TP508 is a 23 amino acid peptide derived from human prothrombin. It helps wound healing in both soft tissues and bones. In our previous study, we have demonstrated that TP508 retains chondrocyte in a less mature differentiation state while expanding the cartilage mass, indicating it may partly help bone healing by expand the cartilage template in the endochondral bone formation stage. In our current study, we want to demonstrate that TP508 also blocks chondrocyte apoptosis. We used rat costochondral growth plate chondrocytes as our model. We first established chelerythrine as an apoptogen in chondrocytes. TP508 is able to block apoptosis caused by chelerythrine. Chelerythrine also causes an increase in NO production, which is known to cause both pathological and physiological apoptosis of chondrocyte, and blocking NO production can in turn block apoptosis caused by them. TP508 is also able to block NO production caused by chelerythrine. Therefore, TP508 may partially block chondrocyte apoptosis by blocking NO production. From all above, we conclude that besides decreasing chondrocyte differentiation, TP508 also blocks their apoptosis, so as to conserve the cartilage template in endochondral bone formation

Knorpeldefekte des Kniegelenks zeichnen sich durch eine sehr begrenzte spontane Heilungstendenz aus und führen im Verlauf häufig zur Arthrose. Trotz intensiver Forschungsbemühungen konnte bisher keine neue Therapieoption eine zufrieden-stellende Alternative zu den bisherigen Therapien hervorbringen. Eine ACI in Kombination mit einem künstlich hergestellten Knorpel-Knochen-Ersatzmaterial scheint jedoch großes Potential für die Therapie von Knorpel-Knochen-Schäden zu besitzen. Im vorliegenden Langzeittierversuch mit Kaninchen wurde eine einzeitige ACI mit einem biphasischen Ersatzmaterial (TruFit®) und platelet-rich-plasma (PRP) kombiniert. Zu diesem Zweck wurde in der medialen Femurkondyle ein critical-size-Defekt mit einem Durchmesser von 4,5 mm gesetzt. In der ersten Versuchsgruppe blieb der Defekt unbehandelt (Leer). Bei der zweiten Gruppe wurde die Defekthöhle mit einem TruFit®-Zylinder aufgefüllt (TFP). Gruppe drei erhielt zusätzlich PRP (TFP+PRP) und Gruppe vier wurde darüber hinaus mit einer einzeitigen ACI kombiniert (TFP+PRP+C), bei der Chondrozyten mit Hilfe eines speziellen Kollagenase-Schnellverdaus isoliert werden konnten. Die Auswertung der Knorpel-Knochen-Regeneration erfolgte nach 12 Monaten durch eine Mikroradiographie, eine intravitale Fluoreszenzmarkierung des Knochens und durch Toluidinblau-O- und Safranin-O-Färbungen. Verwendet wurden die Scores nach Wakitani und O’Driscoll. Dabei konnte gezeigt werden, dass eine TruFit®-Therapie die Knochenregeneration positiv beeinflussen kann. Die Zugabe von PRP bewirkte die Bildung von zahlreichen dünnen Trabekeln mit einer erhöhten Anzahl trabekulärer Verbindungen, allerdings auch eine schlechtere Rekonvaleszenz der subchondralen Knochenschicht. Bezüglich der Knorpelheilung schnitt die Gruppe TFP+PRP+C am besten ab, wobei die Unterschiede nicht signifikant waren. Insgesamt zeigten alle Versuchsgruppen eine unzureichende osteochondrale Regeneration, so dass für die Therapie am Menschen zunächst weitere Studien nötig sind, die sowohl ossär als auch chondral eine verbesserte Heilungspotenz demonstrieren können. Bisher fehlen groß angelegte Studien um Therapieempfehlungen bezüglich des Ersatzmaterials, der genauen Durchführung der einzeitigen ACI und Zusätzen wie Wachstumsfaktoren zu machen.

