Assessment of Protective Effect of Rutin Utilizing Advanced Intranasal Nanoparticulate System in Mice Model after Experimental Challenge with Pasteurella Multocida Isolated from Naturally Infected Sheep

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Abstract
Rutin is a promising nutraceutical with potential antibacterial and prophylactic activities. This work aimed to evaluate the protective effect of Rutin nanovehicle, which were formulated as a mucoadhesive intranasal preparation, on mice experimentally challenged with Pasteurella multocida. The extracted Rutin nanoplatform was laden into different concentrations of mucoadhesive hydroxy propyl methyl cellulose (HPMC). The ex vivo permeability of Rutin nanoformulations across the nasal mucosa was evaluated. The optimal formulation was examined in vivo, and group 1 was not infected or prophylactic. Group 2 was infected with P. multocida-nonprotected, and groups 3 and 4 were prophylactic groups that were protected with free Rutin extract and Rutin nanoformulation, respectively, via the intranasal route. The bacterial count in vital organs was carried out post challenge with P. multocida to determine protective effect of free Rutin extract and Rutin nanoformulation, also pathological changes in the vital organs were examine, level of IgG was determined, Cytokines concentration; Interferon Gamma (INF-γ) and Interlukin-12 (IL -12) was assessment by using sandwich ELISA. The rutin nanoparticulate system containing the lowest amount of HPMC exhibited enhanced permeated amount (311.64± 10.0 μg/cm2) and steady state flux 6.67± 0.10 μg /cm 2/sec across nasal mucosa. The results of the in vivo study revealed that the rutin nanoformulation showed a significant reduction in the bacterial count, histopathological lesions in the examined organs. Also, increased the hormonal response of mice where it increased levels of antibodies IgGs and modulation in the concentration of serum IFN-γ and IL-12. The mucoadhesive intranasal Rutin nanovehicle has remarkable immunomodulatory effects and can be used for protection against Pasteurella multocida in animals.

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