Figure 3
Histological and immunohistochemical analysis of the neo-tissue after 3 months of NM-guided self-generation.
(A) HE, (B) toluidine blue and fast green, and (C) immunohistological staining against Col II showed four typical zones of chondro-osseous transition. (D) In VG and (E) immunohistochemical OC labeling, typical bone tissue structure was observed. (F) Histomorphometrically, trabecular areas were initially reduced in both DBM and NM groups compared to native rib, but significantly increased 3 months postoperatively. Hyaline cartilage tissue stained with (G) Alcian blue and (H) against Col II showed typical hyaline cartilage tissue architecture. Signs of angiogenesis (black arrows) were observed in both groups as seen in (I) HE and (J) immunohistochemical vWF. (K) Despite the increase in newly formed blood vessels between 1.5 and 3 months in both groups, their number was still significantly lower compared to native rib. (L) Bending load strength in the NM group was similar with native rib after 3 months, while this biomechanical parameter rapidly increased in the DBM group. (M) Young’s modulus increased in both groups over time with no significant difference after 3 months. HE: haematoxylin and eosin; Col II: collagen II; VG: Van Gieson; OC: osteocalcin; NM: negative mold; DBM: demineralized bone matrix; vWF: Von Willebrand Factor.
