Fibroblast Cell Line Research Articles

Preparation, characterization, and in vitro evaluation of the anticancer activity of Ce3+ doped CuFe2O4 spinel nanoparticles in MCF-7 cell lines

In the present study, we successfully prepared nanoparticles of Ce-doped Copper ferrite, CuCexFe2-xO4(x = 0.00, 0.03, and 0.05), in the nanoscale using the chemical co-precipitation technique. The structure, composition, optical properties, and morphological information of the obtained ferrite nanoparticles (NPs) were analyzed by making use of powder X-ray diffraction (PXRD) pattern, Field Emission Scanning Electron Microscopic (FE-SEM) studies, Fourier Transform Infrared Spectroscopic (FT-IR) analysis, and Energy-dispersive X-Ray Spectroscopy (EDS), and the UV–Vis-NIR Spectroscopy. The cytotoxicity of CuCexFe2-xO4 nanoparticles (NPs) on the MCF-7 breast cancer cell line was evaluated using the MTT assay. The NPs exhibited dose-dependent cytotoxicity, with lower concentrations (<25 µg/mL) being safe and biocompatible, while higher concentrations reduced cell viability and disrupted L929 fibroblast cell lines. Double staining and flow cytometry experiments revealed that apoptosis is one of the mechanisms by which CuCexFe2-xO4 NPs induce cytotoxicity in MCF-7 cells. An increase in reactive oxygen species (ROS) was found in the presence NPs. To harness the full potential of this nanoparticle formulation, it is recommended that future investigations encompass a wide range of in vitro and in vivo models. These subsequent studies will significantly contribute to the expanding body of knowledge concerning the synthesis of composite nanoparticle formulations for use as anticancer agents.

Multimodal assessment of mitochondrial function in Parkinson's disease.

The heterogenous aetiology of Parkinson's disease is increasingly recognized; both mitochondrial and lysosomal dysfunction have been implicated. Powerful, clinically applicable tools are required to enable mechanistic stratification for future precision medicine approaches. The aim of this study was to characterize bioenergetic dysfunction in Parkinson's disease by applying a multimodal approach, combining standardized clinical assessment with midbrain and putaminal 31-phosphorus magnetic resonance spectroscopy (31P-MRS) and deep phenotyping of mitochondrial and lysosomal function in peripheral tissue in patients with recent-onset Parkinson's disease and control subjects. Sixty participants (35 patients with Parkinson's disease and 25 healthy controls) underwent 31P-MRS for quantification of energy-rich metabolites [ATP, inorganic phosphate (Pi) and phosphocreatine] in putamen and midbrain. In parallel, skin biopsies were obtained from all research participants to establish fibroblast cell lines for subsequent quantification of total intracellular ATP and mitochondrial membrane potential (MMP) as well as mitochondrial and lysosomal morphology, using high content live cell imaging. Lower MMP correlated with higher intracellular ATP (r = -0.55, P = 0.0016), higher mitochondrial counts (r = -0.72, P < 0.0001) and higher lysosomal counts (r = -0.62, P = 0.0002) in Parkinson's disease patient-derived fibroblasts only, consistent with impaired mitophagy and mitochondrial uncoupling. 31P-MRS-derived posterior putaminal Pi/ATP ratio variance was considerably greater in Parkinson's disease than in healthy controls (F-tests, P = 0.0036). Furthermore, elevated 31P-MRS-derived putaminal, but not midbrain Pi/ATP ratios (indicative of impaired oxidative phosphorylation) correlated with both greater mitochondrial (r = 0.37, P = 0.0319) and lysosomal counts (r = 0.48, P = 0.0044) as well as lower MMP in both short (r = -0.52, P = 0.0016) and long (r = -0.47, P = 0.0052) mitochondria in Parkinson's disease. Higher 31P-MRS midbrain phosphocreatine correlated with greater risk of rapid disease progression (r = 0.47, P = 0.0384). Our data suggest that impaired oxidative phosphorylation in the striatal dopaminergic nerve terminals exceeds mitochondrial dysfunction in the midbrain of patients with early Parkinson's disease. Our data further support the hypothesis of a prominent link between impaired mitophagy and impaired striatal energy homeostasis as a key event in early Parkinson's disease.

