Developing a cationic contrast agent for computed tomographic imaging of articular cartilage and synthetic biolubricants for early diagnosis and treatment of osteoarthritis

Osteoarthritis (OA) causes debilitating pain for millions of people, yet OA is typically diagnosed late in the disease process after severe damage to the articular cartilage has occurred and few treatment options exist. Furthermore, destructive techniques are required to measure cartilage biochemical and mechanical properties for studying cartilage function and changes during OA. Hence, research and clinical needs exist for non-destructive measures of cartilage properties. Various arthroscopic (e.g., ultrasound probes) and imaging (e.g., MRI or CT) techniques are available for assessing cartilage less destructively. However, arthroscopic methods are limited by patient anesthesia/infection risks and cost, and MRI is hindered by high cost, long image acquisition times and low resolution. Contrast-enhanced CT (CECT) is a promising diagnostic tool for early-stage OA, yet most of its development work utilizes simplified and ideal cartilage models, and rarely intact, pre-clinical animal or human models. To advance CECT imaging for articular cartilage, this dissertation describes further development of a new cationic contrast agent (CA4+) for minimally-invasive assessment of cartilage biochemical and mechanical properties, including glycosaminoglycan content, compressive modulus, and coefficient of friction. Specifically, CA4+ enhanced CT is compared to these three cartilage properties initially using an ideal bovine osteochondral plug model, then the technique is expanded to examine human finger joints and both euthanized and live mouse knees. Furthermore, CECT attenuations with CA4+ map bovine meniscal GAG content and distribution, signifying CECT can evaluate multiple tissues involved in OA. CECT's sensitivity to critical cartilage and meniscal properties demonstrates its applicability as both a non-destructive research tool as well as a method for diagnosing and monitoring early-stage OA. Additionally, CECT enables evaluation of efficacy for a new biolubricant (2M TEG) for early-stage OA treatment. In particular, CECT can detect the reduced wear on cartilage surfaces for samples tested in 2M TEG compared to samples tested in saline (negative control). With its sensitivity to cartilage GAG content, surface roughness, and mechanical properties, CA4+ enhanced CT will serve as a valuable tool for subsequent in vivo animal and clinical use

