Fluid flow induced calcium response in bone cell network

In our previous work, bone cell networks with controlled spacing and functional intercellular gap junctions had been successfully established by using microcontact printing and self assembled monolayers technologies [Guo, X. E., E. Takai, X. Jiang, Q. Xu, G. M. Whitesides, J. T. Yardley, C. T. Hung, E. M. Chow, T. Hantschel, and K. D. Costa. Mol. Cell. Biomech. 3:95-107, 2006]. The present study investigated the calcium response and the underlying signaling pathways in patterned bone cell networks exposed to a steady fluid flow. The glass slides with cell networks were separated into eight groups for treatment with specific pharmacological agents that inhibit pathways significant in bone cell calcium signaling. The calcium transients of the network were recorded and quantitatively evaluated with a set of network parameters. The results showed that 18 alpha-GA (gap junction blocker), suramin (ATP inhibitor), and thapsigargin (depleting intracellular calcium stores) significantly reduced the occurrence of multiple calcium peaks, which were visually obvious in the untreated group. The number of responsive peaks also decreased slightly yet significantly when either the COX-2/PGE(2) or the NOS/nitric oxide pathway was disrupted. Different from all other groups, cells treated with 18 alpha-GA maintained a high concentration of intracellular calcium following the first peak. In the absence of calcium in the culture medium, the intracellular calcium concentration decreased slowly with fluid flow without any calcium transients observed. These findings have identified important factors in the flow mediated calcium signaling of bone cells within a patterned network.

Analysing the contribution of ATM/ATR pathway activation in establishing the premature senescence of E2F1/E2F2-/- bone-marrow-derived macrophages

E2F1 and E2F2 transcription factors have an important role during the regulation of cell cycle. In experiments done with E2F1/E2F2 knockout mice, it has been described that bone-marrow-derived macrophages (BMDM) undergo an early rapid proliferation event related to DNA hyper-replication. As a consequence, DNA damage response (DDR) pathway is triggered and E2F1/E2F2 knockout macrophages enter premature senescence related to G2/M phase arrest. The exact mechanism trough which DNA hyper-replication leads to DDR in absence of E2F1 and E2F2 remains undiscovered. To determine whether the ATR/ATM pathway, the master regulator of G2/M checkpoint, might be the surveillance mechanism in order to regulate uncontrolled proliferation in the DKO model, we monitored and analysis biochemical properties of BMDM cultures in the presence of caffeine, a potent inhibitor of ATM/ATR activity. Our results show that the addition of caffeine abolishes premature senescence in DKO BMDM, stimulates γ-H2AX accumulation and decreases Mcm2 expression.

Efeitos do tratamento periodontal não cirúrgico sobre a densidade e altura óssea alveolar

O objetivo deste trabalho foi avaliar o efeito do tratamento periodontal não cirúrgico (TPNC), na densidade e na altura óssea alveolar, em pacientes com periodontite, utilizando radiografias digitais diretas. Cento e um sítios, em dezenove pacientes (idade média 36 7.3 anos) foram acompanhados no dia 0, e 90 e 180 dias após TPNC. Os índices clínicos de profundidade de bolsa a sondagem (PBS), nível de inserção clínica, sangramento à sondagem e índice de placa foram registrados e radiografias digitais foram feitas. A densidade foi analisada através de regiões ósseas de interesse colocadas sobre a crista óssea alveolar (ROI I) e sobre o osso medular (ROI II). A altura óssea alveolar foi medida através da distância da crista óssea alveolar até a junção cemento esmalte. Os sítios profundos (PBS ≥ 5mm) apresentaram uma melhora clínica significante (p <0.01), acompanhada de um aumento na densidade da ROI I (p <0.01). A ROI II mostrou um aumento na densidade dos sítios com PBS ≤ 3mm em pacientes com periodontite agressiva (p <0.05). No entanto, houve diminuição nos sítios com PBS ≥ 5mm nesses mesmos pacientes (p <0.03). A altura óssea alveolar não sofreu alteração após TPNC. Após o tratamento periodontal não cirúrgico, observou-se que as radiografias obtidas através da técnica digital direta parecem mostrar um aumento na densidade da crista óssea, nos sítios profundos dos pacientes com periodontite. No entanto, a redução da profundidade de bolsa e do ganho no nível de inserção clínica não foi acompanhada por alterações significantes na altura óssea alveolar nestes sítios.