Sustainable organosolv-lignin coated nanosilver-halloysites reinforced poly (vinyl alcohol) nanocomposites for wound healing application

This study involves the fabrication of innovative poly (vinyl alcohol) nanocomposite films by incorporating silver-embedded clay nanotubes with organosolv-lignin by the solution casting approach. The confirmation of this procedure was achieved through the utilisation of many techniques, including FTIR, PXRD, FE-SEM, and TGA. The aforementioned research have verified the adherence of silver nanoparticles to the surface of raw halloysites. The confirmation of lignin functionalization on these nanotubes has been established. This novel nanofiller was used to make a range of nanocomposite films with varying weight percentages ranging from 0 wt% to 5 wt%. With the increase in the wt% of nanofillers, These nanocomposite films exhibited greater thermal and mechanical stability compared to plain PVA. An investigation was conducted to examine the impact of the films on the cellular behaviour of murine fibroblast (NIH3T3) cell lines. Based on the findings from cell proliferation and scratch testing, it has been determined that these nanocomposite films are not harmful to cells, exhibit a greater rate of cell multiplication (116 ± 1.19), and demonstrate increased migratory capabilities (86.5 ± 0.50). Further investigations of human blood corroborate the evidence that these films are compatible with blood. Nanocomposite films have the potential to serve as wound healers following pre-clinical and clinical testing.

Pigment epithelium-derived factor (PEDF) represses the glucose transporter 1 (GLUT1) mRNA expression and may be a potential therapeutic agent in psoriasis: a case–control and experimental study

We investigated the whole blood GLUT1 mRNA expression and serum pigment epithelium-derived factor (PEDF), interleukin-6 (IL-6), fetuin-A, and pentraxin-3 (PTX3) levels in psoriatic patients and tested their correlations with the severity of psoriasis using the psoriasis area and severity index (PASI) score. Also, we tested the GLUT1 mRNA expression after an in vitro treatment of human skin fibroblast (HSF) cell lines with PEDF. The case–control part of the study recruited 74 participants (44 psoriatic patients and 30 healthy volunteers). Whole blood GLUT1 mRNA fold changes were estimated by RT-PCR, and serum PEDF, IL-6, fetuin-A, and PTX3 levels were measured by ELISA kits. In the experimental part, the HSF cell lines were treated with different concentrations of PEDF for different times to test its effect on the GLUT1 mRNA expression. The whole blood GLUT 1 expression significantly increased in psoriatic patients and correlated positively with serum IL-6, fetuin-A, PTX3 levels and with the severity of psoriasis while negatively with serum PEDF levels. The PEDF-treated HSF cell lines showed a time- and dose-dependent decline in the GLUT 1 mRNA expression. The whole blood GLUT 1 mRNA is a non-invasive biomarker that is associated with the severity of psoriasis. PEDF represses GLUT 1 expression and may be a potential therapeutic agent in psoriasis.Trial registration: ClinicalTrials.gov Identifier: NCT04242082.

Enzymatic production process of capric acid-rich structured lipids: Development of formulation as a new therapeutic approach

The present work reports an optimization of the synthesis of MLM-type (medium, long, medium) structured lipids (SL) through an acidolysis reaction of grape seed oil with capric acid catalyzed by Rhizopus oryzae lipase immobilized. At first, tests were carried out by preparing the biocatalysts using enzyme loadings (0.15 to 1 g of enzymatic powder) for each gram of support. Enzyme loading was used 0.3 g of enzymatic powder, and hydrolytic activity of 1860 ± 23.4 IU/g was reached. Optimized conditions determined by the Central Composite Rotatable Design (CCRD) revealed that the acidolysis reaction reached approximately 59 % incorporation degree (%ID) after 24 h, in addition to the fact that the biocatalyst could maintain the incorporation degree in five consecutive cycles. From this high incorporation degree, cell viability assays were performed with murine fibroblast cell lines and human cervical adenocarcinoma cell lines. Concerning the cytotoxicity assays, the concentration of MLM-SL to 1.75 and 2 % v/v were able to induce cell death in 56 % and 64 % of adenocarcinoma cells, respectively. Human cervical adenocarcinoma cells showed greater sensitivity to the induction of cell death when using emulsions with MLM-SL > 1.75 % v/v compared to emulsions with lower content indicating a potential for combating carcinogenic cells.

