A unique Thy-1-negative immuno-fibroblast population emerges as a key determinant of fibrotic outcomes to biomaterials

Microporous annealed particle (MAP) hydrogels are an exciting new development in biomaterial design. They regulate innate and acquired immunity which has been linked to their ability to evade normal host-material fibrosis. Yet, resident stromal fibroblasts, not immune cells, are the arbiters of the extracellular matrix assembly that characterizes fibrosis. In other idiopathic fibrotic disorders, a fibroblast subpopulation defined by its loss of cell surface Thy-1 expression is strongly correlated with degree of fibrosis. We have previously shown that Thy-1 is a critical αvβ3 integrin regulator that enables normal fibroblast mechanosensing and here, leveraging non-fibrosing MAP gels, we demonstrate that Thy-1-/- mice mount a robust response to MAP gels that remarkably resembles a classical foreign body response. We further find that within the naive, Thy-1+ fibroblast population exists a distinct and cryptic αSMA+ Thy-1- population that emerges in response to IL-1β and TNFα. Employing single-cell RNA sequencing, we find that IL-1β/TNFα-induced Thy-1- fibroblasts actually consist of two distinct subpopulations, both of which are strongly pro-inflammatory. These findings illustrate the emergence of a unique pro-inflammatory, pro-fibrotic fibroblast subpopulation that is central to material-associated fibrosis likely through amplifying local inflammatory signaling.

Growth Inhibition and Apoptotic Effect of Pine Extract and Abietic Acid on MCF-7 Breast Cancer Cells via Alteration of Multiple Gene Expressions Using In Vitro Approach

In vitro anti-proliferative activity of Pinus palustris extract and its purified abietic acid was assessed against different human cancer cell lines (HepG-2, MCF-7 and HCT-116) compared to normal WI-38 cell line. Abietic acid showed more promising IC50 values against MCF-7 cells than pine extract (0.06 µg/mL and 0.11 µM, respectively), with insignificant cytotoxicity toward normal fibroblast WI-38 cells. Abietic acid triggered both G2/M cell arrest and subG0-G1 subpopulation in MCF-7, compared to SubG0-G1 subpopulation arrest only for the extract. It also induced overexpression of key apoptotic genes (Fas, FasL, Casp3, Casp8, Cyt-C and Bax) and downregulation of both proliferation (VEGF, IGFR1, TGF-β) and oncogenic (C-myc and NF-κB) genes. Additionally, abietic acid induced overexpression of cytochrome-C protein. Furthermore, it increased levels of total antioxidants to diminish carcinogenesis and chemotherapy resistance. P. palustris is a valuable source of active abietic acid, an antiproliferative agent to MCF-7 cells through induction of apoptosis with promising future anticancer agency in breast cancer therapy.

Biological activity and its related compounds of Red Jasmine rice extracts linked to normal fibroblast viability for cosmetic product

Red jasmine rice is recognized as a healthy food with high phenolic compounds. These compounds present antibacterial and anti-free radical properties. Moreover, colored rice exhibits a biological activity against anticancer. Objectives of this study are 1) exploring a biological screening and cell viability of 70% ethanol and aqueous extracts of red jasmine rice, 2) investigating cytotoxicity to fibroblast NIH3T3 (IC80) that is one hundred cells were found cell viability 80 cells. Red jasmine rice extracts were dried and transformed into a powder using the freeze-drying method. The extracts were treated with fibroblast NIH3T3 for MTT. The highest of IC50 of red jasmine rice extract to scavenge the DPPH and ABTS radicals was found in ethanol extract (53.20±7.37 and 64.17±5.76, respectively). The experiment showed that the ethanol and aqueous extract of red rice did not show cytotoxicity to fibroblast NIH3T3 (IC80). The extracts of red rice show the biological screening of anti-oxidation with total phenolic compounds and flavonoid contents. Moreover, it does not modify the physical properties of the cream formula. It can be concluded that the red rice extract is highly promising for the value addition.

