Previous studies showed that coculture of primary chondrocytes (PCs) with various sources multipotent cells results in a higher relative amount cartilage matrix formation than cultures containing only chondrocytes. The aim this study was to investigate the mechanism underlying observation. We used pellet models human mesenchymal stem (hMSCs) and PCs or bovine (bPCs) studied fate contribution individual cell populations during coculture. Enhanced deposition confirmed by histology...

There is great interest in genetic modification of bone marrow-derived mesenchymal stem cells (MSC), not only for research purposes but also use (autologous) patient-derived-patient-used transplantations. A major drawback bulk methods modifications (stem) cells, like bulk-electroporation, its limited yield DNA transfection (typically then 10%). This even more when are present at very low numbers, as the case cells. Here we an alternative technology to transfect with high efficiency (>75%),...

We have synthesized and characterized gold nanoparticles (spheres rods) with optical extinction bands within the “optical imaging window.” The intense plasmon resonant driven absorption scattering peaks of these make them suitable as contrast agents for techniques. Further, we conjugated to a mouse monoclonal antibody specific HER2 overexpressing SKBR3 breast carcinoma cells. bioconjugation protocol uses noncovalent modes binding based on combination electrostatic hydrophobic interactions...

Actively steering the chondrogenic differentiation of mesenchymal stromal cells (MSCs) into either permanent cartilage or hypertrophic destined to be replaced by bone has not yet been possible. During limb development, developing long is exposed a concentration gradient oxygen, with lower oxygen tension in region become articular and higher transient cartilage. Here, we prove that metabolic programming MSCs directs chondrogenesis hyaline Human chondrogenically differentiated vitro under...

Osteoarthritis (OA) is a multifactorial disease characterized by gradual degradation of joint cartilage. This study aimed to quantify major pathogenetic factors during OA progression in human Cartilage specimens were isolated from patients and scored 0-5 according the Research Society International (OARSI) guidelines. Protein gene expressions measured immunohistochemistry qPCR, respectively. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays used detect apoptotic...

Upon activation, ERKs translocate from the cytoplasm to nucleus. This process is required for induction of many cellular responses, yet molecular mechanisms that regulate ERK nuclear translocation are not fully understood. We have used a mouse embryo fibroblast ERK1-knock-out cell line expressing green fluorescent protein (GFP)-tagged ERK1 probe spatio-temporal regulation ERK1. Real time fluorescence microscopy and correlation spectroscopy revealed accumulation increased upon serum...

Abstract Introduction Osteoarthritis is, at least in a subset of patients, associated with hypertrophic differentiation articular chondrocytes. Recently, we identified the bone morphogenetic protein (BMP) and wingless-type MMTV integration site (WNT) signaling antagonists Gremlin 1 ( GREM1 ), frizzled-related FRZB ) dickkopf homolog Xenopus laevis DKK1 as cartilage’s natural brakes differentiation. In this study, investigated whether factors implicated osteoarthritis or regulation...

Abstract Osteoarthritis (OA) is a multifactorial degenerative joint disease of which the underlying mechanisms are yet to be fully understood. At molecular level, multiple factors including altered signaling pathways, epigenetics, metabolic imbalance, extracellular matrix degradation, production metalloproteinases, and inflammatory cytokines, known play detrimental role in OA. However, these do not initiate OA, but mediators or consequences disease, while many other causing etiology OA still...

T cell factor/lymphoid enhancer factor (TCF/LEF) transcription factors are downstream effectors of Wnt/β-catenin signaling, which has been implicated in the development and progression osteoarthritis (OA). This study aimed to investigate role TCF/LEF human articular chondrocytes. Primary osteoarthritic cartilage predominantly expressed TCF4 a lesser extent, LEF1 TCF3 mRNA. Overexpression TCF4, but not or LEF1, induced MMP-1, -3, -13 expression generic MMP activity was due potentiating NF-κB...

We constructed a photoactivatable Drosophila histone 2 A variant green fluorescent fusion protein (H2AvD‐paGFP) for tracking chromatin loci in living embryos. Activation of paGFP was achieved by irradiation from single‐photon diode laser at 408 nm, but activated nuclei failed to divide. Photoconversion could also be two‐photon fs pulses the range 780–840 nm. Viability whole‐mount embryos only maintained 820 which we activate, simultaneously track and quantitate mobility multiple loci. This...

Hypertrophic differentiation occurs during in vitro chondrogenesis of mesenchymal stem cells (MSCs), decreasing the quality cartilage construct. Previously we identified WNT pathway antagonists Dickkopf 1 homolog (DKK1) and frizzled-related protein (FRZB) as key factors blocking hypertrophic human MSCs (hMSCs). In this study, investigated role endogenously expressed DKK1 FRZB hMSC chondrocyte redifferentiation preventing cell hypertrophy using three relevant based systems, isolated hMSCs,...

Mesenchymal stem cells (MSCs) are multipotent cells, mainly from bone marrow, and an ideal source of in cartilage tissue engineering. A study the chondrogenic differentiation MSCs is particular interest for MSCs-based regeneration. In this study, we aimed to optimize conditions chrondogenic by regulating WNT signaling using small molecule inhibitor PKF118-310 activator BIO. Human mesenchymal (hMSCs) were isolated marrow aspirates cultured hMSCs proliferation medium. Pellet culture was...

We used human fetal bone marrow-derived mesenchymal stromal cells (hfMSCs) differentiating towards chondrocytes as an alternative model for the growth plate (GP). Our aims were to study gene expression patterns associated with chondrogenic differentiation assess whether derived from hfMSCs are a suitable studying development and maturation of GP. efficiently formed hyaline cartilage in pellet culture presence TGFβ3 BMP6. Microarray principal component analysis applied profiles during...

Nanotechnology has found its potential in every possible field of science and engineering. It offers a plethora options to design tools at the nanometer scale, which can be expected function more effectively than micro‐ macrosystems for specific applications. Although debate regarding safety synthetic nanomaterials clinical applications endures, it is promising technology due augment current treatments. Various materials such as polymer, biopolymers, or naturally occurring proteins peptides...

Interleukin 1 beta (IL1β) and Wingless-Type MMTV Integration Site Family (WNT) signaling are major players in Osteoarthritis (OA) pathogenesis. Despite having a large functional overlap OA onset development, the mechanism of IL1β WNT crosstalk has remained largely unknown. In this study, we have used combination computational modeling molecular biology to reveal direct or indirect between these pathways. Specifically, revealed by which upregulates via downregulating antagonists, DKK1 FRZB....

Computational support is essential in order to reason on the dynamics of biological systems. We have developed software tool ANIMO (Analysis Networks with Interactive MOdeling) provide such computational and allow insight into complex networks signaling events occurring living cells. makes use timed automata as an underlying model, thereby enabling analysis techniques from computer science like model checking. Biology experts are able via a user interface specifically tailored for...

Mesenchymal stem cells (MSC) have the ability to self-renew and differentiate into multiple cell types valuable for clinical treatment of rheumatic pathologies. To study chondrogenic potential MSC identify conditions that recreate native cartilage environment, we used time-of-flight secondary ion mass spectrometry (TOF-SIMS) label-free detection cell-type- environmental-condition-specific molecular profiles. We observed coculture human chondrocytes under standard culture leads improved...

Differentiation of chondrocytes towards hypertrophy is a natural process whose control essential in endochondral bone formation. It additionally thought to play role several pathophysiological processes, with osteoarthritis being prominent example. We perform dynamic analysis qualitative mathematical model the regulatory network that directs this phenotypic switch investigate influence individual factors holistically. To estimate stability SOX9 positive state (associated...