Fabrication of chitosan/fibrin-armored multifunctional silver nanocomposites to improve antibacterial and wound healing activities

A wound healing substitute promotes rapid tissue regeneration and protects wound sites from microbial contamination. The silver-based antiseptic frequently moist skin stains, burns and irritation, penetrates deep wounds and protects against pathogenic infections. Thus, we formulated a novel fibrin/chitosan encapsulated silver nanoparticle (CH:F:SPG-CH:SNP) composites bandage accelerating the polymicrobial wound healing. Electrospinning method was employed to form the nano-porous, inexpensive, and biocompatible smart bandages. The structural, functional, and mechanical properties were analyzed for the prepared composites. The biological capacity of prepared CH:F:SPG-CH:SNP bandage was assessed against NIH-3 T3 fibroblast and HaCaT cell lines. In vitro hemolytic assays using red blood cells were extensively studied and explored the low hemolytic effect (4.5 %). In addition, the improved drug delivery nature captured for the CH:F:SPG-CH:SNP composite bandage. Antibacterial experiments were achieved against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Lactobacillus bulgaricus using zone inhibition method. Moreover, in-vivo wound healing efficacy of fabricated smart bandage was evaluated on the albino Wistar rats which revealed the significant improvement on the postoperative abdomen wounds.

Synthesis of new bis(dimethylamino)benzophenone hydrazone for diabetic management: In-vitro and in-silico approach

Inhibiting α-glucosidase is a reliable method for reducing blood sugar levels in diabetic individuals. Bis(dimethylamino)benzophenone derivatives 1–27 were synthesized from bis(dimethylamino)benzophenone via two-step reaction. Different spectroscopic techniques, including EI-MS and 1H NMR, were employed to characterize all synthetic derivatives. The elemental composition of synthetic compounds was confirmed by elemental analysis and results were found in agreement with the calculated values. The synthetic compounds 1–27 were evaluated for α-glucosidase inhibitory activity, except five compounds all derivatives showed good to moderate inhibitory potential in the range of IC50 = 0.28 ± 2.65 - 0.94 ± 2.20 μM. Among them, the most active compounds were 5, 8, 9, and 12 with IC50 values of 0.29 ± 4.63, 0.29 ± 0.93, 0.28 ± 3.65, and 0.28 ± 2.65, respectively. Furthermore, all these compounds were found to be non-toxic on human fibroblast cell lines (BJ cell lines). Kinetics study of compounds 8 and 9 revealed competitive type of inhibition with Ki values 2.79 ± 0.011 and 3.64 ± 0.012 μM, respectively. The binding interactions of synthetic compounds were also confirmed through molecular docking studies that indicated that compounds fit well in the active site of enzyme. Furthermore, a total of 30ns MD simulation was carried out for the most potent complexes of the series. The molecular dynamics study revealed that compound-8 and compound-12 were stable during the MD simulation.

