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1.
Photochem Photobiol ; 100(1): 225-232, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-37254280

RESUMEN

There are limited data on comparison of pulsed and continuous wave in photobiomodulation therapy (PBM). This study aimed to investigate the effect of PBM with 980 nm laser in pulsed and continuous wave on the proliferation and migration of human gingival fibroblasts (HGF) cells. Cultured HGF were divided into three main groups: (1) irradiated in pulsed mode (frequencies of 50 and 25 KHz; energy densities of 3 and 5 J/cm2 ), (2) irradiated in continuous mode (energy densities of 3.2 and 5.2 J/cm2 ), and (3) no irradiation as control group. HGF proliferation rate was measured by MTT assay at 24, 48, and 72 h post irradiation. In addition, HGF migration rate was measured by scratch test at 24 h post PBM. At 24 h, the group received continuous irradiation at 5.2 J/cm2 showed significantly higher proliferation compared with the control group (p = 0.012). At 48 and 72 h, the groups received continuous, and 50 Hz pulsed irradiation at energy densities of 5.2 and 5 J/cm2 respectively, had significantly higher HGF proliferation rates compared to the control (p < 0.05). Only the continuous irradiations were effective in significant increase of the cell migration. In conclusion, continuous PBM at energy density of 5.2 J/cm2 showed promising effect on HGF proliferation and migration.


Asunto(s)
Terapia por Luz de Baja Intensidad , Humanos , Proliferación Celular/efectos de la radiación , Supervivencia Celular , Rayos Láser , Fibroblastos/efectos de la radiación
2.
Arch Oral Biol ; 159: 105874, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38147800

RESUMEN

OBJECTIVE: Photobiomodulation therapy (PBMT) has proven to reduce inflammation and pain and increase wound healing. Thus, the aim of this study was to analyze the effects of PBMT parameters on migration, proliferation, and gene expression after ionizing radiation and bacterial-induced stress in an in vitro study. DESIGN: Keratinocytes (HaCaT) and Fibroblasts (HGFs) were grown in DMEM with 10 % fetal bovine serum until stressful condition induction with lipopolysaccharide (LPS) of Escherichia coli (1 µg/mL), Porphyromonas gingivalis protein extract (5 µg/mL) and ionizing radiation (8 Gy). Low-laser irradiation (660 nm, 30 mW) was carried out in four sessions, with 6 h intervals, and energy density of 2, 3, 4, and 5 J/cm². Scratch assays, immunofluorescence, and RT-qPCR were performed. RESULTS: Treated fibroblasts and keratinocytes showed significant response in proliferation and migration after scratch assays (p < 0.05). Higher expressions of α-SMA in fibroblasts and F-actin in keratinocytes were observed in cells subjected to 3 J/cm². PI3K-pathway genes expression tended to enhance in fibroblasts, presenting a higher relative expression when compared to keratinocytes. In keratinocytes, PBMT groups demonstrated deregulated expression for all inflammatory cytokines' genes tested while fibroblasts presented a tendency to enhance those genes expression in a dose dependent way. CONCLUSIONS: The present study showed that delivering 660 nm, 30 mW was effective to stimulate cell migration, proliferation and to accelerate wound healing. PBMT can modulate cytokines and pathways involved in wound repair. The different energy densities delivering distinct responses in vitro highlights that understanding laser parameters is fundamental to improve treatment strategies.


Asunto(s)
Terapia por Luz de Baja Intensidad , Fosfatidilinositol 3-Quinasas , Queratinocitos , Fibroblastos/efectos de la radiación , Proliferación Celular/efectos de la radiación , Radiación Ionizante
3.
Lasers Med Sci ; 39(1): 9, 2023 Dec 19.
Artículo en Inglés | MEDLINE | ID: mdl-38110767

RESUMEN

An in vitro study was designed to evaluate the effects of photobiomodulation (PBM) with 915-nm diode laser on human gingival fibroblast (HGF) cells under hyperglycemic condition. The HGF cells were cultured in Dulbecco's modified eagle medium (DMEM) medium containing 30 mM glucose concentration for 48 h to mimic the hyperglycemic condition. Subsequently, the cells received three sessions of PBM (915 nm, continuous emission mode, 200 mW, energy density values of 3.2, 6, and 9.2 J/cm2). Twenty-four hours post-irradiation, cell proliferation, expression of interleukin 6 (IL-6), and vascular endothelial growth factor (VEGF) were assessed with MTT and real-time polymerase chain reaction (PCR) tests, respectively. Also, reactive oxygen species (ROS) production was measured using CM-H2DCFDA fluorimetry. No changes were detected in the cell proliferation rate between the high glucose control group and laser-treated cells, while VEGF and IL-6 gene expression levels increased significantly after PBM in the high glucose-treated cells group. ROS level was significantly decreased in the irradiated cells in high-glucose medium compared with the high glucose control group. Our study revealed the inductive role of 915-nm-mediated PBM on VEGF and the inflammatory response while concurrently reducing reactive oxygen species production in HGF cells in hyperglycemic conditions.


