Journal of Bone Metabolism

"Cell differentiation"

Original Articles

Effects of Bone Morphogenetic Protein Agonist Small Molecule, SB4, on Osteogenic Differentiation of Adipose-Derived Mesenchymal Stem Cells: Fatemeh Naghshnejad, Bahman Zeynali, Iman Shabani, Azita Parvaneh Tafreshi; J Bone Metab 2026;33(1):18-29.
Published online February 28, 2026; DOI: https://doi.org/10.11005/jbm.25.899

Background
Bone morphogenetic protein (BMP) signaling is crucial for osteogenic (OS) differentiation, bone growth, and repair. However, the clinical use of BMPs is limited due to their cost and challenges in delivery. SB4, a potent compound of benzoxazole, is a newly introduced small molecule shown to activate the BMP signaling pathway. In this study, we investigated the effects of SB4 on OS differentiation of adipose-derived mesenchymal stem cells (ADMSCs).
Methods
We first examined whether BMP signaling is active in osteoinduced ADMSCs. ADMSCs were treated with SB4 (5 and 10 μM) for 24 hr, and mRNA expression of BMP direct target genes (ID1 and ID3) was analyzed. ADMSCs were then cultured with or without SB4 (10 μM) for 7, 14, and 21 days, followed by Alizarin red staining and real-time polymerase chain reaction analysis of mRNA expression of OS markers RUNX2, BMP2, alkaline phosphatase (ALP), and Osteopontin. To investigate the most effective time period of SB4 in osteogenesis, SB4 was applied every 3.5 days during the 21-day differentiation period.
Results
After 24 hr of treatment with SB4 (5 and 10 μM), increased mRNA expression of BMP direct target genes (ID1 and ID3) confirmed that SB4 acts as an active BMP agonist. SB4-treated ADMSCs showed enhanced bone matrix production and increased mRNA expression of RUNX2, BMP2, ALP, and Osteopontin. Increased mRNA expression of OS markers, bone matrix formation, and ALP activity during the first 7 days of SB4 treatment suggested the importance of SB4 at the early stage of differentiation.
Conclusions
These results indicate that SB4, with potential BMP-like activity, induces OS differentiation of ADMSCs, leading it to serve as a cost-effective BMP agonist for osteogenesis-related research as well as a potential therapeutic agent for bone repair and regeneration.

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Homogenic Evaluation for Spatial Distribution in Osteoclast Differentiation: Hyun-Sook Lim, Hong-In Shin, Daewon Jeong; J Bone Metab 2022;29(4):265-269.
Published online November 30, 2022; DOI: https://doi.org/10.11005/jbm.2022.29.4.265

Background
Cells have heterogeneous cellular diversity in size, morphology, cell cycle, metabolism, differentiation degree, and spatial distribution. The shift of specific cells towards the desired cells is crucial for maintaining uniform cellular function and can be represented by homogeneity and heterogeneity. Here, we developed a simple and direct method for evaluating the homogeneous distribution of desired cells in a constant region.
Methods
We differentiated osteoclast progenitors into bone-resorbing multinucleated giant osteoclasts in a 2-dimensional culture plate under 2 conditions. Cells were stained with tartrate-resistant acid phosphatase to assess osteoclast differentiation, images were taken using a microscope and divided into sectors, and the number of osteoclasts (≥3 nuclei) in each sector was counted. To assess the homogeneity of the spatial distribution of osteoclasts, the standard deviation (SD) was calculated from the mean number of osteoclasts within each sector.
Results
From the 2 groups, a value with a SD close to 0 indicates high spatial homogeneity while a relatively high SD represents low spatial homogeneity.
Conclusions
Our findings suggest that spatial homogeneity can be represented as SD.

