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Gingival mesenchymal stem cells (GMSCs) are newly developed seed cells for tissue engineering owing to their easy isolation, abundance and high growth rates. Thy-1 is an important regulatory molecule in the differentiation of mesenchymal stem cells (MSCs). In this study, we investigated the function of Thy-1 in the osteogenic differentiation of GMSCs by reducing the expression of Thy-1 using a lentivirus. The results demonstrated that Thy-1 knockdown promoted the osteogenic differentiation of GMSCs in vitro. Validation by RNA-seq revealed an obvious decrease in Vcam1 and Sox9 gene expression with Thy-1 knockdown. Kyoto Encyclopedia of Genes and Genomes pathway analysis suggested that the differentially expressed genes were enriched in the Wnt signalling pathway. We further demonstrated that Thy-1 knockdown promoted osteogenic differentiation of GMSCs by activating the Wnt/ß-catenin signalling pathway. Therefore, Thy-1 has a key regulatory role in the differentiation of GMSCs and maybe a core molecule connecting transcription factors related to the differentiation of MSCs. Our study also highlighted the potential of Thy-1 to modify MSCs, which may help improve their use in tissue engineering.
Assuntos
Células-Tronco Mesenquimais , Osteogênese , Antígenos Thy-1 , beta Catenina/genética , beta Catenina/metabolismo , Diferenciação Celular/genética , Células Cultivadas , Células-Tronco Mesenquimais/metabolismo , Osteogênese/genética , Via de Sinalização Wnt/genética , Antígenos Thy-1/genética , Antígenos Thy-1/metabolismoRESUMO
As industrial informatization progresses, virtual simulation technologies are increasingly demonstrating their potential in industrial applications. These systems utilize various sensors to capture real-time factory data, which are then transmitted to servers via communication interfaces to construct corresponding digital models. This integration facilitates tasks such as monitoring and prediction, enabling more accurate and convenient production scheduling and forecasting. This is particularly significant for flexible or mixed-flow production modes. Bionic optimization algorithms have demonstrated strong performance in factory scheduling and operations. Centered around these algorithms, researchers have explored various strategies to enhance efficiency and optimize processes within manufacturing environments.This study introduces an efficient migratory bird optimization algorithm designed to address production scheduling challenges in an assembly shop with mold quantity constraints. The research aims to minimize the maximum completion time in a batch flow mixed assembly flow shop scheduling problem, incorporating variable batch partitioning strategies. A tailored virtual simulation framework supports this objective. The algorithm employs a two-stage encoding mechanism for batch partitioning and sequencing, adapted to the unique constraints of each production stage. To enhance the search performance of the neighborhood structure, the study identifies and analyzes optimization strategies for batch partitioning and sequencing, and incorporates an adaptive neighborhood structure adjustment strategy. A competition mechanism is also designed to enhance the algorithm's optimization efficiency. Simulation experiments of varying scales demonstrate the effectiveness of the variable batch partitioning strategy, showing a 5-6% improvement over equal batch strategies. Results across different scales and parameters confirm the robustness of the algorithm.
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To promote bone repair, it is desirable to develop three-dimensional multifunctional fiber scaffolds. The densely stacked and tightly arranged conventional two-dimensional electrospun fibers hinder cell penetration into the scaffold. Most of the existing three-dimensional structural materials are isotropic and monofunctional. In this research, a Janus nanofibrous scaffold based on silk fibroin/polycaprolactone (SF/PCL) was fabricated. SF-encapsulated SeNPs demonstrated stability and resistance to aggregation. The outside layer (SF/PCL/Se) of the Janus nanofiber scaffold displayed a structured arrangement of fibers, facilitating cell growth guidance and impeding cell invasion. The inside layer (SF/PCL/HA) featured a porous structure fostering cell adhesion. The Janus fiber scaffold containing SeNPs notably suppressed S. aureus and E. coli activities, correlating with SeNPs concentration. In vitro, findings indicated considerable enhancement in alkaline phosphatase (ALP) activity of MC3T3-E1 osteoblasts and upregulation of genes linked to osteogenic differentiation with exposure to the SF/PCL/HA/Se Janus nanofibrous scaffold. Moreover, in vivo, experiments demonstrated successful critical bone defect repair in mouse skulls using the SF/PCL/HA/Se Janus nanofiber scaffold. These findings highlight the potential of the SF/PCL-based Janus nanofibrous scaffold, integrating SeNPs and nHA, as a promising biomaterial in bone tissue engineering.
Assuntos
Fibroínas , Nanofibras , Camundongos , Animais , Fibroínas/farmacologia , Fibroínas/química , Alicerces Teciduais/química , Osteogênese , Porosidade , Escherichia coli , Staphylococcus aureus , Engenharia Tecidual/métodos , Poliésteres/química , Regeneração Óssea , Nanofibras/química , Seda/químicaRESUMO
Background: Peri-implantitis (PI) is a prevalent complication of implant treatment. Pyroptosis, a distinctive inflammatory programmed cell death, is crucial to the pathophysiology of PI. Despite its importance, the pyroptosis-related genes (PRGs) influencing PI's progression remain largely unexplored. Methods: This study conducted histological staining and transcriptome analyze from three datasets. The intersection of differentially expressed genes (DEGs) and PRGs was identified as pyroptosis-related differentially expressed genes (PRDEGs). Functional enrichment analyses were conducted to shed light on potential underlying mechanisms. Weighted Gene Co-expression Network Analysis (WGCNA) and a pyroptotic macrophage model were utilized to identify and validate hub PRDEGs. Immune cell infiltration in PI and its relationship with hub PRDEGs were also examined. Furthermore, consensus clustering was performed to identify new PI subtypes. Protein-protein interaction (PPI) network, competing endogenous RNA (ceRNA) network, mRNA-mRNA binding protein regulatory (RBP) network, and mRNA-drugs regulatory network of hub PRDEGs were also analyzed. Results: Eight hub PRDEGs were identified: PGF, DPEP1, IL36B, IFIH1, TCEA3, RIPK3, NET7, and TLR3, which are instrumental in the PI's progression. Two PI subtypes were distinguished, with Cluster 1 exhibiting higher immune cell activation. The exploration of regulatory networks provided novel mechanisms and therapeutic targets in PI. Conclusion: Our research highlights the critical role of pyroptosis and identifies eight hub PRDEGs in PI's progression, offering insights into novel immunotherapy targets and laying the foundation for advanced diagnostic and treatment strategies. This contributes to our understanding of PI and underscores the potential for personalized clinical management.