Asunto(s)
Interleucina-6 , Terapia por Luz de Baja Intensidad , Humanos , Interleucina-6/genética , Factor A de Crecimiento Endotelial Vascular/genética , Especies Reactivas de Oxígeno/metabolismo , Glucemia , Fibroblastos/efectos de la radiación , Células Cultivadas
4.
Lasers Med Sci ; 38(1): 27, 2022 Dec 27.
Artículo en Inglés | MEDLINE | ID: mdl-36574084

RESUMEN

Wound treatment, especially for chronic and infected wounds, has been a permanent socio-economical challenge. This study aimed to investigate the ability of red light at 661 nm to accelerate wound healing an in vitro wound model using 3T3 fibroblasts. The purpose is further specified in clarifying the mechanisms of wound closure by means of intracellular ROS production, proliferation and migration of cells, and cellular orientation. Illumination effects of red light from a diode laser (661 nm) at different doses on 3T3 cell viability was assessed via MTT assay and tested in a scratch wound model. Wound closure rates were calculated by image analysis at 0, 24, and 48 h after laser treatment. ROS production was monitored and quantified immediately and 24 h after the treatment by fluorescence microscopy. Cellular orientation was quantified by image analysis. No phototoxic energy doses used and increased cell viability in most of the groups. Scratch assay revealed an energy interval of 3 - 4.5 J/cm2 that promote higher wound healing rate 24 h post treatment. An increase in ROS production was also observed 24 h post irradiation higher in the group with the highest wound healing rate. Also, cellular orientation toward the margin of the wound was observed and quantified after irradiation. Low power laser light at 661 nm activated both the migration and proliferation in the in vitro model used, providing evidence that it could also accelerate wound healing in vivo. Also, ROS production and cellular orientation seem to play an important role in wound healing process.


Asunto(s)
Terapia por Luz de Baja Intensidad , Especies Reactivas de Oxígeno , Terapia por Luz de Baja Intensidad/métodos , Proliferación Celular/efectos de la radiación , Cicatrización de Heridas/efectos de la radiación , Fibroblastos/efectos de la radiación , Láseres de Semiconductores/uso terapéutico
5.
Photochem Photobiol ; 98(5): 1201-1206, 2022 09.
Artículo en Inglés | MEDLINE | ID: mdl-35188978

RESUMEN

5-Fluoroufacil (5FU) is a chemotherapeutic agent indicated for solid tumors but causes oral mucositis, which can be prevented and treated using photobiomodulation therapy (PBMT). It is unknown whether PBMT modifies DNA damage induced by 5FU in oral cells. The aim of this study was to investigate the effect of PBMT on DNA damage and repair and on oxidative stress in gingival fibroblasts exposed to 5FU. Primary gingival fibroblasts were exposed to 5FU and then treated with a laser (660 nm, 100 mW, 1 W cm-2 , 0.09 cm2 spot area) for three different irradiation times (6, 10 or 20 s). Six-second irradiation decreased DNA damage and lipidic peroxidation. All irradiated groups showed low H2AX levels and increased p53 expression. Ten-second irradiation showed a trend to induce high lipidic peroxidation levels and DNA damage than other irradiation groups. In conclusion, the PBMT effect on DNA damage and repair was dependent on the time irradiation: 6 s-time irradiation (6.6 J cm-2 ) protect gingival fibroblasts from 5FU-related genotoxicity and oxidative stress, whereas 10s- and 20s-time irradiations (11.1 J cm-2 and 22.2 J cm-2 , respectively) increased the risk of DNA damage after the 5FU exposure.


Asunto(s)
Terapia por Luz de Baja Intensidad , Daño del ADN , Fibroblastos/efectos de la radiación , Fluorouracilo , Proteína p53 Supresora de Tumor/metabolismo
6.
Photochem Photobiol ; 98(4): 969-973, 2022 07.
Artículo en Inglés | MEDLINE | ID: mdl-34932837

