Rat Aortic Smooth Muscle Cells Cultured on Hydroxyapatite Differentiate into Osteoblast-Like Cells via BMP-2–SMAD-5 Pathway

Nahar‐Gohad, Pranjal; Gohad, Neeraj V.; Tsai, Chen Chih; Bordia, Rajendra K.; Vyavahare, Naren

doi:10.1007/s00223-015-9962-z

Cited by 31 publications

(21 citation statements)

References 37 publications

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“…Further analysis of collagen production showed that both cell types expressed Col1α1 mRNA and protein. In the case of VSMCs, and consistent with earlier work [33], Col1α1 expression was primarily localised in or on the surface of cells. Whilst clearly expressed by VSMCs, no soluble collagen was detected in the culture medium and no deposited collagen fibres were evident in the TEM images.…”

Section: Discussionsupporting

confidence: 89%

Differing calcification processes in cultured vascular smooth muscle cells and osteoblasts

Patel

Bourne

Davies

et al. 2019

Experimental Cell Research

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Arterial medial calcification (AMC) is the deposition of calcium phosphate mineral, often as hydroxyapatite, in the medial layer of the arteries. AMC shares some similarities to skeletal mineralisation and has been associated with the transdifferentiation of vascular smooth muscle cells (VSMCs) towards an osteoblast-like phenotype. This study used primary mouse VSMCs and calvarial osteoblasts to directly compare the established and widely used in vitro models of AMC and bone formation. Significant differences were identified between osteoblasts and calcifying VSMCs. First, osteoblasts formed large mineralised bone nodules that were associated with widespread deposition of an extracellular collagenous matrix. In contrast, VSMCs formed small discrete regions of calcification that were not associated with collagen deposition and did not resemble bone. Second, calcifying VSMCs displayed a progressive reduction in cell viability over time (≤7-fold), with a 50% increase in apoptosis, whereas osteoblast and control VSMCs viability remained unchanged. Third, osteoblasts expressed high levels of alkaline phosphatase (TNAP) activity and TNAP inhibition reduced bone formation by to 90%. TNAP activity in calcifying VSMCs was ∼100-fold lower than that of bone-forming osteoblasts and cultures treated with β-glycerophosphate, a TNAP substrate, did not calcify. Furthermore, TNAP inhibition had no effect on VSMC calcification. Although, VSMC calcification was associated with increased mRNA expression of osteoblast-related genes (e.g. Runx2, osterix, osteocalcin, osteopontin), the relative expression of these genes was up to 40-fold lower in calcifying VSMCs versus bone-forming osteoblasts. In summary, calcifying VSMCs in vitro display some limited osteoblast-like characteristics but also differ in several key respects: 1) their inability to form collagen-containing bone; 2) their lack of reliance on TNAP to promote mineral deposition; and, 3) the deleterious effect of calcification on their viability.

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Section: Discussionsupporting

confidence: 89%

Differing calcification processes in cultured vascular smooth muscle cells and osteoblasts

Patel

Bourne

Davies

et al. 2019

Experimental Cell Research

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“…SMAD5, SMAD6, and BMPR2 are involved in BMP signaling. Studies in the pluripotent cell line C2C12 as well as rat aortic smooth muscle cells have shown that activated SMAD5 induces osteoblast differentiation via RUNX2 [52,53]. SMAD6 plays a role as an inhibitor of BMP signaling [54] and as an important factor for the normal development and tissue specific modulation of vessels [55].…”

Section: Discussionmentioning

confidence: 99%

MicroRNAs 223-3p and 93-5p in patients with chronic kidney disease before and after renal transplantation

Ulbing

Kirsch

Leber

et al. 2017

Bone

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Chronic kidney disease (CKD) is associated with a multifactorial dysregulation of bone and vascular calcification and closely linked to increased cardiovascular mortality and concomitant bone disease. We aimed to investigate specific microRNA (miRNA) signatures in CKD patients to find indicators for vascular calcification and/or bone mineralization changes during CKD and after kidney transplantation (KT). A miRNA array was used to investigate serum miRNA profiles in CKD patients, then selected miRNAs were quantified in a validation cohort comprising 73 patients in CKD stages 3 to 5, 67 CKD patients after KT, and 36 healthy controls. A spectrum of biochemical parameters including markers for kidney function, inflammation, glucose, and mineral metabolism was determined. The relative expression of miR-223-3p and miR-93-5p was down-regulated in patients with CKD stage 4 and 5 compared to healthy controls. This down-regulation disappeared after kidney transplantation even when lower glomerular filtration rates (eGFR) persisted. MiR-223-3p and miR-93-5p were associated with interleukin-6 (IL-6) and eGFR levels, and by trend with interleukin-8 (IL-8), C-peptide, hematocrit, and parathyroid hormone (PTH). This study contributes new knowledge of serum miRNA expression profiles in CKD, potentially reflecting pathophysiological changes of bone and calcification pathways associated with inflammation, vascular calcification, mineral and glucose metabolism. Identified miRNA signatures can contribute to future risk markers or future therapeutic targets in bone and kidney disease.

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“…Phosphorylation of SMAD5 caused by BMPs leads to the formation of a complex of SMAD5, SMAD1, and SMAD8, which further exerts the effect of BMPs after translocation into the nuclei [18, 19]. Previous studies have confirmed its important role in enhancing osteogenic potential [20, 21]. Based on our data with hampered mRNA and protein expression of both pSMAD5 and SMAD5 after overexpression of miR-155, the mechanism of miR-155's effect on SMAD5 was most likely through reducing the stability of SMAD5 mRNA.…”

Section: Discussionmentioning

confidence: 99%

miR-155 Inhibits Mouse Osteoblast Differentiation by Suppressing SMAD5 Expression

Song

et al. 2017

BioMed Research International

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Osteogenesis from preosteoblasts is important for bone tissue engineering. MicroRNAs are a class of endogenous small RNA molecules that potentially modulate osteogenesis. In this study, we found that miR-155 expression was downregulated in a time-dependent manner in cells of the preosteoblast cell line MC3T3-E1 after osteogenic induction using bone morphogenetic protein 2 (BMP2). Transfection with miR-155 decreased alkaline phosphatase (ALP) activity, ALP expression, and the staining intensity of Alizarin Red in MC3T3-E1 cells treated with BMP2, whereas treatment with miR-155 inhibitor promoted BMP2-induced osteoblast differentiation. The luciferase assay confirmed that miR-155 can bind to the 3′ untranslated region of SMAD5 mRNA. miR-155 transfection significantly decreased the expression of SMAD5 protein and mRNA in MC3T3-E1 cells under control media and the p-SMAD5 protein level during osteogenesis. After transfecting cells with the SMAD5 overexpression plasmids, the inhibitory effect of miR-155 on osteogenesis was significantly attenuated. In conclusion, miR-155 inhibited osteoblast differentiation by downregulating the translation of SMAD5 in mouse preosteoblast cells. Inhibition of miR-155 promoted osteogenic potential and thus it can be used as a potential target in the treatment of bone defects.

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Rat Aortic Smooth Muscle Cells Cultured on Hydroxyapatite Differentiate into Osteoblast-Like Cells via BMP-2–SMAD-5 Pathway

Cited by 31 publications

References 37 publications

Differing calcification processes in cultured vascular smooth muscle cells and osteoblasts

Differing calcification processes in cultured vascular smooth muscle cells and osteoblasts

MicroRNAs 223-3p and 93-5p in patients with chronic kidney disease before and after renal transplantation

miR-155 Inhibits Mouse Osteoblast Differentiation by Suppressing SMAD5 Expression

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