Study of resveratrol against bone loss by using in-silico and in-vitro methods / Estudo do resveratrol contra a perda óssea usando métodos in silico e in vitro

Abstract By applying the in-silico method, resveratrol was docked on those proteins which are responsible for bone loss. The Molecular docking data between the resveratrol and Receptor activator of nuclear factor-kappa-Β ligand [RANKL] receptors proved that resveratrol binds tightly to the receptors, showed the highest binding affinities of −6.9, −7.6, −7.1, −6.9, −6.7, and −7.1 kcal/mol. According to in-vitro data, Resveratrol reduced the osteoclasts after treating Marrow-Derived Macrophages [BMM] with Macrophage colony-stimulating factor [MCSF] 20ng / ml and RANKL 50ng / ml, with different concentrations of resveratrol (2.5, 10 μg / ml) For 7 days, the cells were treated with MCSF (20 ng / ml) and RANKL (40 ng / ml) together with concentrated trimethyl ether and resveratrol (2.5, 10 μg / ml) within 12 hours. Which, not affect cell survival. After fixing osteoclast cells with formaldehyde fixative on glass coverslip followed by incubation with 0.1% Triton X-100 in PBS for 5 min and after that stain with rhodamine phalloidin staining for actin and Hoechst for nuclei. Fluorescence microscopy was performed to see the distribution of filaments actin [F.actin]. Finally, resveratrol reduced the actin ring formation. Resveratrol is the best bioactive compound for drug preparation against bone loss.

Resumo Com a aplicação do método in-silico, o resveratrol foi ancorado nas proteínas responsáveis ​​pela perda óssea. Os dados de docking molecular entre o resveratrol e o ligante do receptor ativador do fator nuclear kappa-Β [Receptor Activator of Nuclear Factor kappa-B Ligant (RANKL)] provaram que o resveratrol se liga fortemente aos receptores, mostraram as afinidades de ligação mais altas de −6,9, −7,6, −7,1, −6,9, - 6,7 e -7,1 kcal / mol. De acordo com dados in-vitro, o resveratrol reduziu os osteoclastos após o tratamento de macrófagos derivados da medula óssea [Bone Marrow-derived Macrophage (BMM)] com fator estimulador de colônias de macrófagos [Macrophage Colony-Stimulating Factor (MCSF)] 20ng / ml e RANKL 50ng / ml, com diferentes concentrações de resveratrol (2,5, 10 μg / ml). Durante sete dias, as células foram tratadas com MCSF (20 ng / ml) e RANKL (40 ng / ml) juntamente com éter trimetílico concentrado e resveratrol (2,5, 10 μg / ml) em 12 horas, processo que não afeta a sobrevivência celular. Após a fixação de células de osteoclastos com fixador de formaldeído em lamela de vidro seguido de incubação com 0,1% Triton X-100 em PBS por 5 min, foi realizado posteriormente o procedimento para corar com rodamina faloidina a actina e Hoechst os núcleos. A microscopia de fluorescência foi realizada para ver a distribuição dos filamentos de actina [F.actina]. Finalmente, o resveratrol reduziu a formação do anel de actina. O resveratrol é o melhor composto bioativo para o preparo de medicamentos contra a perda óssea.

Craniomandibular osteopathy in a west highland white terrier

Background: Craniomandibular osteopathy is a bone disorder that occurs in dogs but is not very commonly reported. It is characterized by a non-neoplastic bilaterally symmetrical irregular proliferation of skull or long bones. Immature dogs are most commonly reported with age ranging from three to eight months. The aim of this paper is to report a case of CMO in a West Highland White Terrier.Case: A 5-month-old entire West Highland white terrier was referred to the Veterinary Hospital with sudden history of severe pain and mandibular swelling, dysphagia, lethargy and weight loss. On physical examination the dog was quiet, alert and responsive. There was thickening of the mandibular bodies with pain at palpation and when opening the mouth. Complete blood count was within normal range. Simple radiographs demonstrated discrete and active new bone formation on the ventral aspect of the mandibular body. Carprofen, dypirone and tramadol were prescribed twice daily for 10 days. 30 days later new radiographs showed more extensive areas of periosteal proliferation advancing to the temporomandibular joint and progression of the looseness of the characteristic compact appearance of lamellar bone of the mandible. The dog was more lethargic, not eating well and in pain. Medication was again prescribed but the treatment was unsuccessful. The dog returned for consultation with severe pain...(AU)

