RESUMO
Oxidative stress, in which the amount of oxidants exceeds the capacity of antioxidant defense system, is a well-accepted pathogenesis of several human diseases. Light-emitting diode irradiation (LEDI) is an efficient strategy to counteract this condition. The biological effect of phototherapy, using visible light, has attracted recent attention especially in dermatological practice. However, little is known about the molecular mechanism of the anti-oxidant and anti-inflammatory effects of red light irradiation. We evaluated these effects of LEDI in HaCaT human keratinocyte cells under phorbol-12-myristate-13-acetate (PMA) induced reactive oxygen species (ROS). Microarray analysis revealed changes in 309 genes after LEDI. LEDI at 625â¯nm produced ROS scavenging and anti-inflammatory effects. One of the most important genes identified by microarray analysis was sphingosine kinase-1 (SPHK1), which is a key molecule in sphingolipid metabolism. SPHK1 knock-down drastically reduced ROS scavenging efficiency as well as expression levels of inflammation-related proteins in PMA-treated HaCaT cells. These results not only indicate the potential for the clinical application of 625-nm LEDI in treating skin disorders via ROS and/or inflammation, but also suggest SPHK1 as a potential therapeutic target in phototherapy.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Luz , NF-kappa B/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/antagonistas & inibidores , Proteínas Adaptadoras de Transdução de Sinal/genética , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Citocinas/metabolismo , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/efeitos da radiação , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Humanos , Imuno-Histoquímica , Inflamação/metabolismo , Inflamação/patologia , Queratinócitos/citologia , Queratinócitos/metabolismo , Queratinócitos/efeitos da radiação , Microscopia de Fluorescência , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/efeitos da radiação , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Acetato de Tetradecanoilforbol/farmacologia , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/efeitos da radiaçãoRESUMO
Caffeoylserotonin (CaS), one derivative of serotonin (5-HT), is a secondary metabolite produced in pepper fruits with strong antioxidant activities. In this study, we investigated the effect of CaS on proliferation and migration of human keratinocyte HaCaT cells compared to that of 5-HT. CaS enhanced keratinocyte proliferation even under serum deficient condition. This effect of CaS was mediated by serotonin 2B receptor (5-HT2BR) related to the cell proliferation effect of 5-HT. We also confirmed that both CaS and 5-HT induced G1 progression via 5-HT2BR/ERK pathway in HaCaT cells. However, Akt pathway was additionally involved in upregulated expression levels of cyclin D1 and cyclin E induced by CaS by activating 5-HT2BR. Moreover, CaS and 5-HT induced cell migration in HaCaT cells via 5-HT2BR. However, 5-HT regulated cell migration only through ERK/AP-1/MMP9 pathway while additional Akt/NF-κB/MMP9 pathway was involved in the cell migration effect of CaS. These results suggest that CaS can enhance keratinocyte proliferation and migration. It might have potential as a reagent beneficial for wound closing and cell regeneration. [BMB Reports 2018; 51(4): 188-193].
Assuntos
Queratinócitos/efeitos dos fármacos , Receptor 5-HT2B de Serotonina/metabolismo , Serotonina/metabolismo , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Movimento Celular/fisiologia , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/fisiologia , Humanos , Queratinócitos/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Serotonina/análogos & derivados , Transdução de Sinais/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacosRESUMO
OBJECTIVES: To circumvent the limitations of the current golden standard method, colony-forming unit (CFU) assay, for viability of Bacille Calmette-Guérin (BCG) vaccines, we developed a new method to rapidly and accurately determine the potency of BCG vaccines. METHODS: Based on flow cytometry (FACS) and fluorescein diacetate (FDA) as the most appropriate fluorescent staining reagent, 17 lots of BCG vaccines for percutaneous administration and 5 lots of BCG vaccines for intradermal administration were analyzed in this study. The percentage of viable cells measured by flow cytometry along with the total number of organisms in BCG vaccines, as determined on a cell counter, was used to quantify the number of viable cells. RESULTS: Pearson correlation coefficients of FACS and CFU assays for percutaneous and intradermal BCG vaccines were 0.6962 and 0.7428, respectively, indicating a high correlation. The coefficient of variation value of the FACS assay was less than 7%, which was 11 times lower than that of the CFU assay. CONCLUSION: This study contributes to the evaluation of new potency test method for FACS-based determination of viable cells in BCG vaccines. Accordingly, quality control of BCG vaccines can be significantly improved.
