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目的 慢性牙周炎表现的病理性骨吸收非常常见,牙周膜干细胞(PDLSCs)的外泌体(Exo)对骨吸收的作用和影响尚不明确,本研究分析了该Exo蛋白组分对破骨细胞分化的影响。方法 分别从正畸前拔牙患者和慢性牙周炎患者的牙周膜组织中提取PDLSCs,通过流式细胞术检测表面标记物;通过差速离心分别提取2种细胞的Exo,即Exo-WT和Exo-CP,并通过Western blot、透射电子显微镜(TEM)、蛋白浓度检测、纳米粒径追踪检测Exo特征;通过蛋白质谱检测2种Exo的蛋白组分;通过酶联免疫吸附(ELISA)验证了差异表达蛋白肿瘤坏死因子α(TNF-α)、核因子κB受体活化因子配体(RANKL)、白细胞介素(IL)-1α、转化生长因子β(TGF-β)、骨形态发生蛋白2(BMP-2)表达水平;将10、100、1 000 μg·mL-1的Exo-CP或Exo-WT加入RAW264.7培养基中并于5 d时通过实时荧光定量聚合酶链反应(RT-qPCR)、Western blot、抗酒石酸酸性磷酸酶(TRAP)染色检测破骨分化相关指标表达情况;采用SPSS 24.0软件对实验数据进行统计学分析。结果 Exo-CP富集的差异表达蛋白主要与破骨细胞分化的TNF信号通路、核转录因子κB(NF-κB)信号通路相关;ELISA实验证实了Exo-CP中高表达TNF-α、RANKL、IL-1α,低表达TGF-β1、BMP-2(P<0.05);Exo-CP作用于RAW264.7,显著提高了细胞的破骨分化相关基因及蛋白的表达水平,TRAP染色可见分化的破骨细胞,且呈现浓度依赖性,100、1 000 μg·mL-1浓度Exo-CP对破骨细胞分化的促进作用显著高于10 μg·mL-1浓度组(P<0.05)。结论 慢性牙周炎的病理性骨吸收可能由炎性PDLSCs所分泌的Exo通过促进破骨细胞分化引起,Exo中主要的作用蛋白可能为RANKL和TNF-α,本研究为慢性牙周炎骨吸收的发病机制提供了新视角。 相似文献
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A variety of inflammatory cytokines are involved in spinal cord injury and influence the recovery of neuronal function. In the present study, we established a rat model of acute spinal cord injury by cerclage. The cerclage suture was released 8 or 72 hours later, to simulate decompression surgery. Neurological function was evaluated behaviorally for 3 weeks after surgery, and tumor necrosis factor α immunoreactivity and apoptosis were quantified in the region of injury. Rats that underwent decompression surgery had significantly weaker immunoreactivity of tumor necrosis factor α and significantly fewer apoptotic cells, and showed faster improvement of locomotor function than animals in which decompression surgery was not performed. Decompression at 8 hours resulted in significantly faster recovery than that at 72 hours. These data indicate that early decompression may improve neurological function after spinal cord injury by inhibiting the expression of tumor necrosis factor α. 相似文献
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《Medical engineering & physics》2014,36(3):364-370
A sagittal saw is commonly used for resection of bone during joint replacement surgery. During sawing, heat is generated that can lead to an increase in temperature at the resected surface. The aim of this study was to determine the effect of applied thrust force and blade speed on generating heat. The effect of these factors and their interactions on cutting temperature and bone health were investigated with a full factorial Design of Experiments approach for two levels of thrust force, 15 N and 30 N, and for two levels of blade oscillation rate, 12,000 and 18,000 cycles per minute (cpm). In addition, a preliminary study was conducted to eliminate blade wear as a confounding factor. A custom sawing fixture was used to crosscut samples of fresh bovine cortical bone while temperature in the bone was measured by thermocouple (n = 40), followed by measurements of the depth of thermal necrosis by histopathological analysis (n = 200). An analysis of variance was used to determine the significance of the factor effects on necrotic depth as evidenced by empty lacunae. Both thrust force and blade speed demonstrated a statistically significant effect on the depth of osteonecrosis (p < 0.05), while the interaction of thrust force with blade speed was not significant (p = 0.22). The minimum necrotic depth observed was 0.50 mm, corresponding to a higher level of force and blade speed (30 N, 18,000 cpm). Under these conditions, a maximum temperature of 93 °C was measured at 0.3 mm from the kerf. With a decrease in both thrust force and blade speed (15 N, 12,000 cpm), the temperature in the bone increased to 109 °C, corresponding to a nearly 50% increase in depth of the necrotic zone to 0.74 mm. A predictive equation for necrotic depth in terms of thrust force and blade speed was determined through regression analysis and validated by experiment. The histology results imply that an increase in applied thrust force is more effective in reducing the depth of thermal damage to surrounding bone than an increase in blade speed. 相似文献
