Immunohistochemical analysis of matrix metalloproteinase-9, vascular endothelial growth factor, bone sialoprotein and i-nitric oxide synthase in calvaria vs. iliac crest bone grafts

Objectives The aim of this study was to investigate, in parallel to clinical and histological modifications, the expression of specific proteins involved in different extraoral autologous bone grafts integration in humans. Material and methods Patients needing oral rehabilitation of posterior maxilla, with inadequate bone volume for implant placement, received bone grafts from calvaria (Group 1) and iliac crest (Group 2), respectively. From five patients from each group, with a total of 10 subjects, bone biopsy specimens were collected at two different experimental time points: at bone blocks withdrawal for grafting (T0), from donor sites, and after 4 months, from reconstructed sites. Samples were processed for light microscope and immunohistochemical analyses to evaluate MMP9, VEGF, BSP, iNOS expression. Results Morphological analysis of T0 calvaria evidenced areas of extracellular matrix, uniformly stained and organized in concentric mineralized lamellae edging few vascular canals, while T0 iliac crest showed greater cellularity compared to calvaria, with rare mineralized areas, surrounding wide bone marrow lacunae. In T1, Group 1 samples showed large areas of extracellular matrix, uniformly stained, at the same time as Group 2 samples disclosed few areas of mineralized tissue. Although no significant differences were found in proteins expression among calvaria and iliac crest T0 samples, MMP9, VEGF and BSP expression at T1 were discovered higher in Group 1 samples than in Group 2 ones, while iNOS expression increased in Group 2 samples compared to the others. In any group, molecules expression increased passing from T0 to T1. Conclusion These findings suggested that, even though clinically both extraoral sources of autologous bone could be considered suitable for grafting in case of large oral rehabilitation, some differences might be detected microscopically and biologically. Calvaria bone graft seemed to enhance not only the quantity of bone tissue at the defect site, but also its quality, better than iliac crest bone do. Then, while both grafts appeared to promote a suitable neoangiogenesis, as showed by morphological analysis and by MMP9 and VEGF expression, in terms of new bone formation and lack of occurrence of inflammatory events, calvaria could be considered a more suitable donor site for bone grafts.