Extracellular vesicles derived from human foreskin cells (hFS-Exo) accelerate cell migration and angiogenesis through MAPK pathway: an in vitro study.

BACKGROUND: Wound healing is one of the important processes in the body. Attempts to create new drugs are of interest due to the side effects of natural and chemical wound healing compounds. To overcome this obstacle, stem cells have been used as healing agents. However, both difficulties in collection and risks such as rejection and teratoma in the recipient body have limited the use of stem cells, directly. Since the potential content of the stem cells can be transferred to the recipient cells by vesicles, small extracellular vesicles have recently become prominent agents. METHODS AND RESULTS: The wound-healing effect of extracellular vesicles derived from foreskin cells was investigated in both keratinocyte and endothelial cells. Migration assay, RT-PCR, Col1a1 ELISA and Western Blot experiments were utilized to reveal healing effect of EVs and its possible molecular pathways. EV-treated groups exhibited more proliferative, invasive, and migrative characteristics. When comparing to the control group, new vessel formation was induced in EV groups. An increase in gene levels of growth factors related to wound healing and change in the mitogen-activated protein kinase (MAPK) signaling pathway proteins in EV-treated groups were determined. Possible molecular mechanisms underlying cell movements were associated with the MAPK pathway. It was found that human foreskin cell EVs (hFS-Exo) may have a potential to heal wounds in a short period of time by triggering the MAPK pathway. CONCLUSIONS: hFS-Exo could be a new promising wound healing agent in the future.

Differentiation Potential of Mesenchymal Stem Cells into Pancreatic ß-Cells.

Stem cells having the capability to differentiate into other type of cells and renewing themselves, gained so much importance in recent years. Investigations in stem cells revealed that mesenchymal stem cells can successfully differentiate into other type of cells like adipocytes, hepatocytes, osteocytes, neurocytes and chondrocytes. In addition, these cells can also differentiate into insulin-producing beta cells. Insulin is a crucial hormone for glucose balance of the body. Insufficiency or unavailability of insulin is called diabetes. External insulin intake, as well as pancreas or islet transplantation, is the most basic treatment of diabetes. In vivo and in vitro studies demonstrate that stem cell therapy is also used in the cure of diabetes. Differentiation process of stem cells into beta cells releasing insulin is quite complicated. There are many different reports for the differentiation of stem cells in the literature. The success of differentiation of stem cells into beta cells depends on several factors like the source of stem cells, chemicals added into the differentiation medium and the duration of differentiation protocol. Distinct studies for the differentiation of stem cells into insulin-secreting cells are available in the literature. Moreover, thanks to the superior differentiation capacity of stem cells, they are being preferred in clinical studies. Stem cells were clinically used to heal diabetic ulcer, to increase c-peptide level and insulin secretion in both type 1 and type 2 diabetes. Mesenchymal stem cells having high differentiation potential to insulin-secreting cells are encouraging vehicles for both in vivo and in vitro studies together with clinical trials for diabetes mellitus.

Zooming in across the Skin: A Macro-to-Molecular Panorama.

Maintaining integrity of the skin and its appendages still preserves its top-ranking in priorities of survival for the modern human as it probably once did for the ancient individual, -not only- because it is the primary barrier to external assaults, but also because of social and psychological impact of healthy skin during their life-span. Healing wounds in order to shield off the internal organs from infections and damage, restoring its ability to adapt to various environmental stimuli, and slowing-down and reversing aging of the skin in the quest for an everlasting youth can be named as a few of the main drivers behind the multi-million investments dedicated to the advancement of our understanding of skin's physiology. Over the years, these tremendous efforts culminated in the breakthrough discovery of skin stem cells the regenerative capacity of which accounted for the resilience of the skin through their unique capacity as a special cell type that can both self-renew and differentiate into various lineages. In this review, first we summarize the current knowledge on this amazing organ both at a structural and functional level. Next, we provide a comprehensive -in depth- discussion on epidermal as well as dermal stem cells in terms of the key regulatory pathways as well as the main genetic factors that have been implicated in the orchestration of the skin stem cell biology in regards to the shifts between quiescence and entry into distinct differentiation programs.