A multi-scale imaging approach to understand osteoarthritis development

X-ray phase-contrast imaging is an innovative and advanced imaging method. Contrary to conventional radiology, where the image contrast is primarily determined by X-ray attenuation, phase-contrast images contain additional information generated by the phase shifts or refraction of the X-rays passing through matter. The refractive effect on tissue samples is orders of magnitude higher than the absorption effect in the X-ray energy range used in biomedical imaging. This technique makes it possible to produce excellent and enhanced image contrast, particularly when examining soft biological tissues or features with similar X-ray attenuation properties. In combination with high spatial resolution detector technology and computer tomography, X-ray phase-contrast imaging has been proved to be a powerful method to examine tissue morphology and the evolution of pathologies three-dimensionally, with great detail and without the need of contrast agents. This Thesis work has focused on developing an accurate, multi-scale X-ray-based methodology for imaging and characterizing the early stages of osteoarthritis. X-ray phase-contrast images acquired at different spatial resolutions provide unprecedented insights into cartilage and the development of its degeneration, i.e., osteoarthritis. Other types of X-ray phase-contrast imaging techniques and setups using spatial resolutions ranging from micrometer down to nanometer were applied. Lower spatial resolutions allow large sample coverage and comprehensive representations, while the nanoscale analysis provides a precise depiction of anatomical details and pathological signs. X-ray phase-contrast results are correlated to data obtained, on the same specimens, by standard laboratory methods, such as histology and transmission electron microscopy. Furthermore, X-ray phase-contrast images of cartilage were acquired using different X-ray sources and results were compared in terms of image quality. It was shown that with the use of synchrotron radiation, more detailed images and much faster data acquisitions could be achieved. A second focus in this Thesis work has been the investigation of the reaction of healthy and degenerated cartilage under different physical pressures, simulating the different levels of stress to which the tissue is subject during daily movements. A specifically designed setup was used to dynamically study cartilage response to varying pressures with X-ray phase-contrast micro-computed tomography, and a fully volumetric and quantitative methodology to accurately describe the tissue morphological variations. This study revealed changes in the behavior of the cartilage cell structure, which differ between normal and osteoarthritic cartilage tissues. The third focus of this Thesis is the realization of an automated evaluation procedure for the discrimination of healthy and cartilage images with osteoarthritis. In recent years, developments in neural networks have shown that they are excellently suited for image classification tasks. The transfer learning method was applied, in which a pre-trained neural network with cartilage images is further trained and then used for classification. This enables a fast, robust and automated grouping of images with pathological findings. A neural network constructed in this way could be used as a supporting instrument in pathology. X-ray phase-contrast imaging computed tomography can provide a powerful tool for a fully 3D, highly accurate and quantitative depiction and characterization of healthy and early stage-osteoarthritic cartilage, supporting the understanding of the development of osteoarthritis.Röntgen-Phasenkontrast-Bildgebung ist eine innovative und weiterführende Bildgebungsmethode. Im Gegensatz zu herkömlichen Absorptions-Röntgenaufnahmen, wie sie in der Radiologie verwendet werden, wird der Kontrast bei dieser Methode aus dem Effekt der Phasenverschiebung oder auch Brechung der Röngtenstrahlen gebildet. Der Brechungseffekt bei Gewebeproben ist um ein Vielfaches höher als der Absorptionseffekt des elektromagnetischen Spektrums der Röntgenstrahlen. Diese Methode ermöglicht die Darstellung von großen Kontraste im Gewebe. Unter Verwendung eines hochauflösenden Detektors und in Kombination mit der Computer-Tomographie, ist Phasenkontrast-Bildgebung eine sehr gute Methode um Knorpelgewebe und Arthrose im Knorpel zu untersuchen. Diese Arbeit beschreibt primär ein Verfahren zur Darstellung arthrotischen Knorpels im Anfangsstadium. Die mit verschiedenen Auflösungen und 3D-Phasen-Kontrast-Methoden produzierten Aufnahmen ermöglichen einen noch nie dagewesenen Einblick in den Knorpel und die Entwicklung von Arthrose im Anfangsstadium. Hierbei kam die propagationsbasierte Phasenkontrastmethode mit einer Auflösung im mikrometer Bereich und die (Nano)-Holotomographie-Methode mit einer Auflösung im Submicrometer Bereich zum Einsatz. Durch Auflösung im mikrometer Bereich kann ein großes Volumen im Knorpel gescannt werden, während die Nano-Holotomographie Methode eine sehr große Detailauflösung aufweißt. Die Phasenkontrast-Aufnahmen werden mit zwei anderen wissenschaftlichen Methoden verglichen: mikroskopische Abbildungen histologisch aufgearbeiteter Knorpelproben und Aufnahmen eines Transmissionselektroskop zeigen sehr große Übereinstimmungen zur Röntgen-Phasenkontrast-Bildgebung. Desweiteren wurden Phasenkontrast-Aufnahmen von Knorpel aus unterschiedlichen Röntgenquellen verglichen. Hierbei zeigte sich, dass mit Hilfe des Teilchenbeschleunigers (Synchrotron) detailreichere und schnellere Aufnahmen erzielt werden können. Bilder aus Flüssig-Metall-Quellen zeigen sich durchaus von guter Qualität, erfordern jedoch sehr lange Aufnahmezeiten. In dieser Arbeit wird zudem das Verhalten von Knorpelgewebe, welches ein Anfangsstadium von Arthrose aufweist, unter physikalischem Druck untersucht. Hierfür wurden 3D-Computertomographie-Aufnahmen von komprimiertem Knorpelgewebe angefertig und mit Aufnahmen ohne Komprimierung verglichen. Ein quantitativer Vergleich machte Veränderungen des Verhaltens der Knorpelzellstruktur (Chondronen) sichtbar. Es konnte gezeigt werden, dass Chondrone bei arthrotischem Knorpel ein verändertes Kompressionsverhalten haben. Der dritte Fokus dieser Arbeit liegt auf der automatisierten Auswertung von Aufnahmen gesunden und arthrotischen Knorpelgewebes. Die Entwicklungen im Bereich der Neuronale Netze zeigten in den letzten Jahren, dass diese sich hervoragend für Bildklassifizierungsaufgaben eignen. Es wurde die Methode des transferierenden Lernens angewandt, bei der ein vortrainiertes Neuronales Netz mit Knorpelbildern weitertrainiert und anschließend zur Klassifizierung eingesetzt wird. Dadurch ist eine schnelle, robuste und automatisierte Gruppierung von Bildern mit pathologischen Befunden möglich. Ein derart konstruiertes Neuronales Netz könnte als unterstützendes Instrument in der Pathologie angewandt werden. Röntgen-Phasenkontrast-CT kann ein leistungsstarkes Werkzeug für eine umfassende, hochpräzise und quantitative 3D-Darstellung und Charakterisierung von gesundem Knorpel und athrotischem Knorpel im Frühstadium bieten, um das Verständnis der Entwicklung von Osteoarthritis zu erweitern

