Abstract Objective The differentiation of mesenchymal stem cells (MSCs) into chondrocytes provides an attractive basis for the repair and regeneration articular cartilage. Under clinical conditions, chondrogenesis will often need to occur in presence mediators inflammation produced response injury or disease. purpose this study was examine effects 2 important inflammatory cytokines, interleukin‐1β (IL‐1β) tumor necrosis factor α (TNFα), on chondrogenic behavior human MSCs. Methods Aggregate...

Abstract Elevated levels of PGE2 have been reported in synovial fluid and cartilage from patients with osteoarthritis (OA). However, the functions metabolism not previously studied detail. To do so, we cultured explants, obtained undergoing knee replacement surgery for advanced OA, (0.1–10 μM). inhibited proteoglycan synthesis a dose-dependent manner (maximum 25% inhibition (p < 0.01)). also induced collagen degradation, inhibitable by matrix metalloproteinase (MMP) inhibitor...

The demands of tissue engineering have driven a tremendous amount research effort in 3D culture technology and, more recently, printing. need to use techniques broadly all cell biology is well-recognized, but the transition has been impeded by convenience, effectiveness, and ubiquity 2D materials, assays, protocols, as well lack counterparts these tools. Interestingly, progress discoveries bioprinting may provide technical support needed grow practice culture. Here we investigate an...

Skeletogenesis depends on the activity of bone-forming cells derived from mesenchymal cells. The pathways that control cell differentiation are not well understood. We propose Foxo1 is an early molecular regulator during into osteoblasts. In mouse embryos, expression higher in skeletal tissues, while silencing has a drastic impact skeletogenesis and craniofacial development, specially affecting pre-maxilla, nasal bone, mandible, tibia, clavicle. Similarly, increase under influence osteogenic...

Adult mesenchymal stem cells (MSCs) have the capacity to differentiate into various connective tissues such as cartilage and bone following stimulation with certain growth factors. However, less is known about of these undergo chondrogenesis when proteins are delivered via gene transfer. In this study, we investigated primary, marrow-derived MSCs in aggregate cultures genetic modification adenoviral vectors encoding chondrogenic We found that adenoviral-mediated expression TGF-β1 BMP-2, but...

To determine the effects of antioxidant resveratrol on functions human chondrocytes in osteoarthritis (OA).Chondrocytes and cartilage explants were isolated from OA patients undergoing knee replacement surgery. Effects presence or absence interleukin-1beta (IL-1beta) stimulation assessed by measurement prostaglandin E(2) (PGE(2)) leukotriene B(4) (LTB(4)) synthesis, cyclooxygenase (COX) activity, matrix metalloproteinase (MMP) expression, proteoglycan production. explore mechanisms action...

Because articular cartilage has a poor regeneration capacity, numerous cell-based approaches to therapy are currently being explored. The present study involved the use of gene transfer as means provide sustained delivery chondrogenic proteins primary mesenchymal stem cells (MSCs). In previous work, we found that adenoviral-mediated transforming growth factor-beta1 (TGF-β1) and bone morphogenetic protein 2 (BMP-2), but not insulin-like factor 1 (IGF-1), could be used induce differentiation...

We investigated the role of periostin, an extracellular matrix protein, in pathophysiology osteoarthritis (OA). In OA, dysregulated gene expression and phenotypic changes articular chondrocytes culminate progressive loss cartilage from joint surface. The molecular mechanisms underlying this process are poorly understood. examined periostin by immunohistochemical analysis lesional nonlesional human rodent OA knee cartilage. addition, we used small interfering (si) RNA adenovirus transduction...

In the present study, role of mitogen-activated protein kinases (MAPKs) in chondrocyte mechanotransduction was investigated. We hypothesized that MAPKs participate fluid flow-induced mechanotransduction. To test our hypothesis, we studied cultured chondrocytes subjected to a well-defined mechanical stimulus generated with laminar flow chamber. The extracellular signal-regulated 1 and 2 (ERK1/2) were activated 1.6–3-fold after 5–15 min exposure corresponding chamber wall shear stress 1.6 Pa....

The delivery of anti-arthritic genes to the synovial lining joints is being explored as a strategy for treatment rheumatoid arthritis. In this study, we have investigated use VSV-G pseudotyped, HIV-1-based lentiviral vectors gene articular tissues. Recombinant lentivirus containing β-galactosidase/neomycin resistance fusion under control elongation factor (EF) 1α promoter efficiently transduced human and rat synoviocytes chondrocytes in cell culture. When directly injected into knees rats,...

