The field of nanoparticle related research for the diagnosis and therapy of diseases evolves rapidly. Magnetic nanoparticles in combination with magnetizable implant materials for the treatment of implant related infections present a possible implementation in orthopedics. Magnetic nanoporous silica nanoparticles (MNPSNPs) were developed and equipped with fluorescent dyes. In vitro/in vivo biocompatibility and in vivo biodistribution were examined to appraise their potential applicability. Cell culture tests with NIH-3T3 and HepG2 cell lines indicated a good in vitro biocompatibility. Ferritic and titanium alloy (control) plates were implanted subcutaneously at the hind legs of Balb/c mice. Immediately after i.v. or s.c. injection of MNPSNPs, the caudal half of the mice was placed between the poles of an electro magnet. Exposure to the electromagnetic field of approx. 1.7 T was maintained for 10 minutes. 10 animals each were euthanized at days 0, 1, 7, 21 or 42, respectively. Quantity of MNPSNPs in liver, spleen, kidney, lung and skin/muscle samples was assessed by fluorescent microscopic methods. MNPSNP existence on the implant surface was also appraised after several steps of detachment. MNPSNPs showed a time-dependent accumulation in the organs after i.v. injection with initial accumulation in the lungs followed by redistribution to liver and spleen. After s.c. injection no systemic distribution but local appearance of MNPSNPs could be found. First histological evaluation showed no pathological changes after i.v. injection. With good in vivo biocompatibility, future focus will be laid on increasing circle life time of MNPSNPs and evaluation in an infection model

Tissue engineering and regenerative medicine (TERM) hold the promise to provide therapies for injured tendons despite the challenging cues of tendon niche and the lack of specific factors to guide regeneration. The emerging potential of magnetic responsiveness and magnetic nanoparticles (MNPs) functionalities offers new perspectives to tackle TERM challenges. Moreover, pulsed electromagnetic field (PEMF) is FDA approved for orthopaedics with potential to control inflammation upon injury. We previously demonstrated that magnetic cell-sheets assisted by PEMF trigger the inflammation resolution by modulating cytokine-enriched environments [1]. To further understand the potential of magnetically assisted living patches, we have recently conducted in vivo studies using a rat patellar defect model. After labeling of human adipose stem cells with iron oxide MNPs for 16h, magCSs were cultured up to 3 days in α-MEM medium under non-magnetic or PEMF conditions. MagCSs were evaluated by immunocytochemistry, and real time RT-PCR for tendon markers. Cell metabolic activity was also assessed by MTS and ECM proteins quantified by Sirius Red/Fast Green. The MagCSs effect in ameliorating healing was assessed after implantation in window defects created in the patellar tendon of rats. PEMF was externally applied (3mT, 70Hz) 3d/week for 1h (magnetotherapy). After 4 and 8w, tendons were histologically characterized for immune-detection of tendon and inflammatory markers, and for Perls van Gieson and HE stains. Blood and detoxification organs were screened for inflammatory mediators and biodistribution of MNPs, respectively. In vitro results suggest that PEMF stimulates cellular metabolic activity, influences protein synthesis and the deposition of collagen and non-collagenous proteins is significantly increased compared to non-magnetic conditions. No adverse reactions, as infection or swelling, were observed after surgery or during follow-up. After 8w, magCSs remained at the implantation site and no MNPs were detected on detoxification organs. Plasma levels of IL1α, β, IL6 and TNFα assessed by multiplex assay were below detectable values (<12.5pg/ml). Thus, the combination of cell sheets and magnetic technologies hold promise for the development of living tendon substitutes. Acknowledgement to ERC-COG MagTendon772817, H2020 Achilles 810850, FCT - 2020.01157.CEECIND

Tendons display poor intrinsic healing properties and are difficult to treat[1]. Prior in vitro studies[2] have shown that, by targeting the Activin A receptor with magnetic nanoparticles (MNPs), it is possible to remotely induce the tenogenic differentiation of human adipose stem cells (hASCs). In this study, we investigated the tenogenic regenerative potential of remotely-activated MNPs-labelled hASCs in an in vivo rat model. We consider the potential for magnetic controlled nanoparticle mediated tendon repair strategies. hASCs were labelled with 250 nm MNPs functionalized with anti-Activin Receptor IIA antibody. Using a rapid curing fibrin gel as delivery method, the MNPs-labelled cells were delivered into a Ø2 mm rat patellar tendon defect. The receptor was then remotely stimulated by exposing the rats to a variable magnetic gradient (1.28T), using a customised magnetic box. The stimulation was performed 1 hour/day, 3 days/week up to 8 weeks. Tenogenesis, iron deposition and collagen alignment were assessed by histological staining and IHC. Inflammation mediators levels were assessed by ELISA and IHC. The presence of human cells in tendons after 4 and 8 weeks was assessed by FISH analysis. Histological staining showed a more organised collagen arrangement in animals treated with MNPs-labelled cells compared to the controls. IHC showed positive expression of tenomodulin and scleraxis in the experimental groups. Immunostaining for CD45 and CD163 did not detect leukocytes locally, which is consistent with the non-significant levels of the inflammatory cytokines analysis performed on plasma. While no iron deposition was detected in the main organs or in plasma, the FISH analysis showed the presence of human donor cells in rat tendons even after 8 weeks from surgery. Our approach demonstrates in vivo proof of concept for remote control stem cell tendon repair which could ultimately provide injectable solutions for future treatment. We are grateful for ERC Advanced Grant support ERC No.789119, ERC CoG MagTendon No.772817 and FCT grant 2020.01157.CEECIND

