The role of Porphyromonas gingivalis (P. gingivalis) or its virulence factors, including lipopolysaccharide (LPS) not only has been related with periodontitis but also with endothelial dysfunction, a key mechanism involved in the genesis of atherosclerosis and hypertension that involving systemic inflammatory markers as angiotensin II (Ang II) and cytokines. This study compares the effect of repeated and unique exposures of P. gingivalis W83 LPS and live bacteria on the production and expression of inflammatory mediators and vasoconstrictor molecules with Ang II. Human coronary artery endothelial cells (HCAEC) were stimulated with purified LPS of P. gingivalis (1.0, 3.5 or 7.0 μg/mL) or serial dilutions of live bacteria (MOI 1: 100 - 1:0,1) at a single or repeated exposure for a time of 24 h. mRNA expression levels of AGTR1, AGTR2, IL-8, IL-1β and MCP-1 were determined by RT-qPCR, and IL-6, MCP-1, IL-8, IL-1β and GM-CSF levels were measured by flow cytometry, ELISA determined Ang II levels. Live bacteria in a single dose increased mRNA levels of AGTR1, and repeated doses increased mRNA levels of IL-8 and IL-1β (p < 0.05). Repeated exposure of live-P. gingivalis induced significant production IL-6, MCP-1 and GM-CSF (p < 0.05). Moreover, these MCP-1, IL-6 and GM-CSF levels were greater than in cells treated with single exposure (p < 0.05), The expression of AGTR1 and production of Ang II induced by live-P. gingivalis W83 showed a vasomotor effect of whole bacteria in HCAEC more than LPS. In conclusion, the findings of this study suggest that repeated exposure of P. gingivalis in HCAEC induces the activation of proinflammatory and vasoconstrictor molecules that lead to endothelial dysfunction being a key mechanism of the onset and progression of arterial hypertension and atherosclerosis.

Propolis extracts have been widely studied due to their popularity in traditional medicine, presenting incredible biodiversity. This study aimed to analyze propolis extracts’ phytochemical, physicochemical, and biological activities from four different biogeographic zones of the Huila region (Colombia). The raw material samples were collected by the scraping method and the ethanolic extracts (EEPs) were obtained by cold maceration with ethanol (96%). The physicochemical and sensory characterization was carried out according to the protocols recommended by the Brazilian Ministry of Agriculture and the main components of the EEPs were identified by LC-HRMS analysis. The determination of total phenols and flavonoids was carried out using colorimetric techniques. The antioxidant activity, cytotoxicity, and cell cycle regulation analyses in L929 and HGnF cells were evaluated using DPPH, Alamar Blue, and 7-amino actinomycin D (7-AAD) assays. The propolis samples presented an average yield of 33.1%, humidity between 1.6 and 2.8%, melting point between 54 and 62 °C, ashes between 1.40 and 2.19%, and waxes of 6.6–17.9%, respectively. The sensory characteristics of all samples were heterogeneous, complying with the quality specifications established by international standards. The polyphenolic and total flavonoid content was representative in the samples from Quebradon (255.9 ± 9.2 mg GAE/g, 543.1 ± 8.4 mg QE/g) and Arcadia (543.1 ± 8.4 mg GAE/g, 32.5 ± 1.18 g QE/g) (p < 0.05) that correlated with high antioxidant activity (Quebradon: 37.2 ± 1.2 µmol/g, Arcadia: 38.19 ± 0.7 µmol/g). In the chemical composition analysis, 19 compounds were characterized as phenolic acids and flavonoids, the most representative being chrysoeriol-O-methyl-ether, ellagic acid, and 3,4-O-dimethylcaffeic acid. Regarding biological activity, Quebradon and Arcadia propolis presented low toxicity with IC₅₀ of 2.83 ± 2.3 mg/mL and 4.28 ± 1.4 mg/mL in HGnF cells, respectively, and an arrest of the cell cycle in the G2/M phase of 71.6% and 50.8% compared to the control (11.9%) (p < 0.05). In general, the results of this study contribute to the identification of valid quality criteria to evaluate Colombian propolis, contributing to its study and chemical and biological characterization as a source of raw material for industrial and pharmaceutical use. In addition, Quebradon and Arcadia propolis can be important sources of bioactive molecules for the development of new drugs.

