Induction of cytotoxic effects and changes in DNA methylation-related gene expression in a human fibroblast cell line by the metal-organic framework [H2NMe2]3 [Tb(III)(2,6 pyridinedicarboxylate)3] (Tb-MOF).

Lanthanide metal-organic frameworks (lanthanide MOFs) may be utilized for a variety of environmental and human health applications due to their luminescent properties and high thermal and water stability. However, the cytotoxic and epigenetic effects produced in human cells are not known. Therefore, we evaluated the cytotoxic effects, internalization, and changes in the mRNA abundance of DNA methylation and demethylation enzymes by exposing human fibroblast cells to a metal-organic framework [H2NMe2]3 [Tb(III)(2,6 pyridinedicarboxylate)3] (Tb-MOF). For this purpose, the cells were exposed to six concentrations (0.05 to 1.6 mg/mL) of Tb-MOF for 48 h. Field emission electron microscopy coupled to linear energy dispersive spectroscopy (FESEM‒EDS) and confocal microscopy analysis were performed. The cytotoxicity was determined with crystal violet and MTT assays. The results demonstrated the internalization of Tb-MOF at concentrations as low as 0.05 mg/mL, as well as concentration-dependent toxicity. Additionally, we detected significant changes in the gene expression levels of DNA methyltransferases and demethylases due to the presence of Tb-MOF, suggesting that Tb-MOF could generate epigenetic changes even at low concentrations. The results of our study may establish a foundation for future research attempting to develop and apply secure nanomaterials (e.g., MOFs) to minimize damage to the environment and human health.

Salvia hispanica L. Seed Oil: Effect on Cell Viability in Colon Cancer Line Caco2 and Breast Cancer Line MCF7.

Cancer is one of the main causes of mortality and morbidity worldwide, which does not have a fully effective medical treatment. Therefore, the objective of this study was to evaluate the effect of lipidically characterized and hydrolyzed chia seed (Salvia hispanica L.) oil on in vitro colon cancer (Caco2 cell line) and breast cancer cells (MCF7 cell line) cell viability. Cancer cells were treated with different hydrolyzed oil concentrations (12.5-400 µg/mL) for 48 h; then cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. All chia seed oil concentrations significantly reduced cell viability in the Caco2 line, with the best reduction being 23.88% with the 25 µg/mL concentration. However, in the breast cancer line the highest concentrations (200 and 400 µg/mL) increased the viability of malignant cells. Chia seed oil significantly reduces the viability of Caco2 cells, although at the higher concentrations it can increase the viability of breast cancer cells.

Biocompatibility studies of polyurethane electrospun membranes based on arginine as chain extender.

Electrospun polymers are an example of multi-functional biomaterials that improve the material-cellular interaction and aimed at enhancing wound healing. The main objective of this work is to fabricate electrospun polyurethane membranes using arginine as chain extender (PUUR) in order to test the fibroblasts affinity and adhesion on the material and the polymer toxicity. Polyurethane membranes were prepared in two steps: (i) the polyurethane synthesis, and ii) the electrospinning process. The membranes were characterized by scanning electron microscopy (SEM), Fourier transforms infrared spectroscopy, gel permeation chromatography, and differential scanning calorimetry techniques. The evaluation of PUUR as a scaffolding biomaterial for growing and developing of cells on the material was realized by LIVE/DEAD staining. The results show that the fluorescent surface area of human fibroblasts (hFB), was greater in control dense membranes made from Tecoflex than in electrospun and dense PUUR. From SEM analysis, the electrospun membranes show relatively uniform attachment of cells with a well-spread shape, while Tecoflex dense membranes show a non-proliferating round shape, which is attributed to the fiber's structure in electrospun membranes. The cell morphology and the cell attachment assay results reveal the well spreading of hFB cells on the surface of electrospun PUUR membranes which indicates a good response related to cell adhesion.

Anticancer activity of protein fractions from chia (Salvia hispanica L.).

Salvia hispanica demonstrated to be a source of protein fractions with anticancer activity. The effect of the protein fractions <1, 1-3, and 3-5 kDa, obtained by ultrafiltration of the S. hispanica hydrolysate, was evaluated on the cellular viability of four cancer cell lines (MCF-7, Caco2, PC-3, and HepG2) and on human fibroblasts (hFB) at different concentrations (0.25, 0.5, 0.75, and 1 mg/ml). The protein fractions did not show cytotoxic effects on hFB. The protein fraction <1 kDa at 1 mg/ml showed the highest statistical effect on the cellular viability of all evaluated cancer lines; thus, its amino acid sequence was analyzed. From the multicriteria decision analysis of the peptide sequences obtained by mass spectrometry, the peptide KLKKNL with potential anticancer activity was selected. In conclusion, protein fractions could represent a therapeutic option for cancer treatment. However, further investigations are necessary to establish conclusive arguments. PRACTICAL APPLICATION: The work of this article is based on the background of the increasing potential of peptides for the treatment of chronic diseases. The results of this study present peptides of low molecular weight, obtained from chia seeds, as a potential adjuvant option for cancer treatment.

