Development of non-destructive methods of the characterization of mesenchymal stem cell differentiation in vitro

Real time monitoring of stem cells has been a growing area of interest over the past decade because of new regenerative medicine approaches. Also, the effect of culture composition on stem cell metabolic pathways and their regulation of cellular fate are of increasing importance. In our project, non-destructive metabolomic analysis of stem cells and their physiological status during proliferation and differentiation stages will be investigated. This will be achieved through mass spectrometric analysis of released metabolites by LCMS and GC/MS; therefore stem cells should ideally be cultured in a specialized format to maintain their physiological properties during non-invasive live analysis while being compatible with real time GC/MS. To address this issue, mesenchymal stem cell attachment, proliferation and differentiation were tested in 20ml glass vials required for GC/MS analysis. Environmental conditions in which MSCs can proliferate under sealed condition and at room temperature were also optimized. Our data show the potential of MSCs to attach, proliferate and differentiate in 20ml glass vessels. Furthermore, cells could maintain their metabolic activity for approximately 6hrs not only under hypoxic but also hypothermic conditions. Therefore, live time analysis of their chemical volatiles could be performed in a format compatible for LC-MS and GC/MS which will be performed for undifferentiated MSCs and their differentiated populations (osteogenic and adipogenic lineages) in the next phase of the project. A specific treatment approach has emerged from hMSCs osteogenic study and identified a particular pathway suggested to be involved in chondrogenesis, and linked to the collagen II biosynthesis. The treatment is based on on natural products (L-lysine and ascorbic acid) and when tested on human cell cultures, was observed to significantly increase the morphological and functional markers of chondrogenesis, suggesting its potential to promote de novo cartilage formation in vitro

Non-destructive characterisation of mesenchymal stem cell differentiation using LC-MS-based metabolite footprinting

Bone regeneration is a complex biological process where major cellular changes take place to support the osteogenic differentiation of mesenchymal bone progenitors. To characterise these biological changes and better understand the pathways regulating the formation of mature bone cells, the metabolic profile of mesenchymal stem cell (MSC) differentiation in vitro has been assessed non-invasively during osteogenic (OS) treatment using a footprinting technique. Liquid chromatography (LC)-mass spectrometry (MS)-based metabolite profiling of the culture medium was carried out in parallel to mineral deposition and alkaline phosphatase activity which are two hallmarks of osteogenesis in vitro. Metabolic profiles of spent culture media with a combination of univariate and multivariate analyses investigated concentration changes of extracellular metabolites and nutrients linked to the presence of MSCs in culture media. This non-invasive LC-MS-based analytical approach revealed significant metabolic changes between the media from control and OS-treated cells showing distinct effects of MSC differentiation on the environmental footprint of the cells in different conditions (control vs. OS treatment). A subset of compounds was directly linked to the osteogenic time-course of differentiation, and represent interesting metabolite candidates as non-invasive biomarkers for characterising the differentiation of MSCs in a culture medium

Non-destructive characterisation of mesenchymal stem cell differentiation using LC-MS-based metabolite footprinting

Bone regeneration is a complex biological process where major cellular changes take place to support the osteogenic differentiation of mesenchymal bone progenitors. To characterise these biological changes and better understand the pathways regulating the formation of mature bone cells, the metabolic profile of mesenchymal stem cell (MSC) differentiation in vitro has been assessed non-invasively during osteogenic (OS) treatment using a footprinting technique. Liquid chromatography (LC)-mass spectrometry (MS)-based metabolite profiling of the culture medium was carried out in parallel to mineral deposition and alkaline phosphatase activity which are two hallmarks of osteogenesis in vitro. Metabolic profiles of spent culture media with a combination of univariate and multivariate analyses investigated concentration changes of extracellular metabolites and nutrients linked to the presence of MSCs in culture media. This non-invasive LC-MS-based analytical approach revealed significant metabolic changes between the media from control and OS-treated cells showing distinct effects of MSC differentiation on the environmental footprint of the cells in different conditions (control vs. OS treatment). A subset of compounds was directly linked to the osteogenic time-course of differentiation, and represent interesting metabolite candidates as non-invasive biomarkers for characterising the differentiation of MSCs in a culture medium

Enhancing the characteristics of MHD squeezed Maxwell nanofluids via viscous dissipation impact

