BMP-9 induces the calcification of vascular smooth muscle cells
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Searchable abstracts of presentations at key conferences on calcified tissues ISSN 2052-1219 (online)【Objective】 To determine whether autophagy is involved in the process of vascular smooth muscle cell calcification induced by Ox-LDL. 【Methods】 The in vitro model of vascular calcification was used in this study. Vascular smooth muscle cells were randomly divided into controls,an Ox-LDL-treated group,and a 3MA plus Ox-LDL-treated group. Cells were grown in DMEM supplemented with 10 mM BGP. Calcification was assessed by alizarin red staining. The mRNA and protein expression of cbfa1 and Beclin1 were analyzed by Q-PCR and Western blotting respectively. 【Results】 Ox-LDL promotes vascular smooth muscle cell calcification,up-regulates the mRNA and protein expression of cbfa1 and Beclin1. The specific autophagy inhibitor attenuates vascular calcification,and inhibits the up-regulation of cbfa1 and Beclin1 expression induced by Ox-LDL. 【Conclusion】 The pathogenesis of vascular smooth muscle cell calcification induced by Ox-LDL involves autophagy.
Pathogenesis
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RUNX2
Ex vivo
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Objective To explore the effects of epigallocatechingallate(EGCG) on vascular calcification.Methods Calcific vascular smooth muscular cells(VSMCs) were induced by β-glycerophosphate in rats.Rats were invided into four groups:the control,the calcification,EGCG and EGCG intervention groups(10-7,10-6,10-5 and 10-4 mol·L-1 subgroups).Von Kossa staining,the Ca2+ content and the alkaline phosphatase(ALP) activity in VSMCs were measured.Results Lots of gathered black granules were found among the smooth muscle cells and its matrix by Von Kossa staining in VSMCs calcifacation group;more Calcium contents and ALP activities were observed in calcification group than those in control group;after treated with EGCG 10-7~10-4mol·L-1,the calcification in VSMCs was also attenuated and the total Ca2+ content and ALP activity in VSMCs showed decrease and a concentration-dependent manner.Conclusion The results indicated that EGCG can significantly attenuate the VSMCs calcification.
Von Kossa stain
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Etiology
Intervertebral Disc
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Calcification of cardiovascular tissues and biological substitutes results in abnormal biomechanics, causing stenosis or tissues ruptures. Therefore, the understanding of calcification pathways will facilitate the development of strategies for calcification risk reduction. Even though calcification pathways have been studied since the mid-80s and are widely described in the medical literature, there is no consensus on the cause-and-effect relationships in this pathological process. The current review covers the composition, structural aspects and specific localization of calcific deposits in native heart valves and blood vessels, bioprosthetic heart valves treated with glutaraldehyde. Moreover, experimental data on the composition of in vivo and in vitro calcification are presented. These characteristics of calcific deposits provide new insights in the calcification pathways. According to the results of the literature review, one may conclude that cell death, microfracture of the surrounding tissues with the overall biochemical imbalance may potentiate the calcification process. Moreover, the progression of calcification process is associated with the accumulation of mechanical stress.
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Objective To study the effects of salvianolate on the calcification in rat vascular smooth muscle cells(VSMCs).Methods Calcification of cultured rat VSMCs was prepared by incubation with β-glycerophosphate.Calcification was confirmed by measurement of calcium content and alkaline phosphatase(ALP) activity in VSMCs.The mRNA and protein levels of BMP-2 were detected by using real-time PCR and Western blotting,respectively.Results Compared with control group,3.92-fold calcium content and 10.5-fold increase in ALP activity were observed in calcified VSMCs group,respectively.Levels of mRNA and protein of BMP-2 were significantly increased in calcified group.Salvianolate decreased the calcium and ALP activity in a dose-dependent manner,these data were decreased by 32.3 %,42.2%,60.9%(P 0.05);36.4%,48.8% and 74.7%(P 0.01),respectively.And salvianolate decreased the mRNA and protein levels of BMP-2.Conclusion Salvianolate may have cardiovascular protecting effect by antagonizing vascular calcification,maybe by regulating BMP-2 level.
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Abstract Vascular calcification is characterized by the accumulation of hydroxyapatite crystals, which is a result of aberrant mineral metabolism. Although many clinical studies have reported its adverse effects on cardiovascular morbidity, the molecular mechanism of vascular calcification, especially the involvement of long noncoding RNAs (lncRNAs), is not yet reported. From the transcriptomic analysis, we discovered hundreds of lncRNAs differentially expressed in rat vascular smooth muscle cells (VSMCs) treated with inorganic phosphate, which mimics vascular calcification. We focused on Lrrc75a-as1 and elucidated its transcript structure and confirmed its cytoplasmic localization. Our results showed that calcium deposition was elevated after knockdown of Lrrc75a-as1, while its overexpression inhibited calcium accumulation in A10 cells. In addition, Lrrc75a-as1 attenuated VSMCs calcification by decreasing the expression of osteoblast-related factors. These findings suggest that Lrrc75a-as1 acts as a negative regulator of vascular calcification, and may serve as a possible therapeutic target in vascular calcification.