Boron containing compounds promote the survival and the maintenance of pancreatic ß-cells.

Diabetes mellitus is worldwide disease. The life of diabetic patients are dependent on exogenous insulin. Pancreas or particularly islet transplantations are performed for reducing external insulin dependency. External substances are also used to protect the ß-cells from the death or increase insulin secretion. In the current study, two different boron containing compounds (sodium pentaborate pentahydrate-NaB and boric acid-BA) were investigated for their effect on pancreatic cells in terms of pro-apoptotic and anti-apoptotic markers, genes related to insulin production mechanism, pancreatic development and glucose metabolism, some antioxidant enzymes, and genes for the initiation of diabetes, insulin secretion and antioxidant enzyme activities in vitro. The results revealed that boron containing compounds did not lead to apoptosis. On the contrary, they increased cell viability, antioxidant enzyme activities and the level of genes related to insulin production. Overall evaluation, data in the current study showed that boron containing compounds might be promising therapeutic agents for type 1 diabetes. However, additional investigations are strictly needed to elucidate molecular mechanisms of boron containing compounds.

Stem Cells Derived from Dental Tissues.

Stem cells are undifferentiated cells located in different parts of the body. The major role of stem cells is to restore of injured tissues. Since the discover of stem cells, they gained a big attention due to their differentiation and regeneration capacity. The main source of stem cells was known as bone marrow. However, different sources for obtaining stem cells were discovered. Dental tissues, a new source for stem cells, provide cells having mesenchymal stem cell characteristics such as fibroblast-like structure, expression of surface antigens specific for mesenchymal stem cells, regeneration ability, multilineage differentiation capacity and immunomodulatory features. Dental pulp stem cells (DPSCs), dental follicle progenitor cells (DFPCs), stem cells from apical papilla (SCAP), tooth germ stem cells (TGSCs) and periodontal ligament stem cells (PDLSCs) are stem cells derived from dental tissues as well as stem cells from exfoliated deciduous teeth (SHED). Dental stem cells express mesenchymal stem cell markers like Stro-1, CD146, CD106, CD90, CD73 CD29 and CD13. However, they do not express hematopoietic stem cell markers such as CD11b, CD45 and CD34. Dental stem cells are able to undergo myogenic, chondrogenic, adipogenic, neurogenic, osteogenic and odontogenic differentiation. Thanks to these differentiation ability of dental stem cells, they can easily be manipulated in regenerative medicine. Dental stem cells, that can effortlessly be transfected, can also be used in cell therapy application. Immunomodulatory features of dental stem cells make them suitable candidates for the therapy of immune-related disorders. Dental stem cells with high potentials such as ability of self-renewal, mesenchymal stem cell characteristics, multilineage differentiation and immunomodulation are promising tool for in vitro and in vivo differentiation studies as well as the therapy of immune-related diseases.

Schiff Base-Poloxamer P85 Combination Prevents Prostate Cancer Progression in C57/Bl6 Mice.

BACKGROUND: Prostate cancer which is the second most common cause of death among men has a high incidence in recent years. Current therapeutic regimens should be improved to overcome drug resistance. At the metastatic stage, tumors become refractory to established chemotherapeutic treatments and cause serious problems at the clinics. Development of new drug molecules that are able to transport through the membrane easily and kill tumor cells rapidly is of great interest. METHOD: In the current study, a novel Heterodinuclear copper(II)Mn(II) Schiff base complex combined with P85 was used for prostate cancer treatment in vivo. Tramp-C1 cells injected animals were subjected to chemotherapeutic formulation treatment and results were analyzed by toxicology analysis, tumor volume measurements, and histopathological analysis. 0.5 mg/kg Schiff base was selected and combined with 0.05% P85 according to the toxicology analysis showing the enzyme levels, blood parameters, and multiple organ toxicity. RESULTS: Results demonstrated that Heterodinuclear copper(II)Mn(II) complex-P85 combination decreased tumor formation and tumor volume steadily over the course of experiments. CONCLUSIONS: Overall, Heterodinuclear copper(II)Mn(II) complex-P85 exerted remarkable anti-cancer activity in vivo in C57/B16 mice. Prostate 76:1454-1463, 2016. © 2016 Wiley Periodicals, Inc.