RESUMO
BACKGROUND AND PURPOSE: Cinnamaldehyde, a major component of cinnamon, induces the generation of reactive oxygen species and exerts vasodilator and anticancer effects, but its short half-life limits its clinical use. The present experiments were designed to compare the acute relaxing properties of cinnamaldehyde with those of self-assembling polymer micelles either loaded with cinnamaldehyde or consisting of a polymeric prodrug [poly(cinnamaldehyde)] that incorporates the compound in its backbone. METHODS: Rings of porcine coronary arteries were contracted with the thromboxane A2 receptor agonist U46619 or 40 mM KCl, and changes in isometric tension were recorded. RESULTS: Cinnamaldehyde induced concentration-dependent but endothelium-independent, nitric oxide synthase (NOS)-independent, cyclooxygenase-independent, soluble guanylyl cyclase (sGC)-independent, calcium-activated potassium-independent, and TRPA1 channel-independent relaxations. Cinnamaldehyde also inhibited the contractions induced by 40 mM KCl Ca(2+) reintroduction in 40 mM KCl Ca(2+)-free solution or by the Ca(2+) channel opener Bay K8644. Cinnamaldehyde-loaded control micelles induced complete, partly endothelium-dependent relaxations sensitive to catalase and inhibitors of NOS or sGC, but not cyclooxygenase or TRPA1, channels. Cinnamaldehyde-loaded micelles also inhibited contractions induced by 40 mM KCl Ca(2+) reintroduction or Bay K8644. Poly(cinnamaldehyde) micelles induced only partial, endothelium-dependent relaxations that were reduced by inhibitors of NOS or sGC and by catalase and the antioxidant tiron, but not by indomethacin or TRPA1 channel blockers. CONCLUSION: The present findings demonstrate that cinnamaldehyde-loaded and poly(cinnamaldehyde) micelles possess vasodilator properties, but that the mechanism underlying the relaxation that they cause differs from that of cinnamaldehyde, and thus could be used both to relieve coronary vasospasm and for therapeutic drug delivery.
Assuntos
Acroleína/análogos & derivados , Cálcio/metabolismo , Vasos Coronários/fisiologia , Emulsões/química , Músculo Liso Vascular/fisiologia , Ácido Nítrico/química , Vasodilatação/fisiologia , Acroleína/administração & dosagem , Acroleína/química , Animais , Vasos Coronários/efeitos dos fármacos , Técnicas In Vitro , Micelas , Contração Muscular/efeitos dos fármacos , Contração Muscular/fisiologia , Músculo Liso Vascular/efeitos dos fármacos , Suínos , Vasodilatação/efeitos dos fármacos , Vasodilatadores/administração & dosagem , Vasodilatadores/farmacologiaRESUMO
The safe and effective intracellular delivery of nucleic acids remains the most challenging obstacle to the broad application of gene therapy in clinic. Endosomal escape of nucleic acids is also a major barrier for efficient gene delivery. Ketal linkage is known to readily cleave at the acidic pH of endosomal compartments. Here, we report ketal containing poly(ß-amino ester) (KPAE) as an acid-cleavable non-viral siRNA delivery system. KPAE efficiently condensed siRNA into nanocomplexes with a diameter of ≈ 150 nm, which are stable under neutral conditions but rapidly dissociate to release siRNA at acidic pH. KPAE had a buffering capacity due to the presence of secondary amines in its backbone, confirmed by acid-base titration. Moreover, the studies of confocal fluorescence imaging using calcein and LysoTracker Red revealed that KPAE disrupted endosomes by colloid osmotic mechanism and "proton sponge" effects. Cell culture studies demonstrated that KPAE can deliver tumor necrosis factor-α (TNF-α) siRNA to lipopolysaccharide (LPS)-stimulated macrophages and significantly inhibit the expression of TNF-α. The results demonstrate that acid-cleavable KPAE has great potential as gene delivery systems based on its excellent biocompatibility, pH sensitivity and high gene delivery efficiency.
Assuntos
Técnicas de Transferência de Genes , Polímeros/administração & dosagem , RNA Interferente Pequeno/administração & dosagem , Fator de Necrose Tumoral alfa/genética , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Células HEK293 , Humanos , Camundongos , Células NIH 3T3 , Polímeros/química , RNA Interferente Pequeno/químicaRESUMO
OBJECTIVE: This study is to evaluate the effect of laser activation on the whitening and crystalline structure of enamel surface during whitening treatment with hydrogen peroxide. METHODS: Bovine teeth were treated with whitening gel containing 35% hydrogen peroxide. A whitening gel was applied on the enamel surface for a period of 5 min, and then irradiated using a diode laser (740 nm) during whitening treatment for 0, 30, 60, 120 and 180s for the GL0-W, GL30-W, GL60-W, GL120-W and GL180-W groups, respectively. The total whitening application time was 30 min for all groups. RESULTS: Laser-irradiated enamel groups showed a similar lightness compared to the GL0-W group. The thickness of porous layer observed on the enamel surface of GL0-W group was decreased by increasing the laser irradiation time. While the Ca and P contents of the GL0-W group were lower than those of the non-whitening treated group (GL0-C), the Ca and P contents of the GL180-W group were similar to those of the GL180-C group. The enamel crystallinity was dramatically decreased by whitening treatment without laser irradiation. However, the decrease of crystallinity was protected by laser irradiation during whitening treatment. Raman measurement verified that laser irradiation could prevent the loss of mineral compositions on enamel and maintain its crystalline structure. SIGNIFICANCE: The professional whitening treatment with hydrogen peroxide and diode laser activation improves not only the whitening effect but also protects the change of enamel structure compared to the treatment with only gel.
