Stem cells have the capacity to differentiate into various lineages, and the ability to reliably direct stem cell fate determination would have tremendous potential for basic research and clinical therapy. Nanotopography provides a useful tool for guiding differentiation, as the features are more durable than surface chemistry and can be modified in size and shape to suit the desired application. In this paper, nanotopography is examined as a means to guide differentiation, and its application is described in the context of different subsets of stem cells, with a particular focus on skeletal (mesenchymal) stem cells. To address the mechanistic basis underlying the topographical effects on stem cells, the likely contributions of indirect (biochemical signal-mediated) and direct (force-mediated) mechanotransduction are discussed. Data from proteomic research is also outlined in relation to topography-mediated fate determination, as this approach provides insight into the global molecular changes at the level of the functional effectors.
Previous studies have shown that microgroove-initiated contact guidance can induce bone formation in osteoprogenitor cells (OPGs) and produce changes in the cell proteome. For proteomic analysis, differential in-gel electrophoresis (DIGE) can be used as a powerful diagnostic method to provide comparable data between the proteomic profiles of cells cultured in different conditions. This study focuses on the response of OPGs to a novel nanoscale pit topography with osteoinductive properties compared with planar controls. Disordered nearsquare nanopits with 120 nm diameter and 100 nm depth with an average 300 nm centreto-centre spacing (300 nm spaced pits in square pattern, but with G50 nm disorder) were fabricated on 1!1 cm 2 polycaprolactone sheets. Human OPGs were seeded onto the test materials. DIGE analysis revealed changes in the expression of a number of distinct proteins, including upregulation of actin isoforms, beta-galectin1, vimentin and procollagen-proline, 2-oxoglutarate 4-dioxygenase and prolyl 4-hydroxylase. Downregulation of enolase, caldesmon, zyxin, GRASP55, Hsp70 (BiP/GRP78), RNH1, cathepsin D and Hsp27 was also observed. The differences in cell morphology and mineralization are also reported using histochemical techniques.
IntroductionDental and oral diseases are major concerns in Thailand, affecting the country's health system. Everyone, at any age, is at risk of losing teeth. Tooth loss can lead to many health problems, including difficulty chewing, which increases the risk of poor nutrition. In addition, tooth loss leaves a large gap, leading to plaque accumulation that can cause periodontal disease. Moreover, the remaining teeth start to shift in an attempt to fill in the gap, leading to misalignment of teeth. Losing teeth may also affect personality, including a loss of confidence that may affect work or social activities. Restoration, with dentures or dental implants, is needed to replace the missing teeth.Current research has focused on dental and bone implants capable of promoting cell differentiation Xavier Acasigua et al., 2014;Naddeo et al., 2015). Cells isolated from tooth pulp contain odontoblasts, fibroblasts, immune cells, and undifferentiated mesenchymal cells called dental pulp stem cells (DPSCs). These findings may lead to restoration using dental pulp cells (DPCs), together with dental implants, to replace lost teeth. Various research groups have proposed the possibility of producing bioactive dental implants by adding natural products, such as natural hydroxyapatite/ zircon (Karamian et al., 2014), plant products (Varoni et al., 2012), or herbal extracts (Wang et al., 2012), to dental materials.Snails have been used in medicine since ancient times. Until the late 19th century, the synthetic peptide ziconotide (SNXIII), which is found in the venom of Conus magus, was used as a painkiller following approval by the United States Food and Drug Administration (Bonnemain, 2005). According to reports, snails contain several medically useful substances, including mytimacinlike antimicrobial peptides, which are exclusively found in the mucus of giant African snails (Zhong et al., 2013). In addition, lectin, which acts as an antimicrobial, is also present in the mucus of giant African snails (Ito et al., 2011). High-performance liquid chromatography was used to determine the amount of allantoin and glycolic Abstract: Dental pulp tissue contains stem cells that can be isolated and used for regenerative medicine in tooth restoration or autologous transplantation. The aim of this study was to observe the mineralization and gene expression in dental pulp cells (DPCs) following treatment with snail mucus. Snail mucus was collected from adult Achatina fulica and processed as a dry powder by the freeze-drying technique. The mucus powder was dissolved in a culture medium at a concentration of 15 µg/mL. DPCs were prepared by the outgrowth technique and cultured in a 6-well plate at a density of 5 × 10 4 cells per well. A mucus-supplemented medium (15 µg/mL) was added to each well. Cell culture was maintained for 3 weeks. The results of Alizarin Red S staining indicated that the DPCs cultured in a medium supplemented with snail mucus showed a higher number of mineralized nodules as compared with the control group cultured in a norm...
Background:Sanguisorba Officinalis L. (SO) is a well-known traditional Chinese medicine (TCM), commonly applied to treat complex diseases, such as anticancer, antibacterial, antiviral, anti-inflammatory, anti-oxidant and hemostatic effects. Especially, it has been reported to exert anti-tumor effect in various human cancers. However, its effect and pharmacological mechanism on hepatocellular carcinoma (HCC) remains unclear.Methods: In this study, network pharmacology approach was applied to characterize the underlying mechanism of SO on HCC. Active compounds and potential targets of SO, as well as related genes of HCC were obtained from the public databases, the potential targets and signaling pathways were determined by protein-protein interaction (PPI), gene ontology (GO) and pathway enrichment analyses. And the compound-target and target-pathway networks were constructed. Subsequently, in vitro experiments were also performed to further verify the anticancer effects of SO on HCC.Results: By using the comprehensive network pharmacology analysis, 41 ingredients in SO were collected from the corresponding databases, 12 active ingredients screened according to their oral bioavailability and drug-likeness index, and 258 potential targets related to HCC were predicted. Through enrichment analysis, SO was found to show its excellent therapeutic effects on HCC through several pathways, mainly related to proliferation and survival via the EGFR, PI3K/AKT, NFκB and MAPK signaling pathways. Additionally, in vitro, SO was found to inhibit cell proliferation, induce apoptosis and down-regulate cell migration and invasion in various HCC cells. Moreover, western blot analysis showed that SO treatment down-regulated the expression of p-EGFR, p-PI3K, p-AKT, p-NFκB and p-MAPK proteins in HepG2 cells. These results validated that SO exerted its therapeutic effects on HCC mainly by the regulation of cell proliferation and survival via the EGFR/MAPK and EGFR/PI3K/AKT/NFκB signaling pathways.Conclusion: Taken together, this study, revealed the anti-HCC effects of SO and its potential underlying therapeutic mechanisms in a multi-target and multi-pathway manner.
ABSTRACT. The present study focuses on mechanotransduction in mesenchymal stem cells (MSCs) in response to matrix elasticity. By using photocurable gelatinous gels with tunable stiffness, proteomic profiles of MSCs cultured on tissue culture plastic, soft (3 kPa) and stiff (52 kPa) matrices were deciphered using 2-dimensional differential in-gel analysis (2D-DIGE). The DIGE data, tied to immunofluorescence, indicated abundance and organization changes in the cytoskeletonal proteins as well as differential regulation of important signalingrelated proteins, stress-responsing proteins and also proteins involved in collagen synthesis. The major CSK proteins including actin, tubulin and vimentin of the cells cultured on the gels were remarkably changed their expressions. Significant down-regulation of α-tubulin and β-actin can be observed on gel samples in comparison to the rigid tissue culture plates. The expression abundance of vimentin appeared to be highest in the MSCs cultured on hard gels. These results suggested that the substrate stiffness significantly affects expression balances in cytoskeletal proteins of MSCs with some implications to cellular tensegrity.
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