Boron promotes streptozotocin-induced diabetic wound healing: roles in cell proliferation and migration, growth factor expression, and inflammation.

Acute wounds do not generally require professional treatment modalities and heal in a predictable fashion, but chronic wounds are mainly accompanied with infection and prolonged inflammation, leading to healing impairments and continuous tissue degradation. Although a vast amount of products have been introduced in the market, claiming to provide a better optimization of local and systemic conditions of patients, they do not meet the expectations due to being expensive and not easily accessible, requiring wound care facilities, having patient-specific response, low efficiency, and severe side-effects. In this sense, developing new, safe, self-applicable, effective, and cheap wound care products with broad-range antimicrobial activity is still an attractive area of international research. In the present work, boron derivatives [boric acid and sodium pentaborate pentahydrate (NaB)] were evaluated for their antimicrobial activity, proliferation, migratory, angiogenesis, gene, and growth factor expression promoting effects on dermal cells in vitro. In addition, boron-containing hydrogel formulation was examined for its wound healing promoting potential using full-thickness wound model in streptozotocin-induced diabetic rats. The results revealed that while both boron compounds significantly increased proliferation, migration, vital growth factor, and gene expression levels of dermal cells along with displaying remarkable antimicrobial effects against bacteria, yeast, and fungi, NaB displayed greater antimicrobial properties as well as gene and growth factor expression inductive effects. Animal studies proved that NaB-containing gel formulation enhanced wound healing rate of diabetic animals and histopathological scores. Overall data suggest a potential promising therapeutic option for the management of chronic wounds but further studies are highly warranted to determine signaling pathways and target metabolisms in which boron is involved to elucidate the limitations and extend its use in clinics.

Ultrasensitive electrochemical immunosensor system for determination of autologous SOX2 antibody.

Lung cancer is mainly seen as the cancer type in the world. Lung cancer causes the death of many people. It is classified as large-cell neuroendocrine carcinoma (LCNEC), small-cell lung cancer (SCLC), and adenocarcinoma by the World Health Organization (WHO) in 2015. Small cell lung cancer (SCLC) is a highly aggressive type of cancer, accounting for approximately 20% of all cases. By performing the serological analysis of expression cDNA libraries (SEREX), the humoral immune response of SCLC patients is determined. SEREX of SCLC cell lines using pooled sera of SCLC patients led to the isolation of SOX2 genes. The between SOX2 antigen expression intensity and autologous antibody presence has a significant correlation because SOX2 is the main antigen eliciting anti-SOX responses. Electrochemical biosensors take much attention because of their simplicity, selectivity, and sensitivity in clinical analysis. Antibody-based surface recognizes antibody-specific antigens. This work aims to fabricate an immunosensor for determining autologous SOX2 antibodies using a multi-walled carbon nanotube-modified screen-printed electrode (DRP-MWCNT). All immobilization processes were evaluated with cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The critical parameters were optimized, such as EDC/NHS concentration and time, SOX2 protein concentration and incubation time, BSA ratio, BSA blocking time, and anti-SOX2 antibody incubation time. The developed immunosensor, under optimal conditions, shows a linear response of autologous SOX2 antibody between 0.005 ng.mL-1 and 0.1 ng.mL-1. The limit of detection and quantification were 0.001 and 0.004 ng.mL-1, respectively. The electrode morphologies were examined with a scanning electron microscope (SEM). Lastly, the developed immunosensor was applied to a synthetic serum sample, and the linear range was compared with enzyme-linked immunosorbent assay (ELISA).

Differentiation and neuro-protective properties of immortalized human tooth germ stem cells.