Assuntos
Esmalte Dentário/efeitos da radiação , Peróxido de Hidrogênio/farmacologia , Lasers Semicondutores/uso terapêutico , Clareadores Dentários/farmacologia , Clareamento Dental/métodos , Desmineralização do Dente/prevenção & controle , Animais , Bovinos , Cristalização , Cristalografia por Raios X , Esmalte Dentário/química , Esmalte Dentário/efeitos dos fármacos , Relação Dose-Resposta à Radiação , Propriedades de SuperfícieRESUMO
One hundred Korean adults (50 men, 50 women) were scanned in the upright position using a cone-beam CT (CBCT) scanner. The soft tissue (ST) thicknesses were measured at 31 landmarks, 10 midline and 21 bilateral landmark sites, and the means and standard deviations were obtained for male and female subjects. While 18 of 31 landmarks showed sex differences, the majority showed higher values for male subjects with the exception of a few landmark sites corresponding to the zygoma area, which showed smaller values in men than in women. The mandibular area showed greater differences between the right and left sides. Overall, the ST thickness measurements obtained in this study can be used as a database for the forensic craniofacial reconstruction of Korean adult faces.
Assuntos
Povo Asiático , Bases de Dados Factuais , Face/anatomia & histologia , Face/diagnóstico por imagem , Adulto , Tomografia Computadorizada de Feixe Cônico , Feminino , Antropologia Forense , Humanos , Imageamento Tridimensional , Coreia (Geográfico) , Masculino , Conceitos Matemáticos , Valores de Referência , Caracteres SexuaisRESUMO
The development of biodegradable and biocompatible materials is the basis for tissue engineering and drug delivery. The aims of this study are to develop the poly(oxalate-co-oxamide) (POXAM) and evaluate its physicochemical properties and biocompatibility as the initial step for the development of new biomaterials. POXAM had a molecular weight of ~70,000 Da and rapidly degraded under physiological condition with a half-hydrolysis of ~4 days. POXAM films exhibited relative hydrophilic nature because of the presence of oxamide linkages and induced a higher cell attachment and proliferation compared with poly(lactic-co-glycolic acid) (PLGA) films. In vitro inflammatory responses to POXAM were evaluated using murine macrophage RAW 264.7 cells. POXAM films minimally stimulated the cells to generate less production of tumor necrosis factor-alpha (TNF-α) than PLGA films. We assessed the in vivo inflammatory responses to POXAM films implanted in the dorsal skin of rats. Histological studies revealed that POXAM provoked remarkably reduced inflammatory responses, evidenced by the less accumulation of inflammatory cells and giant cells, thinner fibrotic capsules, in comparison with PLGA. Given its excellent biocompatibility, fast degradation, and very mild inflammatory responses, POXAM has great potential for biomedical applications, such as scaffolds, wound dressing, and fast drug delivery.
