Tibial growth plates (GPs) were fixed overnight in 4% paraformaldehyde (Sigma, United States) followed by 2 weeks of decalcification in 0.5 M EDTA pH 7.4. The samples were then dehydrated and transferred into histoclear (Bar-Naor) and subsequently, embedded in paraffin. Transverse tissue sections of 5 μm were prepared with Leica microtome (Agentec, Israel). For H&E histological staining, sections were deparaffinized and rehydrated (44 (link)), and stained in hematoxylin solution followed by eosin The sections for all histological analyses were dried and mounted with DPX mounting for histology. The thickness of total GP was measured using the Cell A software (Olympus) with a measuring tool feature at 10 selected locations throughout the GP in 4 different samples at each group. For imaging, the stained sections were viewed by the light microscopy Eclipse E400 Nikon. Images were captured by a high-resolution camera (DP71 Olympus), controlled by Cell A software (Olympus) (45 (link)).
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