The in vitro antiproliferative activity of a phenolic-rich extract from Lycium barbarum fruits against head and neck HPV16 squamous cell carcinoma (OSCC) has been demonstrated, indicating for the first time that L. barbarum extract inhibits human papillomavirus (HPV) type 16 cell lines. Ethanol extract of L. barbarum was used for cell viability evaluation on SCC090, CAL27, and HGnF cell lines. After 24 and 48 h, the cell cycle effect of L. barbarum extract (at 1.0, 10, and 100 µg/mL) was measured via flow cytometry. In addition, the mRNA expression on E6/E7 and p53 via RT-PCR and the expression of p16, p53, Ki-67, and Bcl-2 via immunohistochemistry were also determined. Untreated cells, 20 µM cisplatin, and a Camellia sinensis-derived extract were used as negative and positive controls, respectively. We demonstrated that the studied L. barbarum extract resulted in G₀/G₁ arrest and S phase accumulation in SCC090 at 1.0 and 10 μg/mL. A reduction in mRNA levels of E6/E7 oncogenes (p < 0.05) with p53 overexpression was also observed through PCR, while immunohistochemical analyses indicated p16 overexpression (p > 0.05) and a decrease in p53 overexpression. The observed effects were associated with anticancer and immunomodulatory phenolics, such as flavonols/flavan-3-ols and tyramine-conjugated hydroxycinnamic acid amides, identified in the studied extract. These findings revealed that the phenolic-rich extract of L. barbarum fruits has promising properties to be considered further for developing new therapies against oral and oropharyngeal HPV lesions.

In atherosclerosis, the proliferation and migration of endothelial and smooth muscle cells (SMCs) and platelet activation alter endothelial function. Naturally occurring substances, such as caffeic acid (CA) and Trametes versicolor extract (TvE), have medicinal properties and are traditionally used for their antiproliferative, antioxidant, and anti-inflammatory effects. Electrospun 5% and 8% polycaprolactone-loaded CA or TvE was developed as a delivery system. Cytocompatibility was evaluated using human coronary artery endothelial cells (HCAECs), coronary artery SMCs (CASMCs), and platelets. Three types of systems (µF-CA, µF-TvE, and µF-CA/TvE) were developed and microscopically characterized. Analysis with scanning electron microscopy showed multidirectional fibers with diameters of 2–4.5 μm. The µF systems were hydrophobic and low cellular adhesion. The viability of CASMCs decreased with microfibers of 8% PCL and high CA concentration. However, the viability of CASMCs and HCAECs improved with 5% PCL and low CA concentration. Treatment with µF-TvE and µF-CA/TvE increased cell viability. HCAEC proliferation was affected by µF-CA, but incorporating TvE improved it. Platelet viability was unaffected by any µF system, but µF-CA and µF-CA/TvE inhibited the activation and adhesion of platelets. The results suggest that microfibers loaded with CA and TvE play a dual role in modifying HCAEC proliferation and blocking human platelet activation and adhesion. These findings have the potential to mitigate the atherosclerotic process.

Advanced periodontitis potentially contributes to Alzheimer's disease (AD) development and progression by altering the blood-brain barrier microenvironment in the cerebral microvascular endothelium. This results, in cytotoxicity, cell cycle disruption, and increased pro-inflammatory cytokine expression, allowing pathogens to enter the brain and damage the central nervous system (CNS). This study evaluated the effects of Porphyromonas gingivalis W83 infection on pro-inflammatory response, cell viability, and cell cycle regulation in mouse brain endothelial cells (mBECs). mBECs were stimulated with live P. gingivalis at different multiplicity of infection (MOI) values (1:5, 1:10, 1:50, 1:100, 1:200) for 6, 12, 24, and 48 h. Cell viability, cell cycle regulation, and pro-inflammatory cytokine mRNA expression were assessed using the alamarBlue assay, flow cytometry, and reverse transcription quantitative polymerase chain reaction (RT-qPCR), respectively. P. gingivalis reduced cell viability, induced morphological changes in mBECs by >50% after 48 h (p < 0.05) and caused concentration-dependent arrest in the S and G0/G1 phases of the cell cycle at MOI=1:100 and 1:200. The Il6, Il1b, and tumor necrosis factor alpha (Tnf) mRNA expression increased significantly compared to that of the controls (p < 0.05). P. gingivalis reduced cellular metabolism and induced early cell cycle arrest at the G0/G1 phase in MBEC cells. It also increased the pro-inflammatory response, which could be associated with cell death and possible senescence of brain endothelial cells. These results suggested a possible role for P. gingivalis in the pathogenesis of AD. Further studies are required to elucidate these underlying mechanisms.