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BMC Genomics Jun 2024Dynamic metabolic reprogramming occurs at different stages of myogenesis and contributes to the fate determination of skeletal muscle satellite cells (MuSCs)....
Dynamic metabolic reprogramming occurs at different stages of myogenesis and contributes to the fate determination of skeletal muscle satellite cells (MuSCs). Accumulating evidence suggests that mutations in myostatin (MSTN) have a vital role in regulating muscle energy metabolism. Here, we explored the metabolic reprogramming in MuSCs and myotube cells in MSTN and FGF5 dual-gene edited sheep models prepared previously, and also focused on the metabolic alterations during myogenic differentiation of MuSCs. Our study revealed that the pathways of nucleotide metabolism, pantothenate and CoA biosynthesis were weakened, while the unsaturated fatty acids biosynthesis were strengthened during myogenic differentiation of sheep MuSCs. The MSTN and FGF5 dual-gene editing mainly inhibited nucleotide metabolism and biosynthesis of unsaturated fatty acids in sheep MuSCs, reduced the number of lipid droplets in per satellite cell, and promoted the pentose phosphate pathway, and the interconversion of pentose and glucuronate. The MSTN and FGF5 dual-gene editing also resulted in the inhibition of nucleotide metabolism and TCA cycle pathway in differentiated myotube cells. The differential metabolites we identified can be characterized as biomarkers of different cellular states, and providing a new reference for MSTN and FGF5 dual-gene editing in regulation of muscle development. It may also provide a reference for the development of muscle regeneration drugs targeting biomarkers.
Topics: Animals; Myostatin; Muscle Development; Sheep; Gene Editing; Fibroblast Growth Factor 5; Cell Differentiation; Satellite Cells, Skeletal Muscle; Muscle Fibers, Skeletal
PubMed: 38926663
DOI: 10.1186/s12864-024-10494-w -
Anticancer Research Jul 2024This study aimed to investigate the structure and functions of the membrane formed around liquid nitrogen-treated bones in the osteogenesis and revitalization of frozen...
BACKGROUND/AIM
This study aimed to investigate the structure and functions of the membrane formed around liquid nitrogen-treated bones in the osteogenesis and revitalization of frozen bone using a rat model.
MATERIALS AND METHODS
Segmental defects were created in femurs of rats, and resected bones treated with liquid nitrogen [frozen bone (FB) group, n=20] or polymethylmethacrylate (PMMA group; n=20) were implanted as spacers. Histological analysis and quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) of the membrane around each spacer were performed for bone morphogenetic protein 2 (BMP2), transforming growth factor (TGF)-β1, and vascular endothelial growth factor (VEGF). Furthermore, in week 2, spacers were removed from both groups (n=5 each), and autologous cancellous bone (ACB) harvested from the ilium was grafted into the defect. Radiological analysis was performed until bone union was observed.
RESULTS
In week 2, similar two-layered membrane structures were observed in both groups; these matured into fibrous tissues over time. At each evaluation point, qRT-PCR showed higher expression of all factors in the FB than in the PMMA group. In the ACB graft model, the mean period to bone union and new bone volume were significantly shorter and greater, respectively, in the FB. Chondrocytes invaded the osteotomy site from the membrane in the FB, suggesting that endochondral ossification may occur and be related to osteogenesis. Additionally, fibroblasts and capillaries in the membrane invaded the surface of treated bone in week 2, and osteocytes were observed around them in weeks 6 and 8.
CONCLUSION
Fibrous membranous tissue formed around liquid nitrogen-treated bones may be vital for osteogenesis and revitalization of frozen bones.
Topics: Animals; Osteogenesis; Rats; Vascular Endothelial Growth Factor A; Nitrogen; Bone Morphogenetic Protein 2; Male; Bone Transplantation; Transforming Growth Factor beta1; Polymethyl Methacrylate; Femur; Bone and Bones; Rats, Sprague-Dawley
PubMed: 38925839
DOI: 10.21873/anticanres.17101 -
Advanced Healthcare Materials Jun 2024Cell spheroids (esp. organoids) as three-dimensional (3D) culture platforms are popular models for representing cell-cell and cell-extracellular matrix (ECM)... (Review)
Review
Cell spheroids (esp. organoids) as three-dimensional (3D) culture platforms are popular models for representing cell-cell and cell-extracellular matrix (ECM) interactions, bridging the gap between 2D cell cultures and natural tissues. 3D cell models with spatially organized multiple cell types are preferred for gaining comprehensive insights into tissue pathophysiology and constructing in vitro tissues and disease models because of the complexities of natural tissues. In recent years, an assembly strategy using cell spheroids (or organoids) as living building blocks has been developed to construct complex 3D tissue models with spatial organization. Here, we provide a comprehensive overview of recent advances in multi-spheroid assembly studies. The different mechanisms of the multi-spheroid assembly techniques, i.e., automated directed assembly, noncontact remote assembly, and programmed self-assembly, are introduced. The processing steps, advantages, and technical limitations of the existing methodologies are summarized. Applications of the multi-spheroid assembly strategies in disease modeling, drug screening, tissue engineering and organogenesis are reviewed. Finally, this review concludes by emphasizing persistent issues and our perspectives, encouraging researchers to adopt multi-spheroid assembly techniques for generating advanced 3D cell models that better resemble real tissues. This article is protected by copyright. All rights reserved.
