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In vivo Optogenetic Approach to Study Neuron-Oligodendroglia Interactions in Mouse Pups
摘要： Optogenetic and pharmacogenetic techniques have been effective to analyze the role of neuronal activity in controlling oligodendroglia lineage cells in behaving juvenile and adult mice. This kind of studies is also of high interest during early postnatal development since important changes in oligodendroglia dynamics occur during the first two PN weeks. Yet, neuronal manipulation is difficult to implement at an early age because high-level, specific protein expression is less reliable in neonatal mice. Here, we describe a protocol allowing for an optogenetic stimulation of neurons in awake mouse pups with the purpose of investigating the effect of neuronal activity on oligodendroglia dynamics during early PN stages. Since GABAergic interneurons contact oligodendrocyte precursor cells (OPCs) through bona fide synapses and maintain a close relationship with these progenitors during cortical development, we used this relevant example of neuron-oligodendroglia interaction to implement a proof-of-principle optogenetic approach. First, we tested Nkx2.1-Cre and Parvalbumin (PV)-Cre lines to drive the expression of the photosensitive ion channel channelrhodopsin-2 (ChR2) in subpopulations of interneurons at different developmental stages. By using patch-clamp recordings and photostimulation of ChR2-positive interneurons in acute somatosensory cortical slices, we analyzed the level of functional expression of ChR2 in these neurons. We found that ChR2 expression was insufficient in PV-Cre mouse at PN day 10 (PN10) and that this channel needs to be expressed from embryonic stages (as in the Nkx2.1-Cre line) to allow for a reliable photoactivation in mouse pups. Then, we implemented a stereotaxic surgery to place a mini-optic fiber at the cortical surface in order to photostimulate ChR2-positive interneurons at PN10. In vivo field potentials were recorded in Layer V to verify that photostimulation reaches deep cortical layers. Finally, we analyzed the effect of the photostimulation on the layer V oligodendroglia population by conventional immunostainings. Neither the total density nor a proliferative fraction of OPCs were affected by increasing interneuron activity in vivo, complementing previous findings showing the lack of effect of GABAergic synaptic activity on OPC proliferation. The methodology described here should provide a framework for future investigation of the role of early cellular interactions during PN brain maturation.
关键词： optogenetics,developing brain,proliferation,oligodendrocyte precursor cell,GABAergic interneuron,somatosensory cortex
更新于2025-09-23 15:23:52
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Apoptosis-induced Proliferation in UV-Irradiated Human Conjunctival Epithelial Cells
摘要： A pterygium is a benign growth that develops on the conjunctiva and, in some cases, extends to the cornea and interferes with vision. Excessive exposure to ultraviolet (UV) light is one of the causes of pterygium development. We previously reported that UV-induced apoptosis is led by production of reactive oxygen species (ROS) that activate p38 mitogen-activated protein kinase (MAPK) in human conjunctival epithelial (HCE) cells. Also, ROS-dependent induction of interleukin-11 (IL-11) has been reported to upregulate MAPK pathways, which results in compensatory proliferation. In this study, we examined the effect of UV exposure on HCE cells, in terms of change in apoptosis, ROS generation, phosphorylation of c-Jun N-terminal kinase (JNK), levels of IL-11 (a key cytokine in tissue repair and compensatory proliferation), production of activator protein 1 (AP-1), and expression of c-myc, c-fos and c-jun (which provides evidence of healthy cell proliferation). Apoptosis in HCE cells was induced by UV light irradiation (312 nm, 4.94 mW/cm2). Apoptosis was measured using the Muse Annexin V and Dead Cell Assay Kit. ROS generation was measured by using 5-(and 6-) chloromethyl-2?7?-dichlorodihydrofluorescein diacetate, acetyl ester. JNK phosphorylation, IL-11 levels and AP-1 production were measured by enzyme-linked immunosorbent assays (ELISAs). Imnunocytochemical staining was used to measure c-myc, c-fos and c-jun expression. UV irradiation increased ROS generation, phosphorylation of JNK, and apoptotic cell count. IL-11 levels and AP-1 production were significantly increased by UV irradiation. The irradiated cells had increased expression of c-myc, c-fos and c-jun, and treatment of the cells with IL-11 significantly increased expression of c-myc, c-fos and c-jun. These results suggest that the release of IL-11 from UV-induced apoptotic HCE cells and surrounding healthy cells could promote proliferation to maintain homeostasis.
