Diverse Approaches toward Application of Dental Pulp Stem Cells from Human Permanent and Deciduous Teeth in the Treatment of Diabetes.

BACKGROUND: Diabetes Mellitus is defined by hyperglycemia, a condition which is the result of defects in insulin secretion, insulin action, or both. Evidence suggest that islet transplantation is a promising treatment approach, but the shortage of sources of insulin-producing cells is a major problem. Ethical concerns and the limited availability of most stem cells have led scientists to concentrate on mesenchymal stem cells, which are found in stem cells niches of all organs of the body including dental tissues on which dental pulp stem cells (DPSCs) and stem cells from exfoliated deciduous teeth (SHED) are the easiest accessible sources. HIGHLIGHTS: Generally, SHED show characteristics similar to DPSCs; however, its proliferative and clonogenic capacities are higher. It has been proved that these two types of dental mesenchymal stem cells are able to produce islet-like cells capable of insulin secretion. In this review, we discuss various conducted approaches on the application of DPSCs and SHED in the treatment of diseases associated with diabetes such as; pancreatic differentiation cocktails, 2D and 3D culture techniques, factors that affect pancreatic differentiation, in vivo studies (direct administration of DPSCs and SHED, administration of their secretome and encapsulation of their-derived insulin producing cells), clinical trials and future perspectives of these approaches. CONCLUSION: Dental stem cell-based therapy has been considered as a promising therapeutic procedure for treatment of diabetes. Major advances in research on the derivation of insulin producing cells from DPSCs and SHED have enhanced our chance of re-establishing glucose-responsive insulin secretion in patients with diabetes.

Free Fatty Acids from Cow Urine DMSO Fraction Induce Cell Death in Breast Cancer Cells without Affecting Normal GMSCs.

OBJECTIVE: The objective of this study was to explore the biological relevance of free fatty acids derived from cow urine DMSO fraction (CUDF) by employing in vitro and in silico approaches. BACKGROUND: Metabolic heterogeneity at the intra- and intercellular levels contributes to the metabolic plasticity of cancer cells during drug-induced response. Free fatty acid (FFA) availability at intra- and intercellular levels is related to tumor heterogeneity at interpatient and xeno-heterogeneity levels. METHODS: We collected fresh urine from healthy cows and subjected it to fractionation in DMSO using drying, vortexing, and centrifugation. Finally, the sterile filtrate of cow urine DMSO fraction (CUDF) was evaluated for antiproliferative and proapoptotic effects in MCF-7 and ZR-75-1 breast cancer cells using routine cell-based assays. Intracellular metabolites were studied with the help of a novel in-house vertical tube gel electrophoresis (VTGE) method to reveal the nature of CUDF components in MCF-7 cells. Identified intracellular FFAs were studied for their molecular interactions with targeted receptor histone deacetylase (HDAC) using molecular docking and molecular dynamics (MD) simulations. RESULTS: CUDF showed a significant reduction in cell viability and cell death in MCF-7 and ZR-75-1 breast cancer cells. Interestingly, FFAs tetracosanedioic acid, 13Z-docosenoic acid (erucic acid), nervonic acid, 3-hydroxy-tetradecanoic acid, and 3-hydroxcapric acid were found inside the treated MCF-7 cancer cells. These FFAs, including tetracosanedioic acid, indicated a specific affinity to HDAC at their inhibitory sites, similar to trichostatin A, a known inhibitor. CONCLUSIONS: This study reports on FFAs derived from CUDF as potential antiproliferative and pro-cell death agents against breast cancer cells. MD simulations hinted at tetracosanedioic acid and other FFAs as inhibitors of HDAC that could explain the observed effects of FFAs in cancer cells.

Shielding Engineered Islets With Mesenchymal Stem Cells Enhance Survival Under Hypoxia.

In the present study we focused on the improvisation of islet survival in hypoxia.The Islet like cell aggregates (ICAs) derived from wharton's jelly mesenchymal stem cells (WJ MSC) were cultured with and without WJ MSC for 48 h in hypoxia and normoxia and tested for their direct trophic effect on ß cell survival. The WJ MSCs themselves secreted insulin upon glucose challenge and expressed the pancreatic markers at both transcription and translational level (C-peptide, Insulin, Glucagon, and Glut 2). Direct contact of MSCs with ICAs facilitated highest viability under hypoxia as evidenced by fluorescein diacetate/propidium iodide and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cytokine analysis of the co-cultured ICAs revealed amplification of anti-inflammatory cytokine like TGFß and TNFα accompanied by depletion of pro-inflammatory cytokines. The increment in VEGF and PDGFa was also seen showing their ability to vascularize upon transplantation. This was further accompanied by reduction in total reactive oxygen species, nitric oxide, and super oxide ions and down regulation of Caspase3, Caspase8, p53, and up regulation of Bcl2 confirming prevention of apoptosis in ICAs. The western blot analysis confirmed the cytoprotective effect of WJ MSC on ICAs as they enhanced the anti-apoptotic marker BCL2 and reduced the expression of apoptotic markers, Annexin 5 and Caspase 3. There was a significant reduction in the expression of p38 protein in the presence of MSCs making the ICAs responsive to glucose. Taken together our data demonstrate for the first time that the WJ MSC expressed pancreatic markers and their supplementation protected engineered islets against hypoxia and oxidative stress. J. Cell. Biochem. 118: 2672-2683, 2017. © 2017 Wiley Periodicals, Inc.