PubMed: 38924326
DOI: 10.1002/adhm.202400957 -
Journal of Biochemical and Molecular... Jul 2024Osteomyelitis is an invasive bone infection that can lead to severe pain and even disability, posing a challenge for orthopedic surgery. Naringin can reduce bone-related...
Osteomyelitis is an invasive bone infection that can lead to severe pain and even disability, posing a challenge for orthopedic surgery. Naringin can reduce bone-related inflammatory conditions. This study aimed to elucidate the function and mechanism of naringin in a Staphylococcus aureus-induced mouse model of osteomyelitis. Femurs of S. aureus-infected mice were collected after naringin administration and subjected to microcomputed tomography to analyze cortical bone destruction and bone loss. Bacterial growth in femurs was also assessed. Proinflammatory cytokine levels in mouse femurs were measured using enzyme-linked immunosorbent assays. Pathological changes and bone resorption were analyzed using hematoxylin and eosin staining and tartrate-resistant acid phosphatase staining, respectively. Quantitative reverse transcription polymerase chain reaction and western blot analysis were used to quantify the messenger RNA and protein expression of osteogenic differentiation-associated genes in the femurs. The viability of human bone marrow-derived stem cells (hBMSCs) was determined using cell counting kit-8. Alizarin Red S staining and alkaline phosphatase staining were performed to assess the formation of mineralization nodules and bone formation in vitro. Notch signaling-related protein levels in femur tissues and hBMSCs were assessed using western blot analysis. Experimental results revealed that naringin alleviated S. aureus-induced cortical bone destruction and bone loss in mice by increasing the bone volume/total volume ratio. Naringin suppressed S. aureus-induced bacterial growth and inflammation in femurs. Moreover, it alleviated histopathological changes, inhibited bone resorption, and increased the expression of osteogenic markers in osteomyelitic mice. It increased the viability of hBMSCs and promoted their differentiation and bone mineralization in vitro. Furthermore, naringin activated Notch signaling by upregulating the protein levels of Notch1, Jagged1, and Hes1 in the femurs of model mice and S. aureus-stimulated hBMSCs. In conclusion, naringin reduces bacterial growth, inflammation, and bone resorption while upregulating the expression of osteogenic markers in S. aureus-infected mice and hBMSCs by activating Notch signaling.
Topics: Animals; Flavanones; Mice; Osteomyelitis; Staphylococcus aureus; Staphylococcal Infections; Anti-Bacterial Agents; Anti-Inflammatory Agents; Humans; Male; Osteogenesis; Femur
PubMed: 38923626
DOI: 10.1002/jbt.23753 -
Brazilian Journal of Biology = Revista... 2024Despite being valuable for producing a natural sweetener Curculin, Curculigo latifolia has a low growth and difficult to domestificate. So, to solve this problem,...
Despite being valuable for producing a natural sweetener Curculin, Curculigo latifolia has a low growth and difficult to domestificate. So, to solve this problem, propagation on in vitro culture will be an alternative method to propagated this spesies under different cytokinins and light condition. Cytokinins and light has major role in organogenesis, growth and gene expression of many species. Thus, in this study, we aimed to improve the Curculigo latifolia growth on in vitro condition and expression of curculin gene by combining cytokinins addition and different light exposure. Four weeks seedlings were sub-cultured into medium (MS free hormone) containing 3 mg/L benzyladenine (BA) and various concentrations of meta-Topolin (mT) including 0.1 mg/L, 0.5 mg/L, and 5 mg/L. The cultures then incubated under different light types (red, blue, white LED lights and white fluorescence light) with 16-h light/ 18-h dark photoperiod for 14 weeks at 25 ± 2°C. Several parameters, including plant height, leaf number, chlorophyll contents, stomatal structure, and density and curculin expression, were observed every week. Unexpectedly, our results showed that C. latifolia growth displayed significant improvement when it was treated under white LED light without any additional cytokinins. In sum, white LED light further improves plantlets phenotype, such as plant height, leaf number, chlorophyll production, and stomatal number and structure, whereas, red LED light lead to a decreased phenotypes but increase the curculin gene expression.
