353 research outputs found
The Impact of Blue Light Irradiation on Keratinocytes in Vitro
ABSTRACT
Background: This study examined the effects of irradiation with blue
light on HaCaT keratinocytes. As irradiation with blue light is known to be
antimicrobial, it offers a promising alternative therapy for contaminated
wounds. There is evidence that red light promotes wound healing, but
the potential benefits of irradiation with blue light have not yet been ad-
equately investigated.
Methods: The rate of wound closure in sterile and contaminated cells
was measured using an in vitro scratch assay wound-healing model. Ad-
ditionally, cell viability after treatment was determined using a Sulforho-
damine B (SRB) assay.
Results: In both the sterile and contaminated groups, treated cells
showed delayed wound closure when compared with cells not irradiated
with blue light. Additionally, treatment with blue light resulted in poorer
viability in the treatment groups.
Conclusion: Although irradiation with blue light may offer a promising
alternative therapy for reducing bacterial colonization, our data indicate
that re-epithelization may be negatively influenced by blue light. Further
research is needed to clarify possible wound healing applications
Bone Healing Gone Wrong : Pathological Fracture Healing and Non-Unions—Overview of Basic and Clinical Aspects and Systematic Review of Risk Factors
Bone healing is a multifarious process involving mesenchymal stem cells, osteoprogenitor
cells, macrophages, osteoblasts and -clasts, and chondrocytes to restore the osseous tissue. Particularly
in long bones including the tibia, clavicle, humerus and femur, this process fails in 2–10% of all
fractures, with devastating effects for the patient and the healthcare system. Underlying reasons for
this failure are manifold, from lack of biomechanical stability to impaired biological host conditions
and wound-immanent intricacies. In this review, we describe the cellular components involved in
impaired bone healing and how they interfere with the delicately orchestrated processes of bone
repair and formation. We subsequently outline and weigh the risk factors for the development of
non-unions that have been established in the literature. Therapeutic prospects are illustrated and put
into clinical perspective, before the applicability of biomarkers is finally discussed
Subtoxic Concentrations of Hepatotoxic Drugs Lead to Kupffer Cell Activation in a Human In Vitro Liver Model
Drug induced liver injury (DILI) is an idiosyncratic adverse drug reaction
leading to severe liver damage. Kupffer cells (KC) sense hepatic tissue
stress/damage and therefore could be a tool for the estimation of consequent
effects associated with DILI. Aim of the present study was to establish a
human in vitro liver model for the investigation of immune-mediated signaling
in the pathogenesis of DILI. Hepatocytes and KC were isolated from human liver
specimens. The isolated KC yield was cells/g liver tissue with a purity of
>80%. KC activation was investigated by the measurement of reactive oxygen
intermediates (ROI, DCF assay) and cell activity (XTT assay). The initial KC
activation levels showed broad donor variability. Additional activation of KC
using supernatants of hepatocytes treated with hepatotoxic drugs increased KC
activity and led to donor-dependent changes in the formation of ROI compared
to KC incubated with supernatants from untreated hepatocytes. Additionally, a
compound- and donor-dependent increase in proinflammatory cytokines or in
anti-inflammatory cytokines was detected. In conclusion, KC related immune
signaling in hepatotoxicity was successfully determined in a newly established
in vitro liver model. KC were able to detect hepatocyte stress/damage and to
transmit a donor- and compound-dependent immune response via cytokine
production
Comparison of T-cell receptor repertoire restriction in blood and tumor tissue of colorectal cancer patients
Several immunotherapeutic approaches rely on antigen-specific T-cells. Restrictions in the T-cell receptor (TCR) repertoire were reported as indicator of anti-tumor cytotoxic T-lymphocyte (CTL) response in various tumor entities. It is unclear yet whether a TCR restriction in peripheral blood mirrors the tumor compartment. We compared the expression of TCR Vβ-families for the quantification of TCR repertoire alterations in blood and tissue samples from patients with colorectal carcinoma. Blood samples from patients with colorectal carcinoma and healthy volunteers and tissue samples of normal colonic mucosa and colorectal carcinoma were analyzed. Relative Vβ-family quantification was performed based on quantitative reverse transcribed PCR. Standard deviation and average mean of the single families were determined. Two variables describing the degree of Vβ-repertoire restriction were defined. Forty-eight blood samples and 37 tissue samples were analyzed. TCR repertoire restriction was higher in blood of tumor patients than in blood of healthy controls (p < 0.05). No difference in the degree of TCR repertoire restriction was found between carcinoma and unaffected colon tissue. We found no corresponding elevated TCR families among the different compartments blood, normal colon, and carcinoma tissue of the same patient. In conclusion, we observed a repertoire restriction in peripheral blood as well as in tumor tissue of cancer patients. However, in tumor tissue, repertoire alterations were comparable to normal mucosa, suggesting compartment-specific TCR distribution rather than alterations due to tumor-T-cell interaction questioning the presence of highly restricted clonal T-cell expansions in colorectal cancer as they have been described in other, assumingly more immunogenic tumor entities
Human Sterol Regulatory Element-Binding Protein 1a Contributes Significantly to Hepatic Lipogenic Gene Expression