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The use of bone substitute materials is crucial for the healing of large bone defects. Immune response induced by bone substitute materials is essential in bone regeneration. Prior research has mainly concentrated on innate immune cells, such as macrophages. Existing research suggests that T lymphocytes, as adaptive immune cells, play an indispensable role in bone regeneration. However, the mechanisms governing T cell recruitment and specific subsets that are essential for bone regeneration remain unclear. This study demonstrates that CD4+ T cells are indispensable for ectopic osteogenesis by biphasic calcium phosphate (BCP). Subsequently, the recruitment of CD4+ T cells is closely associated with the activation of calcium channels in macrophages by BCP to release chemokines Ccl3 and Ccl17. Finally, these recruited CD4+ T cells are predominantly Tregs, which play a significant role in ectopic osteogenesis by BCP. These findings not only shed light on the immune-regenerative process after bone substitute material implantation but also establish a theoretical basis for developing bone substitute materials for promoting bone tissue regeneration. STATEMENT OF SIGNIFICANCE: Bone substitute material implantation is essential in the healing of large bone defects. Existing research suggests that T lymphocytes are instrumental in bone regeneration. However, the specific mechanisms governing T cell recruitment and specific subsets that are essential for bone regeneration remain unclear. In this study, we demonstrate that activation of calcium channels in macrophages by biphasic calcium phosphate (BCP) causes them to release the chemokines Ccl3 and Ccl17 to recruit CD4+ T cells, predominantly Tregs, which play a crucial role in ectopic osteogenesis by BCP. Our findings provide a theoretical foundation for developing bone substitute material for bone tissue regeneration.
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Substitutos Ósseos , Substitutos Ósseos/farmacologia , Regeneração Óssea , Hidroxiapatitas/farmacologia , Canais de Cálcio , Quimiocinas , Osteogênese , Fosfatos de Cálcio/farmacologiaRESUMO
Quartz crystal microbalance (QCM) resonators are used in a wide range of sensors. Current QCM resonators achieve a simultaneous measurement of multiple physical quantities by analyzing lumped-element equivalent parameters, which are obtained via the introduction of external devices. This introduction of external devices will probably increase measurement error. To realize the measurement of multiple physical quantities while eliminating the measurement error caused by external devices, this paper proposes a measurement method for the lumped-element equivalent parameters of QCM resonators without the need for extra external devices. Accordingly, a numerical method for solving nonlinear equations with fewer data points required and a higher accuracy was adopted. A standard crystal resonator parameter extraction experiment is described. The extracted parameters were consistent with the nominal parameters, which confirms the accuracy of this method. Furthermore, six QCM resonator device samples with different electrode diameters and materials were produced and used in the parameter measurement experiment. The linear relationship between the electrode material conductivity and motional resistance R1 is discussed. The ability of this method to characterize the electrode material and to detect the rust status of the electrode is also demonstrated. These abilities support the potential utility of the proposed method for an electrode quality assessment of piezoelectric devices.
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OBJECTIVE: Biomaterial can be locally applied to promote the osseointegration of dental implants. This study aimed to fabricate an osteogenic inducer (OI) sustained-release system and to evaluate its effects on the adhesion, proliferation, and differentiation of osteoblasts on titanium surfaces. METHODS: First of all, different contents of OI solution were added to the poly (lactic-co-glycolic acid) (PLGA) gel individually to investigate the best physical properties and drug-release rate. Moreover, osteoblasts were isolated from the calvaria of two-month-old New Zealand rabbits through sequential enzymatic digestion. Osteoblasts were seeded onto the surface of Ti disks (control group), Ti coated with PLGA gel (PLGA group), and Ti coated with the OI sustained-release system (PLGA+OI group). Cell adhesion was observed by scanning electron microscopy. Cell proliferation was analyzed by cell counting kit-8. Cell differentiation was tested by alizarin red staining, alkaline phosphatase (ALP) activity and osteogenic-related gene expression. RESULTS: The OI sustained-release system contained 15% OI solution had appropriate physical properties and drug-release rate. The osteoblasts in the PLGA+OI group were in a typical spindle shape with a considerable number indicating the promotion of adhesion and proliferation. The expression of early and late stage osteoblast differentiation genes in the PLGA+OI group were significantly higher than that of the control group and PLGA group at each time point. The PLGA group showed accelerated adhesion and differentiation but reduced proliferation compared with the control. CONCLUSION: The OI sustained-release system promotes the adhesion, proliferation, and differentiation of osteoblasts on titanium surfaces. This system is a cost-effective osteoconductive biomaterial that might be promising for use in dental implantation.