RESUMEN

Photobiomodulation is recognized as an effective method for adjunct therapy in periodontal treatments. Our purpose in this study was to investigate the effects of different energy densities of 915 nm diode laser on the viability and viability capacity of human gingival fibroblast cells. Cell samples were examined in five groups, including four irradiation groups with low-level diode laser 915 nm, 1, 2, 3, 4 J cm-2 and a control group (no Laser irradiation). Cell viability and viability were measured 1, 3 and 5 days after irradiation by MTT and DAPI assay. Statistical differences between groups at any time were analyzed by one-way ANOVA and a post hoc Turkey's test. The cell viability and viability capacity increased on the third day at an energy density of 3 J cm-2 ; (P-value = 0.007) and the fifth day at energy densities of 2, 3 and 4 J cm-2 was recorded compared with the control group (P-value = 0.000). Also, a significant decrease in the viability and viability of irradiated cells with an energy density of 1 J cm-2 was found (P-value = 0.033). According to our results, Photobiomodulation with 915 nm diode laser has a positive stimulating effect on the viability and viability capacity of human gingival fibroblast cells.


Asunto(s)
Terapia por Luz de Baja Intensidad , Proliferación Celular/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Fibroblastos/efectos de la radiación , Encía , Humanos , Láseres de Semiconductores , Terapia por Luz de Baja Intensidad/métodos
7.
Int J Mol Sci ; 22(21)2021 Oct 27.
Artículo en Inglés | MEDLINE | ID: mdl-34769035

RESUMEN

Background: The invention of non-ionizing emission devices revolutionized science, medicine, industry, and the military. Currently, different laser systems are commonly used, generating the potential threat of excessive radiation exposure, which can lead to adverse health effects. Skin is the organ most exposed to laser irradiation; therefore, this study aims to evaluate the effects of 445 nm, 520 nm, and 638 nm non-ionizing irradiation on keratinocytes and fibroblasts. Methods: Keratinocytes and fibroblasts were exposed to a different fluency of 445 nm, 520 nm, and 638 nm laser irradiation. In addition, viability, type of cell death, cell cycle distribution, and proliferation rates were investigated. Results: The 445 nm irradiation was cytotoxic to BJ-5ta (≥58.7 J/cm2) but not to Ker-CT cells. Exposure influenced the cell cycle distribution of Ker-CT (≥61.2 J/cm2) and BJ-5ta (≥27.6 J/cm2) cells, as well as the Bj-5ta proliferation rate (≥50.5 J/cm2). The 520 nm irradiation was cytotoxic to BJ-5ta (≥468.4 J/cm2) and Ker-CT (≥385.7 J/cm2) cells. Cell cycle distribution (≥27.6 J/cm2) of Ker-CT cells was also affected. The 638 nm irradiation was cytotoxic to BJ-5ta and Ker-CT cells (≥151.5 J/cm2). The proliferation rate and cell cycle distribution of BJ-5ta (≥192.9 J/cm2) and Ker-CT (13.8 and 41.3 J/cm2) cells were also affected. Conclusions: At high fluences, 455 nm, 520 nm, and 638 nm irradiation, representing blue, green, and red light spectra, are hazardous to keratinocytes and fibroblasts. However, laser irradiation may benefit the cells at low fluences by modulating the cell cycle and proliferation rate.


Asunto(s)
Fibroblastos/efectos de la radiación , Piel/efectos de la radiación , Ciclo Celular/efectos de la radiación , Muerte Celular/efectos de la radiación , Proliferación Celular/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Células Cultivadas , Humanos , Rayos Láser , Luz , Terapia por Luz de Baja Intensidad/métodos
8.
Biochem Biophys Res Commun ; 580: 41-47, 2021 11 26.
Artículo en Inglés | MEDLINE | ID: mdl-34619551

RESUMEN

Luffa cylindrica stem sap (LuCS) has been traditionally used as a facial cosmetic supplement to enhance the skin condition of Asians. However, LuCS has yet to be described and there is no solid scientific evidence regarding the use of LuCS as an anti-wrinkle agent. In the present study, we have evaluated the functional effect of LuCS and its underlying mechanisms based on scientific evidence. Treatment with LuCS stimulated the growth and migration of human skin fibroblasts. LuCS treatment activated EGFR signaling via the enhanced expression of EGFR and down-regulation of PPARγ in human skin fibroblasts. Exposure to LuCS induced the synthesis of cellular type I procollagen and elastin in consort with the down-regulation of various proteinases including MMP-1, -2 and -9 in human skin fibroblasts. LuCS treatment also reversed the skin damage induced by UV-A irradiation in human skin fibroblasts. 3-bromo-3-methylisoxazol-5-amine was identified as the functional component using UPLC-MS-MS analysis and increased production of cellular type I procollagen. Collectively, these results suggest the efficacy of LuCS supplementation in improving the skin condition via anti-wrinkle effect.