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Deqiang Li Peilai Liu Yuankai Zhang Ming Li 《International journal of clinical and experimental pathology》2015,8(9):10947-10952
Objectives: Avascular necrosis of the femoral head (ANFH) was mainly due to alterations of bone vascularity. And noradrenaline (NA), as the neurotransmitter of the sympathetic nervous system (SNS), leads to the vasoconstriction by activating its α-Receptor. This study was to explore the nerve fiber density of the femoral head in the rabbit model of ANFH. Methods: Twenty New Zealand white rabbits were used in this study. The rabbit model of ANFH was established by the injection of methylprednisolone acetate. The nerve fiber density and distribution in the femoral head was determined using an Olympus BH2 microscope. Results: Significant fewer sympathetic nerve fibers was found in the ANFH intertrochanteric bone samples (P = 0.036) with osteonecrosis. The number of sympathetic nerve fibers was compared between the two groups. And less sympathetic nerve fibers were found in later stage ANFH samples in comparison with those of early stages. Conclusions: ANFH might be preceded by an inflammatory reaction, and an inflammatory response might lead to arthritic changes in tissue samples, which in turn reduces the number of sympathetic nerve fibers. 相似文献
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Lidija Begović Maja Antunovic Igor Matic Ivana Furcic Ana Baricevic Valerija Vojvoda Parcina 《International journal of radiation biology》2016,92(8):475-482
Purpose: Ultraviolet (UV) radiation-induced apoptosis enabled us to study the mechanism of DNA damage and to investigate how cells avoid consequences of damaged DNA. Cells with extensive DNA damage activate extrinsic and intrinsic pathways of apoptosis. The extrinsic pathway is coupled to a FAS-associated protein with death domain (FADD), an adaptor protein molecule necessary for mediating apoptotic signals through the cell.Materials and methods: Viability and apoptosis of wild-type and FADD-deficient mouse embryonic fibroblasts were investigated 1, 3, 24 and 48?h after exposure to three doses (50, 75 and 300 J/m2) of UVC radiation. Morphological changes were observed using DNA binding dyes (Hoechst and propidium iodide) while biochemical changes were monitored using immunodetection of the poly (ADP-ribose) polymerase (PARP) protein cleavage and caspase-3 activity assay.Results: Results showed that the difference in cell death response between wild-type and FADD-deficient cells depended on dose and incubation time after exposure to UVC radiation. FADD-deficient cells are more sensitive to UVC radiation. Even though FADD-deficient cells lack an adapter protein of apoptotic extrinsic pathway, higher doses of UVC triggered their apoptotic response, while wild-type cells die mainly due to necrosis. A different pattern of caspase 3 activity and PARP cleavage was observed 24?h after radiation between two cell lines confirming higher apoptotic response in FADD-deficient cells.Conclusions: Wild-type cells can execute apoptosis via both, the mitochondrial and the receptor-mediated pathway whereas FADD-deficient cells can only activate the intrinsic pathway. There is a difference in UVC radiation response between two cell lines indicating the role of FADD in the selection of cell death modality. 相似文献