Citations

Citations to this article as recorded by

1. Selenoprotein W engages in overactive osteoclast differentiation in multiple myeloma
Hyunsoo Kim, Jiin Oh, Min Kyoung Kim, Kyung Hee Lee, Daewon Jeong
Molecular Biology Reports.2024;[Epub] CrossRef

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Macrophage-Stimulating Protein Enhances Osteoblastic Differentiation via the Recepteur d'Origine Nantais Receptor and Extracellular Signal-Regulated Kinase Signaling Pathway: Byung-Chul Jeong, Sin-Hye Oh, Mi Nam Lee, Jeong-Tae Koh; J Bone Metab 2020;27(4):267-279.
Published online November 30, 2020; DOI: https://doi.org/10.11005/jbm.2020.27.4.267

Background
Macrophage-stimulating protein (MSP; also known as macrophage stimulating 1 and hepatocyte growth factor-like protein) has been shown to play a crucial role in calcium homeostasis and skeletal mineralization in zebrafish. However, the precise role of MSP in osteoblasts has not been elucidated. In this study, we investigated the effect of MSP on osteoblast differentiation of pre-osteoblast cells.
Methods
Osteoblast differentiation upon MSP treatment was evaluated by analyzing the osteogenic gene expression, alkaline phosphatase (ALP) activity, and mineralized nodule formation. To assess changes in the MSP-RON signaling pathway, knockdown of Ron gene was performed using siRNA and pharmacological inhibitor treatment.
Results
Expression of the tyrosine kinase receptor RON, a receptor of MSP, was found to be significantly increased during osteoblast differentiation. MSP treatment significantly upregulated the expression of osteogenic marker genes and remarkably increased ALP activity and mineralized nodule formation. Conversely, knockdown of Ron significantly attenuated the expression of osteogenic marker genes and ALP activity that were induced upon MSP treatment. Mechanistically, MSP treatment significantly enhanced the phosphorylation of extracellular signal-regulated kinase (ERK); however, additional treatment with the selective ERK inhibitor PD98059 attenuated the effect of MSP on osteoblast differentiation.
Conclusions
Altogether, these results indicate that the MSP-RON axis is involved in promoting osteoblast differentiation via activation of the ERK signaling pathway.

Citations

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1. Gut microbiota and blood biomarkers in IBD-Related arthritis: insights from mendelian randomization
Wei Yang, Miao Cui, Peng Yang, Chenlin Liu, Xiuzhen Han, Wenyi Yao, Zhenhua Li
Scientific Reports.2025;[Epub] CrossRef
2. A Multifunctional Therapeutic Strategy Using P7C3 as A Countermeasure Against Bone Loss and Fragility in An Ovariectomized Rat Model of Postmenopausal Osteoporosis
Fei Wei, Megan Hughes, Mahmoud Omer, Christopher Ngo, Abinaya Sindu Pugazhendhi, Elayaraja Kolanthai, Matthew Aceto, Yasmine Ghattas, Mehdi Razavi, Thomas J Kean, Sudipta Seal, Melanie Coathup
Advanced Science.2024;[Epub] CrossRef
3. The microbial metabolite imidazole propionate dysregulates bone homeostasis by inhibiting AMP-activated protein kinase (AMPK) signaling
Suk-Gyun Park, Jung-Woo Kim, Ju Han Song, Seung-Hee Kwon, Sin-Hye Oh, Xianyu Piao, Zhao Wang, Je-Hwang Ryu, Nacksung Kim, Ok-Su Kim, Jeong-Tae Koh
Communications Biology.2024;[Epub] CrossRef
4. A network map of macrophage-stimulating protein (MSP) signaling
Diya Sanjeev, Shobha Dagamajalu, Vineetha Shaji, Mejo George, Yashwanth Subbannayya, T. S. Keshava Prasad, Rajesh Raju, Rex Devasahayam Arokia Balaya
Journal of Cell Communication and Signaling.2023; 17(3): 1113. CrossRef
5. The ATF3–OPG Axis Contributes to Bone Formation by Regulating the Differentiation of Osteoclasts, Osteoblasts, and Adipocytes
Jung Ha Kim, Kabsun Kim, Inyoung Kim, Semun Seong, Jeong-Tae Koh, Nacksung Kim
International Journal of Molecular Sciences.2022; 23(7): 3500. CrossRef
6. Transcription Factor Lmx1b Negatively Regulates Osteoblast Differentiation and Bone Formation
Kabsun Kim, Jung Ha Kim, Inyoung Kim, Semun Seong, Jeong Eun Han, Keun-Bae Lee, Jeong-Tae Koh, Nacksung Kim
International Journal of Molecular Sciences.2022; 23(9): 5225. CrossRef
7. Quantitative Proteomic Analysis of Serum Reveals MST1 as a Potential Candidate Biomarker in Spontaneously Diabetic Cynomolgus Monkeys
Chaoyang Tian, Mingyin Qiu, Haizhou Lv, Feng Yue, Feifan Zhou
ACS Omega.2022; 7(50): 46702. CrossRef