The development of bioengineered cartilage for replacement of damaged articular cartilage has gained momentum in recent years. One such approach has been developed in the Kandel lab, whereby cartilage is formed by seeding primary articular chondrocytes on the top surface of a porous biodegradable calcium polyphosphate (CPP) bone substitute, permitting anchorage of the tissue within the pores of the substrate; however, the interfacial shear properties of the tissue-substrate interface of these biphasic constructs are 1 to 2 orders of magnitude lower than the native cartilage-subchondral bone interface. To overcome this limitation, a strategy was devised to generate a zone of calcified cartilage (ZCC), thereby mimicking the native architecture of the osteochondral junction; however, the ZCC was located slightly above the cartilage-CPP interface. Thus, it was hypothesized that polyphosphate released from the CPP substrate and accumulating in the tissue inhibits the formation of the ZCC at the tissue-substrate interface. Based on this information, a strategy was devised to generate biphasic constructs incorporating a properly located ZCC. This approach involved the application of a thin calcium phosphate film to the surfaces of porous CPP via a sol-gel procedure, thereby limiting the accumulation of polyphosphate in the cartilaginous tissue. This modification to the substrate surface did not negatively impact the quality of the in vitro-formed cartilage tissue or the ZCC. Interfacial shear testing of biphasic constructs demonstrated significantly improved interfacial shear properties in the presence of a properly located ZCC. These studies also led to the observation that chondrocytes produce endogenous polyphosphate and that its levels in deep zone cartilage appear inversely related to mineral deposition within the tissue. Using an in vitro model of cartilage calcification, it was demonstrated that polyphosphate levels are modulated in part by the inhibitory effects of fibroblast growth factor 18 on exopolyphosphatase activity in the tissue. Polyphosphate also appears to act in a feedback loop to control exopolyphosphatase activity. Interestingly, polyphosphate also exhibits positive effects on cartilage matrix accumulation. The potential implication of polyphosphate in the maintenance of articular cartilage homeostasis is intriguing and must be investigated further.

The development of bioengineered cartilage for replacement of damaged articular cartilage has gained momentum in recent years. One such approach has been developed in the Kandel lab, whereby cartilage is formed by seeding primary articular chondrocytes on the top surface of a porous biodegradable calcium polyphosphate (CPP) bone substitute, permitting anchorage of the tissue within the pores of the substrate; however, the interfacial shear properties of the tissue-substrate interface of these biphasic constructs are 1 to 2 orders of magnitude lower than the native cartilage-subchondral bone interface. To overcome this limitation, a strategy was devised to generate a zone of calcified cartilage (ZCC), thereby mimicking the native architecture of the osteochondral junction; however, the ZCC was located slightly above the cartilage-CPP interface. Thus, it was hypothesized that polyphosphate released from the CPP substrate and accumulating in the tissue inhibits the formation of the ZCC at the tissue-substrate interface. Based on this information, a strategy was devised to generate biphasic constructs incorporating a properly located ZCC. This approach involved the application of a thin calcium phosphate film to the surfaces of porous CPP via a sol-gel procedure, thereby limiting the accumulation of polyphosphate in the cartilaginous tissue. This modification to the substrate surface did not negatively impact the quality of the in vitro-formed cartilage tissue or the ZCC. Interfacial shear testing of biphasic constructs demonstrated significantly improved interfacial shear properties in the presence of a properly located ZCC. These studies also led to the observation that chondrocytes produce endogenous polyphosphate and that its levels in deep zone cartilage appear inversely related to mineral deposition within the tissue. Using an in vitro model of cartilage calcification, it was demonstrated that polyphosphate levels are modulated in part by the inhibitory effects of fibroblast growth factor 18 on exopolyphosphatase activity in the tissue. Polyphosphate also appears to act in a feedback loop to control exopolyphosphatase activity. Interestingly, polyphosphate also exhibits positive effects on cartilage matrix accumulation. The potential implication of polyphosphate in the maintenance of articular cartilage homeostasis is intriguing and must be investigated further.

