Localization of VEGF, TGF-ß1, BMP-2, and Apoptosis Factors in Hypertrophic Nonunion of Human Tubular Bones.

We studied localization of VEGF, TGF-ß1, BMP-2, caspase-3, Bcl-2, and TNFα in the callus samples obtained from 5 patients (4 women and 1 man) aged 41-53 years during planned surgery for nonunion and pseudarthrosis of the clavicle (n=1), ulna (n=1), femur (n=1), and tibia (n=2) bones. Two control groups included material of hypertrophied callus (n=3) with consolidated fractures of long bones and samples of intact bones (n=3) obtained by postmortem autopsy of subjects without pathology of the musculoskeletal system. A nonuniform distribution of the studied markers was revealed. Active expression of VEGF was observed in fibroblast-like cells of the fibrous tissue, osteoblasts of the periosteum and osteons. Osteoblasts expressing BMP-2 were localized in the periosteum and the loose connective tissue of the Haversian canals. The number of immunopositive cells expressing TGF-ß1 and TNFα in the callus exceeded that in the control and correlated with the expression of caspase-3 in fibroblast-like cells, osteoblasts, chondroblasts, and microvascular endotheliocytes. The results allow considering fracture nonunion as a result of overproduction of cytotoxic and proapoptotic factors in chronic inflammation and dysfunction of the expression of morphogenetic proteins. The morphochemical patterns of the studied markers open up prospects for the development of new methods of pharmacological correction of fracture repair.

Dynamics of Renewal of Cell Populations of the Bone Tissue on the Surface of Titanium Implants with Bioactive Coating during Fracture Modeling in Rats.

Localization of PCNA, CD44, osteocalcin, Mdm2, p53, and caspase-3 on the surface of implant with calcium phosphate and hydroxyapatite coating was studied by immunocytochemical method in a model of femur fracture in rats. PCNA+, Ost+, CD44+, and Mdm2+ cells were found in the periosteum, in the layer of the outer surrounding plates, and in the connective tissue of the Haversian canals. Cell density increased on day 7 after fracture and then decreased by day 30. The number of p53+ and CASP3+ cells reached a maximum on day 14 (they were predominantly located in the periosteum and bone plates adjacent to it) and decreased by day 30. Calcium phosphate coating stimulated proliferative activity of cells at the early stages of the regeneration phase and apoptotic death at the later stages. Components of coating can be viewed as a positioning clue for differentiation of mesenchymal stromal cells. The effectiveness of reparative osteogenesis is determined by the balance of proliferative and destructive factors at the site of the fracture healing. This process can be optimized with various nanostructured materials with osteoinductive properties, in particular bioresorbable calcium phosphate coatings on titanium implants. However, the influence of these components on the state of cambial cells, their differentiation, and positioning in the repair zone is unknown.

Role of Vascular Endothelial Growth Factor and Transforming Growth Factor-ß2 in Rat Bone Tissue after Bone Fracture and Placement of Titanium Implants with Bioactive Bioresorbable Coatings.

The study established enhanced expression of vascular endothelial growth factor (VEGF) in the subpopulation of osteoblasts located in the regeneration region of femoral bone fracture near the titanium implants with bioactive calcium phosphate and hydroxyapatite coatings and suppressed activity of transforming growth factor-ß2 (TGF-ß2) in chondroblasts during the two weeks after surgery. In the delayed posttraumatic period, the distribution of TGF-ß2 inversely related to its maximal activity. The data revealed the up-regulating effect of bioresorbable coatings on expression of VEGF and TGF-ß2 and their implication in the control over various stages of reparative osteogenesis.