BMP-2 and IL-1ß as Markers of Nasal Mucosa Inflammation in Rhinosinusitis with Nasal Polyps.

Rhinosinusitis with nasal polyps is characterized by chronic inflammation and hyperplasia of the nasal mucosa. The key mechanism for polyp formation is the expression of molecules that regulate proliferation and inflammation. We studied immunolocalization of bone morphogenetic protein-2 (BMP-2) and IL-1ß in the nasal mucosa in patients aged 35-70 years (n=70, mean age 57.4±1.52 years). The typology of polyps was determined depending on the distribution of inflammatory cells, subepithelial edema, the presence of fibrosis and cysts. The immunolocalization of BMP-2 and IL-1ß had the same pattern in edematous, fibrous, and eosinophilic (allergic) polyps. Goblet and connective tissue cells, microvessels and terminal sections of the glands were positively stained. BMP-2+ and IL-1ß+ cells predominated in polyps of the eosinophilic type. BMP-2/IL-1ß can be considered as a specific marker of inflammatory remodeling of the nasal mucosa in refractory rhinosinusitis with nasal polyps.

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.

On-Bone Fixation of Free Gingival Graft Induces an Osteoinductive Effect in Human Alveolar Bone.

We examined alveolar bone samples in the area of on-bone fixation of a free gingival graft performed during surgery in patients aged 37-55 years with a diagnosis of secondary partial adentia of the upper and lower jaws. Six months after fixation of the graft in the alveolar bone, foci of neoosteogenesis were found in the contact zone. They were characterized by the appearance of appositional lines, cords of basophilic osteoblasts, and growing osteons. An immunohistochemical study revealed an increase in the number of CD44+, CD29+, and osteocalcin+ cells in the layer of the outer circumferential lamellae, primary osteons, and the lining of the Haversian canals. TGF-ß1+ cells were located in the intertrabecular reticular tissue and wall of microvessels. The results indicate activation of mesenchymal stem cells in the area of localization of the graft and differentiating osteoblasts. The observed osteoinductive effect of free gingival graft is associated with its participation in reorganization in MSC and induction of morphogenetic molecules.

Osteogenic and Regenerative Potential of Free Gingival Graft.

We studied immunolocalization of CD29, CD44, osteocalcin, and TGF-ß1 in the bone tissue of the mandible of miniature pigs with extra-bone fixation of a free gingival graft. Three months after surgery, neoosteogenesis foci with high expression of the studied markers were found in the contact area of the free gingival graft with the alveolar bone. The markers were localized in the layer of external circumferential lamellae, on the surface of concentric lamellae of the growing osteons, and in the connective tissue of the Haversian canals. TGF-ß1-immunopositive cells predominated in the connective tissue of the Haversian and Volkmann canals and in the adventitia and inner lining of the vascular wall. The established morphochemical patterns of osteogenous cells indicate significant reparative capabilities of a free gingival graft and allows considering it as an effective osteoinductive factor.

Brain-Derived Neurotrophic Factor (BDNF) As a Regulator of Apoptosis under Conditions of Focal Experimental Stroke.

The immunolocalization of apoptotic factors in rat neocortex was studied on the model of permanent occlusion of the middle cerebral artery with administration of exogenous BDNF. We revealed heterogeneous distribution of pro- and anti-apoptotic factors in the stroke area and in the surrounding penumbra, where caspase-3+ and p53+ cells were found. Their number was maximum on day 3 of ischemia. The number of neurons containing anti-apoptotic factors was relatively decreased. Injection of BDNF changed the distribution of apoptotic factors. In the penumbra area, BDNF enhanced the expression of Mdm2 primarily in the pyramid cells of layers V/VI and Bcl-2 in interneurons of layers II and III. Localization of p53 and caspase-3 varied at different terms of the ischemic period and showed an inverse dependence. Considering the selective neuroprotective effect of BDNF, various mechanisms of the formation of ischemic tolerance in neurons are proposed.

[Expression of pro- and anti-apoptotic molecules in the mucous membrane of the nasal cavity with polypous rhinosinusitis]. / Ekspressiya pro- i antiapoptoticheskikh molekul v slizistoi obolochke polosti nosa pri polipoznom rinosinusite.

OBJECTIVE: To study the expression of proapoptotic (p53, p21) and antiapoptotic (MDM2) factors, as well as the distribution of proliferating PCNA-immunoreactive cells in the nasal mucosa in various types of polyposis rhinosinusitis (PRS). MATERIAL AND METHODS: We studied the immunolocalization of proapoptotic (p53, p21) and antiapoptotic (MDM2) factors, as well as the distribution of proliferating PCNA immunoreactive cells in the mucous membrane of the nasal cavity for various types of polypous rhinosinusitis. RESULTS AND DISCUSSION: Comparing with the control group, increased expression of all factors is detected. The main portion of marked cells is located in the loose connective tissue of the lamina propria of mucous membrane, a relatively small number of cells are detected in the epithelial layer. The edematous, eosinophilic (allergic) type of PRS is characterized by the expression of p53 and PCNA and by the reduction of MDM2 immunoreactive cells, while the expression of p53 and p21 is reduced in fibroinflammatory (neutrophilic) type. CONCLUSION: PRS is accompanied by epithelial metaplasia, by formation of inflammatory cell infiltrates, fibrosis and edema. The results are discussed in connection with the data on the regulatory role of apoptosis in the pathomorphism of chronic productive inflammation.

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.

Small molecules regulating apoptosis in the synovium in rheumatoid arthritis.

OBJECTIVES: To study the rate of apoptosis and expression of pro- and anti-apoptotic molecules in the synovial membrane in early and late rheumatoid arthritis (RA). METHODS: Samples of the synovium, cartilage, synovial fluid, and blood serum were obtained from patients with seropositive RA. The localization of Bcl-2-, p53- and TUNEL-immunoreactive cells in the synovial membrane was studied. The level of the soluble Fas (sFas) receptor was determined in the blood serum and synovial fluid using an immunoassay. RESULTS: In early RA, p53-immunoreactive synovial cells of type A were found to form rare aggregates in the intima. In late RA, on the contrary, these cells increased in number and occurred predominantly in the synovial stroma. Bcl-2-immunoreactive synovial cells were observed in lymphocytic infiltrates in the intima. They were found mainly in early RA. In late RA, their number decreased. The apoptotic index determined from the proportion of TUNEL-reactive synovial cell nuclei reached a maximum in late RA. The temporal differences in the rate of apoptosis were correlated with the humoral level of sFas, which increased significantly in late RA. On the contrary, in the synovial fluid, the sFas level decreased monotonically from early to late RA. CONCLUSION: In early RA, in the synovial membrane, the rate of apoptosis and p-53-immunostaining intensity were low, and Bcl-2-immunostaining intensity was high. The sFas level in synovial fluid was high. In late RA, the rate of apoptosis and p-53-immunostaining intensity increased, Bcl-2-immunostaining intensity decreased, as did the sFas level.