Osteoarthritis is a degenerative joint disease characterized by progressive erosion of the articular cartilage. Epidemiological studies have established a relationship between osteoarthritis and menopause suggesting that estrogen may be important in the development of cartilage regeneration therapies. The overall goal of this research project was to advance the field of cartilage tissue regeneration by investigating the role of 17 ß -estradiol (E2), an active estrogen metabolite, on the chondrocyte phenotype. The central hypothesis was that E2 plays an important and sex-specific role in regulating chondrogenesis. Specific Aim-1 focused on establishing and characterizing a primary human articular chondrocyte (HAC) cell source, and then examining the response of the cells in culture to E2. It was demonstrated that the response of HACs to E2 treatment was sex-specific despite both male and females cells expressing estrogen receptors. Female HACs showed changes in proliferation, matrix production, and differentiation while male cells did not. In addition, the female response was regulated through a rapid membrane signaling pathway mediated by protein kinase C. Specific Aim-2 involved establishing an ovariectomized animal model to investigate the effects of E2 on orthopaedic tissue implants. Human demineralized bone matrix (DBM) was implanted intramuscularly into female nude mice and rats. Ovariectomy was shown to reduce the ability of DBM to induce the formation of cartilage and bone tissue. Moreover, the inductive properties of DBM were reestablished with subcutaneous E2 supplementation. Specific Aim-3 entailed developing and characterizing a microencapsulation method for in vitro culture and in vivo delivery of chondrocytes to study the effects of E2 on chondrogenesis. Rat growth plate chondrocytes and HACs were microencapsulated in alginate using an extrusion method in conjunction with high electrostatic potential. Chondrocytes maintained their phenotype in alginate suspension but were unable to form cartilage tissue when implanted into our animal model. Further optimization of the system is required before the role of E2 on chondrogenesis of tissue engineered constructs can be determined. In summary, our results suggest that the successful production of tissue engineered therapies will likely depend on understanding and manipulating the actions of sex hormones in both the in vitro and in vivo environment.

[Truncated abstract] Articular cartilage has no vascular, neural, or lymphatic supply, and hence no intrinsic capacity to self-repair following injury. These physiological limitations, combined with the inability of local chondrocytes to contribute to the repair process, translate to poor structural and functional outcomes in these troublesome defects, and osteoarthritic deterioration with time. Subsequently, many surgical therapies have been trialed to stimulate cartilage repair, but none have produced reliable outcomes. Hence, cartilage repair research has been broadened, with many investigators now focused on cell-based treatment. Smith began a revolution of autologous cell research when in 1965 she isolated chondrocytes from articular cartilage and transplanted them into fresh cartilage nodules (Smith, 1965). Since, new technologies and improved techniques have seen autologous chondrocyte implantation (ACI) widely accepted for use in clinical orthopaedics (Bentley et al., 2003; Brittberg et al., 1994; Grande et al., 1989; Peterson et al., 2002). At present, matrix-induced autologous chondrocyte implantation (MACI) is the most surgically simple form of ACI, boasting clinical outcomes comparable to any technique on the market, and far less complications compared to the first generation of ACI - periosteal ACI (Bartlett et al., 2005; Behrens et al., 2006; Gigante et al., 2006; Henderson et al., 2004; Marlovits et al., 2005; Minas, 2001; Willers et al., 2007; Zheng et al., 2007). But whilst MACI has been adopted by the orthopaedic surgeon for articular cartilage repair, many of the molecular, histological, and clinical factors governing patient outcomes are still largely understudied. Firstly we assessed the bioactivity of fibrin sealant (FS - Tisseel®), a critical component of MACI, on the migration and proliferation of human articular chondrocytes in vitro. We also looked to elucidate the associated molecular mechanisms of thrombin, a key active ingredient in FS, by examining the expression and activation of proteaseactivated receptors (PARs), established thrombin receptors. All four PAR isoforms were detected in human chondrocytes, with PAR-1 being the major isoform expressed. '...' This thesis has demonstrated biological, histological, and clinical features of the MACI technique. Our in vitro has supported the use of fibrin sealant and collagen membrane as the major material components of MACI, illustrating improved chondrocyte proliferation, migration, and chondrogenic differentiation. We have evidenced that MACI stimulates successful production of hyaline-like cartilage by 6 months, while also showing that revised and clinically failed repair tissues are predominantly hyaline-like and fibrocartilage with inferior composition. Clinically, we have documented significant improvements in patient repair structure, function, symptoms, quality of life, and satisfaction, whilst concurrently confirming sentiment within the literature regarding the importance of exercise/ rehabilitation for maximising MACI outcome. In summary, the findings presented in this thesis suggest that MACI is a biologically sound and clinically efficacious cell-based treatment option for repairing articular cartilage defects.

