PREPARASI NANOPARTIKEL KURKUMIN MENGGUNAKAN KITOSAN RANTAI PENDEK DAN TRIPOLIFOSFAT DENGAN VARIASI KONSENTRASI BERBEDA SERTA UJI CELULAR UPTAKE PADA KULTUR SEL HeLa SECARA IN VITRO

Suryani, Dr. rer. nat. Ronny Martien, M.Si.,

2012 | Tesis | S2 Ilmu Farmasi

Kurkumin termasuk golongan senyawa polifenol alam yang memiliki potensi sebagai anti kanker yang terdapat pada kunyit (Curcuma longa L.) dan potensi optimumnya untuk terapi kanker terhambat oleh bioavabilitas yang rendah. Tujuan penelitian ini adalah meningkatkan bioavabilitas kurkumin dengan cara membuat preparasi nanopartikel kurkumin menggunakan kitosan rantai pendek dengan variasi konsentrasi TPP yang berbeda, karakterisasi, uji stabilitas dalam artificial gastric fluid dan artificial intestinal fluid, uji sitotoksisitas untuk mengetahui toksisitas nanopartikel kurkumin terhadap sel kanker (HeLa) dan sel normal (Vero), dan mengkaji kemampuan nanopartikel kurkumin untuk menembus membran sel HeLa secara in vitro. Pada penelitian ini, dilakukan preparasi sistem penghantaran kurkumin yang dienkapsulasi menggunakan metode ionik gelasi dalam nanopartikel kitosan dan tripolifosfat (TPP) pada berbagai variasi konsentrasi, dikarakterisasi termasuk ukuran partikel, zeta potensial, morfologi dan entrapment efficiency, diuji stabilitasnya dalam artificial gastric fluid (AGF) dan artificial intestinal fluid (AIF) dengan cara menghitung jumlah kurkumin yang dilepaskan per waktu, cellular uptake pada sel HeLa diamati menggunakan miskroskop fluorosens dan efek sitotoksisitasnya pada sel HeLa dan sel normal (Vero) dengan menghitung nilai LC50. Hasil penelitian ini menunjukkan bahwa preparasi nanopartikel kurkumin dapat dibuat dengan metode ionik gelasi menggunakan kitosan rantai pendek dan TPP dengan berbagai variasi konsentrasi dengan rata-rata ukuran partikel yang dihasilkan antara 113,9 s.d. 341,2 nm, zeta potensial antara (-) 2,57 s.d. (+) 27,55 mV, nilai entrapment efficiency 73,63 s.d. 85,71%, dan bentuk partikel yang sferik. Nanopartikel kurkumin stabil dalam media artificial gastric fluid (AGF) dan artificial intestinal fluid (AIF). Nanopartikel kurkumin kitosan lebih bersifat sitotoksik terhadap sel HeLa dengan potensi ketoksikan yang berbeda secara signifikan dengan sel Vero. Nanopartikel kurkumin kitosan memiliki kemampuan menembus membran sel HeLa secara in vitro.

Curcumin, a natural polyphenolic compound with an anticancer properties presents in the turmeric (Curcuma longa L.). The optimum potensial for cancer therapy is limited by its poor oral biovability. The objectives of this study were to improve bioavability of curcumin by preparing curcumin nanoparticles using chitosan and tripolyphosphat at various concentrations, to characterize, to evaluate the stability of curcumin nanoparticles in artificial gastric fluid (AGF) and artificial intestinal fluid (AIF), to identify the cytotoxicity effect of curcumin nanoparticles against HeLa cell lines and normal cell lines (Vero) and to investigate the cellular uptake of curcumin nanoparticles. In this study, curcumin nanoparticles prepared by ionic gelation method using chitosan and TPP at various concentrations. The curcumin nanoparticles were then characterized for their physicochemical characterizations include particle size, zeta potensial morphology and entrapment efficiency. Furthermore, curcumin-chitosan nanoparticles were evaluated for their stability in artificial gastric fluid (AGF) and artificial intestinal fluid (AIF) by quantitating the release of curcumin from the nanoparticles as a function of time, cellular uptake of curcumin nanoparticles investigated using fluorescence microscopy and cytotoxic effect against HeLa cell lines and normal cell lines (Vero) by calculating the level of LC50 which was based on the percentage of the cell death following the 24 hours incubation with the nanoparticles. It was demonstrated that curcumin nanoparticles were prepared by ionic gelation technique using low viscous chitosan and TPP at various concentrations. The mean particle size of curcumin nanoparticles were 113,9 to 341, 2 nm, zeta potensial were (-)2,57 to (+)27,55, entrapment efficiency of curcumin in nanoparticles were 73,63 to 85,71% with the observed shapes of nanoparticles was spherical. Curcumin nanoparticles were stable in artificial gastric fluid and artificial intestinal fluid. Curcumin nanoparticles were more cytotoxic to HeLa cell line and there was a statistically significant difference in potensial cytotoxicity in HeLa cell lines in comparison to vero cell lines, cellular uptake studies of curcumin nanoparticles showed that nanoparticles were able to intenalize the HeLa cell lines.

Kata Kunci : preparasi nanopartikel kurkumin, stabilitas, sitotoksik, HeLa, cellular uptake