Cannabis smoke condensate induces human gingival epithelial cell damage through apoptosis, autophagy, and oxidative stress.

OBJECTIVES: This study aims to investigate the effects of cannabis smoke condensate (CSC) on the adhesion, growth, and signaling pathways of human gingival epithelial cells. DESIGN: The effects of CSC on cell shape and adhesion, and viability were evaluated after 30 min, 60 min, 2 h, and 24 h of exposure using microscopic observation, cell metabolic activity, and lactate dehydrogenase activity assays. The effects of CSC on cell apoptosis, necrosis, autophagy, and oxidative stress were determined through flow cytometry, while apoptotic and autophagic gene expression were identified via an RT2-PCR array. Phosphorylated signaling pathway proteins were measured using flow cytometry. RESULTS: CSC deregulated gingival epithelial cell shape and adhesion, decreased cell viability, and increased lactate dehydrogenase release. Its toxic effects included apoptosis, autophagy, and oxidative stress. Moreover, it modulated seven specific apoptotic and six autophagic genes. Furthermore, it decreased phosphorylation in signaling proteins, such as STAT5, ERK12, P38, and nuclear factor κB. CONCLUSIONS: CSC has notable adverse effects on gingival epithelial cells. This finding indicates that cannabis smoke could impair gingival epithelial cell innate immune function, leading to gingivitis and periodontitis. Oral health professionals may need to document observed modifications in the oral cavity of patients who smoke cannabis and consider these potential changes during clinical care.

Effect of Cannabis Smoke Condensate on C. albicans Growth and Biofilm Formation.

The most common use of cannabis is smoking. The oral ecosystem, among other constituents, can be deregulated by the presence of cannabis smoke in the oral cavity. We evaluated the effect of cannabis smoke condensate (CSC) on the behavior of Candida albicans, a common yeast found in the oral cavity. The yeast was first cultured with different concentrations of CSC, and its growth was evaluated. The transition from the blastospore to the hyphal form and the hyphae size were assessed after 3 and 6 h, along with biofilm formation after 72 h of contact with CSC. The response of C. albicans to oxidative (H2O2) stress was also examined. Our results show that CSC contained high amounts of THC (about 1055 ppm), CBN (63 ppm), and CBG (about 47 ppm). The presence of various concentrations of CSC in the culture medium increased C. albicans growth. CSC also contributed to increases in both the hyphal length and biofilm mass. Following oxidative stress (H2O2 at either 100 or 500 µM), CSC prevented the damaging effect of H2O2 on both C. albicans shape and growth. These findings support clinical observations demonstrating that cannabis may promote C. albicans growth and oral candidiasis.

Production of biocompatible and antimicrobial bacterial cellulose polymers functionalized by RGDC grafting groups and gentamicin.

Bacterial cellulose (BC), a three-dimensional fibril, is a natural polymer that can be used for many applications. BC effectiveness may be improved by enhancing surface characteristics contributing to a better physiologic interaction with human and animal cells and to intrinsically present antimicrobial agents. In the present study, gentamicin-activated BC membranes were obtained by chemically grafting RGDC peptides (R: arginine; G: glycine; D: aspartic acid; C: cysteine) using coupling agent 3-aminopropyltriethoxysilane (APTES) followed by covalent attachment of gentamicin onto the surface of the BC membrane network. X-ray photoelectron spectroscopy (XPS) analyses showed that the BC-APTES contained 0.7% of silicon in terms of elemental composition, corresponding to a grafting ratio of 1:12. The presence of silicon and nitrogen in the BC-APTES confirmed the surface functionalization of the BC membrane. Fourier-transform infrared (FTIR) analyses show the formation of the secondary amide as supported by the valence bond C═O (ν(C═O)), a characteristic vibrational transition at 1650 cm(-1) which is particularly intense with the BC-RGDC-gentamicin membrane. Energy-dispersive X-ray (EDX) analyses showed a low level of carbon and nitrogen (C + N) in pure BC but a high level of (C + N) in BC-RGDC-gentamicin confirming the surface modification of the BC membrane by RGDC and gentamicin enrichment. Of great interest, the gentamicin-RGDC-grafted BC membranes are bactericidal against Streptococcus mutans but nontoxic to human dermal fibroblasts and thus may be useful for multiple applications such as improved wound healing and drug delivery systems.

Hydroxyapatite bioactivated bacterial cellulose promotes osteoblast growth and the formation of bone nodules.

The goal of this study was to investigate the feasibility of bacterial cellulose (BC) scaffold to support osteoblast growth and bone formation. BC was produced by culturing Acetobacter xylinum supplemented with hydroxyapatite (HA) to form BC membranes (without HA) and BC/HA membranes. Membranes were subjected to X-ray photoelectron spectroscopy (XPS) analysis to determine surface element composition. The membranes were further used to evaluate osteoblast growth, alkaline phosphatase activity and bone nodule formation. BC was free of calcium and phosphate. However, XPS analysis revealed the presence of both calcium (10%) and phosphate (10%) at the surface of the BC/HA membrane. Osteoblast culture showed that BC alone was non-toxic and could sustain osteoblast adhesion. Furthermore, osteoblast adhesion and growth were significantly (p ≤0.05) increased on BC/HA membranes as compared to BC alone. Both BC and BC/HA membranes improved osteoconductivity, as confirmed by the level of alkaline phosphatase (ALP) activity that increased from 2.5 mM with BC alone to 5.3 mM with BC/HA. BC/HA membranes also showed greater nodule formation and mineralization than the BC membrane alone. This was confirmed by Alizarin red staining (ARS) and energy dispersive X-ray spectroscopy (EDX). This work demonstrates that both BC and BC/HA may be useful in bone tissue engineering.

Farnesol, a fungal quorum-sensing molecule triggers Candida albicans morphological changes by downregulating the expression of different secreted aspartyl proteinase genes.

The aim of this study was to determine the effect of exogenous farnesol in yeast-to-hyphae morphogenesis, and Saps (2, 4, 5 and 6) mRNA expressions by a Candida strain that does not produce endogenous farnesol. C. albicans was cultured in the absence and presence of farnesol at various concentrations (10, 100, and 300 µM), in proteinase induction medium, and then used to determine yeast-to- hyphae changes, Candida ultrastructure and to determine Saps 2, 4, 5 and 6 expressions using q-TR-PCR and ELISA (for Sap2). Data demonstrated that farnesol greatly reduced the yeast-to-hyphae morphogenesis of a Candida strain that does not produce endogenous farnesol. Farnesol induced several ultrastructural alterations, including changes in the cell-wall shape, a visible disconnection between the cell wall and cytoplasm with an electron-lucent zone between them, and the presence of electron-dense vacuoles. Tested on gene expressions, farnesol was able to significantly (p < 0.01) decrease Sap2 secretion and mRNA expression. Farnesol downregulated also Sap4-6 mRNA expression. These results demonstrated for the first time that farnesol modules Candida morphogenesis through a downregulation of Saps 2, 4, 5 and 6 expressions. Overall these data point to the potential use of farnesol as an antifungal molecule.