关键词： conjunctiva,compensatory proliferation,apoptosis,interleukin-11 (IL-11),Ultraviolet (UV)
更新于2025-09-23 15:22:29
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Age-related differences in the prevalence of subtypes of Neovascular age-related macular degeneration in the first diagnosed eye
摘要： Purpose To evaluate age-related differences in the prevalence of subtypes of neovascular age-related macular degeneration (AMD) in the first diagnosed eye. Methods This retrospective, observational study included 1099 eyes of 1099 patients diagnosed with neovascular AMD. The neovascular AMD cases were classified into three subtypes: typical neovascular AMD, polypoidal choroidal vasculopathy (PCV), and type 3 neovascularization. The patients were divided into four groups, according to age: > 50 and < 60 years, ≥ 60 and < 70 years, ≥ 70 and < 80 years, and ≥ 80 years. Difference in the prevalence of three AMD subtypes was evaluated among the four age groups. Results In the age group > 50 and < 60 years, 34 (25.0%) and 102 patients (75.0%) were diagnosed with typical neovascular AMD and PCV, respectively. In the age group ≥ 60 and < 70 years, 90 (28.1%), 206 (64.4%), and 24 patients (7.5%) were diagnosed with typical neovascular AMD, PCV, and type 3 neovascularization, respectively. In the age group ≥ 70 and < 80 years, the corresponding numbers were 200 (41.9%), 197 (41.3%), and 80 (16.8%), respectively; in the age group ≥80 years, the corresponding values were 83 (50.0%), 39 (23.5%), and 44 (26.5%), respectively. A significant difference was observed in the prevalence of the subtypes of neovascular AMD among the four age groups (chi-square test, P < 0.001). Conclusion Subtype prevalence in newly diagnosed neovascular AMD differs significantly according to age. This result suggests that different pathophysiology may be involved in the development of different subtypes of neovascular AMD.
关键词： Retinal angiomatous proliferation,Polypoidal choroidal vasculopathy,Choroidal neovascularization,Type 3 neovascularization,Age-related macular degeneration
更新于2025-09-23 15:22:29
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Effects of Carbon Arc Lamp Irradiation on Wound Healing in a Rat Cutaneous Full-Thickness Wound Model
摘要： Objective: The objective of the present study was to investigate the application of a carbon arc lamp on wound healing in a rat cutaneous full-thickness wound model. Background data: In clinical practice, wound healing has been promoted by irradiation with a carbon arc lamp. However, the corresponding mechanism has not been clearly defined. Methods: A cutaneous full-thickness wound on the back of rats was irradiated using a carbon arc lamp at a wavelength peak range of 620–740 nm with 54 J/cm2. Injured sham-irradiated control rats were used as the control. The rats were euthanized after 7, 14, and 21 days, while wound reepithelialization and healing quality were examined by histological analyses with comparison between groups. Cell proliferation was observed by 5-bromo-2￠-deoxyuridine (BrdU) immunohistochemical staining. Results: Irradiation by the carbon arc lamp significantly accelerated wound healing. The wound-healing rate in the treated group at day 21 was 98.42% – 0.56%, compared with 93.58% – 1.26% in the control group ( p < 0.05). Significant increases in the length of epithelial edges, collagen content, and microvessel density were observed in the wound sites in the treated group at days 7, 14, and 21 ( p < 0.05). Moreover, the number of BrdU-labeled cells increased in the wound edge at days 7 and 14 due to irradiation ( p < 0.05). Conclusions: The results demonstrated that the carbon arc lamp can promote wound healing together with improvement in its quality by stimulating cell proliferation.
关键词： wound healing,cell proliferation,carbon arc lamp
更新于2025-09-23 15:22:29
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Photobiomodulation with 630-nm LED radiation inhibits the proliferation of human synoviocyte MH7A cells possibly via TRPV4/PI3K/AKT/mTOR signaling pathway
摘要： Phototherapy has been used to treat postoperative pain and inflammatory response in rheumatoid arthritis. Confidence in this approach, however, is impaired by lack of understanding of the light-triggered cellular and molecular mechanisms. The purpose of this study was to characterize the response of human synoviocyte MH7A cells to visible LED red light in an attempt to elucidate the associated action mechanism. Human synoviocyte MH7A cells were treated with 630-nm LED light after stimulation of tumor necrosis factor-α (TNF-α). The effects of light radiation on cell proliferation and migration were detected by MTT assay and scratch test. The expressions of inflammatory cytokines were measured using RT-qPCR. This was followed by detection of the levels of extracellular proteins IL-6 and IL-8 after differential radiation. Furthermore, the expression levels and activation of proteins on PI3K/AKT/mTOR signaling pathway were examined with Western blot. In terms of the proliferation and migration, repeated radiation with LED red light (630 nm, 26 and 39 J/cm2) exerted an inhibitory effect on synoviocyte MH7A cells. Expression of inflammatory factors (IL-6, IL-1β, IL-8, and MMP-3) was reduced; meanwhile, the expression of anti-inflammatory factor IL-10 was promoted. At the protein level, treatment with 39 J/cm2 of LED red light could decrease the level of extracellular protein (IL-6 and IL-8) and affect the expression and phosphorylation of proteins on TRPV4/PI3K/AKT/mTOR signaling pathway induced by TNF-α. These results demonstrated that LED red light (630 nm) inhibits proliferation and migration of MH7A cells. The growth-inhibiting effects of LED red light on human synoviocyte MH7A cells appear to be associated with regulation of the TRPV4/PI3K/AKT/mTOR signaling pathway.
