic patients with periodontitis using ELISA technique. Quasi experimental research was conducted to study the response of monocytes and neutrophils in MRP8/MRP14 mRNA expression after both phagocytic cells stimulated with lipopolysaccharide (LPS) E. coli bacteria. Detection of calprotectin expression of this genetic code using RT-PCR technique, was then followed by electrophoresis, and the density of PCR bands with thin layer chromatography (TLC) was analyzed. Test of phagocytosis activity of monocytes and neutrophils was done using latex particles and phagocytosis index was resumed. The results showed an increased calprotectin levels in uncontrolled diabetic patients with periodontitis, both in gingival crevicular fluid, serum, and peripheral blood monocytes and neutrophils. Statistical analysis using Oneway ANOVA showed significant differences (p<0.05) between calprotectin level of periodontitis patients with uncontrolled type 2 diabetes, those with controlled type 2 diabetes, and without diabetes. Meanwhile, test on MRP8/MRP14 mRNA expression of monocytes and neutrophils provided the highest expression in the periodontitis group with uncontrolled type 2 diabetes and lowest expression in non-diabetic periodontitis, which was statistically significant difference (p <0.05). While phagocytosis index indicated the opposite tendency, namely in periodontitis patients with uncontrolled type 2 diabetes phagocytic index of monocytes and neutrophils were precisely lowest compared to periodontitis in controlled type 2 diabetes, and non-diabetic periodontitis with highest phagocytosis index. Result of Oneway ANOVA showed that differences in phagocytic index was significantly different (p <0.05). The conclusion of the research are level calprotetctin and MRP8/14 mRNA expression increased on periodontitis patients with type 2 DM. On the other hand, the phagocytosis activity decreased on those patients. The highest level of calprotectin and MRP8/14 expression was on periodontitis patients with type 2 DM. It was suggested that level of calprotectin and MRP8/14 expression have a role in the regulation of severity diabetic periodontitis.

Kata Kunci : periodontitis, DM, monosit, neutofil, calprotectin, MRP8/14