Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal.

Dizertační práce se zabývá mechanismy mazání v náhradách kyčelního kloubu. Byla provedena systematická studie formování proteinového filmu při zahrnutí různých materiálů a provozních podmínek. Hlavní pozornost je přitom věnována roli jednotlivých proteinů obsažených v synoviální kapalině při současné přítomnosti dalších proteinů. Jelikož metody aplikované v předchozích studiích neumožňovaly separovat jednotlivé složky maziva, byla vyvinuta optická měřící metoda na principu fluorescenční mikroskopie. Z důvodu verifikace metody byly provedeny dvě nezávislé studie zaměřené na měření tloušťky mazacího filmu a dělení maziva na výstupu mazaného kontaktu. Z důvodu určitých limitací fluorescenční mikroskopie byla dále využita i metoda optické interferometrie, jejíž využití je ilustrováno ve studii zabývající se formováním mazacího filmu v náhradách kyčelního klubu při uvažování reálné konformity třecích povrchů. Závěrečná část práce představuje nový metodologický přístup založený na in situ pozorování kontaktní oblasti umožňující popsat roli jednotlivých proteinů ve vztahu k vývoji tloušťky mazacího filmu. Práce obsahuje originální výsledky, které přináší nové poznání v oblasti biotribologie náhrad kyčelního kloubu vedoucí k dalšímu vývoji implantátů při snaze zabránit jejich selhání v důsledku omezené životnosti.

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Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Biotribology, the study of lubrication, wear and friction within the body, has become a topic of high importance in recent times as we continue to encounter debilitating diseases and trauma that destroy function of the joints. A highly successful surgical procedure to replace the joint with an artificial equivalent alleviates dysfunction and pain. However, the wear of the bearing surfaces in prosthetic joints is a significant clinical problem and more patients are surviving longer than the life expectancy of the joint replacement. Revision surgery is associated with increased morbidity and mortality and has a far less successful outcome than primary joint replacement. As such, it is essential to ensure that everything possible is done to limit the rate of revision surgery. Past experience indicates that the survival rate of the implant will be influenced by many parameters, of primary importance, the material properties of the implant, the composition of the synovial fluid and the method of lubrication. In prosthetic joints, effective boundary lubrication is known to take place. The interaction of the boundary lubricant and the bearing material is of utmost importance. The identity of the vital active ingredient within synovial fluid (SF) to which we owe the near frictionless performance of our articulating joints has been the quest of researchers for many years. Once identified, tribo tests can determine what materials and more importantly what surfaces this fraction of SF can function most optimally with. Surface-Active Phospholipids (SAPL) have been implicated as the body’s natural load bearing lubricant. Studies in this thesis are the first to fully characterise the adsorbed SAPL detected on the surface of retrieved prostheses and the first to verify the presence of SAPL on knee prostheses. Rinsings from the bearing surfaces of both hip and knee prostheses removed from revision operations were analysed using High Performance Liquid Chromatography (HPLC) to determine the presence and profile of SAPL. Several common prosthetic materials along with a novel biomaterial were investigated to determine their tribological interaction with various SAPLs. A pin-on-flat tribometer was used to make comparative friction measurements between the various tribo-pairs. A novel material, Pyrolytic Carbon (PyC) was screened as a potential candidate as a load bearing prosthetic material. Friction measurements were also performed on explanted prostheses. SAPL was detected on all retrieved implant bearing surfaces. As a result of the study eight different species of phosphatidylcholines were identified. The relative concentrations of each species were also determined indicating that the unsaturated species are dominant. Initial tribo tests employed a saturated phosphatidylcholine (SPC) and the subsequent tests adopted the addition of the newly identified major constituents of SAPL, unsaturated phosphatidylcholine (USPC), as the test lubricant. All tribo tests showed a dramatic reduction in friction when synthetic SAPL was used as the lubricant under boundary lubrication conditions. Some tribopairs showed more of an affinity to SAPL than others. PyC performed superior to the other prosthetic materials. Friction measurements with explanted prostheses verified the presence and performance of SAPL. SAPL, in particular phosphatidylcholine, plays an essential role in the lubrication of prosthetic joints. Of particular interest was the ability of SAPLs to reduce friction and ultimately wear of the bearing materials. The identification and knowledge of the lubricating constituents of SF is invaluable for not only the future development of artificial joints but also in developing effective cures for several disease processes where lubrication may play a role. The tribological interaction of the various tribo-pairs and SAPL is extremely favourable in the context of reducing friction at the bearing interface. PyC is highly recommended as a future candidate material for use in load bearing prosthetic joints considering its impressive tribological performance.