Asunto(s)
Fibroblastos/efectos de los fármacos , Luffa , Extractos Vegetales/farmacología , Sustancias Protectoras/farmacología , Envejecimiento de la Piel/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Fibroblastos/citología , Fibroblastos/efectos de la radiación , Humanos , Luffa/química , Extractos Vegetales/química , Tallos de la Planta/química , Sustancias Protectoras/química
9.
J Photochem Photobiol B ; 222: 112256, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34330080

RESUMEN

Photobiomodulation (PBM) is a promising medical treatment modality in the area of photodynamic therapy (PDT). In this study, we investigated the effect of combined therapy in a 3D microenvironment using aluminum chloride phthalocyanines (AlClPc) as the photosensitizing agent. Normal human fibroblast-containing collagen biomatrix was prepared and treated with an oil-in-water (o/a) AlClPc-loaded nanoemulsion (from 0.5 to 3.0 µM) and irradiated at a range of fluences (from 0.1 to 3.0 J/cm2) using a continuous-wave light-emitting diode (LED) irradiation system (660 nm). PBM at 1.2 J/cm2 and AlClPc/NE at 0.5 µM modified the fibroblast signaling response under 3D conditions, promoting collagen synthesis, ROS production, MMP-9 secretion, proliferation of the actin network, and facile myofibroblastic differentiation. PBM alone (at 1.2 J/cm2 and 0.3 J/cm2) had no significant effect on any of these parameters. The combined therapy affected myofibroblastic differentiation, inflammatory response, and extracellular matrix pliability, and should thus be examined further in subsequent studies considering that no side effects of PBM have been reported. Even though significant progress has been made in the field of phototherapy in recent years, it is necessary to further elucidate the detailed mechanisms underlying its effects already shown in 2D conditions to increase the acceptance of this beneficial and non-invasive therapeutic approach.


Asunto(s)
Cloruro de Aluminio/farmacología , Fibroblastos/efectos de los fármacos , Indoles/farmacología , Luz , Compuestos Organometálicos/farmacología , Fármacos Fotosensibilizantes/farmacología , Cloruro de Aluminio/química , Técnicas de Cultivo de Célula , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Células Cultivadas , Colágeno/metabolismo , Fibroblastos/citología , Fibroblastos/metabolismo , Fibroblastos/efectos de la radiación , Humanos , Indoles/química , Metaloproteinasa 9 de la Matriz/metabolismo , Compuestos Organometálicos/química , Fármacos Fotosensibilizantes/química , Especies Reactivas de Oxígeno/metabolismo
10.
Sci Rep ; 11(1): 7315, 2021 04 01.
Artículo en Inglés | MEDLINE | ID: mdl-33795767

RESUMEN

Fibrosis occurs when collagen deposition and fibroblast proliferation replace healthy tissue. Red light (RL) may improve skin fibrosis via photobiomodulation, the process by which photosensitive chromophores in cells absorb visible or near-infrared light and undergo photophysical reactions. Our previous research demonstrated that high fluence RL reduces fibroblast proliferation, collagen deposition, and migration. Despite the identification of several cellular mechanisms underpinning RL phototherapy, little is known about the transcriptional changes that lead to anti-fibrotic cellular responses. Herein, RNA sequencing was performed on human dermal fibroblasts treated with RL phototherapy. Pathway enrichment and transcription factor analysis revealed regulation of extracellular matrices, proliferation, and cellular responses to oxygen-containing compounds following RL phototherapy. Specifically, RL phototherapy increased the expression of MMP1, which codes for matrix metalloproteinase-1 (MMP-1) and is responsible for remodeling extracellular collagen. Differential regulation of MMP1 was confirmed with RT-qPCR and ELISA. Additionally, RL upregulated PRSS35, which has not been previously associated with skin activity, but has known anti-fibrotic functions. Our results suggest that RL may benefit patients by altering fibrotic gene expression.


Asunto(s)
Fibroblastos/metabolismo , Fibroblastos/efectos de la radiación , Fototerapia/métodos , Piel/metabolismo , Piel/efectos de la radiación , Transcriptoma , Adulto , Movimiento Celular , Proliferación Celular , Colágeno/metabolismo , Femenino , Fibrosis , Perfilación de la Expresión Génica , Humanos , Masculino , Metaloproteinasa 1 de la Matriz/biosíntesis , Persona de Mediana Edad , Estrés Oxidativo , Oxígeno/metabolismo , RNA-Seq , Especies Reactivas de Oxígeno , Enfermedades de la Piel/metabolismo , Factores de Tiempo , Factores de Transcripción
11.
Oxid Med Cell Longev ; 2021: 6667812, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33628374