A growing number of children are being implanted with magnetically controlled growing rods. Some also have baclofen pumps, a device that delivers a neuromodulating medication using a magnet. There has been no previous testing to determine if the External Remote Controller (ERC) would stall the baclofen pump during magnetic rod lengthening, a potentially life-threatening event. Two baclofen pumps, 20 mL and 40 mL, were placed in varying proximity to the ERC while the magnetic field was generated. The speed (dose) of the baclofen pump, the distance from the ERC, the length of time the ERC was run, the position of the pump in relationship to the magnetic field, and the presence or absence of a stall were recorded. To stall a baclofen pump, the ERC needed to be running continuously for at least 30 mm of lengthening (about 3 minutes). The pump would not stall if it was more than 1 cm away from the ERC, even if the ERC generated a field for 3 minutes. The speed of the pump did not affect the instance of a motor stall. The pump was more likely to stall if it was positioned in front of or behind the magnetic field, as opposed to the center of the field. Baclofen pumps are always more than 1 cm away from the magnetic rods when both are implanted in a child. The typical lengthening interval is under 10 mm, much less than what was required to stall a baclofen pump. The ERC is highly unlikely to stall a baclofen pump during a magnetic rod lengthening, making a life-threatening event almost impossible

Aims. In order to release the contracture band completely without damaging normal tissues (such as the sciatic nerve) in the surgical treatment of gluteal muscle contracture (GMC), we tried to display the relationship between normal tissue and contracture bands by magnetic resonance neurography (MRN) images, and to predesign a minimally invasive surgery based on the MRN images in advance. Methods. A total of 30 patients (60 hips) were included in this study. MRN scans of the pelvis were performed before surgery. The contracture band shape and external rotation angle (ERA) of the proximal femur were also analyzed. Then, the minimally invasive GMC releasing surgery was performed based on the images and measurements, and during the operation, incision lengths, surgery duration, intraoperative bleeding, and complications were recorded; the time of the first postoperative off-bed activity was also recorded. Furthermore, the patients’ clinical functions were evaluated by means of Hip Outcome Score (HOS) and Ye et al’s objective assessments, respectively. Results. The contracture bands exhibited three typical types of shape – feather-like, striped, and mixed shapes – in MR images. Guided by MRN images, we designed minimally invasive approaches directed to each hip. These approaches resulted in a shortened incision length in each hip (0.3 cm (SD 0.1)), shorter surgery duration (25.3 minutes (SD 5.8)), less intraoperative bleeding (8.0 ml (SD 3.6)), and shorter time between the end of the operation and the patient’s first off-bed activity (17.2 hours (SD 2.0)) in each patient. Meanwhile, no serious postoperative complications occurred in all patients. The mean HOS-Sports subscale of patients increased from 71.0 (SD 5.3) to 94.83 (SD 4.24) at six months postoperatively (p < 0.001). The follow-up outcomes from all patients were “good” and “excellent”, based on objective assessments. Conclusion. Preoperative MRN analysis can be used to facilitate the determination of the relationship between contracture band and normal tissues. The minimally invasive surgical design via MRN can avoid nerve damage and improve the release effect. Cite this article: Bone Jt Open 2024;5(9):776–784