The main objective of this diploma thesis is to determine the influence of synovial fluid compodition and the influence of surface microtextures on lubrication of hip join replacements. For all experiments, hip joint simulator based on the pendulum principle was utilized. Metal heads (CoCrMo) manufactured by B Braun company and and tailor made acetabular cups from optical glass were used as contact couples. Acetabulum dimensions were fabricated according to dimensions of real implants. Experiments with surface microtextures simulated walking cycle for 210 s and development of lubrication film over time was observed.Results have shown that all types of tested structures improved lubrication of contact area, where the best results were obtained for square and triangle shapes respectively. Second set of experiments was based on the fact, that the composition of synovial fluid of healthy people differs from the composition of patients with various stages of osteoarthritis. Therefore, experiments with different model fluids were compared, which in their composition correspond to different groups of patients. For the model fluid that corresponds to the largest number of cases, experiments with individual components and combinations thereof were subsequently performed to provide a more detailed description of how these components affect the film-forming mechanism. The results show, that a change in fluid composition of patients with osteoarthritis can have a major negative effect on fluid formation process within the pair.

Dizertační práce se zabývá mechanismy mazání v náhradách kyčelního kloubu. Byla provedena systematická studie formování proteinového filmu při zahrnutí různých materiálů a provozních podmínek. Hlavní pozornost je přitom věnována roli jednotlivých proteinů obsažených v synoviální kapalině při současné přítomnosti dalších proteinů. Jelikož metody aplikované v předchozích studiích neumožňovaly separovat jednotlivé složky maziva, byla vyvinuta optická měřící metoda na principu fluorescenční mikroskopie. Z důvodu verifikace metody byly provedeny dvě nezávislé studie zaměřené na měření tloušťky mazacího filmu a dělení maziva na výstupu mazaného kontaktu. Z důvodu určitých limitací fluorescenční mikroskopie byla dále využita i metoda optické interferometrie, jejíž využití je ilustrováno ve studii zabývající se formováním mazacího filmu v náhradách kyčelního klubu při uvažování reálné konformity třecích povrchů. Závěrečná část práce představuje nový metodologický přístup založený na in situ pozorování kontaktní oblasti umožňující popsat roli jednotlivých proteinů ve vztahu k vývoji tloušťky mazacího filmu. Práce obsahuje originální výsledky, které přináší nové poznání v oblasti biotribologie náhrad kyčelního kloubu vedoucí k dalšímu vývoji implantátů při snaze zabránit jejich selhání v důsledku omezené životnosti.