Anticancer Activity of Moringa peregrina (Forssk.) Fiori.: A Native Plant in Traditional Herbal Medicine of the United Arab Emirates

Moringa peregrina (Forssk.) Fiori. is a native desert tree growing in United Arab Emirates (UAE). The plant is being cultivated in many parts of UAE, owing to its uses in traditional medicinal and food systems. In the present study bioactivities of cultivated M. peregrina species samples are evaluated with cytotoxic studies in the human breast cancer cell line (MCF-7) and human colon adenocarcinoma cell line (Caco-2). Different extracts with hexane, chloroform, acetone and methanol were prepared from tubers, leaves and stem of M. peregrina for estimating their antioxidant contents and anticancer activities. The study was performed at different concentrations and all the extracts showed dose-depended response on both the cell lines. Among the extracts tested, the chloroform extract of stem showed remarkable anti-proliferative/cell death activity (IC50 = 45.53 µg/mL of 48 h incubation and 33.32 µg/mL of 72 h incubation) on MCF-7 cell lines. Whereas the same extract showed comparatively less activity (IC50 = 93.75 µg/mL of 48 h incubation and 87.76 µg/mL of 72 h incubation) on Caco-2 cell lines. The anti-proliferative effect of leaf extract with chloroform showed a drastic change in cell viability from 48 to 72 h incubation, in MCF-7 cells 220 to 87.5 µg/mL and in Caco-2 cells 500.9 to 72.9 µg/mL, respectively. Moreover, less than 200 µg/mL of IC50 values reported in hexane extracts of tubers (188.6 µg/mL for 48 h and 164.3 µg/mL for 72 h), acetone extracts of tubers (167.4 µg/mL for 72 h) and acetone extracts of stem (171.5 µg/mL for 48 h and 101.7 µg/mL for 72 h) on MCF-7 cells. PARP (Poly (ADP-ribose) polymerase) cleavage assay and DNA fragmentation assay performed to understand the cause of cell death. Treatment of extract on the normal fibroblast cell line required more concentration for cytotoxicity compared to the treatment on the cancer cells. This ability of the extract proved the anti-cancer property of the M. peregrina extract from the stem, tuber and leaves. The information provided in the present study enables further studies on the isolation and characterization of an anticancer molecule from the tubers of M. peregrina.

Production and characterization of biocompatible nanofibrous scaffolds made of β-sitosterol loaded polyvinyl alcohol/tragacanth gum composites

Electrospun polyvinyl alcohol and Tragacanth Gum were used to develop nanofibrous scaffolds containing poorly water-soluble beta-sitosterol. Different Concentration and Ratio of Polymeric composite: (10%) of β-S concentration in (PVA) 8 %, (TG) 0.5%, and 1% respectively were added, prepared and electrospun. The methods have included four parameters (Solution concentration, feeding rate, voltage, and distance of the collector to the tip of the needle) for designing and compared the nanofibers' average diameters. The nanofibers collected were identified via SEM, FTIR, and XRD measurements. A contact angle measurement described the hydrophilicity of the scaffold. MTT test was assessed for obtained nanofibers by using L929 normal fibroblast cells. The %age of mechanical strength, porosity, and deterioration of the scaffolds was well discussed. The average nanofibre ranged from 63 ± 20 nm to 97 ± 46 nm in diameters. The nanofibers loaded with β-S were freely soluble in water and displayed a short release lag time. The dissolution was related to an immediate dissolution, submicron-level recrystallization of β-S with sufficient conditions for nanofibers for L929 cell culture that could be used in biomedical applications. It concluded that electrospinning is a promising technique for poorly water-soluble β-S formulations that could be used in biomedical applications.

Topological Analysis of γH2AX and MRE11 Clusters Detected by Localization Microscopy during X-ray-Induced DNA Double-Strand Break Repair

