Axiostar plus 1169 149

Organs (liver, kidney, lung, spleen, heart, brain, ovary and testis) from different groups were fixed in neutral buffered 10% v/v formalin solution for histopathological assessment. After fixation, the organs were dehydrated in graded alcohol (TP1020, semi-enclosed benchtop tissue processor, Leica Biosystems, Germany), embedded in paraffin (EG1130 and EG1150, cold plate for cooling embedding molds and paraffin blocks and modular tissue embedding center, Leica Biosystems, Germany), sectioned into 4 µm thick (RM2255, fully automated rotary microtome, Leica Biosystems, Germany) and stained with Weigert’s Hematoxylin and Eosin (Suvarna et al., 2012 ). Visualization and microphotographs were captured under a light microscope (Axiostar plus 1169-149, Carl Zeiss, Germany) at magnification 10×.

The ultraviolet-visible (UV-vis) spectroscopy (Shimadzu, UV-3101PC UV-vis-NIR spectrophotometer) was used to characterize the nanocurcumin by measuring the absorption ranges from 200–700 nm.
The Fourier transform infrared (FT-IR) spectra of curcumin and nanocurcumin were recorded on a FT-IR spectrophotometer (Varian, 640-IR, USA). The spectra were measured in the wavenumber region of 4000–500 cm⁻¹ at a resolution of 4 cm⁻¹ with a KBr pellet.
The crystalline structure of the dried nanocurcumin powder was recorded by an X-ray diffraction method using two instruments (PANalytical, X'Pert Pro, and Bruker, D-8 Advance).
The mean particle diameter and zeta potential measurements were performed using a particle size molecular size measurement instrument (Malvern Instruments Ltd., NanoSight NS500, Malvern, UK). The sample was prepared by taking 1 mg of the lyophilized nanocurcumin powder in 10 ml of distilled water.
The morphology and particle size of nanocurcumin were observed using transmission electron microscopy (TEM, Jeol JEM-1400, Japan). The images were visualized at 120 kV at 40 000 magnifications.
The nuclear morphology of cells was examined using fluorescence microscopy (Carl Zeiss, Axiostar Plus 1169–149, Germany).