Stem cells are considered to be promising therapeutic options in many neuro-degenerative diseases and injuries to the central nervous system, including brain ischemia and spinal cord trauma. Apart from the gold standard embryonic and mesenchymal origin, human tooth germ stem cells (hTGSCs) have also been shown to enjoy the characteristics of mesenchymal stem cells (MSCs) and the ability to differentiate into adipo-, chondro-, osteo- and neuro-genic cells, suggesting that they might serve as potential alternatives in the cellular therapy of various maladies. Immortalization of stem cells may be useful to avoid senescence of stem cells and to increase their proliferation potential without altering their natural characteristics. This study evaluated the expression of stem cell markers, surface antigens, differentiation capacity, and karyotype of hTGSCs that have been immortalized by human telomerase reverse transcriptase (hTERT) or simian vacuolating virus 40 (SV40) large T antigen. These undying cells were also evaluated for their neuro-protective potential using an in vitro SH-SY5Y neuro-blastoma model treated with hydrogen-peroxide or doxo-rubicin. Although hTGSC-SV40 showed abnormal karyotypes, our results suggest that hTGSC-hTERT preserve their MSC characteristics, differentiation capacity and normal karyotype, and they also possess high proliferation rate and neuro-protective effects even at great passage numbers. These peculiars indicate that hTGSC-hTERT could be used as a viable model for studying adipo-, osteo-, odonto- and neuro-genesis, as well as neuro-protection of MSCs, which may serve as a springboard for potentially utilizing dental waste material in cellular therapy.

The synthesis of boronic-imine structured compounds and identification of their anticancer, antimicrobial and antioxidant activities.

Boronic acid compounds with different substituted groups were handled to synthesize various ligands encoded as B1, B2, B3, B4, B5, B6, B7 and B8. B5 and B7 were tested for the cytotoxic activity against the prostate cancer cells and it was found that the cell viability of cancer cells was decreased while most of the healthy cells could still be viable. 5 µM solutions of B5 and B7 decreased the cell viability to 33% and 44% whereas healthy cells were 71% and 95%, respectively, after treatment. Antimicrobial properties were explored against the bacterial and fungal microorganisms with B1, B5 and B7. The inhibition zones were evaluated for all boronic structures, and the growth inhibition zones were determined in a range of 7-13 mm diameter for different microorganism species. Staphylococcus aureus was the common microorganism that three boronic compounds with imine ligands showed the activity. Antioxidant features of B2, B3, B4, B5, B6, B7 and B8 were investigated by different processes such as Beta-carotene bleaching (BCB), 2,2-diphenyl picryl hydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and CUPric reducing antioxidant capacity (CUPRAC) methods. Significant antioxidant activity was achieved by the phenyl boronic based ligands and these compounds demonstrated as much activity as standards (α-Toc and BHT). In addition, all structures were applied properly without any decomposition during the experiments. They were rather stable both in aqueous media and solid state.

Dose-dependent Effect of Boric Acid on Myogenic Differentiation of Human Adipose-derived Stem Cells (hADSCs).

Boron, a vital micronutrient for plant metabolism, is not fully elucidated for embryonic and adult body development, and tissue regeneration. Although optimized amount of boron supplement has been shown to be essential for normal gestational development in zebrafish and frog and beneficial for bone regeneration in higher animals, effects of boron on myogenesis and myo-regeneration remains to be solved. In the current study, we investigated dose-dependent activity of boric acid on myogenic differentiation of human adipose-derived stem cells (hADSCs) using immunocytochemical, gene, and protein expression analysis. The results revealed that while low- (81.9 µM) and high-dose (819.6 µM) boron treatment increased myogenic gene expression levels such as myosin heavy chain (MYH), MyoD, myogenin, and desmin at day 4 of differentiation, high-dose treatment decreased myogenic-related gene and protein levels at day 21 of differentiation, confirmed by immunocytochemical analysis. The findings of the study present not only an understanding of boron's effect on myogenic differentiation but also an opportunity for the development of scaffolds to be used in skeletal tissue engineering and supplements for embryonic muscle growth. However, fine dose tuning and treatment period arranging are highly warranted as boron treatment over required concentrations and time might result in detrimental outcomes to myogenesis and myo-regeneration.

