RESUMEN

Liposomes is a non-viral vector drug delivery system. Nevertheless, the existing commercial liposomes are quite expensive and not always affordable, particularly in developing countries. To address this challenge, plant-derived nanoparticles offer a cost-effective alternative while maintaining similar drug delivery capabilities. Hence, this study aimed to explore the potential of nanovesicles derived from black cumin (Nigella sativa) as a miRNA delivery system. Gradient sucrose-centrifugation was utilized to separate the nanovesicles derived from black cumin. Subsequently, these isolated nanovesicles, originating from black cumin, underwent centrifugation at a speed of 11,000 rpm. The miRNAs were encapsulated within these nanovesicles through the ethanol injection method. Morphological examinations of the nanovesicles derived from black cumin and DOTAP, as the positive control, were conducted using TEM and SEM. Furthermore, the cytotoxicity of the nanovesicles derived from black cumin was evaluated through the MTT assay on the MCF-7 cell line. Lastly, the process of internalization for both the black cumin-derived nanovesicles and DOTAP was visualized using a confocal microscope. Results demonstrated the successful isolation of nanovesicles from black cumin using the sucrose gradient method. These particles exhibited a spherical shape with diameters ranging from 100 nm to 200 nm, featuring a negative surface charge. When MCF-7 cells were exposed to black cumin-derived nanovesicles at a concentration of 12 mg/mL, cell viability reached 89.8 %, showing no significant difference compared to the positive control (p > 0.05). Furthermore, the MCF-7 cell line effectively internalized the black cumin-derived nanovesicles after a 45-minute incubation period. Notably, the encapsulation of miRNA within these nanovesicles demonstrated an impressive entrapment efficiency of 76.4 %. Subsequent transfection of miRNA-loaded black cumin-derived nanovesicles resulted in a substantial inhibition of MCF-7 cell viability, reducing it to 67 % after 48 h of treatment. These findings underscore the potential of black cumin-derived nanovesicles as potential nanovectors for the encapsulation and delivery of miRNA within drug delivery systems, offering a cost-effective and accessible solution for advanced drug delivery technologies, particularly in developing country.

Asunto(s)

Sistemas de Liberación de Medicamentos , MicroARNs , Nanopartículas , Nigella sativa , Humanos , Células MCF-7 , Sistemas de Liberación de Medicamentos/métodos , Nigella sativa/química , Nanopartículas/química , Supervivencia Celular/efectos de los fármacos , Liposomas , Extractos Vegetales/administración & dosificación , Extractos Vegetales/química

RESUMEN

Introduction: Hepatitis B remains a global problem with no effective treatment. Here, a mucosal vaccine candidate was developed with HBsAg and HBcAg, to provide both prophylactic and therapeutic protection against hepatitis B. The antigens were presented using the P particle of human norovirus (HuNov). As a result, the chimeric HBV - HuNoV P particle can act as a dual vaccine for hepatitis B and HuNoV. Methods: The vaccine candidate was expressed and purified from Escherichia coli BL21 (DE3) cells. HBV-HuNoV chimeric P particles were successfully expressed and isolated, with sizes of approximately 25.64 nm. Then, the HBV-HuNoV chimeric P particles were evaluated for safety and immunogenicity in mice and gnotobiotic (Gn) pigs. After three doses (5 µg/dose in mice and 200 µg/dose in Gn pigs) of intranasal immunization, humoral and cellular immune responses, as well as toxicity, were evaluated. Results: The vaccine candidate induced strong HBV-HuNoV specific IFN-γ producing T-cell responses in the ileum, spleen, and blood of Gn pigs. Serum IgG and IgA antibodies against HBV-HuNoV chimeric P particles also increased significantly in Gn pigs. Increased HBsAg- and HuNoV-specific serum IgG responses were observed in mice and Gn pigs, although not statistically significant. The vaccine candidate did not show any toxicity in mice. Conclusions: In summary, the chimeric HBV-HuNoV P particle vaccine given intranasally was safe and induced strong cellular and humoral immune responses in Gn pig. Modifications to the vaccine structure and dosage need to be evaluated in future studies to further enhance immunogenicity and induce more balanced humoral and cellular responses.