scholarly journals Long Noncoding RNA FAM83H-AS1 Modulates SpA-Inhibited Osteogenic Differentiation in Human Bone Mesenchymal Stem Cells

2019 ◽  
Vol 40 (5) ◽  
Author(s):  
Haojie Wu ◽  
Faqi Cao ◽  
Wu Zhou ◽  
Gang Wang ◽  
Guohui Liu ◽  
...  

ABSTRACT Osteomyelitis, an infection of the bone and bone marrow, imposes a heavy burden on public health care systems owing to its progressive bone destruction and sequestration. Human bone mesenchymal stem cells (hBMSCs) play a key role in the process of bone formation, and mounting evidence has confirmed that long noncoding RNAs (lncRNAs) are involved in hBMSC osteogenic differentiation. Nevertheless, the exact function and molecular mechanism of lncRNAs in osteogenic differentiation during osteomyelitis development remain to be explored. In this study, hBMSCs were treated with staphylococcal protein A (SpA) during osteogenic differentiation induction to mimic osteomyelitis in vitro. The results of lncRNA microarray analysis revealed that FAM83H-AS1 presented the lowest expression among the significantly downregulated lncRNAs. Functionally, ectopic expression of FAM83H-AS1 contributed to osteogenic differentiation of SpA-induced hBMSCs. Additionally, our findings revealed that FAM83H-AS1 negatively regulated microRNA 541-3p (miR-541-3p), and WNT3A was validated as a target gene of miR-541-3p. Mechanically, FAM83H-AS1 elevated WNT3A expression by competitively binding with miR-541-3p. Lastly, it was demonstrated that FAM83H-AS1/miR-541-3p/WNT3A ameliorated SpA-mediated inhibition of the osteogenic differentiation of hBMSCs, which provided a novel therapeutic strategy for patients with osteomyelitis.

RSC Advances ◽  
2019 ◽  
Vol 9 (16) ◽  
pp. 9117-9125
Author(s):  
Ting Ma ◽  
Xi-Yuan Ge ◽  
Ke-Yi Hao ◽  
Xi Jiang ◽  
Yan Zheng ◽  
...  

Titanium discs with simple 3,4-dihydroxy-l-phenylalanine coating enhanced BM-MSC adhesion, spreading, proliferation and differentiation, and upregulated expression of genes involved in focal adhesion in vitro.


Author(s):  
Lan Zhang ◽  
Yan Liu ◽  
Bo Feng ◽  
Li-Gong Liu ◽  
Ying-Cai Zhou ◽  
...  

This study aimed to certify the hypothesis that miR-138-5p is expected to reduced osteodifferentiation of human bone mesenchymal stem cells (hBMSCs) by FOXC1 down-regulation. hBMSCs were separated from bone marrow and osteogenic induction medium was added to stimulate osteogenic differentiation. Flow cytometric analysis was applied to evaluate the expression of cell surface antigens associated with hBMSCs, including CD29, CD44, CD90, CD45 and CD34. qRT-PCR assay and western blot assay were determined to measure the mRNA and protein expression of miR-138-5p, OCN, RUNX2, BSP, ALP and FOXC1. Alkaline phosphatase (ALP) staining assay and Alizarin Red Staining (ARS) assay were determined to validate the osteogenic differentiation. Luciferase assay was applied to test the interaction of miR-138-5p and FOXC1. We demonstrated miR-138-5p is downregulated in osteogenic differentiated hBMSCs. Besides, miR-138-5p overexpression diminished osteodifferentiated markers expression, ALP activity and ARS activity. Furthermore, we revealed that forkhead transcription factor C1 (FOXC1) was a downstream target gene of miR-138-5p and knockdown of miR-138-5p improved the osteogenesis differentiation of hBMSCs by upregulating FOXC1. miR-138-5p knockdown promoted osteogenic differentiation in hBMSCs via directly targeting FOXC1. This study suggested miR-138-5p may be a new target for hBMSCs osteogenic differentiation and the treatment of bone defects.


Molecules ◽  
2011 ◽  
Vol 16 (12) ◽  
pp. 10123-10133 ◽  
Author(s):  
Jun-Jun Fan ◽  
Liang-Guo Cao ◽  
Tao Wu ◽  
De-Xin Wang ◽  
Dan Jin ◽  
...  

2020 ◽  
Author(s):  
Yuli Wang ◽  
Fengyi Lv ◽  
Lintong Huang ◽  
Hengwei Zhang ◽  
Bing Li ◽  
...  