RESUMEN

Chemicals and signaling molecules released by injured cells at the beginning of wound healing prompt inflammation. In diabetes, prolonged inflammation is one of the probable causes for delayed wound healing. Increased levels of cyclooxygenase-2 (cox-2), interleukin-6 (IL-6), and tumour necrosis factor-alpha (TNF-α) are associated with the inflammatory response and in diabetes, and increased levels of these contribute to chronic wounds that do not heal. Rising levels of cox-2, IL-6, and TNF-α have also been associated with increased oxidative stress. Photobiomodulation (PBM) may impact wound healing processes by affecting the signaling pathways and molecules pertinent to tissue repair. In the present study, the effect of PBM (wavelength: 660 nm; energy density: 5 J/cm2) on levels of cox-2, IL-6, and TNF-α was determined in fibroblast cell culture models. Four WS1 models (normal, normal wounded, diabetic, and diabetic wounded) were irradiated at 660 nm, and the culture media was collected at 0, 24, and 48 h postirradiation. Cells that were not irradiated (0 J/cm2) served as the controls. The following parameters were determined postirradiation: cell morphology using light microscopy, cell viability using the Trypan Blue exclusion assay, and levels of the inflammatory markers cox-2, IL-6, and TNF-α were measured using ELISA. Cell migration increased in the wounded groups over the 48 h interval after PBM; viability improved postirradiation in the diabetic wounded groups at 0 and 24 h (P ≤ 0.05 and P ≤ 0.01, respectively); levels of cox-2 decreased in normal and diabetic wounded groups at 0 h (P ≤ 0.001) and increased in the diabetic and diabetic wounded groups at 48 h postirradiation (P ≤ 0.05 and P ≤ 0.01, respectively), while levels of IL-6 decreased in the normal (P ≤ 0.01), diabetic (P ≤ 0.05), and diabetic wounded (P ≤ 0.001) groups at 24 h and in the diabetic and diabetic wounded groups at 48 h (P ≤ 0.05) postirradiation. TNF-α was decreased in the normal wounded groups (P ≤ 0.05) at 48 h. Through its effect on decreased IL-6 levels in diabetic cell models, PBM at 660 nm may be successful at decreasing oxidative stress; however, the present study also found an increase in cox-2 levels at 48 h postirradiation.


Asunto(s)
Ciclooxigenasa 2/metabolismo , Fibroblastos/metabolismo , Fibroblastos/efectos de la radiación , Interleucina-6/metabolismo , Terapia por Luz de Baja Intensidad , Factor de Necrosis Tumoral alfa/metabolismo , Técnicas de Cultivo de Célula , Forma de la Célula/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Humanos
12.
Biol Pharm Bull ; 44(2): 225-231, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33518674

RESUMEN

The dermis is mainly constructed by type I collagen fibers, which provide mechanical strength to the skin by building a frame-like structure, and by elastic fibers, which provide elasticity to respond to movements of the skin. The depletion of collagen fibers and the disappearance of oxytalan fibers, which are a type of elastic fiber, are characteristic changes in photoaged skin. Prostaglandin E2 (PGE2) is one of the chemical mediators involved in inflammation and is responsible for sunburn. Furthermore, it has been reported that PGE2 attenuates the production of collagen and the expression of elastic fiber-related factors in fibroblasts. Tranexamic acid (TXA), which is an anti-inflammatory medicine that inhibits plasmin, reduces the level of PGE2 secreted following UV exposure or after inflammatory stimulation. However, few reports have verified TXA as an anti-skin aging agent. In this study, we examined the potential of TXA as an anti-skin aging agent using repetitively UVA-irradiated fibroblasts as a model for fibroblasts located in chronically sun-exposed dermis. Repetitively UVA-irradiated fibroblasts had higher secretion levels of PGE2. In addition, fibroblasts repetitively irradiated with UVA or treated with PGE2 produced disrupted collagen and fibrillin-1 fibers. Treatment with TXA improved the formation of both types of fibers by repetitively UVA-irradiated fibroblasts by restoring the expression of fiber-related proteins at the mRNA and protein levels. Thus, these results demonstrate that TXA has potential as an anti-photoaging agent.


