The Use of Sequential VEGF- and BMP2-Releasing Biodegradable Scaffolds in Rabbit Mandibular Defects.

PURPOSE: Promising developments have materialized in reconstructive surgical procedures with the applications of tissue engineering. In our study, we used tissue scaffolds fabricated from polylactic acid-polyethylene glycol (PLLA-PEG) copolymers to ensure different release rates of selective growth factors recombinant human bone morphogenetic protein 2 [rhBMP-2] and vascular endothelial growth factor (rhVEGF165) in the repair of mandibular bone defects. MATERIALS AND METHODS: In our experimental study, 54 New Zealand rabbits were used. The rabbits were separated into 4 groups: group I (control group), PLLA-PEG scaffold only; group II, PLLA-PEG scaffold plus rhBMP-2 application; group III, PLLA-PEG scaffold plus VEGF165 application; and group IV, PLLA-PEG scaffold plus rhBMP-2 and VEGF165 applications. The rabbits were killed at 4 and 8 weeks postoperatively, and histopathologic and immunohistochemical assessments were performed. RESULTS: The greatest bone volume was observed in rhBMP-2-containing groups, the greatest vessel volume was observed in VEGF165-containing groups; however, the scaffold containing rhBMP-2 and VEGF165 provided the best outcomes in conjunction with increased remodeling of the new bone. CONCLUSIONS: The use of polymer tissue scaffolds that release rhVEGF165 and rhBMP-2 in coordination and mimic the natural healing process in the regeneration of especially complex tissues, such as bone, is a promising treatment alternative in the field of reconstructive surgery.

Therapeutic Effects of Sildenafil on Experimental Mandibular Fractures.

OBJECTIVES: No previous studies have examined the effect of sildenafil on fracture healing. This study was designed to investigate the effect of sildenafil on the fracture healing process. METHODS: Thirty-six female Sprague-Dawley rats (3-month-old) were used in this study. Animals were randomly divided into 2 groups based on treatment duration (1 week versus 4 weeks) and each group was then divided further into 2 subgroups, control (C) and study (S) groups. Group C (C1, C2) was treated daily with saline solution and group S (S1, S2) was treated daily with 10 mg/kg of sildenafil. Histologic, histomorphometric, radiological, and immunohistochemical analyses were performed at 1 week and 4 weeks after a fracture. RESULTS: The sildenafil group showed a significant increase in fracture healing scores (P = 0.00). The authors observed a transition from fibrous callus to cartilage tissue and immature bone tissue in group S1; and an increased transition of cartilage tissue to completely immature bone tissue in group S2, both of which were administered sildenafil. The strong expression of bone morphogenetic protein 2 and col-1 was observed in the fibrous matrix and osteoblasts within areas of new bone formation, especially in group S1. This group also showed an increase in bone density measurements at 1 week that was statistically significant (P = 0.03). CONCLUSIONS: Sildenafil accelerates fracture healing and can be used as a supporting factor in the improvement of fracture healing under various conditions.

Nobiletin attenuates inflammation via modulating proinflammatory and antiinflammatory cytokine expressions in an autoimmune encephalomyelitis mouse model.

The aim of this study is to investigate the potential preventive and therapeutic effects of nobiletin by evaluating the expression of cytokines associated with inflammatory reactions in an autoimmune encephalomyelitis mouse model. A total of 60 male C57BL/6 mice aged between 8 and 10 weeks were used. Mice were divided into six groups (n = 10 mice per group): control, EAE, low-prophylaxis, high-prophylaxis, low-treatment and high-treatment. Experimental autoimmune encephalomyelitis (EAE) was induced by myelin oligodendrocyte glycoprotein (MOG) and pertussis toxin. Nobiletin was administered in low (25 mg/kg) and high (50 mg/kg) doses, intraperitoneally. The prophylactic and therapeutic effects of nobiletin on brain tissue and spinal cord were evaluated by expression of interleukin-1 beta (IL-1ß), tumor necrosis factor-alpha (TNF-α), interferon gamma (IFNγ), IL-6, IL-10 and transforming growth factor-beta (TGF-ß) using immunohistochemistry and real-time polymerase chain reaction (RT-PCR). Prophylactic and therapeutic use of nobiletin inhibited EAE-induced increase of TNF-α, IL-1ß and IL-6 activities to alleviate inflammatory response in brain and spinal cord. Moreover, nobiletin supplement dramatically increased the IL-10, TGF-ß and IFNγ expressions in prophylaxis and treatment groups compared with the EAE group in the brain and spinal cord. The results obtained from this study show that prophylactic and therapeutic nobiletin modulates expressions of proinflammatory and antiinflammatory cytokines in brain and spinal cord dose-dependent manner in EAE model. These data demonstrates that nobiletin has a potential to attenuate inflammation in EAE mouse model. These experimental findings need to be supported by clinical studies.

Telocytes in the hearts of Saanen goats.

Telocytes, new interstitial cells that have received significant attention in recent years, have been detected in many organs, including the heart. The distinction between telocytes and other interstitial cells can only be made based on their ultrastructural characterization and immunophenotypic features. In this study, we examined the interstitial cells in the healthy heart tissues of Saanen goats to determine whether they are telocytes or not, by using a scanning electron microscope (SEM) and immunohistochemical and immunofluorescence staining methods. The SEM revealed oval and round telocytes with two to four telopodes. Some telopodes also had podoms. The staining for immunohistochemical and immunofluorescence methods used for CD34, c-kit (CD117), and vimentin antibodies. Positive cells were detected in the heart muscle and heart valves by immunohistochemical staining. As these antigens can also be expressed by other non-telocyte cells, we used double immunofluorescence staining with CD34/c-kit and CD34/vimentin antibodies to identify true telocytes. Telocytes were determined in the right atrium and aortic valve. While telocytes were CD34+/c-kit+ and CD34+/vimentin+, fibroblasts were CD34-/vimentin+. These results confirm the presence of telocytes in the hearts of Saanen goats.