Increased Toxicity of Doxorubicin Encapsulated into pH-Responsive Poly(ß-Amino Ester)-Functionalized MCM-41 Silica Nanoparticles.

BACKGROUND: The encapsulation of anti-cancer drugs in stimulus-sensitive release systems may provide advantages such as enhanced drug toxicity in tumour tissue cells due to increased intracellular drug release. Encapsulation may also improve release in targeted tissue due to the response to a stimulus such as pH, which is lower in the tumour tissue microenvironment. Here, we evaluated the in vitro toxicity of the Drug Doxorubicin (DOX) loaded into a release system based on poly(ß-amino ester)- modified MCM-41 silica nanoparticles. METHODS: The MCM-41-DOX-PbAE release system was obtained by loading DOX into MCM-41 nanoparticles amino-functionalized with 3-aminopropyltriethoxysilane (APTES) and then coated with a pH-responsive poly(ß-amino ester) (PbAE). The physicochemical characteristics of the release system were evaluated through TEM, FTIR and TGA. Cytotoxicity assays were performed on the MCM-41- DOX-PbAE system to determine their effects on the inhibition of human MCF-7 breast cancer cell proliferation after 48 h of exposure through crystal violet assay; the investigated systems included MCF-7 cells with MCM-41, PbAE, and MCM-41-PbAE alone. Additionally, the release of DOX and the change in pH in vitro were determined. RESULTS: The physicochemical characteristics of the synthesized MCM-41-PbAE system were confirmed, including the nanoparticle size, spherical morphology, mesoporous ordered structure, and presence of PbAE on the surface of the MCM-41 nanoparticles. Likewise, we demonstrated that the release of DOX from the MCM-41-DOX-PbAE system promoted an important reduction in MCF-7 cell viability (~ 70%) compared to the values obtained with MCM-41, PbAE, and MCM-41-PbAE, as well as a reduction in the viability under treatment with just DOX (~ 50%). CONCLUSION: The results suggest that all the components of the release system are biocompatible and that the encapsulation of DOX in MCM-41-PbAE could allow better intracellular release, which would probably increase the availability and toxic effect of DOX.

Using simple models to describe the kinetics of growth, glucose consumption, and monoclonal antibody formation in naive and infliximab producer CHO cells.

Despite their practical and commercial relevance, there are few reports on the kinetics of growth and production of Chinese hamster ovary (CHO) cells-the most frequently used host for the industrial production of therapeutic proteins. We characterize the kinetics of cell growth, substrate consumption, and product formation in naive and monoclonal antibody (mAb) producing recombinant CHO cells. Culture experiments were performed in 125 mL shake flasks on commercial culture medium (CD Opti CHO™ Invitrogen, Carlsbad, CA, USA) diluted to different glucose concentrations (1.2-4.8 g/L). The time evolution of cell, glucose, lactic acid concentration and monoclonal antibody concentrations was monitored on a daily basis for mAb-producing cultures and their naive counterparts. The time series were differentiated to calculate the corresponding kinetic rates (rx = d[X]/dt; rs = d[S]/dt; rp = d[mAb]/dt). Results showed that these cell lines could be modeled by Monod-like kinetics if a threshold substrate concentration value of [S]t = 0.58 g/L (for recombinant cells) and [S]t = 0.96 g/L (for naïve cells), below which growth is not observed, was considered. A set of values for µmax, and Ks was determined for naive and recombinant cell cultures cultured at 33 and 37 °C. The yield coefficient (Yx/s) was observed to be a function of substrate concentration, with values in the range of 0.27-1.08 × 10(7) cell/mL and 0.72-2.79 × 10(6) cells/mL for naive and recombinant cultures, respectively. The kinetics of mAb production can be described by a Luedeking-Piret model (d[mAb]/dt = αd[X]/dt + ß[X]) with values of α = 7.65 × 10(-7) µg/cell and ß = 7.68 × 10(-8) µg/cell/h for cultures conducted in batch-agitated flasks and batch and instrumented bioreactors operated in batch and fed-batch mode.

Hydrodynamic conditions induce changes in secretion level and glycosylation patterns of Von Willebrand factor (vWF) in endothelial cells.

The effect of low-speed agitation on the level of secretion and glycosylation patterns of the Von Willebrand factor in endothelial cells cultured in 6-cm Petri dishes located in a rotating platform was studied. In this simple system, complex and non-intuitive velocity fields arise. As revealed by simple calculations of tangential velocities and CFD analysis, flow behavior strongly varies as a function of radial distance and rotational velocity. We found significant differences in both secretion level and glycosylation extent and completeness, as a result of relatively small changes in rotational speed. Results obtained support the hypothesis that differences in hydrodynamic conditions may induce important changes in the secretion and assembly of glycoproteins in mammalian cell cultures.