Theoretical and numerical analysis are employed in this study to explore the characteristics of Maxwell squeezed nanofluid flow over a sensor surface, accounting for both the effects of viscous dissipation and an external magnetic field. The objective of this study is to investigate the impact of these two factors on the behavior of the nanofluid as it traverses the sensor surface, with a specific emphasis on the modifications in its physical properties, including thermal conductivity and viscosity. In this study, the theoretical analysis relies on the Navier-Stokes equations and Maxwell's equations, which are numerically solved using a shooting method. According to the findings, the applied magnetic field and viscous dissipation have a notable influence on the nanofluid's physical properties and flow characteristics. The magnetic field induces greater alignment and concentration of nanoparticles in the nanofluid, leading to alterations in the fluid's thermal conductivity and viscosity. The impacts of viscous dissipation are likewise observed to be significant, resulting in a considerable elevation in the fluid temperature as a result of the frictional forces between the fluid and the sensor surface. The values for drag coefficient, heat transfer, and mass transfer rate are organized in a table. Some significant findings were observed in this study, which indicate that the viscosity parameter, the squeezed flow index, and magnetic parameter contribute to a reduction in the temperature distribution across the boundary layer region. Conversely, the thermal conductivity parameter and Eckert number show the opposite trend, resulting in an increase in temperature distribution. Furthermore, the novelty of this investigation can be accentuated by analyzing the flow of squeezed Maxwell nanofluid due to a sensor surface based on the Buongiorno concept. This analysis takes into account external magnetic fields, variable thermal conductivity assumptions, and the phenomenon of viscous dissipation

Dietary knowledge assessment among the patients with type 2 diabetes in Madinah: A cross-sectional study [version 2; peer review: 2 approved]

Background There is a huge burden of nutrition-related non-communicable diseases, and diabetes is one of the leading chronic nutrition-related diseases affecting more than 500 million people globally. Collecting information regarding the awareness of dietary and nutrition knowledge among diabetic patients is the first step to developing a disease prevention program. Thus, this study primarily aims at assessing the dietary awareness of diabetes patients attending the diabetic centre in Madinah governorate, Saudi Arabia. Methods The study was started in November 2020 and ended in October 2021. The study participants (315) were type 2 diabetes mellitus (T2DM) patients attending a diabetic centre in Madinah, Saudi Arabia. A self-prepared dietary knowledge questionnaire (DKQ) was used in this research. The variables include balanced diet, food type, food choice, carbohydrate, protein, and fat. Knowledge score was, and the total score was levelled/categorized into ‘good’, ‘average’, and ‘poor’. Data were analysed by SPSS v.26. Results The study results identified the current knowledge of T2DM patients about different dietary items. The knowledge score of 62.2% of participants showed an average level of dietary knowledge, which is statistically significant. When we separately evaluated their understanding of different dietary components, we found that T2DM patients had poor knowledge of carbohydrates (30.15%), fat, food choices (47.7%), and type (34.6%). However, they had acceptable knowledge of proteins (56.5%). Conclusion Our participants exhibited acceptable knowledge about proteins but poorer knowledge of other food groups. A healthy, well-balanced diet is essential for excellent glycaemic control. Educating and arranging a health education program regarding dietary knowledge is recommended, specially designed for diabetic patients so that patients can opt for a healthier lifestyl

Development of non-destructive methods of the characterization of mesenchymal stem cell differentiation in vitro

Real time monitoring of stem cells has been a growing area of interest over the past decade because of new regenerative medicine approaches. Also, the effect of culture composition on stem cell metabolic pathways and their regulation of cellular fate are of increasing importance. In our project, non-destructive metabolomic analysis of stem cells and their physiological status during proliferation and differentiation stages will be investigated. This will be achieved through mass spectrometric analysis of released metabolites by LCMS and GC/MS; therefore stem cells should ideally be cultured in a specialized format to maintain their physiological properties during non-invasive live analysis while being compatible with real time GC/MS. To address this issue, mesenchymal stem cell attachment, proliferation and differentiation were tested in 20ml glass vials required for GC/MS analysis. Environmental conditions in which MSCs can proliferate under sealed condition and at room temperature were also optimized. Our data show the potential of MSCs to attach, proliferate and differentiate in 20ml glass vessels. Furthermore, cells could maintain their metabolic activity for approximately 6hrs not only under hypoxic but also hypothermic conditions. Therefore, live time analysis of their chemical volatiles could be performed in a format compatible for LC-MS and GC/MS which will be performed for undifferentiated MSCs and their differentiated populations (osteogenic and adipogenic lineages) in the next phase of the project. A specific treatment approach has emerged from hMSCs osteogenic study and identified a particular pathway suggested to be involved in chondrogenesis, and linked to the collagen II biosynthesis. The treatment is based on on natural products (L-lysine and ascorbic acid) and when tested on human cell cultures, was observed to significantly increase the morphological and functional markers of chondrogenesis, suggesting its potential to promote de novo cartilage formation in vitro