关键词： PI3K/AKT/mTOR pathway,TRPV4 channel,Proliferation,630-nm LED,Synoviocyte
更新于2025-09-23 15:19:57
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Long-term effect of low-level diode laser irradiation on proliferation of stem cells from human exfoliated deciduous teeth after cryopreservation protocol
摘要： Low-level laser irradiation (LLLI) could be useful for the stimulation of stem-cell proliferation. LLLI has important physiological properties which increase the mitochondrial activity of cells. This study aimed to evaluate the biological effect of LLLI on cryopreserved stem cell from human exfoliated deciduous teeth (SHEDs). Cryopreserved and characterised SHEDs from a previous study were used for this study. The SHEDs were exposed to GaAlAs diode laser with energy densities of 1.2 (Group 2) and 2 J cm?2 (Group 3). For the control group (Group 1), SHEDs were cultured in culture medium including Dulbecco’s modified Eagle’s Medium supplemented with 15% foetal bovine serum, 1% L-glutamine, 1% penicillin-streptomycin, 1% gentamycin, and amphotericin-B. The cells in the experimental groups were irradiated as previously indicated, and after the irradiation were incubated for 0, 24, 48, and 72 h at 37 °C in 5% CO2. Proliferation of SHEDs was analysed by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay at incubation periods of 0, 24, 48, and 72 h after the single-laser irradiation. The effects of laser irradiation on apoptosis of SHEDs at the last incubation period (72 h) were detected with the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling (TUNEL) method. Group 2 showed increased TUNEL-positive cells compared to Groups 1 and 3, but no significant differences between the three groups were observed (p > 0.05). Comparing Groups 2 and 3 to Group 1, the proliferation of SHEDs significantly increased after 24, 48, and 72 h (p < 0.05), but no significant differences were observed between the groups at 0 h (p > 0.05). In addition to this, at 72 h, Group 3 showed significantly higher proliferative effect on SHEDs compared to Group 2 (p > 0.05). In terms of clinical usage of long-term cryopreserved SHEDs, we can recommend to clinicians and researchers that prior to their use, LLLI may be used to increase the proliferation rate.
关键词： cryopreserved stem cells,low-level diode laser irradiation (LLLI),apoptosis,proliferation
更新于2025-09-23 15:19:57
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Optoelectronic properties analysis of silicon light-emitting diode monolithically integrated in standard CMOS IC
摘要： Objectives: The function of miR‐611 has not yet been reported. We aimed to investigate the effects of miR‐611 on tongue squamous cell carcinoma (TSCC) and the underlying mechanism. Materials and Methods: The expression level of miR‐611 in TSCC tissues was measured using quantitative reverse transcriptase–polymerase chain reaction (RT‐qPCR). Cell proliferation, migration and invasion were examined by performing CCK‐8, IncuCyte and Transwell assays. Bioinformatics analyses and microarrays were used to screen for target genes, which were verified using a luciferase reporter assay, RT‐qPCR and Western blotting. The xenograft model was used to assess the effects of miR‐611 in vivo. Results: miR‐611 was upregulated in TSCC tissues, which was significantly correlated with TNM stage and negatively associated with the overall survival of patients. In addition, upregulation of miR‐611 not only potentiated the proliferation, migration, invasion and epithelial–mesenchymal transition (EMT) of TSCC cells in vitro, but also promoted tumour growth in vivo. FOXN3 was identified as a candidate target gene of miR‐611 and subsequently verified. Finally, miR‐611 induced a malignant phenotype of TSCC, which was rescued by overexpression of FOXN3. Conclusions: Our findings suggest that miR‐611 is a novel therapeutic target for TSCC.
