Abstract

In-situ assessment of collateral ligaments strain could be key to improving total knee arthroplasty outcomes by improving the ability of surgeons to properly balance the knee intraoperatively. Ultrasound (US) speckle tracking methods have shown promise in their capability to measure in-situ soft tissue strain in large tendons but prior work has also highlighted the challenges that arise when attempting to translate these approaches to the in-situ assessment of collateral ligaments strain. Therefore, the aim of this project was to develop and validate an US speckle tracking method to specifically assess in-situ strains of both the MCL and LCL. We hypothesize that coefficients of determination (R²) would be above 0.90 with absolute differences below 0.50% strain for the comparison between US-based and the reference strain, with better results expected for the LCL compared with the MCL.

Five cadaveric legs with total knee implants (NH019 2017-02-03) were submitted to a varus (LCL) and valgus (MCL) ramped loading (0 – 40N). Ultrasound radiofrequency (rf) data and reference surface strains data, obtained with 3D digital image correlation (DIC), were collected synchronously. Prior to processing, US data were qualitatively assessed and specimens displaying substantial imaging artefacts were discarded, leaving five LCL and three MCL specimens in the analysis. Ultrasound rf data were processed in Matlab (The MathWorks, Inc., Natick, MA) with a custom-built speckle tracking approach adapted from a method validated on larger tendons and based on normalized cross-correlation. Digital image correlation data were processed with commercial software VIC3D (Correlated Solutions, Inc., Columbia, SC). To optimize speckle tracking, several tracking parameters were tested: kernel and search window size, minimal correlation coefficient and simulated frame rate. Parameters were ranked according to three comparative measures between US- and DIC-based strains: R², mean absolute error and strains differences at 40N. Parameters with best average rank were considered as optimal.

To quantify the agreement between US- and DIC-based strain of each specimen, the considered metrics were: R², mean absolute error and strain differences at 40N. The LCL showed a good agreement with a high average R² (0.97), small average mean absolute difference (0.37%) and similar strains at 40N (DIC = 2.92 ± 0.10%; US = 2.99 ± 1.16%). The US-based speckle tracking method showed worse performance for the MCL with a lower average correlation (0.55). Such an effect has been observed previously and may relate to the difficulty in acquiring sufficient image quality for tracking the MCL compared to the LCL, which likely arises due to structural or mechanical differences; notably MCL is larger, thinner, more wrapped around the bone and stretches less. However, despite these challenges, the MCL tracking still showed small average mean absolute differences (0.44%) and similar strains at 40N (DIC = 1.48 ± 0.06%; US = 1.44 ± 1.89%).

We conclude that the ultrasound speckle tracking method developed is ready to be used as a tool to assess in-situ strains of LCL. Concerning the MCL strain assessment, despite some promising results in terms of strain differences, further work on acquisition could be beneficial to reach similar performance.

Summary Statement

Calcium phosphate (CaP) particles have attracted great interest as transfection reagents, yet little is known about their mechanism of internalisation. We report live cell time-course tracking of CaP particles during internalisation and the influence of Ca:P ratio on transfection efficiency.