Asunto(s)
Fibroblastos/efectos de los fármacos , Envejecimiento de la Piel/efectos de los fármacos , Piel/efectos de los fármacos , Ácido Tranexámico/farmacología , Línea Celular , Colágeno/metabolismo , Dinoprostona/metabolismo , Evaluación Preclínica de Medicamentos , Fibrilina-1/metabolismo , Fibroblastos/metabolismo , Fibroblastos/efectos de la radiación , Humanos , Piel/citología , Piel/metabolismo , Piel/efectos de la radiación , Envejecimiento de la Piel/efectos de la radiación , Rayos Ultravioleta/efectos adversos
13.
Lasers Med Sci ; 36(6): 1201-1208, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-33037560

RESUMEN

The aim of the present study was to investigate the effects of photobiomodulation (PBM) therapy on the expression of heat shock protein 70 (HSP70) and tissue repair in an experimental model of collagenase-induced Achilles tendinitis. Thirty Wistar rats (aged 12 weeks) were randomly distributed among control group (n = 8), tendinitis group (n = 11), and LED group (n = 11). Tendinitis was induced in the tendinitis and LED groups through a peritendinous injection of collagenase (100 µl). The LED group animals received the first irradiation 1 h after injury. A 630 ± 20 nm, 300-mW continuous wave light-emitting diode (LED), spot size 1 cm2, was placed in contact with the skin. One point over the tendon was irradiated for 30 s, delivering 9 J (9 J/cm2). LED irradiation was performed once daily for 7 days, with the total energy delivered being 63 J. The tendons were surgically removed and expression of the HSP70 protein was calculated using semi-quantitative analyses of immunohistochemistry (HSCORE). Number of fibroblasts and amount of collagen were measured using histological and histochemical analyses. An increase in the mean HSCORE for HSP70, in the number of fibroblasts, and in the amount of collagen were found in the LED group compared with those in the tendinitis and control group (P ≤ 0.05). PBM therapy increased the expression of the HSP70, number of fibroblasts, and amount of collagen in the acute Achilles tendinitis in rats.


Asunto(s)
Tendón Calcáneo/patología , Tendón Calcáneo/efectos de la radiación , Regulación de la Expresión Génica/efectos de la radiación , Proteínas HSP70 de Choque Térmico/metabolismo , Terapia por Luz de Baja Intensidad , Tendinopatía/metabolismo , Tendinopatía/radioterapia , Animales , Modelos Animales de Enfermedad , Fibroblastos/metabolismo , Fibroblastos/efectos de la radiación , Masculino , Ratas , Ratas Wistar , Tendinopatía/patología
14.
Lasers Med Sci ; 36(6): 1169-1175, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32996021

RESUMEN

Although it was demonstrated that curcumin-mediated antimicrobial photodynamic therapy (aPDT) is effective for reducing the viability of microbial cells and the vitality of oral biofilms, the cytotoxicity of this therapeutic approach for host cells has not been yet elucidated. Hence, the aim of this study was to evaluate the cytotoxicity and apoptotic effects of curcumin-mediated aPDT on mouse fibroblasts. Cells were treated with 0.6 or 6 µmol.L-1 curcumin combined with 0.075 or 7.5 J.cm-2 LED at 455 nm. Cytotoxicity was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet (CV) assays, while quantitative reverse transcriptase-PCR (qRT-PCR) was used to assess the expression of Bax, Bad, Bcl-2, VDAC-1, cytochrome C, and Fas-L genes for apoptosis. The differences between groups were detected by Kruskal-Wallis and post hoc Dunn's tests for MTT and CV assays and by ANOVA and post hoc Tukey test for qRT-PCR (P < 0.05). The effect of 0.6 µmol.L-1 curcumin plus 0.075 J.cm-2 LED (minimum parameter) did not differ statistically from control group; however, the combination of 0.6 µmol.L-1 curcumin plus 7.5 J.cm-2 LED reduced viable cells in 34%, while the combinations of 6 µmol.L-1 curcumin plus 0.075 and 7.5 J.cm-2 LED reduced viable cells in 47% and 99%, respectively. aPDT increased significantly the relative expression of Bax/Bcl-2, cytochrome C, VDAC-1, and Fas-L genes, without influence on the ratio Bad/Bcl-2. Therefore, curcumin-mediated aPDT activated Bcl-2 apoptosis signaling pathways in mouse fibroblasts regarding present conditions, reducing the viability of cells with the increase of curcumin concentrations and light energies.


Asunto(s)
Antiinfecciosos/farmacología , Apoptosis/efectos de los fármacos , Curcumina/farmacología , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Fotoquimioterapia , Transducción de Señal/efectos de los fármacos , Animales , Apoptosis/efectos de la radiación , Fibroblastos/efectos de la radiación , Ratones , Transducción de Señal/efectos de la radiación
15.
Lasers Med Sci ; 36(3): 555-562, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-32643032