A critical bone defect may be more frequently the consequence of a trauma, especially when a fracture occurs with wide exposure, but also of an infection, of a neoplasm or congenital deformities. This defect needs to be treated in order to restore the limb function. The treatments most commonly performed are represented by implantation of autologous or homologous bone, vascularized fibular grafting with autologous or use of external fixators; all these treatments are characterized by several limitations. Nowadays bone tissue engineering is looking forward new solutions: magnetic scaffolds have recently attracted significant attention. These scaffolds can improve bone formation by acting as a “fixed station” able to accumulate/release targeted growth factors and other soluble mediators in the defect area under the influence of an external magnetic field. Further, magnetic scaffolds are envisaged to improve implant fixation when compared to not-magnetic implants. We performed a series of experimental studies to evaluate bone regeneration in rabbit femoral condyle defect by implanting hydroxyapatite (HA), polycaprolactone (PCL) and collagen/HA hybrid scaffolds in combination with permanent magnets. Our results showed that ostetoconductive properties of the scaffolds are well preserved despite the presence of a magnetic component. Interestingly, we noticed that, using bio-resorbable collagen/HA magnetic scaffolds, under the effect of the static magnetic field generated by the permanent magnet, the reorganization of the magnetized collagen fibers produces a highly-peculiar bone pattern, with highly-interconnected trabeculae orthogonally oriented with respect to the magnetic field lines. Only partial healing of the defect was seen within the not magnetic control groups. Magnetic scaffolds developed open new perspectives on the possibility to exploiting magnetic forces to improve implant fixation, stimulate bone formation and control the bone morphology of regenerated bone by synergically combining static magnetic fields and magnetized biomaterials. Moreover magnetic forces can be exploited to guide targeted drug delivery of growth factors functionalized with nanoparticles

In orthopedic surgery, implant infections are a serious issue and difficult to treat. The aim of this study was to use superparamagnetic nanoporous silica nanoparticles (MNPSNP) as candidates for directed drug delivery. Currently, short blood circulation half-life due to interactions with the host's immune system hinder nanoparticles in general from being clinically used. PEGylation is an approach to reduce these interactions and to enhance blood circulation time. The effect of PEGylation of the used . 68. Ga-labelled MNPSNP on the distribution and implant accumulation was examined by PET/CT imaging and gamma counting in an implant mouse model. Female Balb/c mice (n=24) received a magnetic implant subcutaneously on the left and a titanium implant on the right hind leg. On day one, 12 of these mice received an additional clodronate®-injection for macrophage depletion. On the second postoperative day, mice were anaesthetized and MNPSNP (native or PEGylated) injected intravenously, followed by a dynamic PET-scan over 60 minutes, a CT- and a static PET-scan at 120 min. As control, 12 mice received only . 68. Ga-MNPSNP (native or PEGylated). Gamma counting of inner organs, urine, blood and implant area was performed as further final analysis. Although PEGylation of the nanoparticles already resulted in lower liver uptakes, both variants of . 68. Ga-labeled MNPSNP accumulated in liver and spleen. Combination of PEGylation with clodronate®-injection led to a highly significant effect whereas clodronate®-injection alone could not reveal significant differences. In gamma counting, a significantly higher %I.D./g was found for the tissue surrounding the magnetic implants compared to the titanium control, although in a low range. PEGylation and/or clodronate®-injection revealed no significant differences regarding nanoparticle accumulation at the implantation site. PEGylation increases circulation time, but MNPSNP accumulation at the implant site was still insufficient for treatment of infections. Additional efforts have to further increase circulation time and local accumulation. Acknowledgements: This work is funded by the German Research Foundation (DFG, project number 280642759)

Dislocation after total hip replacement (THR) is a devastating complication. Risk factors include patient and surgical factors. Mitigation of this complication has proven partially effective. This study investigated a new innovating technique to decrease this problem using rare earth magnets. Computer simulations with design and magnetic finite element analysis software were used to analyze and quantitate the forces around hip implants with embedded magnets into the components during hip range of motion. N52 Neodymium-Iron-Boron rare earth magnets were sized to fit within the existing acetabular shells and the taper of a hip system. Additionally, magnets placed within the existing screw holes were studied. A 50mm titanium acetabular shell and a 36mm ceramic liner utilizing a taper sleeve adapter were modeled which allowed for the use of a 12mm × 5mm magnet placed in the center hole, an 18mm × 15mm magnet within the femoral head, and 10mm × 5mm magnets in the screw holes. Biomechanical testing was also performed using in-vitro bone and implant models to determine retention forces through a range of hip motion. The novel system incorporating magnets generated retentive forces between the acetabular cup and femoral head of between 10 to 20 N through a range of hip motion. Retentive forces were stronger at the extreme position hip range of motion when additional magnets were placed in the acetabular screw holes. Greater retentive forces can be obtained with specially designed femoral head bores and acetabular shells specifically designed to incorporate larger magnets. Mechanical testing validated the loads obtained and demonstrated the feasibility of the magnet system to provide joint stability and prevent dislocations. Rare earth magnets provide exceptional attractive strength and can be used to impart stability and prevent dislocation in THR without the complications and limitations of conventional methods