Influence of hypoxia, high glucose, and low serum on the growth kinetics of mesenchymal stem cells from deciduous and permanent teeth.

The therapeutic potential of mesenchymal stromal cells depends on their ability to survive and proliferate under adverse in vivo scenarios in a particular disease. In most of the sites of injury, especially in diabetic wounds, there can be hypoxia, hyperglycemia, and ischemia, leading to a lack of nutrients. Hence, the aim of our present study was to investigate the influence of hypoxia, high glucose, and low serum concentrations on the growth kinetics and proliferative potential of human dental pulp stem cells from exfoliated deciduous teeth (SHED) and permanent teeth (DPSC). In this study we isolated two types of specialized stem cells from human dental pulp tissues, which were supposedly of neural crest origin, and cultured them in KO-DMEM medium supplemented with 10% fetal bovine serum (FBS). Both SHED and DPSC were characterized for standard CD surface markers, and their ability to differentiate into adipogenic and osteogenic lineages was tested. SHED and DPSC were exposed to either hypoxia or high glucose or low serum conditions, and their growth kinetics and differentiation potentials were compared with those of normal culture conditions. We found that SHED retained their phenotypic expression and differentiation potential under hypoxia, high-glucose, and low-serum conditions and exhibited a higher proliferation in terms of cell yield and a reduced doubling time compared to DPSC. Our findings clearly demonstrate for the first time that SHED are superior to DPSC as evidenced by their enhanced proliferation under adverse culture conditions.

Transplantation of islet-like cell clusters derived from human dental pulp stem cells restores normoglycemia in diabetic mice.

BACKGROUND AIMS: The success of islet transplantation for diabetes depends on the availability of an adequate number of allogeneic or autologous islets. Postnatal stem cells are now considered for the generation of physiologically competent, insulin-producing cells. Our group showed earlier that it is possible to generate functional islets from human dental pulp stem cells by using a serum-free cocktail in a three-step protocol. METHODS: We compared the yield of generated islet-like cell clusters (ICCs) from stem cells from pulps of human exfoliated deciduous teeth (SHED) and dental pulp stem cells from permanent teeth (DPSCs). ICCs derived from SHED were packed in immuno-isolatory biocompatible macro-capsules and transplanted into streptozotocin (STZ)-induced diabetic mice. Non-diabetic and diabetic controls were transplanted with macro-capsules with or without islets. RESULTS: SHED were superior to DPSCs. STZ diabetic mice alone and mice transplanted with empty macro-capsules exhibited hyperglycemia throughout the experiment, whereas mice transplanted with macro-capsules containing ICCs were restored to normoglycemia within 3-4 weeks, which persisted for >60 days. CONCLUSIONS: Our results demonstrate for the first time that ICCs derived from SHED reverse STZ diabetes in mice without immunosuppression and offer an autologous and non-controversial source of human tissue that could be used for stem cell therapy in diabetes.

Who is the culprit for post menopausal syndrome? Uterus/Ovary!

The uterine endometrium of placental mammals in general and human in particular is a highly dynamic, proliferative and regenerative tissue. It undergoes cycles of growth and regression during each menstrual cycle with a growing capacity from 0.5-1mm in the proliferative phase to 5-7 mm during the secretory (leutal) phase. These phases are characterized by cyclic processes of cellular proliferation, differentiation and shedding. Recent studies have revealed that the endometrium harbours a large population of mesenchymal stromal cells. There are several reports indicating the homing of bone marrow stem cells and endothelial progenitor cells in regenerating endometrium. However, it is not clear whether endometrial cells mobilise to participate in the repair and regeneration of vital organs/tissues. We hypothesize that a very small percentage of the endometrial cells may set in circulation during menstrual cycle to facilitate endogenous regeneration of vital organs in the body. These cyclical events may be responsible for providing a protective barrier to women during her child-bearing age. Disappearance of this barrier after menopause probably makes her vulnerable for post menopausal symptoms. There is a circumstantial evidence to vouch for the presence of circulating stem/progenitor cells in peripheral blood which are likely to lodge in injured organs for their possible repair. As the endometrium harbors a large population of mesenchymal stromal cells, it is possible that retention of the uterus through secretion of reparative/growth promoting factors, may provide legitimate stem cells to enter circulation and "set up shop" in other tissues like bone marrow stem cells. In this context we propose uterus to be the culprit for the postmenopausal syndrome.