Investigation of Subchondral Bone Abnormalities associated with Osteoarthritis using Image-Based Biomechanics

Osteoarthritis (OA) is degenerative disease caused by a mechanical failure of bone and cartilage. Common risk factors for developing OA include: being over-weight, female, having joint malalignment, or a history of prior joint injury. Post-traumatic OA is extremely common in the knee as individuals frequently suffer injuries to structures that provide stability to the joint. To enhance our understanding about OA, animal models are employed where the injury can be and monitored in a controlled environment. When used in conjunction with pre-clinical imaging techniques the longitudinal degradation of bone and cartilage can be quantitatively monitored in vivo. Recent evidence has identified cystic lesions within the subchondral bone as the possible source of painful symptoms and accelerated disease progression, but little is known about their etiology. The purpose of this thesis was to improve knowledge regarding the mechanism that causes subchondral cysts. OA was induced in the rodent knee via surgery, and the pathological changes were quantified with micro-CT and MRI. The composition of the cysts was correlated with end-stage histology. Thus, an accurate definition of OA bone cysts was achieved. To assess the effect of cysts in human bone, a study was conducted using a patient data set restrospectively. Using finite element (FE) analysis, higher stress values were found within bone surrounding cysts. Therefore, the probable mechanism of cyst expansion, stress induced resorption, was identified. Finally, the FE models of the bones were combined with soft tissue structures – from a co-registered MRI – to produce comprehensive patient-specific models of the knee

Book of Abstracts 15th International Symposium on Computer Methods in Biomechanics and Biomedical Engineering and 3rd Conference on Imaging and Visualization

In this edition, the two events will run together as a single conference, highlighting the strong connection with the Taylor & Francis journals: Computer Methods in Biomechanics and Biomedical Engineering (John Middleton and Christopher Jacobs, Eds.) and Computer Methods in Biomechanics and Biomedical Engineering: Imaging and Visualization (JoãoManuel R.S. Tavares, Ed.). The conference has become a major international meeting on computational biomechanics, imaging andvisualization. In this edition, the main program includes 212 presentations. In addition, sixteen renowned researchers will give plenary keynotes, addressing current challenges in computational biomechanics and biomedical imaging. In Lisbon, for the first time, a session dedicated to award the winner of the Best Paper in CMBBE Journal will take place. We believe that CMBBE2018 will have a strong impact on the development of computational biomechanics and biomedical imaging and visualization, identifying emerging areas of research and promoting the collaboration and networking between participants. This impact is evidenced through the well-known research groups, commercial companies and scientific organizations, who continue to support and sponsor the CMBBE meeting series. In fact, the conference is enriched with five workshops on specific scientific topics and commercial software.info:eu-repo/semantics/draf

High Resolution Imaging Of Early, Subclinical Cartilage Injuries: Implications For The Diagnosis And Treatment Of Post-Traumatic Osteoarthritis

Osteoarthritis (OA) is a degenerative disease. Currently, there are no treatments to restore damaged cartilage to its native state. Joint injury increases the risk of developing post-traumatic OA (PTOA). Subtle cartilage injury that may progress to PTOA cannot be detected with current clinical imaging modalities. If subtle cartilage injury is detectable, PTOA can be better understood to develop treatments targeting early disease progression. The goal of this dissertation was to evaluate methods for detecting, characterizing, and treating early cartilage injury. Multiphoton microscopy (MPM) can provide high resolution details of live, intact tissue. The purpose of the first study was to validate the use of MPM to detect subtle cartilage damage. The results confirmed the ability of MPM to resolve structural changes and cell death in cartilage immediately after injury. This suggests future application of MPM in the clinic for early diagnosis or in the laboratory to perform longitudinal studies not currently possible due to the necessity of histological processing. In the second study, cartilage and its resiliency to injury were evaluated among eight major joints to determine if there are different susceptibilities to injury in different joints. The structure of articular cartilage and prevalence of OA vary among joints, but typically results from work performed within a single joint are applied to other joints. The results from the second study showed that some joints have more cellular death and/or decrease anabolic gene expression than other joints after receiving the same injury, which suggests the need for joint specific treatments. The third study examined the role of oxygen in the development of PTOA. Cartilage is avascular and has limited oxygen supply. Oxygen tension may be increased in arthritic joints, yet short exposure to high oxygen tension is beneficial to uninjured cartilage. In this final study, the effect of increased oxygen tension on cartilage viability after injury was evaluated. The immediate application of hyperoxic treatment minimized cell death after injury, suggesting that the immediate application of high oxygen following injury may be chondroprotective. These findings have implications in future treatments that could minimize the effect of cartilage injury and development of PTOA