Physical versus Immunological Purification of Mesenchymal Stem Cells

A prerequisite before dealing with any cell type is to identify it and isolate it from the heterogeneous cell population that it belongs to. Mesenchymal stem cells (MSC) can be found in nearly all tissues and are mostly located in perivascular niches

Development of Primary Percutaneous Coronary Intervention as a National Reperfusion Strategy for Patients with ST-Elevation Myocardial Infarction and Assessment of Its Use in Egypt

Objective: Early treatment of acute ischemia of the heart by performing immediate percutaneous coronary intervention (PCI) to restore blood flow in patients with the clinical presentation of an acute coronary syndrome and more specifically with ST-elevation myocardial infarction (STEMI) can save lives. This study aims to identify the mean time (door to balloon time and first contact to balloon time) to primary PCI for STEMI patients and to assess the percentage of primary PCI and its success rate in Egypt. Methods: A registry study of patients presenting to cardiac centers in Egypt was designed, where patients’ basic characteristics, the treatment strategy, and the door to balloon time and the first contact to balloon time were assessed. Results: One thousand six hundred fifty STEMI patients with a mean age of 57 years were included in the study. Immediate transfer for primary PCI was the most used treatment strategy, representing 74.6% of all treatment strategies used. The door to balloon time and the first contact to balloon time were 50 and 60 minutes, respectively, with a primary PCI success rate of 65.1%. Conclusion: The registry study results showed a marked improvement by implementation of the best treatment strategy with respect to the time factor to achieve a better outcome for STEMI patients in Egypt

Cell Culture

Cell culture is one of the major tools used in cellular and molecular biology, delivering an excellent model for studying the normal physiology and biochemistry of cells. This book covers some advanced aspects in cell culture methodologies. The book has four sections discussing different types of cell culture models, including 3D cell culture techniques, their advantages, and limitations in comparison to traditional 2D culturing; cell viability, autophagy, in vitro toxicity tests and live cell imaging; stem cell culture for cell-based therapeutics; and specific applications and methodologies for hybrid cell lines and cancer models. This book provides a comprehensive overview of some of the advanced cell culture methodologies and applications. It serves as a valuable source for scientists, researchers, clinicians and students

Extracellular cystine influences human preadipocyte differentiation and correlates with fat mass in healthy adults

Abstract Plasma cysteine is associated with human obesity, but it is unknown whether this is mediated by reduced, disulfide (cystine and mixed-disulfides) or protein-bound (bCys) fractions. We investigated which cysteine fractions are associated with adiposity in vivo and if a relevant fraction influences human adipogenesis in vitro. In the current study, plasma cysteine fractions were correlated with body fat mass in 35 adults. Strong positive correlations with fat mass were observed for cystine and mixed disulfides ( r  ≥ 0.61, P  < 0.001), but not the quantitatively major form, bCys. Primary human preadipocytes were differentiated in media containing cystine concentrations varying from 10–50 μM, a range similar to that in plasma. Increasing extracellular cystine (10–50 μM) enhanced mRNA expression of PPARG2 (to sixfold ) , PPARG1 , PLIN1 , SCD1 and CDO1 ( P  = 0.042– < 0.001). Adipocyte lipid accumulation and lipid-droplet size showed dose-dependent increases from lowest to highest cystine concentrations ( P  < 0.001), and the malonedialdehyde/total antioxidant capacity increased, suggesting increased oxidative stress. In conclusion, increased cystine concentrations, within the physiological range, are positively associated with both fat mass in healthy adults and human adipogenic differentiation in vitro. The potential role of cystine as a modifiable factor regulating human adipocyte turnover and metabolism deserves further study

The Effect of Bone Marrow-Derived Mesenchymal Stem Cells and Their Conditioned Media Topically Delivered in Fibrin Glue on Chronic Wound Healing in Rats

Bone marrow-derived mesenchymal stem cells (BM-MSCs) represent a modern approach for management of chronic skin injuries. In this work, we describe BM-MSCs application versus their conditioned media (CM) when delivered topically admixed with fibrin glue to enhance the healing of chronic excisional wounds in rats. Fifty-two adult male rats were classified into four groups after induction of large-sized full-thickness skin wound: control group (CG), fibrin only group (FG), fibrin + MSCs group (FG + SCs), and fibrin + CM group (FG + CM). Healing wounds were evaluated functionally and microscopically. Eight days after injury, number of CD68+ macrophages infiltrating granulation tissue was considerably higher in the latter two groups. Although—later—none of the groups depicted a substantially different healing rate, the quality of regenerated skin was significantly boosted by the application of either BM-MSCs or their CM both (1) structurally as demonstrated by the obviously increased mean area percent of collagen fibers in Masson’s trichrome-stained skin biopsies and (2) functionally as supported by the interestingly improved epidermal barrier as well as dermal tensile strength. Thus, we conclude that topically applied BM-MSCs and their CM—via fibrin vehicle—could effectively improve the quality of healed skin in chronic excisional wounds in rats, albeit without true acceleration of wound closure

Hybrid bioactive hydroxyapatite/polycaprolactone nanoparticles for enhanced osteogenesis

