Conditioned medium from bone marrow-derived mesenchymal stem cells inhibits vascular calcification through blockade of the BMP2\u2013Smad1/5/8 signaling pathway

Shuangshuang Wang,Ruochi Zhao,Xiaomin Chen,Siwang Hu,Maoqing Tong,Yahui Liu,Nan Wu,Hanbin Cui,Jian Wang

doi:10.1186/s13287-018-0894-1

Abstract

BackgroundArterial calcification is associated with cardiovascular disease as a complication of advanced atherosclerosis and is a significant contributor to cardiovascular morbidity and mortality. Osteoblastic differentiation of vascular smooth muscle cells (VSMCs) plays an important role in arterial calcification and is characterized by cellular necrosis, inflammation, and lipoprotein and phospholipid complexes, especially in atherosclerotic calcification. The conditioned medium from bone marrow-derived mesenchymal stem cells (MSC-CM) is well known as a rich source of autologous cytokines and is universally used for tissue regeneration in current clinical medicine. Here, we demonstrate that MSC-CM inhibits beta-glycerophosphate (β-GP)-induced vascular calcification through blockade of the bone morphogenetic protein-2 (BMP2)–Smad1/5/8 signaling pathway.MethodsVSMC calcification was induced by β-GP followed by treatment with MSC-CM. Mineral deposition was assessed by Alizarin Red S staining. Intracellular calcium content was determined colorimetrically by the o-cresolphthalein complexone method and alkaline phosphatase (ALP) activity was measured by the para-nitrophenyl phosphate method. Expression of BMP2, BMPR1A, BMPR1B, BMPR2, msh homeobox 2 (Msx2), Runt-related transcription factor 2 (Runx2), and osteocalcin (OC), representative osteoblastic markers, was assessed using real-time polymerase chain reaction analysis while the protein expression of BMP2, Runx2, and phosphorylated Smad1/5/8 was detected by western blot analysis.ResultsOur data demonstrated that MSC-CM inhibits osteoblastic differentiation and mineralization of VSMCs as evidenced by decreased calcium content, ALP activity, and decreased expression of BMP-2, Runx2, Msx2, and OC. MSC-CM suppressed the expression of phosphorylated Smad1/5/8 and the β-GP-induced translocation from the cytoplasm to the nucleus. Further study demonstrated that human recombinant BMP-2 overcame the suppression of VSMC calcification by MSC-CM.ConclusionMSC-CM may act as a novel therapy for VSMC calcification by mediating the BMP2–Smad1/5/8 signaling pathway

Highlights

Arterial calcification is associated with cardiovascular disease as a complication of advanced atherosclerosis and is a significant contributor to cardiovascular morbidity and mortality
Mesenchymal stem cell (MSC)-CM inhibits β-GP-induced calcium deposition in vascular smooth muscle cell (VSMC) To determine the effect of the MSC-CM on VSMC calcification, we investigated the calcium deposition and calcium content in VSMCs
The results show that Noggin inhibited β-GP-induced VSMC calcification, calcium content, and the associated marker genes, such as bone morphogenetic protein-2 (BMP2), msh homeobox 2 (Msx2), Runt-related transcription factor 2 (Runx2), and osteocalcin, suggesting that BMP2 and its downstream signaling are involved in the process of VSMC calcification

Summary

Introduction

Arterial calcification is associated with cardiovascular disease as a complication of advanced atherosclerosis and is a significant contributor to cardiovascular morbidity and mortality. Vascular calcification is a common feature of coronary artery disease as a complication of advanced atherosclerosis and is highly associated with aging, chronic kidney disease, diabetes, hypertension, and dyslipidemia [1, 2]. Whereas medial calcification is a hallmark of diabetes and chronic kidney disease, arterial calcification is clearly associated with the mortality risk in individuals with atherosclerosis and diabetes. Increasing evidence supports that osteo/chondrocytic transformation of vascular smooth muscle cells (VSMCs) and their dedifferentiation from a contractile to a proliferative phenotype are important for the initiation and progression of vascular calcification [1,2,3]. There are currently no ideal therapies directed at the prevention or treatment of vascular calcification to combat this growing problem in clinical practice today

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