After the implantation of endoprotheses or osteosynthesis devices, implant-related infections are one of the major challenges. The surface of implants offers optimal conditions for the formation of a biofilm. Effective carrier systems for the delivery of adequate therapeutics would reduce the concentrations needed for successful treatment and improve cure rates. In cancer diagnosis and therapy, magnetic nanoparticles are concentrated in the target area by an external magnetic field. For orthopaedic applications, in vitro examinations showed that the addition of a magnetic implant in combination with an external magnetic field could increase the amount of MNPSNPs that accumulated in direct vicinity to the implant. The present examinations implemented an electromagnet to increase magnetic field strength and should show if the in vitro set up can be transferred to an in vivo mouse model. Additionally, the loading capacity of the MNPSNPs with enrofloxacin and its release kinetics were determined. Fluorescein-isothiocyanate (FITC) was covalently attached to MNPSNPs. For the in vitro set up, a peristaltic pump was used to establish a closed circuit which contained the MNPSNP dispersion and a magnetic platelet. After 5 minutes fluid samples were taken from the area around the magnetic platelet and analysed using a microplate reader. For the in vivo set up, a BALB/c mouse was implanted subcutaneously with the metallic platelet at the hind leg. The MNPSNP dispersion was injected into the tale vein and the hind leg of the mouse was placed immediately in a magnetic field of 1.9 T. After one week the implant was retrieved and examined by confocal laser scanning microscopy (CLSM). Liver, spleen and kidneys of the mouse were examined by magnetic resonance imaging (MRI). The loading capacity of the MNPs with enrofloxacin was examined by quantification of the enrofloxacin content in the incubation and washing solution after incubation. The release kinetics weres tested in PBS using UV/Vis-spectrometry. The solution in the remaining tube contained no detectable MNPs while the concentration in the vicinity of the platelet was 150 µg/ml. The mouse showed no clinical adverse effects. The CLSM examination revealed a considerable accumulation of the MNPs at the implant surface. MRI could show neither accumulated MNPs nor changes of organ structure. The loading capacity of the MNPs for enrofloxacin was approximately 95 µg/mg. A burst release of nearly a third of the loaded antibiotic occurred within the first 6 hours followed by a further steady release. Conclusion. Loading and release of enrofloxacin showed appropriate results. For future studies antibiotics like rifampicin or vancomycin will be implemented. This first in vivo trial demonstrated an implant-directed targeting of the MNPs and successfully transferred the principle into an in vivo model so that a main study with statistically significant animal numbers has started including histological examinations

Tendon injuries are a worldwide problem affecting several age groups and stem cell based therapies hold potential for tendon strategies guiding tendon regeneration. Tendons rely on mechano-sensing mechanisms that regulate homeostasis and influence regeneration. The mechanosensitive receptors available in cell membranes sense the external stimuli and initiate mechanotransduction processes. Activins are members of the TGF-β superfamily which participate in several tendon biological processes. It is envisioned that the activation of the activin receptor, trigger downstream Smad2/3 pathway thus regulating the transcription of tenogenic genes driving stem cell differentiation. In this work, we propose to target the Activin receptor type IIA (ActRIIA) in human adipose stem cells (hASCs), inducing hASCs commitment towards the tenogenic lineage. Since mechanotransduction can be remotely triggered through magnetic actuation combined with magnetic nanoparticles (MNPs), we stimulated hASCs tagged complexes using a vertical oscillating magnetic bioreactor (MICA Biosystems Ltd). Carboxyl functionalised MNPs (Micromod) were coated with anti-ActRIIA antibody (Abcam) by carbodiimide activation. hASCs were then cultured with MNPs-anti-ActRIIA for 14days with or without magnetic exposure (1Hz, 1h/every other day). hASCs cultured alone in αMEM (negative control) or in αMEM supplemented with ActivinA (R&D systems) (positive control of ActRIIA activation) were used as experimental controls. The tenogenic commitment of hASCs was assessed by real time RT-PCR, immunocytochemistry and quantification of collagen and non-collagenous proteins. Moreover, the phosphorylation of Smad2/3 was also evaluated on hASCs incubated for 2, 10, or 30min under magnetic stimulated (1Hz) and non-stimulated conditions. The increased gene expression of tendon related markers and higher ECM proteins deposition suggests that remote magnetic activation of ActRIIA promotes effectively hASCs tenogenic commitment. Furthermore, the detection of phospho-Smad2/3 proteins by ELISA (Cell Signaling Technology) was significantly more intense after 10min in hASCs under magnetic stimulation and in comparison to the control groups. These outcomes suggest that ActRIIA is a mechanosensitive receptor that can be remotely activated upon magnetic stimulation. In conclusion, remotely activation of MNPs tagged hASCs has potential for modulating tenogenic differentiation of stem cells envisioning successful cell therapies for tendon regeneration. Acknowledgements. FCT/MCTES PD/59/2013 (fellowship PD/BD/113802/2015), FCT post-doctoral grant SFRH/BPD/111729/2015, FCT grant IF/00685/2012, and EU-ITN MagneticFun

