This study investigated whether incorporating organic matrix components (OM) derived from marine sponges into biosilica (BS)-based scaffolds could improve their physicochemical, mechanical, and biological performance for bone tissue engineering. BS and BS/OM scaffolds were fabricated and characterized by Fourier-transform infrared spectroscopy, scanning electron microscopy, porosity analysis, compression testing, and stability assays in phosphate-buffered saline. Their biological performance was evaluated using MC3T3-E1 pre-osteoblastic cells through metabolic activity, dsDNA quantification, reactive oxygen and nitrogen species production, scratch migration, and Alizarin Red S mineralization assays. Genotoxicity was assessed in CHO-K1 cells using comet and micronucleus assays. OM incorporation improved scaffold structural stability and mechanical behavior, increasing compressive modulus and maximum strain while delaying disintegration in aqueous conditions. Both scaffolds were non-cytotoxic and showed no detectable genotoxic effects. However, BS promoted higher metabolic activity, cell proliferation, migration, and extracellular matrix mineralization. BS induced a predominantly ROS-associated response, whereas BS/OM produced higher RNS levels and reduced osteoblastic proliferation and migration. Despite this, BS/OM maintained mineralization levels above the basal control. Marine sponge-derived BS and OM provide complementary functions within bone scaffolds. OM primarily enhances mechanical stability and structural integrity, whereas BS exerts a stronger osteogenic response, highlighting scaffold composition as a key parameter for balancing mechanical and biological performance.
Corneal blindness remains a major global health challenge, and the shortage of donor tissues emphasizes the need for alternative approaches in corneal tissue engineering. This study aimed to evaluate different decellularization methods for tuna corneas (Thunnus albacares) and characterize the selected decellularized matrix as a potential biommetic scaffold for corneal tissue engineering. Four decellularization protocols were evaluated based on their ability to remove cellular components and preserve extracellular matrix (ECM) proteins: (M1) Triton X-100 treatment for 72 h; (M2) five freeze–thaw cycles at − 80 °C;; (M3) freeze–thaw cycles combined with Triton X-100 treatment for 24 h; and (M4) freeze–thaw cycles followed by DNase treatment (1 h). The resulting decellularized tuna cornea (DTC) was characterized regarding morphology and physicochemical properties using scanning electron microscopy (SEM), FTIR, SDS-PAGE, circular dichroism (CD), and thermogravimetric analysis (TGA). Subsequently, DTC was processed into a hydrogel formulation, and its rheological, optical properties and cytocompatibility were evaluated by rheological testing, transmittance analysis and MTT assay, respectively. The freeze–thaw combined with DNase treatment was considered the most suitable approach, effectively removing cellular components while preserving ECM structure. Biochemical analyses demonstrated a 90
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.