Topics: Cytokinins; Light; Curculigo; Plant Growth Regulators; Gene Expression Regulation, Plant
PubMed: 38922193
DOI: 10.1590/1519-6984.280778 -
Cells Jun 2024Resveratrol is a polyphenol known to have metabolic as well as circadian effects. However, there is little information regarding the metabolic and circadian effect of...
Resveratrol is a polyphenol known to have metabolic as well as circadian effects. However, there is little information regarding the metabolic and circadian effect of resveratrol on muscle cells. We sought to investigate the metabolic impact of resveratrol throughout the circadian cycle to clarify the associated signaling pathways. C2C12 myotubes were incubated with resveratrol in the presence of increasing concentrations of glucose, and metabolic and clock proteins were measured for 24 h. Resveratrol led to SIRT1, AMPK and PP2A activation. Myotubes treated with increasing glucose concentrations showed higher activation of the mTOR signaling pathway. However, resveratrol did not activate the mTOR signaling pathway, except for P70S6K and S6. In accordance with the reduced mTOR activity, resveratrol led to advanced circadian rhythms and reduced levels of pBMAL1 and CRY1. Resveratrol increased myogenin expression and advanced its rhythms. In conclusion, resveratrol activates the SIRT1-AMPK-PP2A axis, advances circadian rhythms and induces muscle development.
Topics: Resveratrol; Sirtuin 1; Animals; Mice; Muscle Fibers, Skeletal; Protein Phosphatase 2; AMP-Activated Protein Kinases; Circadian Rhythm; Signal Transduction; Cell Line; Glucose; Muscle Development; TOR Serine-Threonine Kinases
PubMed: 38920697
DOI: 10.3390/cells13121069 -
Cells Jun 2024Successful heart development depends on the careful orchestration of a network of transcription factors and signaling pathways. In recent years, in vitro cardiac...
Successful heart development depends on the careful orchestration of a network of transcription factors and signaling pathways. In recent years, in vitro cardiac differentiation using human pluripotent stem cells (hPSCs) has been used to uncover the intricate gene-network regulation involved in the proper formation and function of the human heart. Here, we searched for uncharacterized cardiac-development genes by combining a temporal evaluation of human cardiac specification in vitro with an analysis of gene expression in fetal and adult heart tissue. We discovered that (CARdiac DEvelopment Long non-coding RNA; LINC00890; SERTM2) expression coincides with the commitment to the cardiac lineage. knockout hPSCs differentiated poorly into cardiac cells, and hPSC-derived cardiomyocytes showed faster beating rates after controlled overexpression of during differentiation. Altogether, we provide physiological and molecular evidence that expression contributes to sculpting the cardiac program during cell-fate commitment.
Topics: Humans; RNA, Long Noncoding; Cell Differentiation; Heart; Homeostasis; Myocytes, Cardiac; Gene Expression Regulation, Developmental; Pluripotent Stem Cells; Cell Lineage; Organogenesis
PubMed: 38920678
DOI: 10.3390/cells13121050 -
Cells Jun 2024Mesenchymal stem cells (MSCs) of placental origin hold great promise in tissue engineering and regenerative medicine for diseases affecting cartilage and bone. However,...
