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Noggin suppression decreases BMP-2-induced osteogenesis of human bone marrow-derived mesenchymal stem cells in vitro
Chao Chen1, Hasan Uludağ, Zhixiang Wang
1Department of Surgery, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada.
Abstract:
Numerous studies with rodent cells and animal models indicate that noggin inhibits osteogenesis by antagonizing bone morphogenetic proteins (BMPs); however, the effect of noggin on osteogenesis of human cells remains ambiguous. This study aims to examine the effects of noggin suppression on viability and BMP-2-induced osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (MSCs) in vitro. Noggin expression in human MSCs was suppressed by noggin-specific small interfering RNA (siRNA), and viability of human MSCs was determined by measuring the mitochondrial dehydrogenase activity, cellular DNA content and protein amount. The BMP-2-induced osteogenic differentiation of human MSCs was assessed by analyzing the expression levels of several osteoblastic genes, enzymatic alkaline phosphatase (ALP) activity and calcification. Our study showed that noggin suppression significantly decreased human MSC metabolism and DNA content on Days 3 and 6, and decreased total protein amount on Day 14. Noggin suppression also reduced the expression levels of osteoblastic genes, ALP, integrin-binding sialoprotein (IBSP), muscle segment homeobox gene (MSX2), osteocalcin (OC), osteopontin (OPN), and runt-related transcription factor-2 (RUNX2). Significantly decreased enzymatic ALP activity in noggin-suppressed group was evident. Moreover, noggin suppression decreased calcium deposits by BMP-2-induced osteoblasts. Collectively, this study showed that noggin suppression decreased viability and BMP-2-induced osteogenic differentiation of human MSCs, which suggests that noggin is stimulatory to osteogenesis of human MSCs.
Insights
Noggin suppression reduces human mesenchymal stem cell (MSC) viability and their ability to differentiate into bone cells. This suggests noggin stimulates osteogenesis in human MSCs.
Area of Science:
- Cell Biology
- Stem Cell Biology
- Bone Biology
Background:
- Noggin is known to inhibit osteogenesis in rodent models by antagonizing bone morphogenetic proteins (BMPs).
- The role of noggin in human osteogenesis is not well understood.
- Human bone marrow-derived mesenchymal stem cells (MSCs) are crucial for bone regeneration.
Purpose of the Study:
- To investigate the effect of noggin suppression on human MSC viability.
- To determine the impact of noggin suppression on BMP-2-induced osteogenic differentiation in human MSCs.
Main Methods:
- Noggin expression in human MSCs was suppressed using noggin-specific small interfering RNA (siRNA).
- Cell viability was assessed by measuring mitochondrial dehydrogenase activity, DNA content, and protein levels.
- Osteogenic differentiation was evaluated by analyzing osteoblastic gene expression, alkaline phosphatase (ALP) activity, and calcium deposition.
Main Results:
- Noggin suppression significantly decreased MSC metabolism, DNA content, and total protein.
- Suppression of noggin reduced the expression of key osteoblastic genes (ALP, IBSP, MSX2, OC, OPN, RUNX2).
- Noggin suppression led to decreased ALP activity and reduced calcium deposits in BMP-2-treated cells.
Conclusions:
- Noggin suppression impairs human MSC viability and BMP-2-induced osteogenic differentiation.
- These findings indicate that noggin plays a stimulatory role in human MSC osteogenesis.
- Targeting noggin could be a potential strategy for modulating bone formation.