RESUMEN

Delayed wound healing is one of the most challenging complications of diabetes mellitus (DM) in clinical medicine, and it is related to the excessive generation of reactive oxygen species (ROS). Photobiomodulation (PBM) can promote wound healing in many ways, so it can be used as a method for the treatment of delayed healing of DM wounds. In this study, we investigated the effect of PBM on ROS homeostasis in human embryonic skin fibroblast cells (CCC-ESFs) cultured in high glucose concentrations. The CCC-ESFs were cultured in vitro and divided into two groups, including the control group and the 635 nm laser irradiation group. After 2 days of high glucose treatment, the experimental group was irradiated with different doses of laser for 3 days. First, we measured the cellular proliferation, and the results showed that laser irradiation could promote cellular proliferation. Then, we measured the generation of ROS, the activities of total superoxide dismutase (SOD), and total antioxidant capacity (TAC) of the cells; the results showed that high glucose destroyed cells by inducing high concentration of ROS, the balance of oxidation, and antioxidation cause oxidative stress damage to cells. PBM can increase the antioxidant capacity of cells, reducing the high concentration of ROS induced by high glucose. Finally, we measured the levels of mitochondrial membrane potential (∆ψm) and the secretion of nuclear factor kappa-B (NF-κB), tumor necrosis factor-α (TNF-α), and interleukin-1ß (IL-1ß); the results showed that PBM can reduce apoptosis and regulate the inflammatory state. We conclude that PBM can maintain the ROS homeostasis, increase the TAC of cells, and trigger the cellular proliferation, and the response of CCC-ESFs to PBM was dose-dependent.


Asunto(s)
Medios de Cultivo/química , Embrión de Mamíferos/citología , Fibroblastos/efectos de la radiación , Glucosa/farmacología , Terapia por Luz de Baja Intensidad , Especies Reactivas de Oxígeno/metabolismo , Piel/citología , Animales , Antioxidantes/metabolismo , Proliferación Celular/efectos de los fármacos , Proliferación Celular/efectos de la radiación , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Humanos , Interleucina-1beta/metabolismo , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Potencial de la Membrana Mitocondrial/efectos de la radiación , FN-kappa B/metabolismo , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/efectos de la radiación , Superóxido Dismutasa/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Cicatrización de Heridas/efectos de los fármacos , Cicatrización de Heridas/efectos de la radiación
16.
Lasers Med Sci ; 36(2): 365-374, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-32483750

RESUMEN

A disrupted wound repair process often leads to the development of chronic wounds, and pose a major physical, social and economic inconvenience on patients and the public health sector. Chronic wounds are a common complication seen in diabetes mellitus (DM), and often the severity necessitates amputation of the lower limbs. Recently, there has been increasing evidence that photobiomodulation (PBM) initiates wound healing, including increased protein transcription for cell proliferation, viability, migration and tissue reepithelialisation. Here, the hypothesis that PBM at a wavelength of 660 nm and energy density of 5 J/cm2 regulates wound repair in diabetic wounded and hypoxic diabetic wounded fibroblasts by enhancing cell migration and survival was investigated. PBM increased migration and survival in diabetic wounded and hypoxic diabetic wounded fibroblasts. Our findings suggest that PBM enhances migration and survival in diabetic wounded and hypoxic diabetic wounded fibroblasts, indicating that this therapeutic method may be beneficial against chronic wounds in diabetic patients.


Asunto(s)
Movimiento Celular/efectos de la radiación , Diabetes Mellitus/patología , Fibroblastos/patología , Fibroblastos/efectos de la radiación , Hipoxia/patología , Terapia por Luz de Baja Intensidad , Animales , Apoptosis/efectos de la radiación , Proliferación Celular/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Humanos , Cicatrización de Heridas/efectos de la radiación
17.
Environ Toxicol ; 36(4): 607-619, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33270331

RESUMEN

Ultraviolet (UV) irradiation is a crucial factor that leads to skin photoaging and results in increased DNA damage, oxidative stress, and collagen degradation. Jasmine flowers have been utilized as a traditional medicine in Asia to treat various diseases, including dermatitis, diarrhea, and fever. Furthermore, the fermented broth of Lactobacillus rhamnosus has been reported to exert protective effects on the skin. In the present study, jasmine flower extract was fermented with L. rhamnosus. We investigated the antioxidant and collagen-promoting effects on UVB/H2 O2 -induced HS68 dermal fibroblast cell damage. The results indicated that treatment with the fermented flower extracts of Jasminum sambac (F-FEJS) could enhance the viability of HS68 cells. Furthermore, the UVB/H2 O2 -induced excessive production of reactive oxygen species, degradation of collagen, activation of MAPKs, including P38, ERK, and JNK, and premature senescence were remarkably attenuated by F-FEJS in dermal fibroblast cells. The nuclear accumulation of p-c-jun, which is downstream of MAPK, and the inactivation of p-smad2/3, which is one of the crucial transcription factors that enhance collagen synthesis, were reversed in response to F-FEJS treatment in UVB/H2 O2 -exposed cells. Notably, the expression of antioxidant genes, such as HO-1, and the nuclear translocation of Nrf2 were further enhanced by F-FEJS in UVB/H2 O2 -treated cells. Interestingly, the F-FEJS-induced increase in ARE luciferase activity indicated the activation of Nrf2/ARE signaling. In conclusion, our findings demonstrated that F-FEJS can effectively ameliorate UVB/H2 O2 -induced dermal cell aging and may be considered a promising ingredient in skin aging therapy.


