Mechanical loading influences osteocytes and further regulates osteoblast metabolism and osteogenic differentiation(miRNA Array)
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ABSTRACT:
PROVIDER: PRJNA742690 | ENA |
REPOSITORIES: ENA
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Project description:A previous study indicated that mechanical tensile strain of 2500 microstrain (με) at 0.5 Hz for 8 h promoted osteogenesis and corresponding mechanoresponsive microRNAs (miRs) were identified in osteoblasts. However, in osteocytes, it has not been identified which miRs respond to the mechanical strain, and it is not fully understood how the mechanoresponsive miRs regulate osteoblast differentiation. Mouse MLO-Y4 osteocytes were applied to the same mechanical tensile strain in vitro.MiR microarray and reverse transcription-quantitative polymerase chainreaction(RT-qPCR) assays were applied to select and validate differentially expressed miRs, and the target genes of these miRs were then predicted. In addition, the differentially expressed miRNAs in mechanically strained osteocytes in vitro were compared with those in treadmill exercise of the bone tissues .
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Project description:Aim: To identify differentially expressed genes by mapping the response of human primary osteocytes in their native matrix to mechanical loading using RNA sequencing.
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Project description:Inherited bone disorders account for about 10% of documented Mendelian disorders and are associated with high financial burden. Their study requires osteoblasts which play a critical role in regulating the development and maintenance of bone tissue. However, bone tissue is not always available from patients. We developed a highly efficient platelet lysate-based approach to directly transdifferentiate skin-derived human fibroblasts to osteoblast-like cells. We extensively characterized our in vitro model by examining the expression of osteoblast-specific markers during the transdifferentiation process both at the mRNA and protein level. The transdifferentiated osteoblast-like cells showed significantly increased expression of a panel of osteogenic markers. Mineral deposition and ALP activity were also shown, confirming their osteogenic properties. RNA-seq analysis allowed the global study of changes in the transcriptome of the transdifferentiated cells. The transdifferentiated cells clustered separately from the fibroblasts with regard to the significantly upregulated genes indicating a distinct transcriptome profile; transdifferentiated osteoblasts also showed significant enrichment in gene expression related to skeletal development and bone mineralization. Our presented in vitro model may potentially contribute to the prospect of studying osteoblast-dependent disorders in patient-derived cells.
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Project description: Not available
| S-EPMC6700265 | biostudies-literature
Project description:Type of Experiment: 1) Profiles of Osteoblast vs. osteocyte in vitro; 2) Profiles of osteoblast low density vs. confluency in vitro; 3) Profiles of osteocyte with gap junction vs. without gap junction. Experimental factors: 1) 2T3 osteoblast cells at low density expressed extensive filopodia, reminiscent of early osteoblast precursors and similar to MLO-Y4 dendritic processes. 2) MLO-Y4 osteocytes at low vs. high density represent the genes that are changed in a highly connected network vs. low connected network. The number of hybridizations performed: Triplicate hybridizations for each status. Keywords = Osteoblast Keywords = Osteocyte
2010-06-09 | E-GEOD-2234 | biostudies-arrayexpress
Project description:Type of Experiment: 1) Profiles of Osteoblast vs. osteocyte in vitro; 2) Profiles of osteoblast low density vs. confluency in vitro; 3) Profiles of osteocyte with gap junction vs. without gap junction. Experimental factors: 1) 2T3 osteoblast cells at low density expressed extensive filopodia, reminiscent of early osteoblast precursors and similar to MLO-Y4 dendritic processes. 2) MLO-Y4 osteocytes at low vs. high density represent the genes that are changed in a highly connected network vs. low connected network. The number of hybridizations performed: Triplicate hybridizations for each status. Keywords = Osteoblast Keywords = Osteocyte Keywords: parallel sample