Objectives. The purpose of this study was to assess N-acetyl aspartate changes in the thalamus in patients with osteoarthritis of the hip using proton magnetic resonance spectroscopy. Methods. Nine patients with osteoarthritis of the hip (symptomatic group, nine women; mean age 61.4 years (48 to 78)) and nine healthy volunteers (control group, six men, three women; mean age 30.0 years (26 to 38)) underwent proton magnetic resonance spectroscopy to assess the changes of N-acetyl aspartate in the thalamus. . Results. The ratio of N-acetyl aspartate to creatine plus phosphocreatine in the thalamus contralateral to the symptomatic hip in patients with osteoarthritis of the hip was significantly lower than the ratio of N-acetyl aspartate to creatine plus phosphocreatine in the thalamus in the control group (1.611 (1.194 to 1.882) vs 1.355 (1.043 to 1.502), p < 0.001). And, a strong negative correlation was detected between the ratio of N-acetyl aspartate to creatine plus phosphocreatine in the thalamus contralateral to the symptomatic hip in patients with osteoarthritis of the hip and pain duration (r = -0.83, p = 0.018). Conclusions. We evaluated the ratio of N-acetyl aspartate to creatine plus phosphocreatine in the thalamus of patients with osteoarthritis of the hip by using proton magnetic resonance spectroscopy. We concluded that the ratio of N-acetyl aspartate to creatine plus phosphocreatine in the thalamus contralateral to the symptomatic hip in patients with osteoarthritis of the hip were significantly lower than those in the thalamus of the control group, and that pain duration was strongly related to the decrease of the ratio of N-acetyl aspartate to creatine plus phosphocreatine

Aim. The aim of this study was to investigate the metabolomic profile of synovial fluid in periprosthetic joint infection (PJI) cases regarding a possible diagnostic approach. Also, further information about the metabolic composition of synovial fluid in PJI may point to future diagnostic and therapeutic approaches. Method. Patients with a clinical suspicion of a prosthesis infection who underwent a joint puncture in our outpatient department or ward were included. After sample preparation, the nuclear magnetic resonance (NMR) experiments were performed at 310 K on an AVANCE™ NeoBruker Ultrashield 600 MHz spectrometer. Bruker Topspin version 4.0.2 was used for NMR data acquisition. The spectra for all samples were automatically processed (exponential line broadening of 0.3 Hz), phased, and referenced using TSP at 0.0 ppm. In total, 37 metabolites were analysed using a volume of 200 µl per synovial sample. The PJI and aseptic cases were assigned according to the EBJIS criteria. Results. In total, 76 samples were included in the final analysis with 48 PJI cases and 28 aseptic cases. Five measured metabolites have shown an area under the curve (AUC) over 0.8, with Taurine (AUC 0.8558, p<0.0001) and Glutamine (AUC 0.8333, p<0.0001) showing the best diagnostic performance. When combining two metabolites, the AUC indicated even higher diagnostic performance: Glucose/Glycogen (AUC 0.9073, p<0.0001), Taurine/Mannose (AUC 0.9073, p<0.0001), Mannose/Glycogen (AUC 0.8992, p<0.0001) and Taurine/Glucose (AUC 0.8956, p<0.0001). Conclusions. While NMR as a method in PJI diagnostics is currently not broadly available for daily clinical work, our results indicate that certain synovial metabolites and their combinations can be used for PJI diagnosis

MSCs (mesenchymal stem cells) are bone marrow-derived cells capable of replication and differentiation in-vitro into several tissues including bone, cartilage, stroma, fat, muscle and tendon. MSCs can be isolated by relatively simple procedures and then expanded without losing the ability to differentiate into multiple lineages. As such, these cells have immense clinical potential in regenerative medicine and in orthopaedics for repair or replacement of damaged tissues. In this work we investigated the interaction between magnetic carbon nanotubes (CNTs) and MSCs and their ability to guide these cells injected intravenously in living mice by using an external magnetic field. CNTs did not affect cell viability and their ability to differentiate. Both the CNTs and the magnetic field did not alter cell growth rate, phenotype and cytoskeletal conformation. CNTs, when exposed to magnetic fields, are able to shepherd MSCs towards the magnetic source in vitro. Moreover, the application of a magnetic field alters the biodistribution of CNT-labelled MSCs after intravenous injection into rats. We demonstrated that CNTs hold the potential for use as nano-devices to improve therapeutic protocols for transplantation and homing of stem cells in vivo. This could pave the way for the development of new strategies for manipulation/guidance of MSCs in regenerative medicine and cell transplantation for the treatment of many orthopaedic diseases