Cytotoxicity of dihydropyridone and dihydropyrimidone curcumin derivatives against colon (HCT-116) and breast (MDA-MB-231) cancer cell lines

Breast cancer is the most common invasive cancer and colorectal cancer (CRC) the second most diagnosed malignancy in women worldwide. In men CRC is the third most common cancer. Despite the recent advances in targeted therapy, the clinical efficacy is often limited, noncurative, with a high toxicity profile, and exorbitant costs. Therefore, there is a growing interest in identifying natural compounds that are safe and affordable as adjunctive treatments to the conventional therapy currently offered for these patients. Curcumin from the roots of the Curcuma longa, is one such compound that has become one of the leading and most studied natural medicines for its role in cancer prevention and powerful antioxidant activity. However, its fast metabolism, low bioavailability, and the lack of specificity call for curcumin analogues to be synthesized with increased potency and higher specificity. In search of lead compounds in the present study the cytotoxic effects of curcumin and 14 heterocyclic curcumin derivatives have been screened using the MTT assay on two cancer cell lines HCT-116 and MDA-MB-231 and also on the normal GM08402 (human fibroblast) cell line. Electrochemical oxidation potentials were determined for selected compounds to reveal their electron-donating capacity and as a general indicator of their radical scavenging ability. Two dihydropyridone lead compounds have been identified which compared to curcumin have higher cytotoxicity on both cancer cell lines and at the same time with reduced cytotoxicity on the normal cell line.

Rare A360T Mutation Alters GSK3β(Ser9) Binding in the Cytosolic Loop of Presenilin 1, Influencing β-Catenin Nuclear Localization and Pro-Death Gene Expression in Alzheimer’s Disease Case

Presenilin 1 (PS1) forms, via its large cytosolic loop, a trimeric complex with N-cadherin and β-catenin, which is a key component of Wnt signaling. PS1 undergoes phosphorylation at 353 and 357 serines upon enhanced activity and elevated levels of the GSK3β isoform. PS1 mutations surrounding these serines may alter the stability of the β-catenin complex. Such mutations are found in some cases of familial early-onset Alzheimer’s disease (fEOAD), but their functional impact remains obscure. One of such variants of PS1, the A360T substitution, is located close to GSK3β-targeted serine residues. This variant was recently demonstrated in the French population, but more detail is needed to understand its biological effects. To assess the significance of this variant, we employed functional studies using a fibroblast cell line from an Alzheimer’s disease case (a female proband) carrying the A360T mutation. Based on functional transcriptomic, cellular, and biochemical assays, we demonstrated atypically impaired β-catenin/GSK3β signaling in the A360T patient’s fibroblasts. In detail, this was characterized by a decreased level of active cytosolic β-catenin and bound by PS1, an increased level of nuclear β-catenin, an increased level of inhibited GSK3β phosphorylated on Ser9, and enhanced interaction of GSK3β(Ser9) with PS1. Based on the transcriptomic profile of the A360T fibroblasts, we proposed a dysregulated transcriptional activity of β-catenin, exemplified by increased expression of various cyclin-dependent kinases and cyclins, such as cyclin D1, potentially inducing neurons’ cell cycle re-entry followed by apoptosis. The A360T cells did not exhibit significant amyloid pathology. Therefore, cell death in this PS1 cytosolic loop mutation may be attributed to impaired β-catenin/GSK3β signaling rather than amyloid deposition per se. We further estimated the biological and clinical relevance of the A360T variant by whole exome sequencing (WES). WES was performed on DNA from the blood of an A360T female proband, as well as an unrelated male patient carrying the A360T mutation and his mutation-free daughter (both unavailable for the derivation of the fibroblast cell lines). WES confirmed the highest-priority AD causality of the A360T variant in PS1 and also profiled the pathways and processes involved in the A360T case, highlighting the greatest importance of altered Wnt signaling.

Comprehensive Stereoselectivity Assessment of Toxicokinetics, Tissue Distribution, Cytotoxicity, and Environmental Fate of Chiral Pesticide Propiconazole.