DNA double-strand breaks (DSBs), known as the most severe damage in chromatin, were induced in breast cancer cells and normal skin fibroblasts by 2 Gy ionizing photon radiation. In response to DSB induction, phosphorylation of the histone variant H2AX to γH2AX was observed in the form of foci visualized by specific antibodies. By means of super-resolution single-molecule localization microscopy (SMLM), it has been recently shown in a first article about these data that these foci can be separated into clusters of about the same size (diameter ~400 nm). The number of clusters increased with the dose applied and decreased with the repair time. It has also been shown that during the repair period, antibody-labeled MRE11 clusters of about half of the γH2AX cluster diameter were formed inside several γH2AX clusters. MRE11 is part of the MRE11–RAD50–NBS1 (MRN) complex, which is known as a DNA strand resection and broken-end bridging component in homologous recombination repair (HRR) and alternative non-homologous end joining (a-NHEJ). This article is a follow-up of the former ones applying novel procedures of mathematics (topology) and similarity measurements on the data set: to obtain a measure for cluster shape and shape similarities, topological quantifications employing persistent homology were calculated and compared. In addition, based on our findings that γH2AX clusters associated with heterochromatin show a high degree of similarity independently of dose and repair time, these earlier published topological analyses and similarity calculations comparing repair foci within individual cells were extended by topological data averaging (2nd-generation heatmaps) over all cells analyzed at a given repair time point; thereby, the two dimensions (0 and 1) expressed by components and holes were studied separately. Finally, these mean value heatmaps were averaged, in addition. For γH2AX clusters, in both normal fibroblast and MCF-7 cancer cell lines, an increased similarity was found at early time points (up to 60 min) after irradiation for both components and holes of clusters. In contrast, for MRE11, the peak in similarity was found at later time points (2 h up to 48 h) after irradiation. In general, the normal fibroblasts showed quicker phosphorylation of H2AX and recruitment of MRE11 to γH2AX clusters compared to breast cancer cells and a shorter time interval of increased similarity for γH2AX clusters. γH2AX foci and randomly distributed MRE11 molecules naturally occurring in non-irradiated control cells did not show any significant topological similarity.

Green Fabrication of Zinc Oxide Nanoparticles Using Phlomis Leaf Extract: Characterization and In Vitro Evaluation of Cytotoxicity and Antibacterial Properties

Green nanoparticle synthesis is an environmentally friendly approach that uses natural solvents. It is preferred over chemical and physical techniques due to the time and energy savings. This study aimed to synthesize zinc oxide nanoparticles (ZnO NPs) through a green method that used Phlomis leaf extract as an effective reducing agent. The synthesis and characterization of ZnO NPs were confirmed by UV-Vis spectrophotometry, Fourier Transform Infrared Spectroscopy (FTIR), X-Ray Diffraction (XRD), Dynamic light scattering (DLS), Zeta potential, and Field Emission Scanning Electron Microscope (FESEM) techniques. In vitro cytotoxicity was determined in L929 normal fibroblast cells using MTT assay. The antibacterial activity of ZnO nanoparticles was investigated using a disk-diffusion method against S. aureus and E. coli, as well as minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) content concentrations. XRD results confirmed the nanoparticles’ crystalline structure. Nanoparticle sizes were found to be around 79 nm by FESEM, whereas the hydrodynamic radius of nanoparticles was estimated to be around 165 3 nm by DLS. FTIR spectra revealed the formation of ZnO bonding and surfactant molecule adsorption on the surface of ZnO NPs. It is interesting to observe that aqueous extracts of phlomis leave plant are efficient reducing agents for green synthesis of ZnO NPs in vitro, with no cytotoxic effect on L929 normal cells and a significant impact on the bacteria tested.

Erlotinib Promotes Ligand-Induced EGFR Degradation in 3D but Not 2D Cultures of Pancreatic Ductal Adenocarcinoma Cells

The epithelial growth factor receptor (EGFR) is a tyrosine kinase receptor that participates in many biological processes such as cell proliferation. In addition, EGFR is overexpressed in many epithelial cancers and therefore is a target for cancer therapy. Moreover, EGFR responds to lots of stimuli by internalizing into endosomes from where it can be recycled to the membrane or further sorted into lysosomes where it undergoes degradation. Two-dimensional cell cultures have been classically used to study EGFR trafficking mechanisms in cancer cells. However, it has been widely demonstrated that in 2D cultures cells are exposed to a non-physiological environment as compared to 3D cultures that provide the normal cellular conformation, matrix dimensionality and stiffness, as well as molecular gradients. Therefore, the microenvironment of solid tumors is better recreated in 3D culture models, and this is why they are becoming a more physiological alternative to study cancer physiology. Here, we develop a new model of EGFR internalization and degradation upon erlotinib treatment in pancreatic ductal adenocarcinoma (PDAC) cells cultured in a 3D self-assembling peptide scaffold. In this work, we show that treatment with the tyrosine kinase inhibitor erlotinib promotes EGFR degradation in 3D cultures of PDAC cell lines but not in 2D cultures. We also show that this receptor degradation does not occur in normal fibroblast cells, regardless of culture dimensionality. In conclusion, we demonstrate not only that erlotinib has a distinct effect on tumor and normal cells but also that pancreatic ductal adenocarcinoma cells respond differently to drug treatment when cultured in a 3D microenvironment. This study highlights the importance of culture systems that can more accurately mimic the in vivo tumor physiology.