Vascular tissue
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The lifetime of bioprosthetic heart valves is limited by calcification. To investigate the calcification behavior of bioprostheses and gain insight into the etiology of valve calcification, a test protocol for accelerated valve calcification was developed. This protocol includes a pulsatile valve tester, a synthetic calcification fluid, and non-destructive radiographic assessment of calcification sites. About 40 porcine bioprostheses from different manufacturers have been investigated previously using this test protocol and showed that valves exhibited different calcification patterns and even different degrees of calcification within their leaflets. A positive correlation of calcification versus tissue anomalies/stress concentrations (r = 0.72; n = 29 valves) and lipid deposits (r = 0.81) was found. In the present study, bovine pericardial valves were investigated in comparison with porcine valves.Four bovine pericardial and two porcine mitral valves (Baxter) with a tissue annulus diameter (TAD) of 29 mm (one 27 mm) were investigated in parallel under identical test conditions. The valves were cyclically loaded at 300 per min with a delta p of 110 mmHg at 37 degrees C for up to 19 x 10(6) cycles. The synthetic calcification fluid was changed weekly. Sites of calcification were assessed by microradiography. Radiographs were analyzed by PC images processing with respect to the degree of calcification, defined as calcified surface area in relation to total leaflet surface area.This analysis showed that, for bovine pericardial valves, the mean degree of calcification increased by 14% and 20% after 12 and 19 x 10(6) cycles, respectively. Under identical conditions, the mean degree of calcification of porcine valves increased by 28% and 37%.Pericardial valves appear less prone to calcification than porcine valves. Further studies must be performed in order to prove this finding since, as recognized previously in porcine valves, other factors such as tissue or manufacturing anomalies may be as important as the tissue source itself.
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Although the clinical and biological importance of calcification is well recognized for the extracerebral vasculature, its role in cerebral vascular disease, particularly, intracranial aneurysms (IAs), remains poorly understood. Extracerebrally, 2 distinct mechanisms drive calcification, a nonatherosclerotic, rapid mineralization in the media and a slower, inflammation driven, atherosclerotic mechanism in the intima. This study aims to determine the prevalence, distribution, and type (atherosclerotic, nonatherosclerotic) of calcification in IAs and assess differences in occurrence between ruptured and unruptured IAs. Approach and Results: Sixty-five 65 IA specimens (48 unruptured, 17 ruptured) were resected perioperatively. Calcification and lipid pools were analyzed nondestructively in intact samples using high resolution (0.35 μm) microcomputed tomography. Calcification is highly prevalent (78%) appearing as micro (<500 µm), meso (500 µm-1 mm), and macro (>1 mm) calcifications. Calcification manifests in IAs as both nonatherosclerotic (calcification distinct from lipid pools) and atherosclerotic (calcification in the presence of lipid pools) with 3 wall types: Type I-only calcification, no lipid pools (20/51, 39%), Type II-calcification and lipid pools, not colocalized (19/51, 37%), Type III-calcification colocalized with lipid pools (12/51, 24%). Ruptured IAs either had no calcifications or had nonatherosclerotic micro- or meso-calcifications (Type I or II), without macro-calcifications.
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Objective To analyze and evaluate the size, shape and distribution characteristics of calcification in breast masses by high-resolution sonography for the differentiation of the benign masses from the malignant ones. Methods Fifty-one cases with calcification in which included 30 malignant and 21 benign masses in breast masses were evaluated and analyzed by high-resolution sonography . All cases had pathological results. Results According to the sizes, calcifications were classified: type 1, micro calcification; type 2, coarse calcification and type 3, curved calcification. Furthermore, micro calcifications were divided into three types due to their shapes and distributions: a, spot calcification, b, pearl calcification, c, clustered calcification. Of the 30 malignant cases, 29 were micro calcifications and only one coarse calcification, whereas 13 cases were micro calcifications, 4 were coarse calcification and 4 curved calcification in the 21 benign cases. Micro calcifications were more likely to be found in malignant diseases, and coarse calcification, curved calcification were more likely to be in benign diseases. There is a significant difference between the two groups (χ 2=12.22, P 0.001). Moreover, of the 42 micro calcifications, the calcification features of malignant masses tended to be appeared clustered distributions (22/42), and benign calcifications tended to be appeared single or loose distributions with pearl or spot micro calcifications (12/42). Statistic significance was different between malignant and benign masses (χ 2=16.44, P 0.001). Additionally, 7 malignant masses (7/42) appeared loose distributions of spot micro calcifications. Conclusion Clustered calcification can be characters of breast cancer. Most of pearl micro calcifications, coarse and curved calcifications with single or loose distributions are benign lesions. Spot micro calcification can be appeared in both malignant and benign lesions in which single or loosely sparse distributions are more likely to be benign lesions. Relatively accumulated distribution with two or more spot calcifications, combining with gray-scale and color Doppler sonographic features can improve the diagnostic accordance of breast cancer.
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