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Methanol Extract of Croton Pycnanthus Benth. Inhibits Osteoclast Differentiation by Suppressing the MAPK and NF-κB Signaling Pathways: Jiyeon Lee, Hong-Hee Kim; J Bone Metab 2014;21(4):269-275.
Published online November 30, 2014; DOI: https://doi.org/10.11005/jbm.2014.21.4.269

Background

Osteoclasts are differentiated from monocytes/macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor-kappa B (NF-κB) ligand (RANKL). Croton pycnanthus Benth. (CPB) is a herbal plant that belongs to Euphorbiaceae family. The aim of this study was to investigate the effects of CPB on osteoclastogenesis and RANKL-dependent signaling pathways.

Methods

Methanol extract of CPB was obtained from International Biological Material Research Center. Osteoclast differentiation was achieved by culturing mouse bone marrow-derived macrophages (BMMs) with M-CSF and RANKL. Osteoclast numbers were evaluated by counting multinuclear cells positive for tartrate-resistant acid phosphatase (TRAP). mRNA and protein levels were analyzed by real-time polymerase chain reaction (PCR) and Western blotting, respectively. The activation of signaling molecules were assessed after acute stimulation of cells with high dose of RANKL by Western blotting with phospho-specific antibodies.

Results

CPB reduced the generation of TRAP-positive multinucleated cells and the activation of mitogen-activated protein kinase (MAPK) and NF-κB signaling pathways. The induction of the expression of c-Fos, nuclear factor-activated T cells c1 (NFATc1) and dendritic cell-specific transmembrane protein (DC-STAMP) by RANKL was also suppressed.

Conclusions

CPB exerts negative effects on osteoclast differentiation in response to the RANKL. The inhibitory mechanism involves the suppression of MAPK and NF-κB signaling pathways and subsequently the down-regulation of c-Fos and NFATc1 transcription factors.

Citations

Citations to this article as recorded by

1. Inhibitory Effect of Rosae Multiflorae Fructus Extracts on the Receptor Activator of NF-κB Ligand-Induced Osteoclastogenesis through Modulation of P38- and Ca2+-Mediated Nuclear Factor of Activated T-Cells Cytoplasmic 1 Expression
Keun Ha Park, Dong Ryun Gu, Min Seuk Kim, Seoung Hoon Lee
Journal of Bone Metabolism.2020; 27(1): 53. CrossRef
2. Inhibitory effect of Chaenomelis Fructus ethanol extract on receptor activator of nuclear factor-kappa B ligand-mediated osteoclastogenesis
Geun Ha Park, Dong Ryun Gu, Seoung Hoon Lee
International Journal of Oral Biology.2020; 45(1): 15. CrossRef
3. Effects of light-emitting diode irradiation on RANKL-induced osteoclastogenesis
HongMoon Sohn, Youngjong Ko, Mineon Park, Donghwi Kim, Young Lae Moon, Yeon Joo Jeong, Hyeonjun Lee, Yeonhee Moon, Byung-Chul Jeong, Okjoon Kim, Wonbong Lim
Lasers in Surgery and Medicine.2015; 47(9): 745. CrossRef
4. ZIP4 silencing improves bone loss in pancreatic cancer
Qiang Zhang, Xiaotian Sun, Jingxuan Yang, Hao Ding, Drake LeBrun, Kai Ding, Courtney W. Houchen, Russell G. Postier, Catherine G. Ambrose, Zhaoshen Li, Xiaohong Bi, Min Li
Oncotarget.2015; 6(28): 26041. CrossRef