关键词： miR‐611,FOXN3,tongue squamous cell carcinoma,epithelial–mesenchymal transition,proliferation
更新于2025-09-23 15:19:57
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Irradiation with blue light-emitting diode enhances osteogenic differentiation of stem cells from the apical papilla
摘要： This study aimed to evaluate the effects of low-energy blue LED irradiation on the osteogenic differentiation of stem cells from the apical papilla (SCAPs). SCAPs were derived from human tooth root tips and were irradiated with 0 (control group), 1 J/cm2, 2 J/cm2, 3 J/cm2, or 4 J/cm2 blue light in osteogenic induction medium. Cell proliferation was analyzed using the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. Osteogenic differentiation activity was evaluated by monitoring alkaline phosphatase (ALP), alizarin red staining, and real-time polymerase chain reaction (RT-PCR). The results of the MTT assay indicated that SCAPs in the LED groups exhibited a lower proliferation rate than those in the control group, and there were statistically differences between the 2 J/cm2, 3 J/cm2, and 4 J/cm2 groups and the control group (P < 0.05). The results of the ALP and alizarin red analyses showed that blue LED promoted osteogenic differentiation of the SCAPs. And 4 J/cm2 blue light upregulates the expression levels of the osteogenic/dentinogenic genes ALP, dentin sialophosphoprotein (DSPP), dentin matrix protein-1 (DMP-1), and osteocalcin (OCN) in SCAPs. Our results confirmed that low-energy blue LED at 1 J/cm2, 2 J/cm2, 3 J/cm2, and 4 J/cm2 could inhibit the proliferation of SCAPs and promotes osteogenic differentiation of SCAPs. Further in vitro studies are required to explore the mechanisms of the effects by low-energy blue LED.
关键词： Stem cells from apical papilla,Mesenchymal stem cells,Proliferation,Osteogenic differentiation,LED
更新于2025-09-23 15:19:57
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Regulation of NADPH-dependent Nitric Oxide and reactive oxygen species signalling in endothelial and melanoma cells by a photoactive NADPH analogue
摘要： Nitric Oxide (NO) and Reactive oxygen species (ROS) are endogenous regulators of angiogenesis-related events as endothelial cell proliferation and survival, but NO/ROS defect or unbalance contribute to cancers. We recently designed a novel photoactive inhibitor of NO-Synthases (NOS) called NS1, which binds their NADPH site in vitro. Here, we show that NS1 inhibited NO formed in aortic rings. NS1-induced NO decrease led to an inhibition of angiogenesis in a model of VEGF-induced endothelial tubes formation. Beside this effect, NS1 reduced ROS levels in endothelial and melanoma A375 cells and in aorta. In metastatic melanoma cells, NS1 first induced a strong decrease of VEGF and blocked melanoma cell cycle at G2/M. NS1 decreased NOX4 and ROS levels that could lead to a specific proliferation arrest and cell death. In contrast, NS1 did not perturb melanocytes growth. Altogether, NS1 revealed a possible cross-talk between eNOS- and NOX4 – associated pathways in melanoma cells via VEGF, Erk and Akt modulation by NS1 that could be targeted to stop proliferation. NS1 thus constitutes a promising tool that modulates NO and redox stresses by targeting and directly inhibiting eNOS and, at least indirectly, NADPH oxidase(s), with great potential to control angiogenesis.
关键词： ROS,endothelium,NADPH analogue,Angiogenesis,melanoma,cell proliferation,Cellular signaling
更新于2025-09-19 17:15:36
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The Effect of Laser Structuring of Carbon Nanotubes on the Proliferation of Chondroblasts and Mesenchymal Stem Cells
摘要： The density of cartilage cells (chondroblasts) proliferating on a silicon substrate coated with vertically oriented arrays of multi-walled carbon nanotubes (MWCNTs) was shown to be higher than on a pure silicon substrate. Electron microscopy showed that the cells in a nutrient medium affected the vertical position of the nanotubes in the array. A method for structuring the MWCNT arrays by 100-ns laser pulse scanning and abrasive water processing on planar substrates was developed. As a result of the structuring of the MWCNTs, the arrays become resistant to bending under the influence of the nutrient medium with mesenchymal stem cells. Structured MWCNT arrays were shown to have no toxic or pathological effect on the viability and morphology of stem cells. Thus, such materials can be suggested for use in cell-adhesive components of biomedical devices.
关键词： carbon nanotubes,laser structuring,mesenchymal stem cells,chondroblasts,cell proliferation
更新于2025-09-19 17:13:59