Propiconazole (PRO) has been widely used in the treatment of fungal infection in fruits, vegetables, cereals, and seeds. In this study, a newly established chiral liquid chromatography tandem mass spectrometry method was applied to the systemic stereoselectivity evaluation of PRO enantiomers, including toxicokinetics, tissue distributions, cytotoxicity, accumulation, and degradation. Our results showed that both trans (+)-2S,4S-PRO and cis (-)-2S,4R-PRO had lower Cmax and AUC0-∞ and higher CLz/F values in plasma and lower accumulation concentrations in the liver, heart, and brain. In cytotoxic assays, cis (-)-2S,4R-PRO exhibited the lowest cytotoxicity in PC12 neuronal, N9 microglia, SH-SY5Y neuroblastoma, and MRC5 lung fibroblast cell lines. Moreover, the Eisenia fetida incubation experiment revealed that the accumulations of both trans (+)-2S,4S-PRO and cis (-)-2S,4R-PRO were higher than those of their antipodes in E. fetida. In summary, our findings first suggested that the application of cis (-)-2S,4R-PRO for agriculture would hugely reduce the environmental risk.

Designing and fabrication of colloidal nano-phytosomes with gamma-oryzanol and phosphatidylcholine for encapsulation and delivery of polyphenol-rich extract from pomegranate peel

Nano-carriers are well-known delivery systems to encapsulate different bioactive compounds and extracts. Such nano-systems are used in various food and drug areas to protect active ingredients, increase bioavailability, control the release, and deliver bioactive substances. This study aimed to design and fabricate a stable colloidal nano-delivery system to better preserve the antioxidant properties of pomegranate peel extract (PPE) and protect its sustained release in a gastrointestinal model. To achieve this goal, a nano-phytosomal system was fabricated with plant-based, cost-effective, and food-grade compounds, i.e., phosphatidylcholine (PC) and gamma-oryzanol (GO) for encapsulation of PPE. To fabricate the nano-phytosomes, thin film hydration/sonication method was used. The parameters of particle size, zeta potential, polydispersity index (PDI), loading capacity (LC), and encapsulation efficiency (EE) were investigated to evaluate the efficiency of the produced nano-system. In summary, the size, zeta potential, PDI, LC, and EE of homogenous spherical PC-GO-PPE nano-phytosomes (NPs) in the ratio of 8:2:2 % w/w were achieved as 60.61 ± 0.81 nm, −32.24 ± 0.84 mV, 0.19 ± 0.01, 19.13 ± 0.30 %, and 95.66 ± 1.52 %, respectively. Also, the structure of NPs was approved by Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscope (SEM). The optimized NPs were stable during one month of storage at 4 °C, and changes in the size of particles and PPE retention rate were insignificant (p > 0.05). The nano-encapsulation of PPE significantly decreased the loss of its antioxidant activity during one month of storage at 4 °C. The optimized NPs exhibited prolonged and sustained release of PPE in a gastrointestinal model, so that after 2 h in simulated gastric fluid (SGF) and 4 h in simulated intestinal fluid (SIF), 22.66 ± 2.51 % and 69.33 ± 4.50 % of initially loaded PPE was released, respectively. Optimized NPs had considerable cytotoxicity against the Michigan Cancer Foundation-7 cell line (MCF7) (IC50 = 103 μg/ml), but not against Human Foreskin Fibroblast cell line (HFF-2) (IC50 = 453 μg/ml). In conclusion, spherical PC-GO-PPE NPs were identified as a promising delivery system to efficiently encapsulate PPE, as well as protect and preserve its bioactivity, including antioxidant and cytotoxicity against cancer cell line.

Whole stromal fibroblast signature is linked to specific chemokine and immune infiltration patterns and to improved survival in NSCLC