A perda ou falha de um órgão ou tecido é um dos problemas mais severos da saúde humana. A engenharia de tecidos, definida como o cultivo e adesão de células humanas in vitro em um scaffold ou arcabouço, surge como uma alternativa viável para reposição de órgãos e tecidos. Estas células proliferam, migram e se diferenciam num tecido específico enquanto produzem os componentes de matriz extracelular (ECM) necessários para criar este tecido. A obtenção de scaffolds fibrosos a partir da blenda polimérica de Poli (Ácido Láctico-co-Glicólico) (PLGA) e Poliisopreno (PI) é proposta como uma alternativa à engenharia de tecidos moles. Este material foi processado como estrutura fibrosa por meio de métodos de gotejamento (FD) e electrospinning (FS). Caracterização físico-química foi aplicada à blenda e às fibras geradas. Também foi averiguada a viabilidade das fibras em culturas de mioblastos murinos, fibroblastos dérmicos humanos, condrócitos bovinos e hepatocarcinomas. Nota-se que o processo de obtenção da blenda não apresentou alterações na estrutura química dos polímeros, sendo apontada também a imiscibilidade entre eles. A ductilidade do material foi apontada como efeito da presença de PI na blenda, embora esta composição apresente similar molhabilidade entre a mistura e os polímeros puros. As fibras geradas por electrospinning geraram um scaffold com menor porosidade do que as fibras obtidas por gotejamento, mesmo apresentando um diâmetro menor e uma orientação paralela entre fibras. As fibras obtidas por gotejamento apresentaram fibras emaranhadas de maior diâmetro, mas maior tammanho de poros, gerando scaffolds de maior porosidade. As propriedades mecânicas de ambos scaffolds indicam sua aplicação enquanto substitutos de tecidos moles. Ensaios de viabilidade celular condenaram o uso das fibras FS, uma vez que estas apresentaram solvente residual no interior da fibra, causando indesejada lise celular. As fibras FD apresentaram resultados de adesão e proliferação adequados para mioblastos, fibroblastos e condrócitos, porém os resultados foram considerados impróprios para hepatócitos. / The lost or failure of an organ or tissue is one of the most severe problems in human health. Tissue engineering, defined as the seeding and adhesion of human cells in vitro over a scaffold, arises as an viable alternative for reproduction of organs and tissues. These cells proliferate, migrate and differentiate into a specific tissue while producing extracellular matrix components. The obtaining of fibrous scaffolds from a polymeric blend of Poly (Lactic-co-Glycolic Acid) (PLGA) and Polyisoprene (PI) is proposed as an alternative to soft tissue engineering. This material was processed as a fibrous structure through dripping (FD) and electrospinning (FS) methods. Physical-chemical characterization was applied to the blend and to the generated fibres. Fibres viability was also observed for murine myoblasts, human dermal fibroblasts, bovine chondrocytes and hepatocellular carcinoma cultures. It was noticed that the blending process didn't have any influence over polymer's chemical structure, being observed the immiscibility between the raw materials. Blend's ductile behaviour was pointed out as an effect of PI presence, although this mixture presents similar wettability to the one presented by these raw polymers. Fibres obtained by electrospinnig generated a scaffold with smaller porosity, even presenting fibres with smaller diameter and a parallel organized topography. The fibres obtained by dripping presented a tangled structure of thicker fibres, but assembling a scaffold with higher porosity and inner space. Mechanical properties of both scaffolds indicate their applicability as soft tissue substitutes. Cell viability assays condemn the use of FS fibres, seen that they present residual solvent trapped into the fibre, causing undesirable cell lysis. On the other hand, FD fibres presented positive adhesion and proliferation results for myoblasts, fibroblasts and chondrocytes cell lines, however the results were consider inappropriate for hepatocytes.