Common tendon injuries impair healing, leading to debilitation and an increased re-rupture risk. The impact of oxygen-sensing pathways on repair mechanisms, vital in regulating inflammation and fibrosis, remains unclear despite their relevance in tendon pathologies. Recent studies show that pulsed electromagnetic field (PEMF) reduce inflammation in human tendon cells (hTDCs) and in hypoxia-induced inflammation. We investigated the hypoxia's impact (1% and 2% oxygen tension) using magnetic cell sheet constructs (IL-1β-magCSs) primed with IL-1β. IL-1β-magCSs were exposed to low OT (1h, 4h,6h) in a hypoxic chamber. To confirm the role of PEMF (5Hz, 4mT, 50% duty cycle) on hypoxia modulation, IL-1β-magCSs, previously exposed to OT, were 1h-stimulated with PEMF. Our results show a significant increase in HIF- 1a and HIF-2a expression on IL-1β-magCSs after exposure to 2%-OT at all time points, compared to 1%- OT and normoxia. TNFa, IL-6, and IL-8 expression increased after 6 hours of 1%-OT exposure. PEMF stimulation of hypoxic IL-1β-magCSs led to decreased pro-inflammatory genes and increased anti-inflammatory (IL-4,IL-10) expression compared to unstimulated magCSs. IFN-g, TNF-α, and IL-6 release increased after 6 hours, regardless of %-OT, while IL-10 levels tended to rise after PEMF stimulation at 2%-OT. Also, NFkB expression was increased on IL-1β-magCSs exposed to 4 h and 6 h of 2%-OT, suggesting a link between NFkB and the production of pro-inflammatory factors. Moreover, PEMF stimulation showed a significantly decreased NFkB level in IL-1β-magCSs. Overall, low OT enhances expression of hypoxia-associated genes and inflammatory markers in IL-1β-magCSs with the involvement of NFkB. PEMF modulates the response of magCSs, previously conditioned to hypoxia and to inflammatory triggers, favouring expression of anti-inflammatory genes and proteins, supporting PEMF impact in pro-regenerative tendon strategies. Acknowledgements: ERC CoG MagTendon(No.772817), FCT under the Scientific Employment Stimulus-2020.01157.CEECIND. Thanks to Hospital da Prelada for providing tendon tissue samples (Portugal), and TERM. RES Hub (Norte-01-0145-FEDER-022190)

Abstract. Objectives. Investigate Magnetic Resonance Imaging (MRI) as an alternative to Computerised Tomography (CT) when calculating kinematics using Biplane Video X-ray (BVX) by quantifying the accuracy of a combined MRI-BVX methodology by comparing with results from a gold-standard bead-based method. Methods. Written informed consent was given by one participant who had four tantalum beads implanted into their distal femur and proximal tibia from a previous study. Three-dimensional (3D) models of the femur and tibia were segmented (Simpleware Scan IP, Synopsis) from an MRI scan (Magnetom 3T Prisma, Siemens). Anatomical Coordinate Systems (ACS) were applied to the bone models using automated algorithms. 1. The beads were segmented from a previous CT and co-registered with the MRI bone models to calculate their positions. BVX (60 FPS, 1.25 ms pulse width) was recorded whilst the participant performed a lunge. The beads were tracked, and the ACS position of the femur and tibia were calculated at each frame (DSX Suite, C-Motion Inc.). The beads were digitally removed from the X-rays (MATLAB, MathWorks) allowing for blinded image-registration of the MRI models to the radiographs. The mean difference and standard deviation (STD) between bead-generated and image-registered bone poses were calculated for all degrees of freedom (DOF) for both bones. Using the principles defined by Grood and Suntay. 2. , 6 DOF kinematics of the tibiofemoral joint were calculated (MATLAB, MathWorks). The mean difference and STD between these two sets of kinematics were calculated. Results. The absolute mean femur and tibia ACS position differences (Table 1) between the bead and image-registered poses were found to be within 0.75mm for XYZ, with all STD within ±0.5mm. Mean rotation differences for both bones were found to be within 0.2º for XYZ (Table 1). The absolute mean tibiofemoral joint translations (Table 1) were found to be within ±0.7mm for all DOF, with the smallest absolute mean in compression-distraction. The absolute mean tibiofemoral rotations were found to be within 0.25º for all DOF (Table 1), with the smallest mean was found in abduction-adduction. The largest mean and STD were found in internal-external rotation due to the angle of the X-rays relative to the joint movement, increasing the difficulty of manual image registration in that plane. Conclusion. The combined MRI-BVX method produced bone pose and tibiofemoral kinematics accuracy similar to previous CT results. 3. This allows for confidence in future results, especially in clinical applications where high accuracy is needed to understand the effects of disease and the efficacy of surgical interventions. Acknowledgements: This research was supported by the Engineering and Physical Sciences Research Council (EPSRC) doctoral training grant (EP/T517951/1). 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

