In this paper, a human synthetic liver was constructed using adipose-derived stem cells (ADSCs) on a PCL/sodium alginate/gelatin hydrogel scaffold. First, a 3D scaffold was fabricated using polycaprolactone (PCL), sodium alginate, and gelatin hydrogel via electrospinning. During scaffold fabrication and testing, human adipose-derived stem cells (ADSCs) were transformed into hepatocyte cells using a mesenchymal process for 21 days. Human placenta extract (HPE) was used in cell culture to increase cell growth, proliferation, and differentiation. ADSC cells with and without a scaffold were applied as 2D and 3D models, respectively. Here, 4 basic groups were constructed and used for the study, including 2D ADSC cells, 2D ADSC cells plus HPE, 3D ADSC cells, and 3D ADSC cells with HPE. The scaffold results revealed that the tensile elastic modulus of the scaffold was 85.5 ± 15.5 kPa. The contact angle and porosity values were 67.7^^∘ and 71.351
In the preclinical testing of novel drugs cell-based in vitro models can be used to replace animal testing. The cell line CI-pNaEC is a promising candidate for a suitable in vitro model of the nasal epithelium but suffers from low ciliation and inconsistent epithelial architecture. In this study, the influence of Notch and ROCK inhibition on differentiation benchmarks in CI-pNaEC was investigated. Ciliation was monitored via immunofluorescence for acetylated α-tubulin, mucus expression was visualized and quantified using alcian blue, barrier properties were assessed using TEER measurements and permeation studies, while histology was evaluated by light microscopy. Treatment with low-micromolar concentrations of DAPT markedly increased the proportion of ciliated cells and promoted a more uniform, pseudostratified epithelial layer, while mucus secretion remained comparable to untreated controls. However, DAPT also induced a slight increase in barrier properties, resulting in permeation coefficients below ex vivo nasal tissue benchmarks for some markers. In contrast, Y-27632 alone had a visible effect on ciliation and histological uniformity, with minimal effects on mucus expression, and its combination with DAPT did not further improve differentiation. The results demonstrate that targeted Notch inhibition at low doses significantly enhances ciliation and epithelial organization in CI-pNaEC without compromising mucin production, although barrier function is modestly affected.
Phlomis cashmeriana is a perennial medicinal herb endemic to the Kashmir Himalaya. This study is the first attempt to investigates the phytochemical profile, anticancer, and antioxidant activities of P. cashmeriana seed oil using in vitro approaches. Phytochemical analysis was carried through HR-LCMS and GC-MS techniques that detected presence of alkaloids, triterpenoids, glycosides, fatty acids, and esters. Antioxidant activities were assessed using DPPH (IC50 = 114.39 µg/mL), ABTS (IC50 = 137.90 µg/mL), and FRAP (IC50 = 102.35 µg/mL) assays. The seed oil exhibited dose and time dependent anticancer activity, with the highest antiproliferative potential against HeLa cells (GI50 = 35.55 µg/mL) and the least against LN-18 cells (GI50 = 65.39 µg/mL).Changes in cellular morphological aspects through fluorescence microscopy against different cancer cells revealed potent apoptotic activity of the seed oil, with minimal toxicity towards normal fibroblast L-929 cells (GI50 = 339.52 µg/mL). Elemental analysis using MPAES and amino acid profiling with Agilent 1260 Series HPLC indicated high nutritional value of P. cashmeriana seeds. The identified elements included as Na, Al, Mg, Ca, Zn, and Fe, and amino acids detected were aspartic acid, glutamic acid, glycine, threonine, and arginine. Concludingly P. cashmeriana seed oil possess potential as a natural chemo preventive source, food supplement, and nutraceutical.
Bone tumor resection often results in critical-sized defects that standard treatments, such as PMMA cement, fail to regenerate effectively due to their bioinert nature. Furthermore, systemic chemotherapy, used to treat remaining cancer cells after tumor resection, lacks specificity and induces severe systemic toxicities. This study aimed to develop and characterize a personalized, 3D-printed composite scaffold made of gelatin methacryloyl (GelMA) and hydroxyapatite (HAp) as a potential dual-action platform for bone tissue engineering and localized carboplatin delivery. Scaffolds were fabricated using extrusion-based 3D printing. Comprehensive rheological assessments were performed to evaluate printability. The carboplatin release profile was analyzed using different kinetic models, and in vitro bioactivity was assessed in Simulated Body Fluid (SBF). All experiments were performed in triplicate (n = 3). Rheological characterization showed that HAp incorporation improved shape recovery (from 49 to 90
This study explores the thrombolytic potential of recombinant tissue plasminogen activator (rtPA) immobilized on chitosan-coated magnetic nanoparticles (rtPA-CS-MNPs) in comparison with free rtPA and commercialized MNPs (CMNPs). The objective was to evaluate whether MNPs immobilization enhances stability, prolongs enzymatic activity, and improves clot dissolution efficiency under in vitro conditions. rtPA was immobilized onto CS-coated MNPs via covalent bonding. Characterization was performed using dynamic light scattering (DLS) for size and zeta potential (ZP) analysis, Fourier-transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM). In vitro thrombolysis assays were conducted using fibrin clots prepared from human plasma. Comparative analysis was performed between rtPA-CS-MNPs, rtPA-CMNPs, and free rtPA. rtPA-CS-MNPs demonstrated sustained enzymatic activity with consistently higher OD values compared to free rtPA and CMNPs. DLS and ZP of rtPA-CS-MNPs were around 152 and − 9, respectively. FTIR results confirmed that CS successfully coated MNPs as well as rtPA immobilized into CS-MNPs. TEM images exhibited mainly spherical MNPs in the 20–50 nm range. The findings suggested that MNPs, CS, and the administered dose of rtPA are safe for potential in vivo applications. The immobilized formulation demonstrated prolonged stability, reduced degradation, and enhanced clot-dissolution efficacy over 24 h. Blank controls confirmed that activity was attributable to rtPA immobilization rather than nanoparticle interference. The findings indicate that rtPA-CS-MNPs provide superior thrombolytic activity compared to free rtPA, with advantages in stability and sustained release. This comparative study highlights the potential of CS-coated MNPs as a promising platform for controlled and efficient thrombolytic therapy.