Background/Aims:Sterol regulatory element-binding protein (SREBP) 1, the master regulator of lipogenesis, was shown to be associated with non-alcoholic fatty liver disease, which is attributed to its major isoform SREBP1c. Based on studies in mice, the minor isoform SREBP1a is regarded as negligible for hepatic lipogenesis. This study aims to elucidate the expression and functional role of SREBP1a in human liver. Methods: mRNA expression of both isoforms was quantified in cohorts of human livers and primary human hepatocytes. Hepatocytes were treated with PF-429242 to inhibit the proteolytic activation of SREBP precursor protein. SREBP1a-specific and pan-SREBP1 knock-down were performed by transfection of respective siRNAs. Lipogenic SREBP-target gene expression was analyzed by real-time RT-PCR. Results: In human liver, SREBP1a accounts for up to half of the total SREBP1 pool. Treatment with PF-429242 indicated SREBP-dependent auto-regulation of SREBP1a, which however was much weaker than of SREBP1c. SREBP1a-specific knock-down also reduced significantly the expression of SREBP1c and of SREBP-target genes. Regarding most SREBP-target genes, simultaneous knock-down of both isoforms resulted in effects of only similar extent as SREBP1a-specific knock-down. Conclusion: We here showed that SREBP1a is significantly contributing to the human hepatic SREBP1 pool and has a share in human hepatic lipogenic gene expression. Copyright (C) 2015 S. Karger AG, Base
A systems biology approach to dynamic modeling and inter-subject variability of statin pharmacokinetics in human hepatocytes
A dynamic model for the biotransformation of atorvastatin has been developed using quantitative metabolite measurements in primary human hepatocytes. The model comprises kinetics for transport processes and metabolic enzymes as well as population liver expression data allowing us to assess the impact of inter-individual variability of concentrations of key proteins. Application of computational tools for parameter sensitivity analysis enabled us to considerably improve the validity of the model and to create a consistent framework for precise computer-aided simulations in toxicology
Posttranslational Modification of Vesicular Stomatitis Virus Glycoprotein, but Not JNK Inhibition, Is the Antiviral Mechanism of SP600125
Vesicular stomatitis virus (VSV), a negative-sense single-stranded-RNA rhabdovirus, is an extremely promising oncolytic agent for cancer treatment. Since oncolytic virotherapy is moving closer to clinical application, potentially synergistic combinations of oncolytic viruses and molecularly targeted antitumor agents are becoming a meaningful strategy for cancer treatment. Mitogenactivated protein kinase (MAPK) inhibitors have been shown to impair liver cell proliferation and tumor development, suggesting their potential use as therapeutic agents for hepatocellular carcinoma (HCC). In this work, we show that the impairment of MAPK in vitro did not interfere with the oncolytic properties of VSV in HCC cell lines. Moreover, the administration of MAPK inhibitors did not restore the responsiveness of HCC cells to alpha/beta interferon (IFN-α/β). In contrast to previous reports, we show that JNK inhibition by the inhibitor SP600125 is not responsible for VSV attenuation in HCC cells and that this compound acts by causing a posttranslational modification of the viral glycoprotein
EGF and HB-EGF enhance the proliferation of programmable cells of monocytic origin (PCMO) through activation of MEK/ERK signaling and improve differentiation of PCMO-derived hepatocyte-like cells
Differentiation of In Vitro–Modified Human Peripheral Blood Monocytes Into Hepatocyte–like and Pancreatic Islet-like Cells
BACKGROUND & AIMS:
Adult stem cells provide a promising alternative for the treatment of diabetes mellitus and end-stage liver diseases. We evaluated the differentiation potential of human peripheral blood monocytes into hepatocyte-like and pancreatic islet-like cells.
METHODS:
Monocytes were treated with macrophage colony-stimulating factor and interleukin 3 for 6 days, followed by incubation with hepatocyte and pancreatic islet-specific differentiation media. Cells were characterized by flow cytometry, gene-expression analysis, metabolic assays, and transplantation for their state of differentiation and tissue-specific functions.
RESULTS:
In response to macrophage colony-stimulating factor and interleukin 3, monocytes resumed cell division in a CD115-dependent fashion, which was associated with a down-regulation of the PRDM1 and ICSBP genes. These programmable cells of monocytic origin were capable of differentiating into neohepatocytes, which closely resemble primary human hepatocytes with respect to morphology, expression of hepatocyte markers, and specific metabolic functions. After transplantation into the liver of severe combined immunodeficiency disease/nonobese diabetic mice, neohepatocytes integrated well into the liver tissue and showed a morphology and albumin expression similar to that of primary human hepatocytes transplanted under identical conditions. Programmable cells of monocytic origin-derived pancreatic neoislets expressed beta cell-specific transcription factors, secreted insulin and C peptide in a glucose-dependent manner, and normalized blood glucose levels when xenotransplanted into immunocompetent, streptozotocin-treated diabetic mice. Programmable cells of monocytic origin retained monocytic characteristics, notably CD14 expression, a monocyte-specific methylation pattern of the CD115 gene, and expression of the transcription factor PU.1.
CONCLUSIONS:
The ability to reprogram, expand, and differentiate peripheral blood monocytes in large quantities opens the real possibility of the clinical application of programmable cells of monocytic origin in tissue repair and organ regeneration
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