A perda ou falha de um órgão ou tecido é um dos problemas mais severos da saúde humana. A engenharia de tecidos, definida como o cultivo e adesão de células humanas in vitro em um scaffold ou arcabouço, surge como uma alternativa viável para reposição de órgãos e tecidos. Estas células proliferam, migram e se diferenciam num tecido específico enquanto produzem os componentes de matriz extracelular (ECM) necessários para criar este tecido. A obtenção de scaffolds fibrosos a partir da blenda polimérica de Poli (Ácido Láctico-co-Glicólico) (PLGA) e Poliisopreno (PI) é proposta como uma alternativa à engenharia de tecidos moles. Este material foi processado como estrutura fibrosa por meio de métodos de gotejamento (FD) e electrospinning (FS). Caracterização físico-química foi aplicada à blenda e às fibras geradas. Também foi averiguada a viabilidade das fibras em culturas de mioblastos murinos, fibroblastos dérmicos humanos, condrócitos bovinos e hepatocarcinomas. Nota-se que o processo de obtenção da blenda não apresentou alterações na estrutura química dos polímeros, sendo apontada também a imiscibilidade entre eles. A ductilidade do material foi apontada como efeito da presença de PI na blenda, embora esta composição apresente similar molhabilidade entre a mistura e os polímeros puros. As fibras geradas por electrospinning geraram um scaffold com menor porosidade do que as fibras obtidas por gotejamento, mesmo apresentando um diâmetro menor e uma orientação paralela entre fibras. As fibras obtidas por gotejamento apresentaram fibras emaranhadas de maior diâmetro, mas maior tammanho de poros, gerando scaffolds de maior porosidade. As propriedades mecânicas de ambos scaffolds indicam sua aplicação enquanto substitutos de tecidos moles. Ensaios de viabilidade celular condenaram o uso das fibras FS, uma vez que estas apresentaram solvente residual no interior da fibra, causando indesejada lise celular. As fibras FD apresentaram resultados de adesão e proliferação adequados para mioblastos, fibroblastos e condrócitos, porém os resultados foram considerados impróprios para hepatócitos. / The lost or failure of an organ or tissue is one of the most severe problems in human health. Tissue engineering, defined as the seeding and adhesion of human cells in vitro over a scaffold, arises as an viable alternative for reproduction of organs and tissues. These cells proliferate, migrate and differentiate into a specific tissue while producing extracellular matrix components. The obtaining of fibrous scaffolds from a polymeric blend of Poly (Lactic-co-Glycolic Acid) (PLGA) and Polyisoprene (PI) is proposed as an alternative to soft tissue engineering. This material was processed as a fibrous structure through dripping (FD) and electrospinning (FS) methods. Physical-chemical characterization was applied to the blend and to the generated fibres. Fibres viability was also observed for murine myoblasts, human dermal fibroblasts, bovine chondrocytes and hepatocellular carcinoma cultures. It was noticed that the blending process didn't have any influence over polymer's chemical structure, being observed the immiscibility between the raw materials. Blend's ductile behaviour was pointed out as an effect of PI presence, although this mixture presents similar wettability to the one presented by these raw polymers. Fibres obtained by electrospinnig generated a scaffold with smaller porosity, even presenting fibres with smaller diameter and a parallel organized topography. The fibres obtained by dripping presented a tangled structure of thicker fibres, but assembling a scaffold with higher porosity and inner space. Mechanical properties of both scaffolds indicate their applicability as soft tissue substitutes. Cell viability assays condemn the use of FS fibres, seen that they present residual solvent trapped into the fibre, causing undesirable cell lysis. On the other hand, FD fibres presented positive adhesion and proliferation results for myoblasts, fibroblasts and chondrocytes cell lines, however the results were consider inappropriate for hepatocytes.