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Project description:Human adult mesenchymal stromal cells (hMSC) have the potential to differentiate into chondrogenic, adipogenic or osteogenic lineages, providing a potential source for tissue regeneration. An important issue for efficient bone regeneration is to identify factors that can be targeted to promote the osteogenic potential of hMSCs. Using transcriptomic analysis, we found that integrin alpha5 (ITGA5) expression is upregulated during dexamethasone-induced hMSCs osteoblast differentiation. Gain-of-function studies showed that ITGA5 promotes the expression of osteoblast phenotypic markers as well as in vitro osteogenesis in hMSCs. Downregulation of endogenous ITGA5 using shRNA blunted osteoblast marker expression and osteogenic differentiation. Pharmacological and molecular analyses showed that the enhanced hMSCs osteoblast differentiation induced by ITGA5 was mediated by activation of FAK/ERK1/2-MAPKs and PI3K signaling pathways. Remarkably, activation of ITGA5 using a specific antibody that primes the integrin or a peptide that specifically activates ITGA5 was sufficient to enhance ERK1/2-MAPKs and PI3K signaling and to promote osteoblast differentiation and osteogenic capacity of hMSCs. We also demonstrate that hMSCs engineered to over-express ITGA5 exhibited a marked increase in their osteogenic potential in vivo. These findings not only reveal that ITGA5 is required for osteoblast differentiation of adult human MSCs but also provide a novel targeted strategy using ITGA5 agonists to promote the osteogenic capacity of hMSCs, which may be used for tissue regeneration in bone disorders where the recruitment or capacity of MSCs is compromised. Keywords: Time course of osteogenic differentiation processes
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Project description:Gene expression profiles were examined in mineralizing-osteoblast specific androgen receptor knockout (mOBL-ARKO) and WT male mice to identify genes regulated by the AR in mineralizing osteoblasts and osteocytes.
2012-05-15 | E-MTAB-1123 | biostudies-arrayexpress
Project description:Human adult mesenchymal stromal cells (hMSC) have the potential to differentiate into chondrogenic, adipogenic or osteogenic lineages, providing a potential source for tissue regeneration. An important issue for efficient bone regeneration is to identify factors that can be targeted to promote the osteogenic potential of hMSCs. Using transcriptomic analysis, we found that integrin alpha5 (ITGA5) expression is upregulated during dexamethasone-induced hMSCs osteoblast differentiation. Gain-of-function studies showed that ITGA5 promotes the expression of osteoblast phenotypic markers as well as in vitro osteogenesis in hMSCs. Downregulation of endogenous ITGA5 using shRNA blunted osteoblast marker expression and osteogenic differentiation. Pharmacological and molecular analyses showed that the enhanced hMSCs osteoblast differentiation induced by ITGA5 was mediated by activation of FAK/ERK1/2-MAPKs and PI3K signaling pathways. Remarkably, activation of ITGA5 using a specific antibody that primes the integrin or a peptide that specifically activates ITGA5 was sufficient to enhance ERK1/2-MAPKs and PI3K signaling and to promote osteoblast differentiation and osteogenic capacity of hMSCs. We also demonstrate that hMSCs engineered to over-express ITGA5 exhibited a marked increase in their osteogenic potential in vivo. These findings not only reveal that ITGA5 is required for osteoblast differentiation of adult human MSCs but also provide a novel targeted strategy using ITGA5 agonists to promote the osteogenic capacity of hMSCs, which may be used for tissue regeneration in bone disorders where the recruitment or capacity of MSCs is compromised. Experiment Overall Design: Gene expression profiles were generated from bone marrow MSC before and 1, 3 and 7 days after stimulation with 10E-7M dexamethasone to study the early molecular processes of osteogenic differentiation. 3 replicates per timepoint.
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Project description:hMSCs (Lonza) were differentiated towards adipocytes with three cycles of adipocyte differentiation induction and maintenance. hMSCs were differentiated towards osteocytes with osteogenic differentiation medium for 3 days and for 7 days. ArrayCGH experiments were performed before differentiation induction, after 2 and 3 cycles of adipogenic differentiation induction and after 3 days and 7 days of osteogenic differentiation induction.
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