Effect of Bone Morphogenetic Protein-2 (BMP-2) on Matrix Production, Other BMPs, and BMP Receptors in Rat Intervertebral Disc Cells

Modern therapeutic strategies for intervertebral disc repair mainly focus on targeting molecular pathways of extracel-lular matrix degeneration. Anabolic strategies for regeneration are aimed to increase production of major extracellular molecules. Members of TGF-b superfamily proteins, particularly the bone morphogenetic proteins (BMP) have a high regenerative potential regarding the mesenchymal cells. The goal of this study is to study production of proteoglycans by the intervertebral disc cells under the influence of bone morphogenetic protein 2.Material and methods. The experiment was carried out on the cell cultures derived from the annulus fibrosis cells and nucleus pulposus cells of the human intervertebral disc. We studied cell livability, metabolic activity and proteoglycan expression. Cell livability was assessed using the trypan blue staining. Alamar blue test was used for the estimation of metabolic activity. Amount of sulfated glycosaminoglycans was assessed using the assay based on the reaction with 1,9-Dimethylmethylene Blue.Results. Cultivation with bone morphogenetic protein 2 in different concentrations did not decrease livability of the cells. Study cell cultures with application of bone morphogenetic protein 2 in different concentrations showed significant increase in metabolic activity and proteoglycan synthesis by the annulus fibrosis cells. Despite the relative increase in the number of the nucleus pulposus cells treated with the bone morphogenetic protein 2, the differences in metabolic and synthetic activity compared with control group was not significant. Conclusion. The bone morphogenetic protein 2 has an anabolic effect towards the intervertebral disc cells, particularly in the production of proteoglycans by the annulus fibrosis cells.

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Bone morphogenetic protein-7 protects human intervertebral disc cells in vitro from apoptosis

The Spine Journal ◽

10.1016/j.spinee.2007.04.021 ◽

2008 ◽

Vol 8 (3) ◽

pp. 466-474 ◽

Cited By ~ 57

Author(s):

Aiqun Wei ◽

Helena Brisby ◽

Sylvia A. Chung ◽

Ashish D. Diwan

Keyword(s):

Intervertebral Disc ◽

Bone Morphogenetic Protein ◽

Bone Morphogenetic Protein 7 ◽

Morphogenetic Protein ◽

Disc Cells

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Bone Morphogenetic Protein-2 Induces Non-Canonical Inflammatory and Oxidative Pathways in Human Retinal Endothelial Cells

Frontiers in Immunology ◽

10.3389/fimmu.2020.568795 ◽

2021 ◽

Vol 11 ◽

Author(s):

Mohamed Al-Shabrawey ◽

Khaled Hussein ◽

Fang Wang ◽

Ming Wan ◽

Khaled Elmasry ◽

...

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Bone Morphogenetic Protein ◽

P38 Mapk ◽

High Glucose ◽

Bone Morphogenetic Protein 2 ◽

Barrier Dysfunction ◽

Retinal Endothelial Cells ◽

Bmp Receptors ◽

Morphogenetic Protein

The mechanisms of diabetic retinopathy (DR), are not yet fully understood. We previously demonstrated an upregulation of retinal bone morphogenetic protein-2 (BMP2) in experimental diabetes and in retinas of diabetic human subjects. The purpose of current study was to investigate the role of non-canonical inflammatory pathway in BMP2-induced retinal endothelial cell (REC) barrier dysfunction. For this purpose, we used RT-PCR and western blotting to evaluate the levels of BMP2 signaling components (BMP2, BMP4, BMP receptors), VEGF, phosphorylated p38 MAPK and NFκB, and oxidative stress markers in cultured human retinal endothelial cells (HRECs) subjected to BMP2 (50ng/ml) for up to 24 h. Also, effect of high glucose (HG, 30mM D-glucose) on the expression of BMP2 and its downstream genes was examined in HRECs. H2-DCF is a fluorogenic dye that measures the levels of cellular reactive oxygen species (ROS) was used to measure the pro-oxidative effect of BMP2. Moreover, we evaluated the effect of inhibiting p38 and VEGF signaling on BMP2-induced HRECs barrier dysfunction by measuring the trans-endothelial cell electrical resistance (TER) using electric cell-substrate impedance sensing (ECIS). We also tested the effect of HG on the integrity of HRECs barrier in the presence or absence of inhibitors of BMP2 signaling. Our data reveals that BMP2 and high glucose upregulates BMP components of the BMP signaling pathway (SMAD effectors, BMP receptors, and TGFβ ligand itself) and induces phosphorylation of p38 MAPK and NFκB with nuclear translocation of NFκB. Inhibition of p38 or NFκB attenuated BMP2-induced VEGF expression and barrier dysfunction in HRECs. Also, inhibition of VEGFR2 attenuated BMP2-induced barrier dysfunction. Moreover, BMP2 induces generation of ROS and endothelial nitric oxide synthase (eNOS) expression and activity in HRECs. Finally, HG upregulated BMP2 and its downstream genes (SMAD, BMP4, ALKs, and TGF-β) in HRECs and BMP2 inhibitors attenuated HG-induced HRECs barrier dysfunction. Our results suggest that in addition to the regular canonical SMAD signaling BMP2 induces non-canonical inflammatory pathway in HRECs via activation of p38/NFκB pathway that causes the upregulation of VEGF and the disruption of HRECs. Inhibition of BMP2 signaling is a potential therapeutic intervention to preserve endothelial cell barrier function in DR.

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Bone Morphogenetic Protein-2 Conjugated to Quantum Dot®s is Biologically Functional

Nanomaterials ◽

10.3390/nano10061208 ◽

2020 ◽

Vol 10 (6) ◽

pp. 1208

Author(s):

Daniel Halloran ◽

Vrathasha Vrathasha ◽

Hilary W. Durbano ◽

Anja Nohe

Keyword(s):

Quantum Dot ◽

Bone Morphogenetic Protein ◽

Fluorescent Dyes ◽

Fluorescein Isothiocyanate ◽

Biologically Active ◽

Bone Morphogenetic Protein 2 ◽

Bmp Receptors ◽

Morphogenetic Protein ◽

Long Time ◽

Neural Growth

Quantum Dot®s (QDot®s) are novel, semi-conductive nanostructures that emit a certain fluorescence when excited by specific wavelengths. QDot®s are more photostable, brighter, and photobleach less than other fluorescent dyes. These characteristics give them the potential to be used in many biological applications. The shells of QDot®s are coated with functional groups, such as carboxylate and organic groups, allowing them to couple to peptides/proteins and be used for real-time imaging and high-resolution microscopy. Here, we utilize Quantum Dot®s and Bone Morphogenetic Protein-2 (BMP-2) to create a BMP-2-QDot®s conjugate. BMP-2 is a growth factor that drives many processes such as cardiogenesis, neural growth, and osteogenesis. Despite its numerous roles, the trafficking and uptake of BMP-2 into cells is not well-established, especially during progression of diseases. The results presented here demonstrate for the first time a fluorescent BMP-2 analog that binds to the BMP-receptors (BMPRs), remains biologically active, and is stable for long time periods. Previous attempts to develop a biological BMP-2 analog with Fluorescein isothiocyanate (FITC) or nanodiamonds lacked data on the analog’s stability. Furthermore, these analogs did not address whether they can signal within the cell by binding to the BMPRs or were mediated by non-stable conjugates.

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