In Vitro Evaluation of ProRoot MTA, Biodentine, and MM-MTA on Human Alveolar Bone Marrow Stem Cells in Terms of Biocompatibility and Mineralization.

INTRODUCTION: Stem cell technology has been a great hope for the regeneration of cells of pulp-dentin complex and dental structures together with surrounding bone and periodontium. The main challenge in the regeneration process is a successful combination of stem cells and efficient inductors such as inductive biomaterials. In this regard, today, manufacturers propose novel tooth filling materials. The current study was aimed to compare the effect of ProRoot MTA (Dentsply Tulsa Dental, Tulsa, OK), Biodentine (Septodont, Saint Maur des Fossés, France), and MM-MTA (Micro-Mega, Besançon Cedex, France) on the cell viability, hard tissue deposition capacity, and osteogenic differentiation of human bone marrow stem cells (hBMSCs) derived from mandibular bone. METHODS: Dental materials were packed into Teflon rings (Grover Corp, Milwaukee, WI) and placed on Transwell inserts (Corning, Corning, NY) to determine the toxicity of tooth filling materials by the 3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H tetrazolium assay on days 1, 3, 7, and 14; 20% dimethyl sulfoxide (DMSO) was used as a positive control for the toxicity assay. hBMSCs were characterized by their surface markers with mesenchymal stem cell antibodies. Teflon rings were cocultured with hBMSCs followed by the induction of osteogenic differentiation. The osteogenic differentiation of hBMSCs and hard tissue formation of the materials were evaluated by analyzing the messenger RNA expression levels of osteonectin, Runt-related transcription factor 2, and collagen type 1A by real-time polymerase chain reaction expression analysis, measurement of alkaline phosphatase activity, and visualization of calcium deposits by alizarin red staining. RESULTS: MTA, Biodentine, and MM-MTA did not exhibit a cytotoxic effect on hBMSCs after 14 days in culture. Even though all the materials significantly stimulate (P < .05) osteogenic differentiation of hBMSCs compared with the negative control, ProRoot MTA showed greater osteoinductivity than Biodentine or MM-MTA according to the messenger RNA expression, alkaline phosphatase, immunocytochemistry, and alizarin red staining data. CONCLUSIONS: All of the dental materials used in this study show the osteogenic differentiation potential of hBMSCs. Therefore, newly introduced MM-MTA can also be used as a material of choice in routine dental treatment.

Bmp 2 and bmp 7 induce odonto- and osteogenesis of human tooth germ stem cells.

Bone morphogenetic proteins (BMPs) initiate, promote, and maintain odontogenesis and osteogenesis. In this study, we studied the effect of bone morphogenic protein 2 (BMP 2) and bone morphogenic protein 7 (BMP 7) as differentiation inducers in tooth and bone regeneration. We compared the effect of BMP 2 and BMP 7 on odontogenic and osteogenic differentiation of human tooth germ stem cells (hTGSCs). Third molar-derived hTGSCs were characterized with mesenchymal stem cell surface markers by flow cytometry. BMP 2 and BMP 7 were transfected into hTGSCs and the cells were seeded onto six-well plates. One day after the transfection, hTGSCs were treated with odontogenic and osteogenic mediums for 14 days. For confirmation of odontogenic and osteogenic differentiation, mRNA levels of BMP2, BMP 7, collagen type 1 (COL1A), osteocalsin (OCN), and dentin sialophosphoprotein (DSPP) genes were measured by quantitative real-time PCR. In addition to this, immunocytochemistry was performed by odontogenic and osteogenic antibodies and mineralization obtained by von Kossa staining. Our results showed that the BMP 2 and BMP 7 both promoted odontogenic and osteogenic differentiation of hTGSCs. Data indicated that BMP 2 treatment and BMP 7 treatment induce odontogenic differentiation without affecting each other, whereas they induce osteogenic differentiation by triggering expression of each other. These findings provide a feasible tool for tooth and bone tissue engineering.