Macroanatomic, light, and electron microscopic examination of pecten oculi in the seagull (Larus canus).

The present study was conducted to determine macroanatomic characteristic as well as light and electron microscopic examination (SEM) of pecten oculi and totally 20 bulbus oculi belonging to 10 seagulls (Larus canus) were used. Pecten oculi formations consisted of 18 to 21 pleats and their shape looked like a snail. Apical length of the pleats forming pecten oculi were averagely measured as 5.77 ± 0.56 mm, retina-dependent base length was 9.01 ± 1.35 mm and height was measured as 6.4 ± 0.62 mm. In pecten oculi formations which extend up to 1/3 of the bulbus oculi, two different vascular formations were determined according to thickness of the vessel diameter. Among these, vessels with larger diameters which are less than the others in count were classified as afferent and efferent vessels, smaller vessels which are greater in size were classified as capillaries. Furthermore, the granules which were observed intensely in apical side of the pleats of pecten oculi were observed to distribute randomly along the plica.

Biological significance of the overexpression of HSP70 and alpha B-crystallin in rat substantia nigra exposed to different doses of permethrin.

The aim of this study was to investigate the possible role of the Heat Shock Protein 70 (HSP70) and Alpha B-crystallin (αBC) in the substantia nigra of rats exposed to permethrin at different doses on the apoptotic cell status. The orogastric gavage method was used to administer the different doses of permethrin (75 mg kg-1 in Group I, 150 mg kg-1 in group II, 300 mg kg-1 in group III) to the rats. Using the Western blot test, all the permethrin-treated groups showed a dose-dependent increase in the expression of HSP70 and αBC when compared to the control group. TUNEL positive apoptotic cells were not detected in the dopaminergic neurons of the substantia nigra after treatment with permethrin; however, upon immunofluorescent staining, intense positive reactions for HSP70 and αBC were observed in all of the treated groups. No immunopositive cells were detected in the tissue sections of the control group. These results suggest that the different administered doses of permethrin did not cause apoptotic cell death in the substantia nigra dopaminergic neurons; however, they did induce an increase in HSP70 and αBC expression. Thus, it appears that HSP70 and αBC could play a neuroprotective role in permethrin-induced neurotoxicity.

Alpha basic crystallin expression in canine mammary tumors.

The aim of this study was to evaluate prognostic and/or diagnostic factors of canine mammary tumors by immunohistochemically analyzing the expression of alpha basic crystallin (αB-c). For this, formalin-fixed, paraffin-embedded blocks of 51 naturally-occurring canine mammary tumors (11 benign and 40 malignant) were used. Tissue from eight normal canine mammary glands were served as a control. Immunohistochemically, in the control mammary tissues, a few luminal epithelial cells were αB-c positive but myoepithelial cells were negative. In benign or simple type malignant tumors, αB-c expression was observed in luminal epithelial cells while the myoepithelial basal cells were negative. In benign or complex type malign tumors, positive staining was predominantly found in the cytoplasm of epithelial cells. Immunoreactivity of αB-c was also observed in neoplastic myoepithelial cells. Statistically, the number of cells immunolabeled with αB-c was found to be significantly different among tissues from normal canine mammary glands, benign lesions, and malignant tumors (p < 0.05). αB-c immunoreactivity was higher in malignant tumors than the control mammary tissues (p < 0.001). Data obtained in the current study revealed a strong association between high expression levels of αB-c and primary mammary gland tumors in canines.

The expressions of HSP70 and αB-crystallin in myocarditis associated with foot-and-mouth disease virus in lambs.

This study describes the expression of heat shock protein70 (HSP70) and alpha-basic-crystallin (α-BC) and their association with apoptosis and some related adaptor proteins in the pathogenesis of foot-and-mouth disease virus (FMDV)-induced myocarditis in lambs. HSP70 was generally overexpressed in the myocardial tissues and inflammatory cells of FMDV-induced myocarditis with differential accumulation and localization in same hearts when compared to non-foot-and-mouth disease control hearts. α-BC immunolabeling showed coarse aggregations in the Z line of the cardiomyocytes in FMDV-infected hearts in contrast to control hearts. Overall, the results of this study show that the anti-apoptotic proteins, HSP70 and α-BC, were overexpressed with increased apoptosis in FMDV-infected heart tissues. Both proteins failed to protect the cardiomyocytes from apoptosis as defense mechanisms to the FMDV during the infection, suggesting that the virus is able to increase apoptosis via both downregulation and/or upregulation of these anti-apoptotic proteins.

Aromatase expression in the cerebellum of the dog infected with canine distemper virus.

Aromatase is the enzyme that catalyzes the biosynthesis of estrogens. It is implicated in neuroprotection.The present study investigated aromatase expression in the cerebellum of dogs infected with canine distemper virus (CDV), a disease characterized by demyelination in the white matter of the cerebellum. The presence of CDV infection was confirmed on the basis of histopathology and immunohistochemical localization of CDV antigen in glial cells of the white matter.The number of aromatase immunoreactive astrocytes were significantly (p < 0.05) higher in CDV-infected dogs compared to control dogs. The results suggest that astrocytes respond to invasion and persistence of CDV by means of increased estrogen production.The results also suggest that the high level of estrogen expression is maintained similarly throughout all stages of the disease since the number of aromatase immunoreactive astrocytes did not vary during the different stages of CDV infection.