A perda ou falha de um órgão ou tecido é um dos problemas mais severos da saúde humana. A engenharia de tecidos, definida como o cultivo e adesão de células humanas in vitro em um scaffold ou arcabouço, surge como uma alternativa viável para reposição de órgãos e tecidos. Estas células proliferam, migram e se diferenciam num tecido específico enquanto produzem os componentes de matriz extracelular (ECM) necessários para criar este tecido. A obtenção de scaffolds fibrosos a partir da blenda polimérica de Poli (Ácido Láctico-co-Glicólico) (PLGA) e Poliisopreno (PI) é proposta como uma alternativa à engenharia de tecidos moles. Este material foi processado como estrutura fibrosa por meio de métodos de gotejamento (FD) e electrospinning (FS). Caracterização físico-química foi aplicada à blenda e às fibras geradas. Também foi averiguada a viabilidade das fibras em culturas de mioblastos murinos, fibroblastos dérmicos humanos, condrócitos bovinos e hepatocarcinomas. Nota-se que o processo de obtenção da blenda não apresentou alterações na estrutura química dos polímeros, sendo apontada também a imiscibilidade entre eles. A ductilidade do material foi apontada como efeito da presença de PI na blenda, embora esta composição apresente similar molhabilidade entre a mistura e os polímeros puros. As fibras geradas por electrospinning geraram um scaffold com menor porosidade do que as fibras obtidas por gotejamento, mesmo apresentando um diâmetro menor e uma orientação paralela entre fibras. As fibras obtidas por gotejamento apresentaram fibras emaranhadas de maior diâmetro, mas maior tammanho de poros, gerando scaffolds de maior porosidade. As propriedades mecânicas de ambos scaffolds indicam sua aplicação enquanto substitutos de tecidos moles. Ensaios de viabilidade celular condenaram o uso das fibras FS, uma vez que estas apresentaram solvente residual no interior da fibra, causando indesejada lise celular. As fibras FD apresentaram resultados de adesão e proliferação adequados para mioblastos, fibroblastos e condrócitos, porém os resultados foram considerados impróprios para hepatócitos. / The lost or failure of an organ or tissue is one of the most severe problems in human health. Tissue engineering, defined as the seeding and adhesion of human cells in vitro over a scaffold, arises as an viable alternative for reproduction of organs and tissues. These cells proliferate, migrate and differentiate into a specific tissue while producing extracellular matrix components. The obtaining of fibrous scaffolds from a polymeric blend of Poly (Lactic-co-Glycolic Acid) (PLGA) and Polyisoprene (PI) is proposed as an alternative to soft tissue engineering. This material was processed as a fibrous structure through dripping (FD) and electrospinning (FS) methods. Physical-chemical characterization was applied to the blend and to the generated fibres. Fibres viability was also observed for murine myoblasts, human dermal fibroblasts, bovine chondrocytes and hepatocellular carcinoma cultures. It was noticed that the blending process didn't have any influence over polymer's chemical structure, being observed the immiscibility between the raw materials. Blend's ductile behaviour was pointed out as an effect of PI presence, although this mixture presents similar wettability to the one presented by these raw polymers. Fibres obtained by electrospinnig generated a scaffold with smaller porosity, even presenting fibres with smaller diameter and a parallel organized topography. The fibres obtained by dripping presented a tangled structure of thicker fibres, but assembling a scaffold with higher porosity and inner space. Mechanical properties of both scaffolds indicate their applicability as soft tissue substitutes. Cell viability assays condemn the use of FS fibres, seen that they present residual solvent trapped into the fibre, causing undesirable cell lysis. On the other hand, FD fibres presented positive adhesion and proliferation results for myoblasts, fibroblasts and chondrocytes cell lines, however the results were consider inappropriate for hepatocytes.