Le cartilage articulaire est un tissu avasculaire et non innervé, ce qui lui confère des capacités de réparation très limitées. Les traitements chirurgicaux actuels ne permettent pas d’obtenir un tissu de réparation similaire au cartilage natif. Les recherches s’orientent depuis de nombreuses années vers l’ingénierie cellulaire et tissulaire du cartilage selon le type de lésions à traiter, focale ou diffuse. Les cellules souches mésenchymateuses (CSMs) constituent une source cellulaire intéressante pour l’ingénierie du cartilage. Elles sont facilement accessibles et ont des potentialités de différenciation chondrogénique. Les CSMs issues de la moelle osseuse sont les plus étudiées et constituent la référence. D’autres sources de CSMs sont également très prometteuses. Notre choix s’est porté sur les CSMs issues de la membrane et du liquide synovial. Ces deux tissus articulaires présentent l’avantage de pouvoir être prélevés facilement lors d’un examen arthroscopique et leurs CSMs sont adaptées au microenvironnement (hypoxie, inflammation) de l’articulation. Ce travail a porté sur l’étude de deux sources cellulaires d’origine synoviale dans le traitement des lésions focales et diffuses du cartilage. Ces CSMs d’origine synoviale ont d’abord été caractérisées selon leurs phénotypes et leurs capacités de différenciation vers les voies ostéogénique, adipogénique et chondrogénique, par rapport aux CSMs issues de la moelle osseuse. Ensuite, les capacités chondrogéniques de ces CSMs synoviales destinées à produire un substitut cartilagineux consacré aux traitements des lésions focales du cartilage articulaire ont été étudiées. Les CSMs ont été ensemencées dans un biomatériau collagénique et différentes conditions environnementales (facteurs de croissance et oxymétrie) ont été évaluées afin de définir les conditions de culture les plus appropriées. La chondrogenèse a été induite avec succès par l’utilisation de facteurs de croissance tels que TGF-1 et TGF-3 seuls ou en association avec la BMP-2. L’hypoxie n’a pas montré d’effet bénéfique sur la synthèse matricielle au sein des substituts cartilagineux. Enfin, nous avons évalué les capacités des CSMs issues du liquide synovial à traiter les lésions diffuses du cartilage induites par un modèle de section du ligament croisé antérieur chez le rat athymique. Les deux injections intra-articulaires de CSMs issues du liquide synovial à 1 et 2 semaines après chirurgie n’ont pas permis de prévenir les lésions arthrosiques. / Joint cartilage is avascular and not innervated, which gives it very limited repair capabilities. Current surgical treatments do not provide repair tissue similar to native cartilage. For many years, research has been focused on cellular and tissue engineering of cartilage depending on the type of lesions to be treated, focal or diffuse. Mesenchymal stem cells (MSCs) are an interesting cellular source for cartilage engineering. They are easily accessible and have the potential for chondrogenic differentiation. MSCs from bone marrow are the most studied and are the gold-standard. Other MSCs sources of are also very promising. We chose MSCs from the synovial membrane and synovial fluid. These both joint tissues have the advantage of being easily retrievable during arthroscopic examination and their MSCs are adapted to the microenvironment (hypoxia, inflammation) of the joint. This thesis work focused on the study of two cellular sources of synovial origin in the treatment of focal and diffuse cartilage lesions. These synovial-derived MSCs were first characterized according to their phenotypes and their ability to differentiate to the osteogenic, adipogenic and chondrogenic pathways, compared to bone marrow derived MSCs. Then, the chondrogenic capacities of these synovial MSCs to produce a cartilage substitute for the treatment of focal lesions of joint cartilage were studied. The MSCs were seeded in a collagenic biomaterial and different environmental conditions (growth factors and oximetry) were evaluated to define the most appropriate culture conditions. Chondrogenesis has been induced with success by the use of growth factors such as TGF-β1 or TGF-β3 alone or in combination with BMP-2. Hypoxia has not exerted a beneficial effect on matrix synthesis in cartilage substitutes.Finally, we evaluated the ability of CSMs from human synovial fluid to treat diffuse cartilage lesions induced by an anterior cruciate ligament section model in athymic rats. The two intra-articular injections of synovial fluid MSCs, 1 and 2 weeks after surgery did not prevent osteoarthritic lesions.

La bioinspiration consiste à analyser les systèmes naturels qui se sont adaptés parfaitement à leurs environnements pour développer des solutions ingénieuses. Ce projet de thèse aborde la thématique de la lubrification articulaire dans le but de développer un traitement contre l'ostéoarthrite (OA). Nous nous sommes inspirés des articulations synoviales, systèmes tribologiques très performants grâce aux interactions synergiques entre la structure unique du cartilage et les molécules lubrifiantes (ML) du fluide synovial (SF). Cependant, lors de l'OA des mécanismes inflammatoires et d'érosion mécanique aboutissent à la dégénérescence progressive du cartilage et la dégradation spécifique des ML du SF (aggrécane et lubricine). Des mimes des ML du SF ont été synthétisés reprenant leur structure particulière dite en écouvillon moléculaire (BB), structure responsable de la lubrification. Des tests tribologiques (SFA, tribomètre) ont montré que les BB garantissent à la fois une faible friction et une résistance à l'usure sur des surfaces dures de mica. Ceci est dû à la présence, sur nos EM, de groupements d'ancrage spécifiques assurant l’adsorption sur la surface de mica et à la formation d'enchevêtrements et d’interactions intermoléculaires avec l'acide hyaluronique de haut poids moléculaire, composant essentiel du SF. Des mimes de cartilage à base d'hydrogels de chitosane multicouches ont été également réalisés reprenant les principales propriétés architecturales du cartilage. En combinaison avec nos EM, ces hydrogels, matériaux poroélastiques fragiles, sont capables d’être lubrifiés avec une friction dans la gamme physiologique et une nette amélioration de leur usure / Bioinspiration consists in the design of materials inspired by biological systems which have developed ingenious solutions to suit their environment. This project deals with bioinspiration for joint lubrication and in particular for the development of treatments for patients suffering from osteoarthritis (OA). To do so, we took our inspiration from joints which are amongst the most efficient aqueous tribological systems. Their unique properties arise from the complex synergistic interactions between cartilage structure and the lubricant macromolecules of the synovial fluid (SF). However, during OA, inflammatory mechanisms as long as mechanical erosion result in the degeneration of cartilage and lubricant macromolecules (aggrecan and lubricin). Polymeric mimes of the SF have been synthesized based on the bottle-brush (BB) architecture of LUB and AGG which is responsible for the joint lubrication. Tribological tests (SFA, tribometer) showed that BB polymers provided mica surfaces with a low friction and a wear protection up to several megapascals, typically in the range of natural joints. This wear protection was essentially due to the incorporation of anchoring groups specific to mica tribopairs on the BB polymers and the intermolecular bridging and entanglements emerging between BB polymers and high molecular weight HA, another main SF component. Cartilage mimes composed of multilayered chitosan hydrogels were designed to mimic the basic features of cartilage. Along with our BB polymers, the hydrogels, which are poroelastic and fragile materials, provided a low friction and a great decrease of wear