To investigate the use of a scaffold seeded with genetically modified meniscal cells or mesenchymal stem (MSCs) for healing lesions, primary meniscus and bone marrow–derived MSCs were isolated from bovine calves transduced first-generation adenoviral vectors encoding green fluorescent protein, luciferase, transforming growth factor (TGF)-β1 complementary deoxyribonucleic acid (cDNA). The in type I collagen–glycosaminoglycan (GAG) matrices transplanted into tears avascular zone menisci. After...

Clinical translation of gene-based therapies for arthritis could be accelerated by vectors capable efficient intra-articular gene delivery and long-term transgene expression. Previously, we have shown that lentiviral transduce rat synovium efficiently in vivo. Here, evaluated the functional capacity expression provided lentiviral-mediated to joint. To do this, measured ability a vector containing cDNA human interleukin-1 receptor antagonist (LV-hIL-1Ra) suppress responses IL-1beta. Groups...

The complete exon-intron organization of the human aggrecan gene has been defined, and exon compared with individual domains protein core. A yeast artificial chromosome containing was selected from Centre d'Etude du Polymorphisme Humaine library. cosmid sulibrary created this, direct sequencing cosmids used to provide organization. found be composed 19 exons ranging in size 77 4224 bp. Exon 1 is non-coding, whereas 2-19 code for a core 2454 amino acids calculated mass 254379 Da. Intron at...

The inability of the ruptured anterior cruciate ligament (ACL) knee joint to heal spontaneously presents numerous clinical problems. Here we describe a novel, gene-based approach augment ACL healing. It is based upon migration cells from ends into collagen hydrogel laden with recombinant adenovirus. Cells entering gel become transduced by vector, which provides basis for local synthesis gene products that aid repair. Monolayers bovine were readily first-generation, adenovirus, and transgene...

In osteoarthritis (OA) articular chondrocytes undergo phenotypic changes culminating in the progressive loss of cartilage from joint surface. The molecular mechanisms underlying these are poorly understood. Here we report enhanced (approximately 7-fold) expression F-spondin, a neuronal extracellular matrix glycoprotein, human OA (P<0.005). OA-specific up-regulation F-spondin was also demonstrated rat knee following surgical menisectomy. treatment explants caused 2-fold increase levels active...

Local gene therapy for chronic joint diseases requires prolonged transgenic expression, but this has not been reliably achieved in animal models. Using normal and immunocompromised animals, we examined the capacity of various cell types tissues to maintain express exogenous transgenes after direct intra-articular delivery. We found that expression could persist lifetime required precise immunological compatibility between vector, transgene product, host. It was dependent on vector...

Anakinra, the recombinant form of IL-1 receptor antagonist (IL-1Ra), has been approved for clinical use in treatment rheumatoid arthritis as drug Kineret™, but it must be administered daily by subcutaneous injection. Gene transfer may offer a more effective means delivery. In this study, using prostaglandin E2 production measure stimulation, we quantitatively compared ability anakinra, well that IL-1Ra delivered gene transfer, to inhibit biologic actions IL-1β. Human synovial fibroblast...

F-spondin is a pericellular matrix protein upregulated in developing growth plate cartilage and articular during osteoarthritis. To address its function bone vivo, we generated mice that were deficient for the gene, Spon1. Spon1−/− viable developed normally to adulthood with no major skeletal abnormalities. At 6 months, femurs tibiae of exhibited increased mass, evidenced by histological staining micro CT analyses, which persisted up 12 months. In contrast, abnormalities observed at any age...

Leiomyosarcoma (LMS) is an aggressive, often poorly differentiated cancer of the smooth muscle (SM) lineage for which molecular drivers transformation and progression are understood. In microRNA (miRNA) profiling studies, miR-130b was previously found to be upregulated in LMS vs. normal SM, down-regulated during differentiation mesenchymal stem cells (MSCs) into suggesting a role tumor progression. present study, effects on human tumorigenesis were investigated. Stable overexpression...

Articular cartilage forms the load-bearing surfaces of diarthrodial joints. Its dense extracellular matrix, composed primarily water, collagen, and proteoglycans is maintained by a resident population chondrocytes. These all combine to form highly organized structure that provides tissue with its unique biomechanical properties. However, partly because avascular nature, has poor regenerative properties, damage from disease or injury will often lead an inferior repair ultimately fails. One...