ABSTRACT Cancer associated fibroblasts (CAF) are known to orchestrate multiple components of the tumor microenvironment, whereas the influence of the whole stromal-fibroblast compartment is less understood. Here, an extended stromal fibroblast signature was investigated to define its impact on immune cell infiltration. The lung cancer adenocarcinoma (LUAD) data set of the cancer genome atlas (TCGA) was used to test whole stroma signatures and cancer-associated fibroblast signatures for their impact on prognosis. 3D cell cultures of the NSCLC cancer cell line A549 together with the fibroblast cell line SV80 were used in combination with infiltrating peripheral blood mononuclear cells (PBMC) for in-vitro investigations. Immune cell infiltration was assessed via flow cytometry, chemokines were analyzed by immunoassays and RNA microarrays. Results were confirmed in specimens from NSCLC patients by flow cytometry or immunohistochemistry as well as in the TCGA data set. The TCGA analyses correlated the whole stromal-fibroblast signature with an improved outcome, whereas no effect was found for the CAF signatures. In 3D microtumors, the presence of fibroblasts induced infiltration of B cells and CD69+CD4+ T cells, which was linked to an increased expression of CCL13 and CXCL16. The stroma/lymphocyte interaction was confirmed in NSCLC patients, as stroma-rich tumors displayed an elevated B cell count and survival in the local cohort and the TCGA data set. A whole stromal fibroblast signature was associated with an improved clinical outcome in lung adenocarcinoma and in vitro and in vivo experiments suggest that this signature increases B and T cell recruitment via induction of chemokines.

Bioactive Fe Foam for Degradable Bone Graft Cages

Fe foams when combined with a proper surface coating can be effectively used as a biodegradable metal with improved biocompatibility and bioactivity. Herein, Fe foams prepared via powder metallurgy route coated with two different bioactive coatings namely polyvinyl alcohol (PVA)/bioglass (BG) and polylactic acid (PLA)/BG are investigated. The effect of hydrophilic PVA and hydrophobic PLA, in conjunction with BG, has been examined. The study of biomineralization, degradation, and weight loss of the composite‐coated material over a period of 28 days is compared with that of the uncoated Fe foam samples in simulated body fluid. The composite coatings on the Fe foam effectively tailor the degradation with enhanced biomineralization. The composite‐coated material exhibits viability toward the L929 fibroblast cell line. The work reveals the great potential of Fe foams to be used as a biodegradable metallic material for temporary implant applications such as bone graft cages.

Synergistic impacts of rifampicin and doxorubicin against thioacetamide-induced hepatocellular carcinoma in rats

Background and aimsCombination therapy is a promising new strategy that has been proposed to increase the efficacy of cancer treatment. We aimed to investigate the anti-cancer activity of rifampicin monotherapy and its combination with doxorubicin against hepatocellular carcinoma (HCC). Materials and methodsThe in vitro half maximal inhibitory concentration (IC50) and selectivity index (SI) of the drugs under investigation against HepG2 and human lung fibroblast (WI38) cell lines were determined. For the in vivo experiment, male Sprague-Dawley albino rats were injected with thioacetamide at 200 mg/kg twice a week for 90 days; HCC development was confirmed histopathologically. Following HCC induction, the rats were treated with intraperitoneal doxorubicin, rifampicin, or their combination for 45 or 90 days. After sacrifice, the livers were examined histopathologically. The levels of aminotransferases, albumin, bilirubin, malondialdehyde, superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (TAC), and nitric oxide were measured by spectrophotometry. Alpha-fetoprotein, cancer antigen 19-9, tumor necrosis factor-alpha, interleukin-6, Bcl-2-associated X protein, caspase 3, caspase 8, and p53 were estimated using ELISA. ResultsIn vitro, the combination of doxorubicin and rifampicin showed the highest SI of 3.43. In vivo, among the measured markers, the levels of TAC, CAT, SOD, and p53 decreased (P < 0.001) and the rest of the measured marker levels increased (P < 0.001) in the HCC-bearing rats; after treatment in all groups, all these changes improved toward normal in a time-dependent manner. The combination of doxorubicin and rifampicin optimized the effects of the two individual drugs and exerted the best antioxidant effects. ConclusionsIn general, compared with rifampicin or doxorubicin alone, combination therapy has favorable outcomes. Based on our results, the combination of rifampicin and doxorubicin might be applicable for HCC chemotherapy.