En bakteriell ledinfektion kallas för septisk artrit och utan snabb behandling kan tillståndet leda till irreversibla ledskador och flera allvarliga komplikationer. Snabb diagnostik är viktigt för en god prognos och med nuvarande metod för odling av ledvätska tar det upp till fem dygn för detektion av mikroorganismer. Metoden inkluderar odling på fasta substrat och i blododlingsflaskor. Med en ny metod som använder multiplex PCR (polymerase chain reaction) skulle analystiden kunna minskas till en timme. BioFire Joint Infection (JI) Panel är ett test som kan detektera flera mikroorganismer samtidigt genom amplifiering av DNA. Mikroorganismer detekteras och identifieras genom cellysering, DNA-rening, PCR-reaktioner i två steg och smältanalys. Syftet med arbetet var att utvärdera BioFire JI Panel för detektion av mikroorganismer i ledvätska och jämföra metodens känslighet med standardiserad odling respektive odling i buljong, och svara på frågeställningen hur känslig JI-panelen är jämfört med standardiserad odlingsmetod. Prov konstruerades genom att suspendera målbakterierna Staphylococcus aureus, Escherichia coli, Streptococcus pyogenes, Neisseria gonorrhoeae, Haemophilus influenzae, Kingella kingae och Finegoldia magna i natriumklorid. Suspensionerna seriespäddes och odlades ut på fasta substrat, inokulerades i blododlingsflaskor och buljonger samt analyserades med JI-panelen för att identifiera metodernas detektionsgränser. Fasta substrat, blododlingsflaskor och buljonger visade likvärdig känslighet för majoriteten av bakterieisolaten medan JI-panelen generellt visade en sämre eller likvärdig känslighet i jämförelse med standardmetod. Panelen visade potential för detektion av svårodlade bakterier och skulle vara ett bra komplement till nuvarande odlingsmetod. Ytterligare studier hade behövts men den snabba analystiden som möjliggör snabbare behandling framhävde metodens kliniska potential. / Bacterial joint infections are called septic arthritis and without rapid diagnosis and treatment the condition could lead to irreversible joint damage and other serious complications. With the current method for synovial fluid cultivation, which includes both solid substrates and culture vials, detection of microorganisms takes up to five days. With a new method utilizing multiplex polymerase chain reaction (PCR), the time to detection could decrease to an hour. BioFire Joint Infection (JI) Panel can detect multiple microorganisms simultaneously through amplification of DNA. Microorganisms are detected and identified by cell lysis, DNA purification and two consecutive PCR reactions with subsequent melting curve analysis. The aim of the study was to evaluate the BioFire JI Panel for detection of microorganisms in synovial fluid and compare the method’s sensitivity to standard and broth cultivation. Samples were constructed by suspending target bacteria, Staphylococcus aureus, Escherichia coli, Streptococcus pyogenes, Neisseria gonorrhoeae, Haemophilus influenzae, Kingella kingae and Finegoldia magna, in sodium chloride. The suspensions were serially diluted and cultivated on solid substrates, in culture vials, broth and analyzed with the JI panel to find the detection limits for each method. Solid substrates, culture vials and broth exhibited similar sensitivity while the JI panel generally had a lower or similar sensitivity compared to standard methods. The panel showed potential detecting difficult to grow bacteria and would be a good complement to current methods. Further studies are required but the quick analysis enabling faster treatment highlighted the clinical potential.