Abstract Background and aim: Periodontitis is a chronic inflammatory disease inducing the absorption of alveolar bone and leading to tooth loss. Human amnion–derived mesenchymal stem cells (HAMSCs) have been used for studying inflammatory processes. This study aimed to explore the role of long noncoding RNA (lncRNA) antisense noncoding RNA in the INK4 locus (ANRIL) in HAMSC-driven osteogenesis in lipopolysaccharide (LPS)-induced human bone marrow mesenchymal stem cells (HBMSCs).Methods: The cells were incubated with a co-culture system. Reactive oxygen species (ROS) level and superoxide dismutase (SOD) activity were used to detect the oxidative stress level. Flow cytometry was performed to determine cell proliferation. The alkaline phosphatase (ALP) activity, Alizarin red assay, cell transfection, and rat mandibular defect model were used to evaluate the osteogenic differentiation. Quantitative real-time reverse transcription–polymerase chain reaction (RT-PCR), Western blot analysis, dual-luciferase reporter assay, and immunofluorescence staining were used to evaluate the molecular mechanisms.Results: This study showed that HAMSCs promoted the osteogenesis of LPS-induced HBMSCs, while the ANRIL level in HBMSCs decreased during co-culture. ANRIL had no significant influence on the proliferation of LPS-induced HBMSCs. However, its overexpression inhibited the HAMSC-driven osteogenesis in vivo and in vitro, whereas its knockdown reversed these effects. Mechanistically, this study found that downregulating ANRIL led to the overexpression of microRNA-125a (miR-125a), and further contributed to the competitive binding of miR-125a and adenomatous polyposis coli (APC), thus significantly activating the Wnt/β-catenin pathway.Conclusion: The study indicated that HAMSCs promoted the osteogenic differentiation of LPS-induced HBMSCs via the ANRIL/miR-125a/APC axis, and offered a novel approach for periodontitis therapy.


2016 ◽  
Vol 38 (1) ◽  
pp. 40-48 ◽  
Author(s):  
Jianhou Huang ◽  
Yake Meng ◽  
Yan Liu ◽  
Yu Chen ◽  
Haisong Yang ◽  
...  

Background/Aims: Human bone marrow-derived mesenchymal stem cells (hMSCs) are a promising cell source for bone engineering owing to their high potential to differentiate into osteoblasts. The bone morphogenetic protein-inducible gene homeobox a10 (HOXA10) is a critical regulator of osteogenesis. The objective of the present study was to identify microR-NAs (miRNAs) targeting HOXA10 and examine the effects on the osteogenic differentiation of hMSCs. Methods: Based on in silico analysis, HOXA10-targeting miRNAs were selected and their regulatory roles in osteoblast differentiation were investigated. Results: Six HOXA10-targeting miRNAs were identifIed by computational analysis, of which miR-320a was selected for further analysis because it was downregulated during osteogenic induction. Overexpression of miR-320a downregulated HOXA10 and significantly inhibited osteogenesis in hMSCs, as determined by the downregulation of the osteogenic markers Runx2, ALP, and OC and the inhibition of ALP activity and matrix mineralization, whereas miR-320a inhibition had the opposite effects. Furthermore, ectopic expression of HOXA10 (not including 3′-UTR) rescued the effects of miR-320a on osteogenic differentiation. Conclusion: These results suggest that miR-320a acts as a critical regulator of osteogenic differentiation of hMSCs by repressing its target HOXA10.


2020 ◽  
Author(s):  
Yuli Wang ◽  
Fengyi Lv ◽  
Lintong Huang ◽  
Hengwei Zhang ◽  
Bing Li ◽  
...  

Abstract Background and aim: Periodontitis is a chronic inflammatory disease inducing the absorption of alveolar bone and leading to tooth loss. Human amnion–derived mesenchymal stem cells (HAMSCs) have been used for studying inflammatory processes. This study aimed to explore the role of long noncoding RNA (lncRNA) antisense noncoding RNA in the INK4 locus (ANRIL) in HAMSC-driven osteogenesis in lipopolysaccharide (LPS)-induced human bone marrow mesenchymal stem cells (HBMSCs). Methods: The cells were incubated with a co-culture system. Reactive oxygen species (ROS) level and superoxide dismutase (SOD) activity were used to detect the oxidative stress level. Flow cytometry was performed to determine cell proliferation. The alkaline phosphatase (ALP) activity, Alizarin red assay, cell transfection, and rat mandibular defect model were used to evaluate the osteogenic differentiation. Quantitative real-time reverse transcription–polymerase chain reaction (RT-PCR), Western blot analysis, dual-luciferase reporter assay, and immunofluorescence staining were used to evaluate the molecular mechanisms.Results: This study showed that HAMSCs promoted the osteogenesis of LPS-induced HBMSCs, while the ANRIL level in HBMSCs decreased during co-culture. ANRIL had no significant influence on the proliferation of LPS-induced HBMSCs. However, its overexpression inhibited the HAMSC-driven osteogenesis in vivo and in vitro, whereas its knockdown reversed these effects. Mechanistically, this study found that downregulating ANRIL led to the overexpression of microRNA-125a (miR-125a), and further contributed to the competitive binding of miR-125a and adenomatous polyposis coli (APC), thus significantly activating the Wnt/β-catenin pathway. Conclusion: The study indicated that HAMSCs promoted the osteogenic differentiation of LPS-induced HBMSCs via the ANRIL/miR-125a/APC axis, and offered a novel approach for periodontitis therapy.


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