Osteoarthritis (OA) is a progressive joint disorder affecting over 250 million people globally and is characterized by chronic pain and disability. Among its key pathogenic mechanisms are mitochondrial dysfunction and elevated reactive oxygen species (ROS), often triggered by inflammatory mediators such as lipopolysaccharide (LPS). This study evaluates the protective effects of curcumin on mitochondrial function, autophagy, and apoptosis in an in vitro model of OA. Human bone marrow-derived mesenchymal stem cells (BMSCs) were differentiated into chondrocytes using MesenCult™-ACF medium. Differentiation was confirmed by histological staining for Type II Collagen, Alcian Blue, and Toluidine Blue. LPS was used to induce an OA-like inflammatory response. Mitochondrial membrane potential (ΔΨm) was assessed using Rhodamine 123 staining. Autophagy and apoptosis were evaluated using Acridine orange and propidium iodide staining, respectively. Western blotting was performed to analyze the expression of pro-caspase-3, Bcl-2, Beclin-1, LC3-I/II, and GAPDH. LPS significantly impaired mitochondrial function, limited autophagy, and enhanced apoptotic signaling (reduced pro-caspase-3). Curcumin (25 µM and 100 µM) restored ΔΨm, increased Beclin-1 and LC3-II, and maintained pro-caspase-3 expression, with Bcl-2 showing a non-monotonic response (higher at 25 µM than at 100 µM). Curcumin exerted cytoprotective effects in inflamed chondrocytes by stabilizing ΔΨm, promoting autophagy, and attenuating apoptotic activation, supporting its multi-target therapeutic potential in OA.

Sturge-Weber-Syndrome (SWS) is an uncommon, non-genetic neuro-vascular disorderconsidered by nonstandard blood vessel development in the eye, skin and brain. People with SWS oftenpresent with dermal capillary malformations, recognized as port-wine-birthmarks (PWBs), along withleptomeningeal vascular malformations visible on contrast-enhanced MRI, abnormal eye blood vessels,health risk and glaucoma. Those affected experience reduced brain blood flow, increasing their risk forvenous stroke, stroke-like episodes, seizures, as well as mechanical and rational impairments. The R183-Q GNAQ somatic mutation, which activates this gene, is the most communal mutation linked to SWS;however, new exploration also points to somatic mutations in GNA11 and GNB2 as contributors. Currentretroactive studies recommend that low dose aspirin and Vitamin-D are effective treatments for

Background: Osteoarthritis (OA) is a degenerative joint disorder marked by chronic inflammation, extracellular matrix (ECM) breakdown, and subchondral bone remodeling, ultimately leading to loss of cartilage integrity and joint function. Hyaluronic acid (HA), a key component of synovial fluid, contributes to joint lubrication and has been implicated in the regulation of inflammatory and anabolic processes. High-molecular-weight (HMW) HA (> 1 MDa) is reported to exert anti-inflammatory and matrix-preserving effects; however, its role in multicellular systems remains incompletely understood. In this study, we investigated the effects of HMW HA using an in vitro chondrocyte–osteoclast co-culture model designed to reflect key features of the osteoarthritic microenvironment. Methods: Bone marrow-derived mesenchymal stem cells were differentiated into chondrocytes, while RAW 264.7 macrophages were induced into osteoclast-like cells using M-CSF and RANKL. Cells were co-cultured at a 1:1 ratio and stimulated with lipopolysaccharide (LPS; 1 µg/mL) to induce inflammatory stress, followed by HA treatment (50–500 µg/mL) for 24–48 h. Cell viability, morphology, and TRAP staining were assessed. Expression of inflammatory and matrix-associated markers was analysed by Western blot, qPCR, and ELISA. Cell-cycle distribution and DNA integrity were evaluated using flow cytometry and agarose gel electrophoresis. Results: LPS exposure reduced metabolic activity, altered morphology, and was associated with increased expression of NF-κB-related inflammatory markers and elevated expression of IL-1β, TNF-α, MMP-13, and ADAMTS-5, alongside reduced COL2A1 and ACAN levels. HA treatment improved metabolic activity and preserved cellular morphology in a concentration-dependent manner, with the most pronounced effects observed at 200–500 µg/mL. HA-treated cultures showed lower levels of inflammatory and catabolic markers together with increased expression of ECM-associated genes. In addition, HA treatment was associated with a more balanced cell-cycle profile and reduced DNA fragmentation. Conclusion: HMW HA was associated with coordinated modulation of inflammatory and matrix-related markers in an LPS-induced co-culture system. These findings suggest that HA may contribute to maintaining cellular homeostasis under inflammatory conditions in vitro. Further studies are needed to clarify the underlying mechanisms and translational relevance.

Osteoarthritis (OA) is a progressive joint disease characterized by cartilage degeneration, chronic low-grade inflammation, and disruption of tissue homeostasis. Although the mechanosensitive ion channel Piezo1 has been implicated in OA pathogenesis, its expression may also be modulated by inflammatory stimuli independently of applied mechanical loading. This study established a scaffold-free three-dimensional co-culture organoid model comprising human bone marrow-derived mesenchymal stem cell-derived chondrocyte-like cells and M-CSF/RANKL-differentiated RAW264.7-derived osteoclast-like cells to investigate Piezo1-associated molecular responses, inflammatory signaling, and mitophagy-related markers under lipopolysaccharide (LPS)-induced inflammatory conditions. Osteoclast-like differentiation was validated in parallel monolayer cultures by tartrate-resistant acid phosphatase staining and the presence of multinucleated cells before the corresponding differentiated cultures were used for organoid generation. Histological staining, immunofluorescence, CellTiter-Glo 3D viability assay, lactate dehydrogenase cytotoxicity assay, RT-qPCR, and Western blotting were used to evaluate extracellular matrix formation and inflammatory, catabolic, and mitochondrial quality-control-associated markers. LPS stimulation increased the expression of Piezo1, HIF-1α, phosphorylated CaMKII, NLRP3, cleaved Caspase-1, and MMP13, together with alterations in mitophagy- and autophagy-associated markers. Among the evaluated compounds, curcumin produced the greatest improvement in viability relative to the LPS-treated group and was selected for subsequent molecular analyses. Curcumin treatment was associated with reduced inflammatory and catabolic marker expression and partial preservation of cartilage-associated matrix markers. These findings demonstrate inflammation-associated changes in Piezo1 expression and related molecular markers but do not establish mechanically activated Piezo1 signaling or Piezo1-dependent causality. The organoid system therefore represents an exploratory LPS-induced inflammatory model exhibiting selected OA-relevant molecular and matrix-associated features.