Hydroxyapatite nanoparticles (HApN) are largely employed as osteogenic inorganic material. Inorganic/polymeric hybrid nanostructures can provide versatile bioactivity for superior osteogenicity, particularly as nanoparticles. Herein, we present hybrid biomaterial-based hydroxyapatite/polycaprolactone nanoparticles (HAp/PCL NPs) realized using simple preparation techniques to augment HApN osteogenicity. Using wet chemical precipitation, we optimized HApN crystalline properties utilizing a 23-factorial design. Optimized HApN exhibited typical Ca/P elemental ratio with high reaction yield. Surface area analysis revealed their mesoporous nature and high surface area. Hybrid HAp/PCL NPs prepared using direct emulsification-solvent evaporation maintained HApN crystallinity with no observed chemical interactions. To the best of our knowledge, we are the first to elaborate the biocompatibility and osteogenicity of nanoparticulate hybrid HAp/PCL. Hybrid HAp/PCL NPs outperformed HApN regarding mesenchymal cell proliferation and osteodifferentiation with reduction of possible cytotoxicity. Unlike HApN, hybrid HAp/PCL NPs presented moderate expression of early osteogenic markers, Runx-2 and osteopontin and significantly elevated expression of the late osteogenic marker, bone sialoprotein after 10-day culture. Our results indicate that hybrid bioactive HAp/PCL NPs could offer a more prominent osteogenic potential than plain HApN for bone regenerative applications as a standalone nanoplatform or as part of complex engineered systems

An Improved Oral Nutraceutical-Based Intervention for Management of Obesity: Pterostilbene Loaded Chitosan Nanoparticles - Fig S2.tif

       Aim: To formulate and assess the oral   anti-obesity effect of polymeric-based Pterostilbene(PS)-nanoparticles. Methods: Pterostilbene-hydroxypropyl   beta-cyclodextrin inclusion complex-loaded in chitosan-nanoparticles (PS/HPβCD-NPs) were   prepared and characterized in-vitro. Cytotoxicity, pharmacokinetics   and anti-obesity effects were assessed on Caco-2 cell line and high fat   diet-induced obesity. In-vivo   assessment included histological examination, protein and gene expression of   obesity biomarkers in adipose tissues. Results: Safe PS/HPβCD-NPs were successfully prepared with improved   bioavailability compared to free PS. PS/HPβCD-NPs showed improved anti-obesity   effect supported by histological examination, lipid profile, UCP1 gene   expression and protein expression of SIRT-1, COX-2, IL-6 and leptin.  Conclusion: Orally administered PS   nanoparticles is a new and promising anti-obesity strategy owing to its   sustainable weight loss and minimal side effects which is of great   socio-economic impact.        Supplementary figure 1:  Transmission   electron microscope of a) Unloaded NP and b) Pt/HPβCD loaded NP at magnification 25,000x. The scale bar   represents 200 nm. Supplementary   figure 2: a) Cellular uptake of free   C6, C6/HPßCD complex and C6/HPßCD-loaded NP (100 ng/mL C-6) in caco-2 cells using confocal laser microscopy and b)   Quantitative analysis of fluorescence intensity using Image J. Statistical   significance was shown at ***p≤0.001 and ****p≤0.0001 when compared to free   C-6. Supplementary figure 3: a) Serum   lipid profile in rats of different experimental groups where b) VLDL-C , c)   LDL/HDL ratio and d) Total cholesterol were calculated and plotted against   time (4, 8 and 16 weeks). Each point represents the   mean ± SD of 8   samples. Statistical significance is shown where ****p≤0.0001,   ***p≤0.001, **p≤0.01 and *p≤0.05 when samples when compared to untreated HFD   groups.</p

An Improved Oral Nutraceutical-Based Intervention for Management of Obesity: Pterostilbene Loaded Chitosan Nanoparticles - Fig S3.tif

       Aim: To formulate and assess the oral   anti-obesity effect of polymeric-based Pterostilbene(PS)-nanoparticles. Methods: Pterostilbene-hydroxypropyl   beta-cyclodextrin inclusion complex-loaded in chitosan-nanoparticles (PS/HPβCD-NPs) were   prepared and characterized in-vitro. Cytotoxicity, pharmacokinetics   and anti-obesity effects were assessed on Caco-2 cell line and high fat   diet-induced obesity. In-vivo   assessment included histological examination, protein and gene expression of   obesity biomarkers in adipose tissues. Results: Safe PS/HPβCD-NPs were successfully prepared with improved   bioavailability compared to free PS. PS/HPβCD-NPs showed improved anti-obesity   effect supported by histological examination, lipid profile, UCP1 gene   expression and protein expression of SIRT-1, COX-2, IL-6 and leptin.  Conclusion: Orally administered PS   nanoparticles is a new and promising anti-obesity strategy owing to its   sustainable weight loss and minimal side effects which is of great   socio-economic impact.        Supplementary figure 1:  Transmission   electron microscope of a) Unloaded NP and b) Pt/HPβCD loaded NP at magnification 25,000x. The scale bar   represents 200 nm. Supplementary   figure 2: a) Cellular uptake of free   C6, C6/HPßCD complex and C6/HPßCD-loaded NP (100 ng/mL C-6) in caco-2 cells using confocal laser microscopy and b)   Quantitative analysis of fluorescence intensity using Image J. Statistical   significance was shown at ***p≤0.001 and ****p≤0.0001 when compared to free   C-6. Supplementary figure 3: a) Serum   lipid profile in rats of different experimental groups where b) VLDL-C , c)   LDL/HDL ratio and d) Total cholesterol were calculated and plotted against   time (4, 8 and 16 weeks). Each point represents the   mean ± SD of 8   samples. Statistical significance is shown where ****p≤0.0001,   ***p≤0.001, **p≤0.01 and *p≤0.05 when samples when compared to untreated HFD   groups.</p