Exploring NiFe2O4 nanoparticles: Electrochemical analysis and evaluation of cytotoxic effects on normal human dermal fibroblasts (HDF) and mouse melanoma (B16-F10) cell lines

Nanobiotechnology facilitates the development of novel materials with potential applications in human health, the environment, and a variety of industries. We used X-ray diffraction (XRD) to characterize NiFe2O4 nanoparticles (NPs), confirming a cubic crystal structure with crystallite sizes of 19 nm and 21 nm. Synthesized NPs spherical shape morphology was confirmed by field emission-scanning electron microscopy (FE-SEM) and an average diameter (prepared NPs) of 20 nm and 23 nm was revealed by Transmission electron microscopy (TEM). The chemical state was determined using X-ray photoelectron spectroscopy (XPS). A vibrating sample magnetometer (VSM) was used to measure the magnetic characteristics. The electrochemical characteristics of prepared electrodes could be useful for supercapacitor applications. Changing the surfactants, reaction temperature, and heating duration considerably enhanced electrochemical performance, according to our findings. At 700 °C, specific capacitance (Cs) attained an outstanding value of 67.2 F/g. The prepared NPs cytotoxicity was performed on normal human dermal fibroblast (HDF) and mouse melanoma (B16-F10) cell lines. The current findings show that these NPs are non-toxic at concentrations ranging from 0 to 1000 µg/mL. These findings point to prospective biomedical uses for NiFe2O4 NPs.

Preparation and in vitro evaluation of biological agents based on Zinc-laponite- curcumin incorporated in alginate hydrogel

Due to their outstanding structures and properties, three-dimensional (3D) hydrogels and nanoparticles have been widely studied and indicated a very high potential for medical, therapeutic, and diagnostic applications. However, hydrogels and nanoparticles systems have particular drawbacks that limit their widespread applications. In recent years, the incorporation of nanostructured systems into hydrogel has been developed as a novel way for the formation of new biomaterials with various functions to solve biomedical challenges. In this study, alginate-loaded Zinc- laponite—curcumin (Zn/La/Cur) nanocomposites were fabricated via ionic cross-linking. The prepared nanocomposite hydrogels were characterized via FTIR and FE-SEM. Moreover, energy dispersive x-ray spectroscopy (EDX) was used to study the elements of the Zn/La/Cur nanocomposite. The NIH3T3 fibroblast cell line was utilized for the MTT assay to determine the cell viability of the fabricated alginate-loaded Zn/La/Cur nanocomposites. MTT results demonstrated that there was no evidence of toxicity in the samples. These outcomes suggest that applying Al/Zn/La/Cur nanocomposite as a biological agent could be a novel tissue engineering strategy for treating soft tissue disorders.Graphical

Rapidly derived equimolar Ca: P phasic bioactive glass infused flexible gelatin multi-functional scaffolds – A promising tissue engineering

The study aims to design and fabricate an ultra-easier multi-functional biomedical polymeric scaffold loaded with unique equimolar Ca:P phasic bioactive glass material (BG). Gelatin (G) – 45S5 bioactive glass (BG) scaffolds were synthesized via a simple laboratory refrigerator with higher biocompatibility and cytocompatibility. The results proved that BG has enhanced bio-mineralization of the scaffolds and results support that the G: BG (1:2) ratio is the more appropriate composition. Brunauer-Emmett-Teller (BET) study confirms the higher surface area for pure Gelatin and G: BG (1:2). Scanning Electron Microscopic images display the precipitation of hydroxycarbonate apatite layer over the scaffolds on immersing it in simulated body fluid. Alkaline phosphate activity proved that G: BG (1:2) scaffold could induce mitogenesis in MG-63 osteoblast cells, thus helping in hard tissue regeneration. Sirius red collagen deposition showed that higher content bioactive glass incorporated Gelatin polymeric scaffold G: BG (1:2) could induce rapid collagen secretion of NIH 3T3 fibroblast cell line that could help in soft tissue regeneration and earlier wound healing. The scaffolds were also tested for cell viability using NIH 3T3 fibroblast cell lines and MG 63 osteoblastic cell lines through methyl thiazolyl tetrazolium (MTT) assay. Thus, the study shows a scaffold of appropriate composition G: BG (1:2) can be a multifunctional material to regenerate hard and soft tissues.

