Studies on the Superovulatory Response of Rfsh Eluting Nanosuspension in Laboratory Rats

Abstract

In the present study Wistar rats with similar age and body weight were administered with rhFSH chitosan gold nano particle to induce superovulation. The phases of estrous cycle of rats were characterized by visual and vaginal cytology methods. Morphological changes noticed during proestrous were: initiation of vaginal opening, moist and reddish pink vaginal tissues and longitudinal folds at dorsal and ventral vulvar lips. During estrus vaginal opening was similar to proestrus but tissues appeared lighter pink and less moist and longitudinal folds were more pronounced than proestrus. During metestrus swelling on the dorsal lip reduced and tissues appeared pale and dry. During diestrus vaginal tissues appeared moist and vaginal closure was observed in most of the rats. During vaginal cytology the proestrus was characterized by the predominance of small nucleated cells. The estrus was characterized by the presence of predominantly anucleated cornified cells. The Metestrus phase was characterized by the presence of abundant leucocytes with few anucleated cornified cells. The diestrus phase was characterized by predominance of PMN cells. The nano particles were prepared by ionic gelation method and PDI was 0.335 to 0.45. The size of gold, chitosan gold and rhFSH chitosan gold nano particle was 9.0 nm, 320 nm and 680 nm, respectively. The zeta potentials of chitosan gold and rhFSH chitosan gold nano particle were 41.0 mV and 38.4 mV, respectively. Following administration of rhFSH chitosan gold nano suspension during diestrus phase, rats were euthanized during successive diestrus, ovaries were collected, sectioned, stained and observed under microscope. The number of post ovulatory follicles and corpora lutea per ovary in groups 1, 2, 3 and 4/ control were 30.14±0.57, 34.71±1.04, 33.21±0.96 and 7.71±0.42, respectively. Highly significant difference (P≤0.01) was found between the means (±SE) of the groups. There was non-significant difference (P≤0.05) in the means (±SE) of groups 2 and 3. The dose response relationship was not found after increasing FSH to very high levels as observed in group 1.