Assuntos
Materiais Biocompatíveis/química , Materiais Biocompatíveis/metabolismo , Oxalatos/química , Oxalatos/metabolismo , Ácido Oxâmico/análogos & derivados , Animais , Linhagem Celular , Implantes Experimentais , Ácido Láctico/química , Ácido Láctico/metabolismo , Macrófagos/citologia , Macrófagos/imunologia , Teste de Materiais , Camundongos , Estrutura Molecular , Ácido Oxâmico/química , Ácido Oxâmico/metabolismo , Ácido Poliglicólico/química , Ácido Poliglicólico/metabolismo , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Ratos , Ratos WistarRESUMO
CONTEXT: A growing body of evidence shows that compounds of plant origin have the ability to prevent cancer. The fruit of gardenia, Gardenia jasminoides Ellis (Rubiaceae), has long been used as a food additive and herbal medicine, and its pharmacological actions, such as protective activity against oxidative damage, cytotoxic effect, and anti-inflammatory and anti-tumor activity, have already been reported. OBJECTIVE: The purpose of the present study was to investigate the presence of DNA topoisomerase 1 inhibitor in various solvent fractions of Gardenia extract and examine the induction of oral cancer cell death upon treatment with Gardenia extract. MATERIALS AND METHODS: The methanol extract of Gardenia was partitioned with n-hexane, dichloromethane, ethyl acetate, n-butanol, and water. RESULTS: In the DNA topoisomerase 1 assay, n-hexane and dichloromethane fractions inhibited topoisomerase 1 and led to a decrease in the cell viability of KB cells. The dichloromethane fraction (0.1 mg/mL) also showed 77% inhibition of cell viability in KB cells compared with HaCaT cells. Treatment with dichloromethane fraction led to apoptotic cell death as evidenced by flow cytometric analysis and morphological changes. In addition, treatment with Gardenia extract dichloromethane fraction led to the partial increase of caspase-3, caspase-8 and caspase-9 activities and the cleavage of poly (ADP-ribose) polymerase. CONCLUSION: Taken together, these results suggest that the dichloromethane fraction from Gardenia extract induces apoptotic cell death by DNA topoisomerase 1 inhibition in KB cells. These findings suggest the possibility that Gardenia extract could be developed as an anticancer modality.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Gardenia/química , Neoplasias Bucais/tratamento farmacológico , Extratos Vegetais/farmacologia , Antineoplásicos Fitogênicos/isolamento & purificação , Apoptose/efeitos dos fármacos , Caspases/efeitos dos fármacos , Caspases/metabolismo , Linhagem Celular , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , DNA Topoisomerases Tipo I/efeitos dos fármacos , Citometria de Fluxo , Frutas , Humanos , Células KB , Cloreto de Metileno/química , Neoplasias Bucais/patologia , Poli(ADP-Ribose) Polimerases/efeitos dos fármacos , Poli(ADP-Ribose) Polimerases/metabolismo , Solventes/químicaRESUMO
BACKGROUND: There is no accepted landmark for the mechanical axis of the femoral axis in sagittal plane in conventional total knee arthroplasty. METHODS: As palpable anatomic landmarks of the femur, lateral epicondyle, and anterior margin of the greater trochanter were identified. The line connecting these two landmarks was defined as the "palpable sagittal axis". The mechanical axis of the femur was compared with the palpable sagittal axis and the distal femoral anterior cortex axis. These axes were also compared with sagittal bowing of the femur. RESULTS: The distal femoral anterior cortex axis and the palpable sagittal axis were flexed by 4.1 degrees and 2.4 degrees more than the sagittal mechanical axes, respectively (p < 0.05). However, the palpable sagittal axis was not correlated with sagittal bowing of the femur (Spearman's rs, 0.17; p = 0.14). CONCLUSIONS: The palpable sagittal axis showed a consistent relationship with the sagittal mechanical femoral axes regardless of the severity of the sagittal bowing of the femur.
Assuntos
Artroplastia do Joelho/métodos , Fêmur/cirurgia , Idoso , Idoso de 80 Anos ou mais , Fenômenos Biomecânicos , Feminino , Fêmur/anatomia & histologia , Humanos , Articulação do Joelho/anatomia & histologia , Articulação do Joelho/cirurgia , Masculino , Pessoa de Meia-IdadeRESUMO
OBJECTIVE: The objective of this study is to investigate the effect of intracellular photosensitizer distribution on tumor cell death after photodynamic therapy (PDT). BACKGROUND DATA: The photosensitizer accumulates in tumor tissue during PDT, and generates intracellular reactive oxygen species (ROS), resulting in tumor cell death. MATERIALS AND METHODS: This study was carried out to elucidate the effects of PDT in a KB oral cancer cell line using hematoporphyrin with irradiation at 635 nm and 5 mW/cm(2). After irradiation, the MTT reduction method, agarose gel electrophoresis, flow cytometry, and Diff-Quick staining were performed. The intracellular ROS level was measured by DCF-DA. Intracellular hematoporphyrin was monitored with a confocal microscope, and Western blot and caspase activity assays were performed. RESULTS: In our study, cell survival was reduced by about 50% after 3 h of hematoporphyrin incubation time. In DNA fragmentation, flow cytometry, and Diff-Quick assay, necrosis was identified within 12 h and apoptosis soon thereafter. Confocal microscopy revealed that hematoporphyrin was localized in the cell membrane, cytoplasm, and nucleus as time passed. The quantities of intracellular ROS correlated with the time of hematoporphyrin accumulation. Additionally, Western blot analysis of Bcl-2/Bax, the release of cytochrome C, and activity of caspase-3 and caspase-9 showed that apoptosis followed the mitochondria-dependent pathway. CONCLUSION: PDT with hematoporphyrin in the KB cell line showed morphological changes of cell necrosis and apoptosis, which were associated with the time of distribution and localization of hematoporphyrin. Also, the apoptosis evoked followed the mitochondria-dependent pathway.