Investigating Cell Surface Markers and Differentiation Potential of Compact Bone-Derived Mesenchymal Stem Cells

Background: The differentiation potential of mesenchymal stem cells (MSCs) derived from the bone-tissue to multiple lineages is not clear. Objective: This study was conducted to investigate the surface antigen expression and multilineage stem cell potential of the cells derived from culture of collagenase digested marrow-free compact bones of C57BL/6 mouse. Materials & Methods: Long bones of C57BL/6 mouse (n=6) were collected aseptically and bone marrow was flushed out. Collagenase-digested bone fragments were washed and cultured in plastic flasks. The plastic-adherent fibroblast-like spindle-shaped cells were cultured sequentially in multiple passages in low-glucose DMEM (Dulbecco’s Modified Eagle’s Medium) supplemented with 15% FBS (Foetal Bovine Serum) and antibiotics in a 37°C incubator with 5% CO2. Immunophenotyping for cell surface markers was done using flow cytometry. The cells were differentiated into the osteoblastic, adipogenic and chondrogenic lineages. Results: The culture of the adherent cells exhibited active proliferation and multiplication in consequent passages. The cultured cells revealed evidence of adipogenic and osteogenic differentiation confirmed by staining with oil red O and von Kossa stains. Under flow cytometry observation, a significant proportion of cultured cells expressed CD29 and stem cell antigen (Sca-1). Only 9.8% cells showed expression of CD105. These MSCs exhibited low ability in chondrogenic differentiation, which can potentially be attributed to their lack of CD105 expression. Lack of expression of CD45 showed evidence of absence of hematopoietic stem cells. Conclusion: This study showed that murine compact bone-chip culture can yield MSCs with significant proliferation capacity. The cells displayed the ability to differentiate into osteoblast and adipocyte lineages

TOpic: rare and special cases, the real "Strange cases"

Introduction: The bladder hernia represents approximately 1-3% of all inguinal hernias, where patients aged more than 50 years have a higher incidence (10%). Many factors contribute to the development of a bladder hernia, including the presence of a urinary outlet obstruction causing chronic bladder distention, the loss of bladder tone, pericystitis, the perivesical bladder fat protrusion and the obesity

Synergistic Effects of Curcumin and Piperine as Potent Acetylcholine and Amyloidogenic Inhibitors With Significant Neuroprotective Activity in SH-SY5Y Cells via Computational Molecular Modeling and in vitro Assay

Hallmarks of Alzheimer's disease (AD) pathology include acetylcholine (ACh) deficiency and plaque deposition. Emerging studies suggest that acetylcholinesterase (AChE) may interact with amyloid β (Aβ) to promote aggregation of insoluble Aβ plaques in brains of patients. Current therapeutic options available for AD patients, such as AChE inhibitors, provide only symptomatic relief. In this study, we screened four natural compounds believed to harbor cognitive benefits-curcumin, piperine, bacoside A, and chebulinic acid. In the first section, preliminary screening through computational molecular docking simulations gauged the suitability of the compounds as novel AChE inhibitors. From here, only compounds that met the in silico selection criteria were selected for the second section through in vitro investigations, including AChE enzyme inhibition assay, 3-(4,5-dimenthylthiazol-2-yl)-2,5-dimethyltetrazolium bromide (MTT) assay, Thioflavin T (ThT) assay, and biochemical analysis via a neuronal cell line model. Of the four compounds screened, only curcumin (-9.6 kcal/mol) and piperine (-10.5 kcal/mol) showed favorable binding affinities and interactions towards AChE and were hence selected. In vitro AChE inhibition demonstrated that combination of curcumin and piperine showed greater AChE inhibition with an IC50 of 62.81 ± 0.01 μg/ml as compared to individual compounds, i.e., IC50 of curcumin at 134.5 ± 0.06 μg/ml and IC50 of piperine at 76.6 ± 0.08 μg/ml. In the SH-SY5Y cell model, this combination preserved cell viability up to 85%, indicating that the compounds protect against Aβ-induced neuronal damage (p < 0.01). Interestingly, our results also showed that curcumin and piperine achieved a synergistic effect at 35 μM with an synergism quotient (SQ) value of 1.824. Synergistic behavior indicates that the combination of these two compounds at lower concentrations may provide a better outcome than singularly used for Aβ proteins. Combined curcumin and piperine managed to inhibit aggregation (reduced ThT intensity at 0.432 a.u.; p < 0.01) as well as disaggregation (reduced ThT intensity at 0.532 a.u.; p < 0.01) of fibrillar Aβ42. Furthermore, combined curcumin and piperine reversed the Aβ-induced up-regulation of neuronal oxidative stress (p < 0.01). In conclusion, curcumin and piperine demonstrated promising neuroprotective effects, whereas bacoside A and chebulinic acid may not be suitable lead compounds. These results are hoped to advance the field of natural products research as potentially therapeutic and curative AD agents

Bacopa monnieri, a Neuroprotective Lead in Alzheimer Disease: A Review on Its Properties, Mechanisms of Action, and Preclinical and Clinical Studies

Alzheimer disease is a neurodegenerative disease that is signified by cognitive decline, memory loss, and erratic behavior. Till date, no cure for Alzheimer exists and the current Alzheimer medications have limited effectiveness. However, herbal medicines may slow down the disease's progression, which may hopefully reduce the number of cases in the years to come. Numerous studies have been done on characterizing the neuroprotective properties from plants belonging to Scrophulariaceae family, particularly Bacopa monnieri and its polyphenolic compounds known as bacosides. This review presents the findings on bacosides in therapeutic plants and their impact on Alzheimer disease pathology. These reports present data on the clinical, cellular activities, phytochemistry, and biological applications that may be used in new drug treatment for Alzheimer disease

Investigating Cell Surface Markers and Differentiation Potential of Compact Bone-Derived Mesenchymal Stem Cells

Background: The differentiation potential of mesenchymal stem cells (MSCs) derived from the bone-tissue to multiple lineages is not clear. Objective: This study was conducted to investigate the surface antigen expression and multilineage stem cell potential of the cells derived from culture of collagenase digested marrow-free compact bones of C57BL/6 mouse. Materials & Methods: Long bones of C57BL/6 mouse (n=6) were collected aseptically and bone marrow was flushed out. Collagenase-digested bone fragments were washed and cultured in plastic flasks. The plastic-adherent fibroblast-like spindle-shaped cells were cultured sequentially in multiple passages in low-glucose DMEM (Dulbecco’s Modified Eagle’s Medium) supplemented with 15% FBS (Foetal Bovine Serum) and antibiotics in a 37°C incubator with 5% CO2 . Immunophenotyping for cell surface markers was done using flow cytometry. The cells were differentiated into the osteoblastic, adipogenic and chondrogenic lineages. Results: The culture of the adherent cells exhibited active proliferation and multiplication in consequent passages. The cultured cells revealed evidence of adipogenic and osteogenic differentiation confirmed by staining with oil red O and von Kossa stains. Under flow cytometry observation, a significant proportion of cultured cells expressed CD29 and stem cell antigen (Sca-1). Only 9.8% cells showed expression of CD105. These MSCs exhibited low ability in chondrogenic differentiation, which can potentially be attributed to their lack of CD105 expression. Lack of expression of CD45 showed evidence of absence of hematopoietic stem cells. Conclusion: This study showed that murine compact bone-chip culture can yield MSCs with significant proliferation capacity. The cells displayed the ability to differentiate into osteoblast and adipocyte lineages