Nurul Hafizah Mohd Nor, Nur Azhani Zainol Abidin, Nur Liyana Mohd Fozi, Nur Sulaihah Omar & Farahidah MohamedPharmaceutical Technology Department, Kulliyyah of Pharmacy, International Islamic University Malaysia

Email: farahidah@iium.edu.my

1. To develop an oil-in-water (o/w) Nigella sativa microemulsion (ME)

2. To improve ME’s aqueous solubility and uptake by PC12 cell line

3. To investigate in vitro effect of N. sativa in exerting neurite outgrowth on aqueous environment of PC12 cell line

OBJECTIVES

ABSTRACT

The aims of this project were to develop the oil-in-water (o/w) Nigella sativa microemulsion in order to improve its aqueous solubility and hence uptake by PC12 cell lines. Such system was envisaged to be feasibly used to see in vitro effect of N. sativa in exerting neurite outgrowth on aqueous environment of PC12. The microemulsion (ME), prepared by spontaneous emulsification method, composed of varying pre-determined volumes of the N. sativa oil and sorbitan-based surfactant blends, were titrated against water until either turbidity or clear translucent phase were formed. Corresponding volumes of the 3 components were plotted into a phase diagram. All translucent mixtures were characterized for their viscosity and droplet size. Further stability tests on the microemulsions were also conducted following storage at room temperature, heating and centrifugation. Results showed that droplet diameters of ME 12 (9.09% oil, 72.73% Tween20:Tween80 (6:4), 18.18% water), ME 13 (10.71% oil, 71.43% Tween20:Tween80 (6:4), 17.86% water) and ME 14 (7.41% oil, 74.07% Tween20:Tween80 (6:4), 18.52% water) were 24.18 nm, 14.96 nm and 46.86 nm respectively and gave transparent solution with no turbidity after being subjected to vortex mixer. Subsequent analysis will involve loading of these microemulsion into PC12 cell line to see the effect of N. sativa oil in exerting neurite extension. This approach seems to be potentially useful for in vitro rapid screening study of lipophilic agent that involves aqueous cell lines.

The authors would like to thank Ministry of Science, Technology and Innovation Malaysia (MOSTI) for the funds, IIUM and all laboratory staffs.

CONCLUSIONThis approach seems to be potentially used for in vitro rapid screening study of lipophilic agent that involves aqueous cell lines. Stable N. Sativa oil-in-water microemulsion had been successfully produced by appropriate blending of surfactant types with the N. sativa oil and water. The findings also suggest that range of HLB ratio for similar surfactant groups could be utilized as guideline to find correct ratio between the ternary components

RESULT

Table 1 Table shows the evaluation of N. sativa oil in water-based media microemulsion formulations

Figure 1 It shows the pseudoternary diagram of N. sativa microemulsions

 Table 2 Table shows the physical characteristic of N. sativa oil in water-based media microemulsion formulations

Table 3 Table shows the stress-testing for phase separation of N. sativa microemulsions.

Figure 2 It shows the neurite outgrowth of PC12 cell line 96H post-treatment with ME12, ME13 and ME14

Translucent phase were

formed

Spontaneous emulsification

method

Figure 3 It shows the neurite extension of PC12 cell line 96H post-treatment with ME12, ME13 and ME14

ACKNOWLEDGEMENT

REFERENCE

DISCUSSION

1. Bagwe, R.P., Kanicky, J.R., Palla J., Patanjali, P.K., Shah, D.O., Improved Drug  Delivery  Using  Microemulsion:  Rationale,  Recent  Progress,  and New  Horizons,  Critical  Reviews  in  Therapeutic  Drug  Carrier  Systems, 18(1), pp. 77-140, 2001.

ME formulation of 12, 13 and 14 were closely spotted near to the upper left of the phase region, concluding that these formulations were oil in water ME corresponding to the hypothetical phase pseudoternary diagram. They also exhibited less than 100 nm range of particle size. After stress testing, no significant changes in the phase separation, meaning no changes  in  the structures of  the non-ionic  surfactants, thus ME remained stable. For neurite outgrowth; at low concentration, no changes were observed. At high concentration, bipolar & multipolar neurite extensions appeared longer in length grown individually, indicating ME stimulated neurite entension.

PREPARATION AND CHARACTERIZATION OF NIGELLA SATIVA MICROEMULSION INTENDED FOR NEURITE OUTGROWTH STUDY ON

PC12 CELL LINE

METHODOLOGY