Uji Sitotoksik dan Apoptosis ekstrak Saponin Getah Kamboja (Plumeria acuminata Ait) pada kultur sel Fibroblast Pulpa

SUSANTO, Hendri, dr. Harijadi, Sp PA

2004 | Tesis | S2 Ilmu Kedokteran Gigi

Aplikasi getah kamboja (Plumeria acuminatae Ait) pada kavitas gigi Molar satu rahang bawah Marmot (Cavia cabaya) menyebabkan gigi devital selama beberapa hari. Hal ini menunjukkan adanya saponin pada getah kamboja kemungkinan dapat dipakai sebagai bahan devitalisasi pulpa alternatif Arsen Trioksida yang memiliki efek samping merugikan. Saponin diketahui bersifat sitotoksik dan menyebabkan apoptosis sel. Tujuan penelitian ini adalah untuk mengetahui sitotoksik ekstrak saponin getah kamboja terhadap kultur sel fibroblas pulpa, adanya apoptosis sel fibroblas pulpa dan hubungannya dengan konsentrasi ekstrak saponin getah kamboja dan waktu paparan Penelitian dilakukan pada 66 sumur sampel kultur sel fibroblas pulpa gigi manusia. Enam puluh enam sampel tersebut yang berisi 2x104 sel/100 μl dibagi menjadi 4 kelompok terdiri atas kelompok ekstrak saponin getah kamboja 0,01; 0,1; 1; 15,76; 100; 1000 μg/ml, kontrol pelarut DMSO 0,25%, kontrol positif Arsen Trioksida 1; 7,5; 15,76 μg/ml, dan kelompoks sel tanpa pelakuan. Setiap perlakuan terdiri atas 3 sumur sampel dengan waktu paparan, yaitu 24 jam dan 48 jam. Uji sitotoksik dilakukan mengunakan metode dye exclusion test. Pemeriksaan sel apoptosis dilakukan dengan pemeriksaaan imunohistokimiawi ekspresi protein CAS (Cellular Apoptosis Susceptibility) pada kultur sel fibroblast pulpa yang diberi ekstrak saponin getah kamboja dengan menggunakan antibodi monoklonal terhadap ekspresi CAS. Aktivitas sitotoksik ditunjukkan dengan nilai IC50 dan apoptosis sel fibroblas pulpa ditunjukkan dengan prosentase jumlah sel yang mengekspresikan CAS. Hasil penelitian ini menunjukkan bahwa ekstrak saponin getah kamboja memiliki aktivitas sitotoksik dengan menghasilkan nilai IC50 42,48 μg/ml (24 jam) dan 22,47 μg/ml (48 jam). Ekstrak saponin getah kamboja menyebabkan apoptosis sel fibroblas pulpa dan jumlah sel fibroblas apoptosis memiliki hubungan negatif (p<0,05) dengan konsentrasi ekstrak saponin getah kamboja dan waktu paparan. Kesimpulan penelitian ini adalah ekstrak saponin getah kamboja memiliki aktivitas sitotoksik terhadap kultur sel fibroblas pulpa dan menyebabkan apoptosis sel fibroblas pulpa serta terdapat hubungan negatif antara konsentrasi ekstrak saponin getah kamboja dan waktu paparan dengan jumlah sel fibroblas yang apoptosis.

The use of Plumeria acuminate Ait secretion in Cavia cabaya lower first Molar cavity resulted a devital tooth for several days. It meant there was possibility saponin of Plumeria acuminatae Ait secretion can be used as an alternative devitalization agent of the Arsenic Trioxide that has been proved giving adverse side effect. Saponin has been known having cytotoxic activity and induce apoptosis. The aims of this study were investigate the cytotoxic effect of saponin of Plumeria acuminatae Ait secretion in cultured pulp fibroblast cell, apoptotic number of pulp fibroblast cell which induced by saponin of Plumeria acuminate Ait secretion and the relationship between saponin of Plumeria acuminatae Ait secretion concentration and exposure time with apoptotic number of pulp fibroblast cells. Sixty six cultured human pulp fibroblast cell sample wells used for this study. Each well contains 2x104/100 μL. All samples were divided into four groups consist of saponin of Plumeria acuminatae Ait secretion (0.01; 0.1; 1; 15.76; 100; 1000 μg/ml) groups, Arsenic Trioxide (1; 7.5; 15.76 μg/ml) groups as positive control, negative control group without any treatment, DMSO 0,25% control group. Each treatment consists of three sample wells with 24 hours and 48 hours exposure time. Dye exclution test used for Cytotoxic study. Monoclonal antibody to protein CAS (Cellular Apoptosis Susceptibility) expression immunohistochemistrically on cultured pulp fibroblast cell samples treated by saponin of Plumeria acuminatae Ait secretion used for Apoptotic cell examination. Cytotoxic activity is showed by IC50 of saponin of Plumeria acuminatae Ait secretion in cultured pulp fibroblast cell. Apoptotic pulp fibroblast cell showed by CAS expression of pulp fibroblast cells percentage. The result of this study showed that saponin of Plumeria acuminatae Ait secretion has cytotoxic activity in cultured pulp fibroblast cell. The IC50 of saponin of Plumeria acuminatae Ait secretion were 42.48 μg/ml (24 hrs) and 22.47 μg/ml (48 hrs). There were apoptosis of pulp fibroblast cells and a negative relationship by exposure Plumeria acuminatae Ait. secretion in level of concentration and exposure time (p<0,05). This study concluded that saponin of Plumeria acuminatae Ait secretion has cytotoxic activity in cultured pulp fibroblast cell and induce apoptosis of pulp fibroblast cell. There was a negative relationship between saponin of Plumeria acuminatae Ait secretion concentration and exposure time with apoptotic number of pulp fibroblast cells.

Kata Kunci : Gigi,Kultur Sel Fibroblast Pulpa,Sitotoksik Ekstrak Saponin Getah Kamboja, saponin of Plumeria acuminatae Ait secretion, cultured pulp fibroblast cell, cytotoxic, apoptosis