Significant challenges remain to accomplishing the development of fully functional tendon tissue substitutes that can lead to clinically effective and successful applications. Scaffolding materials must meet demanding requirements such i) mimic the hierarchical and anisotropically aligned structure of tendon tissues from the nano- up to the macroscale, ii) meet tendon mechanical requirements and non-linear biomechanical behaviour, iii) provide the necessary biophysical/biochemical cues and mechanical responsiveness to induce the tenogenic differentiation of stem cells and potentiating the effects of biochemical supplementation. On the other side, tenogenic differentiation of stem cells is still to be established, as well as the role of such cells (either naïve or pre-differentiated) in promoting tissue regeneration. We have recently found evidences that magnetic actuation can provide means of mechanically stimulating cells in a contact-free manner and, more interestingly, can also modulate inflammatory response, a critical issue for achieving tissue regeneration instead of repair. In summary, synergies of scaffold design and magnetic responsiveness can impact significantly cells behaviour as well as in vivo response and thus widen the therapeutically range of cell-laden tissue engineered constructs in tendon regeneration

Introduction. Diaphyseal bone defect represents a significant problem for orthopaedic surgeons and patients. Bone is a complex tissue whose structure and function depend strictly on ultrastructural organization of its components: cells, organic (extracellular matrix, ECM) and inorganic components. The purpose of this study was to evaluate bone regeneration in a critical diaphyseal defect treated by implantation of a magnetic scaffold fixed by hybrid system (magnetic and mechanical), supplied through nanoparticle-magnetic (MNP) functionalized with Vascular Endothelial-Growth-Factor-(VEGF) and magnetic-guiding. Methods. A critical long bone defect was created in 8 sheep metatarsus diaphysis: it was 20.0 mm in length; the medullary canal was reamed till 8.00 mm of inner diameter. Then a 8.00 mm diameter magnetic rod was fitted into proximal medullary canal (10 mm in length). After that a scaffold made of Hydroxyapatite (outer diameter 17.00 mm) that incorporates magnetite (HA/Mgn 90/10) was implanted to fill critical long bone defect. A magnetic rod (6.00 mm diameter) was firmly incorporated at proximal side into the scaffold. Both magnets had 10 mm length. To give stability to the complex bone-scaffold-bone a plate was used as a bridge; it was fixed proximally by 2 screws and distally by 3 screws. Scaffolds biocompatibility was previously assessed in vitro using human osteoblast-like cells. Magnetic forces through scaffold were calculated by finite element software (COMSOL Multiphysics, AC/DC Model). One week after surgery, magnetic nanoparticles functionalized with VEGF were injected at the mid portion of the scaffold using a cutaneous marker positioned during surgery as reference point in 4 sheep; other sheep were used as control group. After sixteen weeks, sheep were sacrificed to analyze metatarsi. Macroscopical, radiological and microCT examinations were performed. Results. Samples obtained didn't show any inflammatory tissue around the scaffold and revealed bone tissue formation inside pores of the scaffolds and we could see also complete coverage of the scaffolds. Formation of new bone tissue was more evident at magnetized bone-scaffold interface. X-rays showed a good integration of the scaffold with a good healing process of critical bone defect: new cortical bone formation seemed to be present, recreating continuity of metatarsus diaphysis. No signs of scaffold mobilization was showed (Fig. 1). All these datas were confirmed by the microCT: new bone formation inside the scaffolds was evident, in particular at proximal bone-scaffold interface, where permanent magnet were present (Fig. 2). These preliminary results lead our research to exploiting magnetic forces to stimulate bone formation, as attested in both in vitro and in vivo models and to improve fixation at bone scaffold interface, as calculated by finite element software, and moreover to guide targeted drug delivery without functionalized magnetic nanoparticles dissemination in all body

Introduction. Motorised intramedullary lengthening nails are considered more expensive than external fixators for limb lengthening. This research aims to compare the cost of femoral lengthening in children using the PRECICE magnetic lengthening nail with external fixation. Materials and Methods. Patients: Retrospective analysis of 50 children who underwent femoral lengthening. One group included patients who were treated with PRECICE lengthening nails, the other group included patients who had lengthening with external fixation. Each group included 25 patients aged between 11–17 years. The patients in both groups were matched for age. Cost analysis was performed following micro-costing and analysis of the used resources during the different phases of the treatments. Results. : Each group's mean patient age was 14.7 years. Lengthening nails were associated with longer operative times compared to external fixators, both for implantation and removal surgery (P-value 0.007 and <0.0001 respectively). Length of stay following the implantation surgery, frequency of radiographs, frequency of outpatient department appointments were all more favourable with lengthening nails. The overall cost of lengthening nails was £1393 more than external fixators, although this difference was not statistically significant (P-value 0.088). Conclusions. The cost of femoral lengthening with lengthening nails was not significantly higher than the external fixators’ cost. Further research to review the effectiveness of the devices and the quality of life during the lengthening process is crucial for robust health economic evaluation

