Peran ekstrak etanolik daun teh (Camellia sinensis) terhadap respon imun pada rongga mulut :: Kajian laboratoris dan klinis

HANDAJANI, Juni, Promotor Prof.Dr.drg. Al. Supartinah S., SU.,SpKGA

2007 | Disertasi | S3 Ilmu Kedokteran Gigi

Teh (Camellia sinensis) sebagai salah satu jenis tanaman yang telah diketahui memiliki berbagai khasiat antara lain sebagai bahan anti bakteri dan anti inflamasi. Penelitian ini bertujuan untuk untuk mengkaji peran ekstrak etanolik daun teh terhadap respon imun rongga mulut melalui penelitian secara laboratoris dan klinis. Penelitian laboratoris dilakukan pada 30 ekor tikus Wistar jenis kelamin jantan. Sampel dibagi menjadi 3 kelompok, masing-masing berjumlah 10 ekor. Kelompok I diberikan ekstrak etanolik daun teh konsentrasi 2%, Kelompok II diberikan Epigallocathechin gallate (EGCG) dalam akuabides konsentrasi 0,1% sebagai kontrol positif dan Kelompok III diberikan akuabides sebagai kontrol negatif. Tikus diberikan 1 ml bahan sesuai kelompok perlakuan secara per oral selama 14 hari. Mulai hari ke-7 sampai dengan hari ke-14, semua tikus diolesi bakteri A. actinomycetemcomitans pada regio anterior rahang bawah. Pada hari ke-15 dilakukan pengambilan darah tikus dari plexus retro orbitalis untuk isolasi makrofag. Pengujian aktivitas fagositosis makrofag terhadap bakteri A. actinomycetemcomitans. Uji sekresi IL-1β makrofag menggunakan ELISA kit (R&D Systems Inc, USA). Gingiva regio labial anterior rahang bawah dieksisi untuk pengujian ekspresi CD4+ dengan metode immunohistokimia menggunakan monoklonal antibodi anti CD4+ (Santa Cruz Biotechnology, California). Pewarnaan menggunakan ABC Staining System Kit (Santa Cruz Biotechnology, California). Subyek penelitian klinis adalah anak perempuan usia 8 – 10 tahun (Kelas II-IV SD) sebanyak 45 anak. Subyek dibagi menjadi 3 kelompok, masing-masing terdiri dari 15 anak dengan kriteria karies (kelompok I), gingivitis sedang (kelompok II) dan bebas karies dan gingivitis (kelompok III). Pada masing-masing kelompok dibagi menjadi 3 sub kelompok sesuai bahan pasta yang diberikan yaitu pasta ekstrak etanolik daun teh 2%, pasta EGCG 0,1% dan bahan dasar pasta. Penggunaan pasta gigi sehari 2 kali pada pagi hari dan malam hari selama 21 hari. Pemeriksaan klinis dilakukan pengukuran indeks gingiva, indeks PHP, status karies gigi dan kadar sIgA total saliva. Pengukuran sIgA total saliva menggunakan Anti-human sIgA ELISA kit (Salimetrics, LLC, California USA). Pemeriksaan dilakukan pada sebelum perlakuan, hari ke-7, 14 dan ke-21 setelah menggunakan pasta. Data dianalisis menggunakan ANAVA dan LSD. Hasil penelitian menunjukkan ekstrak etanolik daun teh secara laboratoris dapat meningkatkan aktivitas fagositosis makrofag, sekresi IL-1β dan ekspresi CD4+ sel makrofag tikus Wistar. Penggunaan pasta gigi kandungan ekstrak etanolik daun teh secara klinis dapat menurunkan kadar sIgA saliva dan status gingivitis tetapi tidak terdapat perbedaan bermakna (p > 0,05) terhadap status karies gigi. Berdasarkan hasil penelitian dapat disimpulkan bahwa penggunaan ekstrak etanolik daun teh 2% secara sistemik dapat meningkatkan respon imun untuk mencegah terjadinya infeksi bakteri. Secara lokal, ekstrak etanolik daun teh 2% dapat menyembuhkan kondisi gingivitis yang ditandai dengan penurunan kadar sIgA.

Tea (Camellia sinensis) is one of plantation that has been known as many beverages for years. It has an antibacterial and anti-inflammatory activity. The aim of this study was to evaluate the effect of tea leaf ethanolic extract on immune response in the oral cavity by laboratory and clinical method. Thirty male Wistar rats were used and divided into 3 groups based on the material. They were 2% tea leaf ethanolic extract used as treatment (Group I), 0.1% EGCG used as positive control (Group 2) and akuabides used as negative control (Group III). Each group consisted of 10 rats and was administered 1 ml of the material during 14 consecutive days. On the 7th to 14th days, A. actinomycetemcomitans bacteria was applied on the regio anterior lower jaw of all rats. On the fifteenth day, the blood from plexus retro orbitalis was collected by hematokrit. Macrophage from blood was collected by Histopaque (Sigma). Then, the mandibles were cut. Phagocytosis activity was tested to A. actinomycetemcomitans bacteria. IL-1β ELISA kit (R&D Systems Inc, USA) was used to find out the secretion IL-1β of macrophage. The gingival tissue from labial mandibles was disected, and then the expression of CD4+ was determined using immunohistochemistry by adding Primary Monoclonal Antibody CD4+ (Santa Cruz Biotechnology, California) and applying ABC Staining System Kit (Santa Cruz Biotechnology, California). In clinical method, subjects were 45 girls, aged range 8 – 10 years old, divided into 3 groups based on caries status (Group I), gingivitis status (Group II) and healthy (Group III). Each group consisted of 15 subjects, and divided into 3 sub groups based on toothpaste. Tea leaf ethanolic extract toothpaste was used as treatment, EGCG toothpaste was used as positive control and base material toothpaste was used as negative control. The subjects were instructed to brush their teeth during 21 consecutive days in the morning and at night. Clinical evaluation were done by Gingival Index, PHP Index, caries severity index, and total sIgA level. The sIgA level was tested using sIgA ELISA kit (Salimetrics, LLC, California USA). Data were taken before treatment, the 7th, the 14th, the 21st day. Then, they were analyzed using ANOVA and LSD tests. The statistical analysis showed that the effects of 2% tea leaf ethanolic extract were significantly different on phagocytosis activity of macrophage, secretion IL-1β of macrophage, and expression of CD4+. There were significant differences on the effects of tea leaf ethanolic extract toothpaste on the gingival index, PHP index and sIgA saliva level, but there were no significant differences on the caries severity index. This study indicated that the application of 2% tea leaf ethanolic extract could sistematically increase the immune respons. Tea extract toothpaste could decrease the gingivitis effectively indicating by decreasing of sIgA level.

Kata Kunci : Respon Imun Rongga Mulut, Ekstrak Etanolik Daun Teh, Tea (Camellia sinensis), EGCG, oral immune response.