Mesenchymal stem cells (MSCs) of placental origin hold great promise in tissue engineering and regenerative medicine for diseases affecting cartilage and bone. However, their utility has been limited by their tendency to undergo premature senescence and phenotypic drift into adipocytes. This study aimed to explore the potential involvement of a specific subset of aging and antiaging genes by measuring their expression prior to and following in vitro-induced differentiation of placental MSCs into chondrocytes and osteoblasts as opposed to adipocytes. The targeted genes of interest included the various transcript variants (lamin A, lamin C, and lamin A∆10), sirtuin 7 (SIRT7), and SM22α, along with the classic aging markers plasminogen activator inhibitor 1 (PAI-1), p53, and p16. MSCs were isolated from the decidua basalis of human term placentas, expanded, and then analyzed for phenotypic properties by flow cytometry and evaluated for colony-forming efficiency. The cells were then induced to differentiate in vitro into chondrocytes, osteocytes, and adipocytes following established protocols. The mRNA expression of the targeted genes was measured by RT-qPCR in the undifferentiated cells and those fully differentiated into the three cellular lineages. Compared to undifferentiated cells, the differentiated chondrocytes demonstrated decreased expression of SIRT7, along with decreased PAI-1, lamin A, and SM22α expression, but the expression of p16 and p53 increased, suggesting their tendency to undergo premature senescence. Interestingly, the cells maintained the expression of lamin C, which indicates that it is the primary lamin variant influencing the mechanoelastic properties of the differentiated cells. Notably, the expression of all targeted genes did not differ from the undifferentiated cells following osteogenic differentiation. On the other hand, the differentiation of the cells into adipocytes was associated with decreased expression of lamin A and PAI-1. The distinct patterns of expression of aging and antiaging genes following in vitro-induced differentiation of MSCs into chondrocytes, osteocytes, and adipocytes potentially reflect specific roles for these genes during and following differentiation in the fully functional cells. Understanding these roles and the network of signaling molecules involved can open opportunities to improve the handling and utility of MSCs as cellular precursors for the treatment of cartilage and bone diseases.
Topics: Humans; Mesenchymal Stem Cells; Female; Placenta; Cell Differentiation; Chondrogenesis; Pregnancy; Osteogenesis; Biomarkers; Cellular Senescence; Chondrocytes; Aging; Lamin Type A
PubMed: 38920652
DOI: 10.3390/cells13121022 -
Cells Jun 2024Bone formation is a complex process regulated by a variety of pathways that are not yet fully understood. One of the proteins involved in multiple osteogenic pathways is...
Bone formation is a complex process regulated by a variety of pathways that are not yet fully understood. One of the proteins involved in multiple osteogenic pathways is TID (DNAJA3). The aim of this work was to study the association of TID with osteogenesis. Therefore, the expression profiles of the splice variants (, ) and their protein products were analyzed during the proliferation and differentiation of bone marrow mesenchymal stromal cells (B-MSCs) into osteoblasts. As the reference, the hFOB1.19 cell line was used. The phenotype of B-MSCs was confirmed by the presence of CD73, CD90, and CD105 surface antigens on ~97% of cells. The osteoblast phenotype was confirmed by increased alkaline phosphatase activity, calcium deposition, and expression of ALPL and SPP1. The effect of silencing the gene on the expression of and was also investigated. The TID proteins and the expression of splice variants were detected. After differentiation, the expression of and increased 5-fold and 3.7-fold, respectively, while their silencing resulted in increased expression of . Three days after transfection, the expression of increased 7.6-fold and 5.6-fold in B-MSCs and differentiating cells, respectively. Our preliminary study demonstrated that the expression of and changes under differentiation of B-MSCs into osteoblasts and may influence the expression of . However, for better understanding the functional association of these results with the relevant osteogenic pathways, further studies are needed.
Topics: Humans; Osteoblasts; Mesenchymal Stem Cells; Cell Differentiation; Osteogenesis; Protein Isoforms; Alkaline Phosphatase; Bone Marrow Cells; Cell Proliferation
PubMed: 38920651
DOI: 10.3390/cells13121021 -
Cells Jun 2024Circular RNAs (circRNAs) have emerged as pivotal regulators of gene expression with diverse roles in various biological processes. In recent years, research into... (Review)
Review
Circular RNAs (circRNAs) have emerged as pivotal regulators of gene expression with diverse roles in various biological processes. In recent years, research into circRNAs' involvement in bone biology has gained significant attention, unveiling their potential as novel regulators and biomarkers in bone-related disorders and diseases. CircRNAs, characterized by their closed-loop structure, exhibit stability and resistance to degradation, underscoring their functional significance. In bone tissue, circRNAs are involved in critical processes such as osteogenic differentiation, osteoclastogenesis, and bone remodeling through intricate molecular mechanisms including microRNA regulation. Dysregulated circRNAs are associated with various bone disorders, suggesting their potential as diagnostic and prognostic biomarkers. The therapeutic targeting of these circRNAs holds promise for addressing bone-related conditions, offering new perspectives for precision medicine. Thus, circRNAs constitute integral components of bone regulatory networks, impacting both physiological bone homeostasis and pathological conditions. This review provides a comprehensive overview of circRNAs in bone biology, emphasizing their regulatory mechanisms, functional implications, and therapeutic potential.
Topics: Humans; RNA, Circular; Bone and Bones; Animals; Bone Diseases; Osteogenesis; Biomarkers; MicroRNAs; Gene Expression Regulation
PubMed: 38920630
DOI: 10.3390/cells13120999