Introduction: The anterior drawer test for anterior cruciate ligament (ACL) deficiency, requires a subjective assessment of joint movement, as the tibia is pulled forward. The aim of this study was to objectively quantify this movement using a magnetic tracking device. Materials and Methods: Ten patients aged 24 to 44 years were assessed as having unilateral ACL deficiency with conventional clinical tests. These patients were then re-assessed using a magnetic tracking device (Polhemus Fastrak). Patients had magnetic sensors attached around their femurs and tibias using elasticated Velcro straps. The Anterior Drawer test was then performed with the patient lying within range of the system’s magnetic source. The test was performed three times on the normal and injured knees of each patient, using a spring balance to apply a standard 20 lb (=89 N) force. During the tests, sensor position and orientation data was collected with an accuracy better than 1 mm and 1 degree, respectively. The data was sampled at 10Hz and stored on a computer for post-test analysis. This analysis deduced the tibial displacement resulting from each anterior drawer. Results: During the anterior drawer test the supine patient’s knee is in 90 degrees flexion, with the foot planted on the examination couch. As the tibia is pulled anteriorly, it rotates upwards from the foot and the femur experiences a corresponding rotation from the hip. These complex coupled movements are best quantified in terms of absolute displacement of the tibia from the femur. In the normal knees, the mean displacement of the tibia from the femur was 4.2 mm (SD=1.6). In comparison the ACL deficient knees had a mean displacement of 6.3 mm (SD=2.9). This is 50 % more. A paired t test of this data showed a highly significant difference, with P = 0.005. Conclusion: This study has quantified the movement produced during the Anterior Draw test for ACL deficiency. The tracker’s lightweight sensors caused minimal disturbance to the established clinical test. The system therefore provides objective measurement data to augment the clinicians subjective assessment

Introduction: Proton magnetic resonance spectroscopy (. 1. H MRS) is a powerful non-invasive technique used to identify and quantify chemical compounds. In a recent study, the early histopathologic findings of osteonecrosis showed marrow edema and hemorrhage, and the late findings were fibrous marrow transformation with new bone formation. The purpose of this study was to apply . 1. H MRS to the bone marrow of osteonecrosis and bone marrow edema syndrome by measuring the amount of lipid relative to water of the femoral head and greater trochanter. Materials and Methods: Magnetic Resonance (MR) imaging and MR spectroscopy were performed in twenty-five patients (male: female = 17:8, age = 29–69 years) who were diagnosed with osteonecrosis and bone marrow edema syndrome and compared with three normal control patients. Twenty-three cases were osteonecrosis and two were bone marrow edema syndrome. Ficat stages of osteonecrosis in the femoral heads were 1 patient with stage I, 8 patients with stage II, and 14 patients with stage III disease. Osteonecrosis developed in 14 patients with steroid therapy after kidney transplantation, in 6 alcoholics, and 3 were idiopathic. After routine hip MRI, spectroscopy was obtained from T2 weighted images by the 3-dimensional localization technique. Locations of voxels were the center of the osteonecrotic zone verified by T2-weighted MR images and from the fat marrow in the greater trochanter of femur. The values of the [Lipid/Water] ratios were calculated for all patients. Results: The average Lipid/Water ratio of the osteonecrotic area was 3.15, those of the greater trochanter was 6.45, compared with 10.28 in the normal control group. The MRS pattern for osteonecrosis could be divided into 4 patterns: A, Lipid/Water > 10 ; B, 3 < Lipid/Water ≤ 10; C, 0.3 ≤ Lipid/Water < 3; and D, Lipid/Water ≤ 0.3. The numbers of cases for each pattern were 1 in A, 6 in B, 10 in C, and 6 in D. Interestingly, in one patient with Ficat stage I osteonecrosis diagnosed by only bone scintigraphy, . 1. H MR spectroscopy revealed a much higher lipid/water peak ratio compared to the normal control group. The average Lipid/Water ratio of the bone marrow edema syndrome patients was 0.71. Discussion: This study revealed the decreasing pattern of lipid content as osteonecrosis progresses, which correlates with histopathologic results. In bone marrow edema syndrome, a reverse pattern to the normal control group was found. Further study is needed for the change of Lipid/Water ratio in early change of osteonecrosis