Craniomandibular osteopathy in a west highland white terrier

Background: Craniomandibular osteopathy is a bone disorder that occurs in dogs but is not very commonly reported. It is characterized by a non-neoplastic bilaterally symmetrical irregular proliferation of skull or long bones. Immature dogs are most commonly reported with age ranging from three to eight months. The aim of this paper is to report a case of CMO in a West Highland White Terrier.Case: A 5-month-old entire West Highland white terrier was referred to the Veterinary Hospital with sudden history of severe pain and mandibular swelling, dysphagia, lethargy and weight loss. On physical examination the dog was quiet, alert and responsive. There was thickening of the mandibular bodies with pain at palpation and when opening the mouth. Complete blood count was within normal range. Simple radiographs demonstrated discrete and active new bone formation on the ventral aspect of the mandibular body. Carprofen, dypirone and tramadol were prescribed twice daily for 10 days. 30 days later new radiographs showed more extensive areas of periosteal proliferation advancing to the temporomandibular joint and progression of the looseness of the characteristic compact appearance of lamellar bone of the mandible. The dog was more lethargic, not eating well and in pain. Medication was again prescribed but the treatment was unsuccessful. The dog returned for consultation with severe pain...

Aqueous-ozone irrigation of bone monocortical wounds in hyperglycemic rats

PURPOSE: To analyze the effects of aqueous ozone irrigation over bone healing in hyperglycemia-induced rats. METHODS: Forty-eight male Wistar rats were allocated into Group H (hyperglycemic) or Group N (control). Monocortical bone wound were performed on femurs' anterolateral face. Wounds were treated with a trans-operatory single irrigation of 100ml of aqueous ozone [0.004mg/ml] whereas control groups received 100ml of pure water (Milli-Q®). Histomorphological and histomorphometrical analyses were accomplished after seven, 14 and 21 days. Kruskal-Wallis and Mann-Whitney statistical tests were applied for bone neoformation quantification and assessment. RESULTS: Aqueous ozone wounds irrigated revealed diffuse hemorrhage and increased neoformed of blood vessels number. There was no statistical significant difference in bone trabeculae neoformation. After seven and 14 days, the number of osteoclasts was higher in aqueous ozone groups than in those treated with pure water. CONCLUSION: Independently of blood glucose levels, aqueous ozone allowed an increase in blood vessels neoformation and osteoclast migration, without affect bone trabeculae neoformation.(AU)

Osteodistrofia fibrosa nutricional em caprinos / Nutritional fibrous osteodystrophy in goats

Seven out of 25 goats from a southern Brazilian flock developed nutritional fibrous osteodystrophy. Affected animals were younger than 1 year of age and were confined in stalls and fed a concentrate ration containing 1:6 calcium:phosphorus ratio. The remaining flock (35 goats) was managed at pasture and showed no disease. Clinical signs were characterized by mandibular and maxillary enlargements, varying degrees of mouth opening and protruding tongue, dyspnea, apart of abnormalities of prehension and mastication. Affected animals had increased seric levels of phosphorus and parathormone, as well as higher alkaline phosphatase activity. Postmortem examination on three succumbed goats revealed bilateral enlargement of the maxilla and mandibula, and loose teeth, apart of multiple incomplete rib fractures in one of them. Severe diffuse proliferation of loose connective tissue surrounded the osteoid trabeculae, many of which were partially or completely non-mineralized. Mineralized osteoid trabeculae showed osteoclasts in the Howship's lacunae.(AU)

Sete de um total de 25 caprinos jovens (menos de um ano de idade) de um rebanho no sul do Brasil desenvolveram osteodistrofia fibrosa nutricional. Os animais afetados eram confinados em baias e alimentados com concentrado, cuja relação Ca:P era 1:6. O restante do rebanho (35 cabras) era mantido na pastagem e não desenvolveu a doença. Os sinais clínicos se caracterizaram por aumento de volume da mandibula e maxila, vários graus de abertura de boca com protrusão da língua, em associação com dispneia e anormalidades de apreensão de alimento e mastigação. Os animais afetados apresentaram os níveis séricos de fósforo e paratormônio aumentados, bem como maior atividade de fosfatase alcalina. Três caprinos foram necropsiados e os achados de necropsia incluíram aumento bilateral da maxi-la e mandíbula, dentes frouxos, além de múltiplas fraturas incompletas de costelas em um caprino. Microscopicamente, havia intensa proliferação de tecido conjuntivo frouxo ao redor de trabéculas ósseas, muitas das quais estavam parcial ou completamente não mineralizadas. Trabéculas ósseas mineralizadas apresentaram osteoclastos em lacunas de Howship.(AU)