Asunto(s)
Antioxidantes/farmacología , Senescencia Celular , Fibroblastos/efectos de los fármacos , Jasminum/química , Lacticaseibacillus rhamnosus/metabolismo , Extractos Vegetales/farmacología , Antioxidantes/aislamiento & purificación , Antioxidantes/metabolismo , Línea Celular , Senescencia Celular/efectos de los fármacos , Senescencia Celular/efectos de la radiación , Fermentación , Fibroblastos/metabolismo , Fibroblastos/efectos de la radiación , Flores/química , Humanos , Peróxido de Hidrógeno/toxicidad , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal , Envejecimiento de la Piel/efectos de los fármacos , Envejecimiento de la Piel/efectos de la radiación , Rayos Ultravioleta
19.
Arch Oral Biol ; 121: 104968, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33221565

RESUMEN

OBJECTIVE: Oral biofilms burden host responses by induction of inflammatory mediators, exacerbating periodontal inflammation. Photobiomodulation Therapy (PBMT) has been shown to decrease levels of pro-inflammatory cytokines and chemokines. However, optimal wavelengths and exposure doses have not been established. This study investigated the effects of PBMT on human periodontal ligament fibroblasts (hPDLFs) stimulated with inflammatory mediators (LPS, TNF-α, and IL-1ß). METHODS: Cytotoxic effects of laser wavelengths 660 nm and 810 nm were assessed by measuring their effects on cellular dehydrogenase activity. The study was expanded to include 980 nm, 660 nm + 810 nm, and 810 nm + 980 nm. P.g. LPS, TNF-α, and/or IL-1ß were added one hour before irradiation, then exposed to laser irradiation to determine the most appropriate stimulus. The levels of INF-γ, IL-6, IL-8, IL-17A/F, and MCP-1 production in stimulated hPDLFs were measured and analyzed. RESULTS: P.g. LPS was a poor stimulus for hPDLFs, while TNF-α and IL-1ß significantly elevated the analytes. The 660 nm laser treatment induced pro-inflammatory cytokines when stimulated, while 810 nm exhibited significant suppression. IL-1ß was the stimulus of choice and the 810 nm wavelength alone exhibited anti-inflammatory effects for all analytes except IL-8, while the 810 nm in combination with 660 nm and/or 980 nm exhibited effects similar to 810 nm alone. CONCLUSIONS: The downregulation of inflammatory mediators by the combination or individual treatment with 810 nm wavelength shows promise for the management of periodontal inflammation. PBMT may lead to the development of a novel approach in the management of periodontal disease.


Asunto(s)
Fibroblastos/efectos de la radiación , Inmunomodulación , Terapia por Luz de Baja Intensidad , Células Cultivadas , Citocinas/inmunología , Fibroblastos/inmunología , Humanos , Interleucina-1beta/farmacología , Lipopolisacáridos , Ligamento Periodontal/citología , Factor de Necrosis Tumoral alfa/farmacología
20.
J Photochem Photobiol B ; 214: 112100, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33316625

RESUMEN

Photobiomodulation (PBM) is reported to impart a range of clinical benefits, from the healing of chronic wounds to athletic performance enhancement. The increasing prevalence of this therapy conflicts with the lack of understanding concerning specific cellular mechanisms induced by PBM. Herein, we systematically explore the literature base, specifically related to PBM (within the range 600-1070 nm) and its influence on dermal fibroblasts. The existing research in this field is appraised through five areas: cellular proliferation and viability; cellular migration; ATP production and mitochondrial membrane potential; cellular protein expression and synthesis; and gene expression. This review demonstrates that when fibroblasts are irradiated in vitro within a set range of intensities, they exhibit a multitude of positive effects related to the wound healing process. However, the development of an optimal in vitro framework is paramount to improve the reliability and validity of research in this field.


Asunto(s)
Fibroblastos/efectos de la radiación , Terapia por Luz de Baja Intensidad/métodos , Cicatrización de Heridas/efectos de la radiación , Línea Celular , Movimiento Celular/efectos de la radiación , Proliferación Celular/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Fibroblastos/citología , Regulación de la Expresión Génica/efectos de la radiación , Humanos , Técnicas In Vitro , Proteínas/genética , Proteínas/metabolismo , Piel
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