Search

Knee Osteoarthritis (OA) is a highly prevalent condition affecting knee joint that causes loss of physical function and pain. Clinical treatments are mainly focused on pain relief and limitation of disabilities; therefore, it is crucial to find new paradigms assessing cartilage conditions for detecting and monitoring the progression of OA. The goal of this paper is to highlight the predictive power of several features, such as cartilage density, volume and surface. These features were extracted from the 3D reconstruction of knee joint of forty-seven different patients, subdivided into two categories: degenerative and non-degenerative. The most influent parameters for the degeneration of the knee cartilage were determined using two machine learning classification algorithms (logistic regression and support vector machine); later, box plots, which depicted differences between the classes by gender, were presented to analyze several of the key features' trend. This work is part of a strategy that aims to find a new solution to assess cartilage condition based on new-investigated features.

For the observation of human joint cartilage, X-ray, computed tomography (CT) or magnetic resonance imaging (MRI) are the main diagnostic tools to evaluate pathologies or traumas. The current work introduces a set of novel measurements and 3D features based on MRI and CT data of the knee joint, used to reconstruct bone and cartilages and to assess cartilage condition from a new perspective. Forty-seven subjects presenting a degenerative disease, a traumatic injury or no symptoms or trauma were recruited in this study and scanned using CT and MRI. Using medical imaging software, the bone and cartilage of the knee joint were segmented and 3D reconstructed. Several features such as cartilage density, volume and surface were extracted. Moreover, an investigation was carried out on the distribution of cartilage thickness and curvature analysis to identify new markers of cartilage condition. All the extracted features were used with advanced statistics tools and machine learning to test the ability of our model to predict cartilage conditions. This work is a first step towards the development of a new gold standard of cartilage assessment based on 3D measurements.

Rolling circle amplification (RCA) of a synthetic nucleic acid target is detected using magnetic nanoparticles (MNPs) combined with an optomagnetic (OM) readout. Two RCA assays are developed with on-chip detection of rolling circle products (RCPs) either at end-point where MNPs are mixed with the sample after completion of RCA or in real time where MNPs are mixed with the sample during RCA. The plastic chip acts as a cuvette, which is positioned in a setup integrated with temperature control and simultaneous detection of four parallel DNA hybridization reactions between functionalized MNPs and products of DNA amplification. The OM technique probes the small-angle rotation of MNPs bearing oligonucleotide probes complementary to the repeated nucleotide sequence of the RCPs. This rotation is restricted when MNPs bind to RCPs, which can be observed as a turn-off of the signal from MNPs that are free to rotate. The amount of MNPs bound to RCPs is found to increase in response to the amplification time as well as in response to the synthetic DNA target concentration (2-40 pM dynamic range). We report OM real-time results obtained with MNPs present during RCA and compare to relevant end-point OM results for RCPs generated for different RCA times. The real-time approach avoids opening of tubes post-RCA and thus reduces risk of lab contamination with amplification products without compromising the sensitivity and dynamic range of the assay.

Rolling circle amplification (RCA) is a linear isothermal amplification technique that is widely applied in biomolecular assays due to its high specificity. Handling of a target sample using magnetic microbeads (MMBs) in a multi-step assay is appealing as the MMBs enable separation and transportation using an external magnet. Detection of amplicons using optomagnetic measurements of the rotational diffusion properties of magnetic nanoparticles (MNPs) is also appealing as it can be performed on any transparent sample container. Two strategies are described for integration of MMB sample handling in an RCA assay with on-chip optomagnetic detection of the amplification products. The first strategy relies on selective and irreversible release of the amplicons from the MMBs so that the binding of functionalized MNPs to the amplicons can be detected optomagnetically. The second strategy relies on the incorporation of MNPs into RCA products during RCA, followed by their separation on MMBs and subsequent optomagnetic detection upon release from the RCA products. Using MMB handling of RCA steps, the limits of detection (LODs) for a synthetic DNA target representative of Victoria Influenza type B were found to be between 4 and 20 pM with total assay times between 2 and 2.5 h. Without magnetic microbead sample handling, the LOD was 200 fM. The findings provide deeper insight into the use of magnetic microbeads as solid substrates to handle a DNA target for integration of RCA as well as other DNA-based assays. Graphical Abstract Schematic illustration of magnetic microbeads transporting a DNA target through the steps in a rolling circle amplification assay. Optomagnetic measurements detect the binding of magnetic nanoparticles to amplicons released from microbeads (top) or the pH-induced release of magnetic nanoparticles trapped in amplicons (bottom).