Photodynamic therapy provides the formation of reactive oxygen species that are capable of inducing cell death. Human laryngeal carcinoma (HEp-2) cells have been evaluated in this study under PDT treatment. Cells were treated with photosensitizer aluminum phthalocyanine tetrasulfonate (AlPcS4) and irradiated with a Biopdi/Irrad-Led5 660 LED with 660 nm wavelength, intensity of delivered light of 25 mW/cm2, power of 70 mW, fluence of 5 J/cm2for 24 h and 48 h, and then evaluated. Cell population was not increased by PDT treatment after the tested period. The apoptosis assay demonstrated that control groups exhibit approximately 60% of living cells in the 24 h and 48 h periods, however. A significant increase in apoptotic cells was observed after the photodynamic therapy treatments, for both 24 and 48 h groups. Over 50% of cells were under apoptosis after photodynamic therapy, evidencing a death process generated from the oxidative damage of the treatment. Comet assay and micronucleus assessments, both of which evaluate genotoxicity, demonstrated favorable results to damages caused by the photodynamic therapy treatment. Thus, photodynamic therapy is proposed to damage nuclear cells and the subcellular structure of carcinogenic cells.Impact statement: Recently, the use of photodynamic therapy grows as an alternative treatment for cancer, since it has a noninvasive characteristic and affinity to the tumor tissue. Accordingly, understanding the therapy’s foci of action is important for the technique improvement. This work aims to understand the genotoxic effect triggered by the therapy action, thus evidencing the permanent changes caused to the genetic material of the tumor cell after the treatment. Therefore, to increase the knowledge in this study field, the methodology of the comet assay and count of micronucleus formed after the therapy was adopted in order to understand if the damage caused to the DNA of tumor cell makes its replication process unfeasible in future generations. The study allows a better therapeutic approach to the cancer treatment, making the process of association between therapies a more effective option during the disease treatment.
This study evaluates the antimicrobial activity of the glycolic extract of G. sylvestre against anaerobic pathogens, along with its cytotoxicity, genotoxicity, anti-inflammatory activity, antioxidant effects, and phytochemical composition. Phytochemical analysis was conducted using high-performance liquid chromatography and liquid chromatography–mass spectrometry, while the antioxidant effect was assessed through a DPPH assay. Antimicrobial action was tested on planktonic cultures and biofilms of Porphyromonas gingivalis, Porphyromonas endodontalis, Parvimonas micra, and Fusobacterium nucleatum. Cytotoxicity was evaluated using mouse macrophages (RAW 264.7), rat fibroblasts (L929), and human keratinocytes (HaCaT). Anti-inflammatory effects were measured by an immunoenzymatic assay (ELISA) on RAW 264.7 cells. Statistical analysis was performed using a one-way ANOVA and Tukey’s test. Phytochemical analysis revealed the presence of phenolic compounds and flavonoids. The extract demonstrated a reduction of over 95% in biofilms of P. gingivalis, P. micra, and F. nucleatum within 5 min of treatment. Cell viability (HaCaT) remained above 80%. Antioxidant activity showed an EC50 of 353.43 µg/mL, achieving a 50% reduction in free radicals. A significant decrease in TNF-α (a pro-inflammatory cytokine) and an increase in IL-10 (an anti-inflammatory cytokine) were observed. In conclusion, the extract of G. sylvestre exhibits promising potential as a therapeutic agent for treating anaerobic infections, inflammation, and oxidative stress.
Proteínas de choque térmico (HSP) são moléculas intracelulares multifuncionais que, eventualmente, podem estar envolvidas na malignização celular. Terapia fotodinâmica (TFD) pode levar à redução do tumor e vasos sanguíneos ao redor. Objetivo foi avaliar ação da TFD sobre as HSPs em células neoplásicas de carcinoma de laringe humana (HEp-2). As células foram irradiadas com LED a 660 nm, 5 J/cm2, 70 mW, por 3 minutos e 20 segundos; incubadas por períodos de 24, 48 e 72 horas; após os períodos de incubação foi realizada a extração de proteínas e corrido gel de poliacrilamida para avaliação das proteínas por Western-Blotting. As células foram imunomarcadas com anticorpos anti-HSP27, anti-HSP70 e anti-HSP90 e analisadas no microscópio confocal. A viabilidade celular foi avaliada pelo teste de cristal violeta. No gel de poliacrilamida foram identificadas bandas próximas a 27 kDa, 70 kDa e 90 kDa. Nas fotomicrografias observou-se redução do número de células após TFD, comprovado por teste de viabilidade (cristal violeta); e intensa marcação de HSPs após TFD, principalmente próximas ao núcleo. Concluiu-se que HSP27, HSP70 e HSP90 são muito produzidas em células HEp-2. TFD foi eficaz devido à redução do número de células e considerada opção viável para o tratamento dessa doença. Embora seja relatada a participação das HSPs 27, 70 e 90 como protetoras das células tumorais, nossos resultados indicam que a TFD ativa tais proteínas para a redução das células tumorais.
Background/Objectives: Colorectal neoplasms rank as the third most prevalent cancer globally and stand as the second leading cause of cancer-related mortality. Its etiology is multifaceted, pointing to the role of microorganisms within the human microbiota in its development. Notably, the high prevalence of oral pathogens like Fusobacterium nucleatum and Parvimonas micra is implicated in inducing gut dysbiosis and stimulating the proliferation and metastasis of cancer cells. Therefore, this study aimed to evaluate in vitro the biological effects of extracts from Juglans regia and Pfaffia paniculata. Methods: Phytochemical analysis was carried out by HPLC, and the antioxidant effect was determined by DPPH. Antimicrobial activity was investigated on F. nucleatum and P. micra planktonic and biofilms. Metabolic activity and genotoxicity were performed. Results:J. regia and P. paniculata expressed CE50 37.26 and 1367.57 mcg, respectively. The extracts exhibited a minimum bactericidal concentration of 1.73 and 0.48 mg/mL for J. regia and P. paniculata, respectively. Reduction superiorly 90% of P. micra biofilms. Metabolic activity was varied proportionally to the extract concentration, and no genotoxic effects were observed. Conclusions: The J. regia extract has great antioxidant activity and could be used as an alternative in combating pathogens associated with the onset of dysbiosis and tumor progression in colorectal neoplasms. Nevertheless, further studies are needed to validate their clinical applicability.
6 February 2022 in Research, Society and Development
Introduction: Cancer is one of the diseases with the highest incidence globally and that associated with the patient's emotional state, can act positively or negatively in the treatment. Cortisol is a principal primary stress hormone in the human body. The corticoids can increase cell proliferation and reactive oxygen species that contribute to DNA damage. Prolonged exposure to stress can contribute to tissues becoming insensitive to cortisol, the primary human stress hormone. Objective: This study explores cortisol's influence on tumor cell development, particularly in human cells of carcinoma of the human laryngeal (HEp-2). Methodology: HEp-2 cells were exposed to increasing cortisol (hydrocortisone) concentrations for 24 or 48 hours, and cytotoxicity (MTT assay) proliferation assay (crystal violet assay), and immunolabeled 3D culture for fibronectin and FAK were analyzed. Results: The group treated with hydrocortisone showed a significant increase in mitochondrial activity, as for the evaluation by the violet crystal, the treated group showed similar behavior to the control. The 3D culture showed dispersed cells within 24 hours with reduced FAK labeling; however, no changes were observed within 48 hours. Conclusion: Although some cases favored corticosteroid use in cancer patients, a more detailed analysis is necessary before prescribing them.