Orthopedic Device-Related Infections (ODRIs) are a major medical challenge, particularly due to the involvement of biofilm-encased and multidrug-resistant bacteria. Current treatments, based on antibiotic administration, have proven to be ineffective. Consequently, there is a need for antibiotic-free alternatives. Antimicrobial peptides (AMPs) are a promising solution due to their broad-spectrum of activity, high efficacy at very low concentrations, and low propensity to induce resistance. We aim to develop a new AMP-based chitosan nanogel to be injected during orthopedic device implantation to prevent ODRIs. Chitosan was functionalized with norbornenes (NorChit) through the reaction with carbic anhydride and then, a cysteine-modified AMP, Dhvar5, a peptide with potent antibacterial activity, even against methicillin-resistant Staphylococcus aureus (MRSA), was covalently conjugated to NorChit (NorChit- Dhvar5), through a thiol-norbornene photoclick chemistry (UV= 365 nm). For NorChit-Dhvar5 nanogels production, the NorChit-Dhvar5 solution (0.15% w/v) and Milli-Q water were injected separately into microfluidic system. The nanogels were characterized regarding size, concentration, and shape, using Transmission Electron Microscopy (TEM), Nanoparticle Tracking Analysis (NTA) and Dynamic light scattering (DLS). The nanogels antibacterial properties were assessed in Phosphate Buffer (PBS) for 6 h, against four relevant microorganisms (Pseudomonas aeruginosa, S. aureus and MRSA, and in Muller- Hinton Broth (MHB), 50% (v/v) in PBS, supplemented with human plasma (1% (v/v)), for 6 and 24 h against MRSA. The obtained NorChit-Dhvar5 nanogels, presented a round-shaped and ∼100 nm. NorChit- Dhvar5 nanogels in a concentration of 10. 10. nanogels/mL in PBS were capable of reducing the initial inoculum of P. aeruginosa by 99%, S. aureus by 99%, and MRSA by 90%. These results were corroborated by a 99% MRSA reduction, after 24 h in medium. Furthermore, NorChit-Dhvar5 nanogels do not demonstrate signs of cytotoxicity against MC3T3-E1 cells (a pre-osteoblast cell line) after 14 days, having high potential to prevent antibiotic-resistant infection in the context of ODRIs

Abstract. Objectives. Neonatal motor development transitions from initially spontaneous to later increasingly complex voluntary movements. A delay in transitioning may indicate cerebral palsy (CP). The general movement optimality score (GMOS) evaluates infant movement variety and is used to diagnose CP, but depends on specialized physiotherapists, is time-consuming, and is subject to inter-observer differences. We hypothesised that an objective means of quantifying movements in young infants using motion tracking data may provide a more consistent early diagnosis of CP and reduce the burden on healthcare systems. This study assessed lower limb kinematic and muscle force variances during neonatal infant kicking movements, and determined that movement variances were associated with GMOS scores, and therefore CP. Methods. Electromagnetic motion tracking data (Polhemus) was collected from neonatal infants performing kicking movements (min 50° knee extension-flexion, <2 seconds) in the supine position over 7 minutes. Tracking data from lower limb anatomical landmarks (midfoot inferior, lateral malleolus, lateral knee epicondyle, ASIS, sacrum) were applied to subject-scaled musculoskeletal models (Gait2354_simbody, OpenSim). Inverse kinematics and static optimisation were applied to estimate lower limb kinematics (knee flexion, hip flexion, hip adduction) and muscle forces (quadriceps femoris, biceps femoris) for isolated kicks. Functional principal component analysis (fPCA) was carried out to reduce kicking kinematic and muscle force waveforms to PC scores capturing ‘modes’ of variance. GMOS scores (lower scores = reduced variety of movement) were collected in parallel with motion capture by a trained operator and specialised physiotherapist. Pearson's correlations were performed to assess if the standard deviation (SD) of kinematic and muscle force waveform PC scores, representing the intra-subject variance of movement or muscle activation, were associated with the GMOS scores. Results. The study compared GMOS scores, kinematics, and muscle force variances from a total of 26 infants with a mean corrected gestational age of 39.7 (±3.34) weeks and GMOS scores between 21 and 40. There was a significant association between the SD of the PC scores for knee flexion and the GMOS scores (PC1: R = 0.59, p = 0.002; PC2: R = 0.49, p = 0.011; PC3: R = 0.56, p = 0.003). The three PCs captured variances of the overall flexion magnitude (66% variance explained), early-to-late kick knee extension (20%), and continual to biphasic kicking (6%). For hip flexion, only the SD of PC1 correlated with GMOS scores (PC1: R = 0.52, p = 0.0068), which captured the variance of the overall flexion magnitude (81%). For the biceps femoris, the SD of PC1 and PC3 associated with GMOS scores (PC1: R = 0.50, p = 0.002; PC3: R = 0.45, p = 0.03), which captured the variance of the overall bicep force magnitude (79%) and early-to-late kick bicep activation (8%). Conclusions. Infants with reduced motor development as scored in the GMOS displayed reduced variances of knee and hip flexion and biceps femoris activation across kicking cycles. These findings suggest that combining objectively measured movement variances with existing classification methods could facilitate the development of more consistent and accurate diagnostic tools for early detection of CP. Declaration of Interest. (b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project