The Wound-Healing Potential of the Endemic Plant Helianthemum canum (L.) Baumg: Preclinical Studies Supported with Phytochemical Profiling.

The study's objective is to clarify the probable mechanisms underlying the wound-healing properties of Helianthemum canum L. (Cistaceae), a traditional anti-inflammatory and wound-healing medicine. LC/MS-MS was used to perform phytochemical analyses on a 70 % methanol extract of the plant's aerial parts. In vivo, linear incision and circular excision models were used to evaluate the wound healing activity. For anti-inflammatory effect, in vivo acetic acid capillary permeability assay and in vitro Interleukin 1, Interleukin 6, and Interferon ɣ levels in LPS-induced FR skin fibroblast cell line were also evaluated. The extract significantly improved wound healing in experimental models, with tensile strength values of 27.8 % and a contraction value of 35.09 %. Histopathological examinations, hydroxyproline estimation, hyaluronidase, collagenase, and elastase enzyme inhibitory assays confirmed wound healing potential. Inflammatory cytokines were significantly inhibited in the LPS-induced FR cell line, with the highest effect seen on IL-6 (34.5±2.12 pg/mL). This study offered the first concrete proof that H. canum can be used to treat wounds by suggesting that the myricetin and quinic acid content identified by LCMS-MS analysis may be accountable for the effect of H. canum on wound contraction and hydroxyproline production.

Machine learning-based morphological quantification of replicative senescence in human fibroblasts.

Although aging has been investigated extensively at the organismal and cellular level, the morphological changes that individual cells undergo along their replicative lifespan have not been precisely quantified. Here, we present the results of a readily accessible machine learning-based pipeline that uses standard fluorescence microscope and open access software to quantify the minute morphological changes that human fibroblasts undergo during their replicative lifespan in culture. Applying this pipeline in a widely used fibroblast cell line (IMR-90), we find that advanced replicative age robustly increases (+28-79%) cell surface area, perimeter, number and total length of pseudopodia, and nuclear surface area, while decreasing cell circularity, with phenotypic changes largely occurring as replicative senescence is reached. These senescence-related morphological changes are recapitulated, albeit to a variable extent, in primary dermal fibroblasts derived from human donors of different ancestry, age, and sex groups. By performing integrative analysis of single-cell morphology, our pipeline further classifies senescent-like cells and quantifies how their numbers increase with replicative senescence in IMR-90 cells and in dermal fibroblasts across all tested donors. These findings provide quantitative insights into replicative senescence, while demonstrating applicability of a readily accessible computational pipeline for high-throughput cell phenotyping in aging research.

Synthesis, Characterization and Evaluation of Antimicrobial Activity, Phytotoxicity, Molecular Docking &amp; Dynamic Simulations of Benzothiazole Functionalized C−C Linked Pyrazolyl‐Thiazoles

AbstractSynthesis of phenyl/benzothiazole functionalized C−C linked pyrazolyl‐thiazoles has been achieved from non‐lachrymatory α‐tosyloxy pyrazolylketones with different thioamides. The pyrazolyl‐thiazoles were characterized by spectroscopic studies and evaluated for antimicrobial activity and phytotoxicity. Compound 6 n displayed better activity with MIC value of 0.0045 μmol/mL against Gram‐positive bacteria B. cereus in comparison to the standard drug Ciprofloxacin (0.0047 μmol/mL). Compound 6 i displayed better activity against fungal strain C. albicans (MIC=0.0152 μmol/mL) in comparison to the standard drug Fluconazole (0.0204 μmol/mL). Compound 6 o diplayed 95 % cell viability against Mouse Fibroblast cell line and 100 % plant seed germination. Docking studies at the topoisomerase II DNA gyrase B (PDB ID: 1KZN) and C. albicans (PDB ID: 1IYL) were used to investigate potential interactions between the most active compound and the receptor protein. The stability of the compounds with 1KZN and 1IYL by molecular dynamic simulations showed that 6 r, 6 n and 6 i present leading structures for next drug development because of their simple synthesis and useful bioactivity.