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Fig 4. Histology, Immunohistology, and Micro-CT Images of Spike Formation from Animals. (A) and (B) Histology and Micro-CT. Red Arrows: Porous calcified tissue growth or old bone growth remodeling. Signs of osteoblast and osteoclast reconstruction can be observed in the tissues dense structure that is filled with marrow pockets. Orange Arrow: Bony callus formation, this is typically connected to the old bone around the amputation site and is hollow. Black Arrows: Formation of hyaline cartilage behind the old bone. Green Arrow: Connection site of old marrow cavity to new tissue growth. Brown Arrow: Signs that the old marrow cavity did not connect with the new tissue growth. Purple Arrow: Invasion of tissue into the cartilage spike core. (C) and (D) Immunohistological Stains. Green: Acetyl- N-Alpha Tubulin (α-Tubulin) (Nerve) Red: Smooth Muscle Actin (SMA) (Vasculature) Yellow: Collagen type II (Col2) (cartillage type II only found in abundance in hyaline cartilage) Purple: Phosphorylated Mothers Against Decapentaplegic Homolog (pSMAD 5/9) (TGF-β activator, signs of proliferation and differentiation/ not cartilage behavior) Orange Arrow: Location where protein would be typically found in spike. White Arrow: Location where protein would not be typically found in spike.
http://dx.doi.org/10.1371/journal.pone.0155618.g004 |