L’arthrose est la pathologie articulaire la plus fréquente, caractérisée par une destruction progressive du cartilage articulaire et impliquant une communication anormale entre l'os sous-chondral et le cartilage. Notre équipe a identifié la protéine 14-3-3ε comme un médiateur soluble secrété par l’os et capable d’altérer l'homéostasie du cartilage en stimulant l’expression de MMP-3 et MMP-13, deux métalloprotéases impliquées dans la dégradation du cartilage au cours de l’arthrose. CD13/APN, récepteur potentiel pour cette protéine, a été mis en évidence à la surface des chondrocytes murins et humains. Le but de cette étude était d’étudier son implication dans la réponse des chondrocytes à 14-3-3ε. L’invalidation de CD13/APN par des siRNA ou des anticorps bloquants, réduit significativement l’effet catabolique chondrocytaire induit par 14-3-3ε. Les chondrocytes articulaires possèdent une activité APN mais elle n’est pas modifiée en présence de 14-3-3ε. Nous avons mis en évidence la présence d’une interaction directe entre 14-3-3ε et CD13/APN grâce à la technologie SPR (Surface Plasmon Resonance) et le système Biacore. En utilisant 14-3-3ε marquée à la biotine, nous avons montré que 14-3-3ε est capable de se lier à la surface des chondrocytes via CD13/APN. Nous avons donc eu recours ensuite aux études de modélisation in silico qui ont permis d’identifier le résidu Y582 phosphorylé appartenant à la séquence E579FNYVW584 de CD13/APN, comme résidu indispensable pour sa liaison à 14-3-3ε. Ce travail de thèse permet de mieux comprendre le mécanisme d’action de 14-3-3ε et propose l’interaction entre 14-3-3ε et CD13 comme une nouvelle cible thérapeutique dans l’arthrose. / Osteoarthritis (OA) is a whole-joint disease characterized by progressive destruction of articular cartilage involving abnormal communication between subchondral bone and cartilage. Our team identified 14-3-3ε protein as a subchondral bone soluble mediator altering cartilage homeostasis. This protein acts as a potent stimulatory factor of MMP-3 and MMP-13 involved in the degradation of cartilage matrix in OA. CD13/APN, potential receptor of this protein, was identified on the surface of chondrocytes. The aim of this study was to investigate its involvement in chondrocytes response to 14-3-3ε. CD13/APN invalidation, using the siRNA strategy and blocking antibodies, reduces significantly the catabolic effect induced by 14-3-3ε in chondrocytes. APN activity was identified in chondrocytes but found unchanged following stimulation with 14-3-3ε. Then, we have revealed the presence of a direct interaction between 14-3-3ε and CD13/APN through the SPR (Surface Plasmon Resonance) and the Biacore system. Using biotin-labeled 14-3-3ε, we have shown that 14-3-3ε is able to bind to the surface of chondrocytes in a manner that is dependent on CD13/APN. It was then necessary to identify the putative motifs involved in this interaction. We therefore used in silico modelling studies which have identified the phosphorylated residue Y582, belonging to the E579FNYVW584 sequence of CD13/APN, as a critical residue for its binding to 14-3-3ε. This thesis work suggest that CD13 plays its receptor role to bind 14-3-3ε and transmit its signal in chondrocytes to induce a catabolic phenotype similar to that observed in OA. Thus, 14-3-3ε-CD13 interaction could be a novel therapeutic target in OA.