In the heart, unidirectional blood flow depends on proper heart valve function. As, in mammals, regulatory mechanisms of early heart valve and bone development are shown to contribute to adult heart valve pathologies, we used the animal model zebrafish (ZF, Danio rerio) to investigate the microarchitecture and differentiation of cardiac valve interstitial cells in the transition from juvenile (35 days) to end of adult breeding (2.5 years) stages. Of note, light microscopy and immunohistochemistry revealed major differences in ZF heart valve microarchitecture when compared with adult mice. We demonstrate evidence for rather chondrogenic features of valvular interstitial cells by histological staining and immunodetection of SOX-9, aggrecan, and type 2a1 collagen. Collagen depositions are enriched in a thin layer at the atrial aspect of atrioventricular valves and the ventricular aspect of bulboventricular valves, respectively. At the ultrastructural level, the collagen fibrils are lacking obvious periodicity and orientation throughout the entire valve. (J Histochem Cytochem 67:361–373, 2019)

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No description available.

Expression studies have implicated Fibronectin Leucine Rich Transmembrane protein 2 (FLRT2) in cranial neural crest cell migration and pre-chondrogenic cell condensation during craniofacial skeletogenesis. This aim of this study was to characterize the expression of FLRT2 and its relationship to the extracellular matrix (ECM) in ATDC5 chondroprogenitor cells. Immunofluorescence studies localized FLRT2 to the cell membrane as well as exracellularly, where it colocalized with fibronectin. FLRT2 was identified in the ATDC5-derived ECM after cell extraction. Further to its colocalization with fibronectin, FLRT2 associated with fibronectin-coated beads in cell cultures. Co-immunoprecipitation confirmed that FLRT2 and fibronectin interact, either directly or indirectly. Blocking fibronectin fibril formation in ATDC5 cell cultures demonstrated a concomitant decrease in extracellular FLRT2 accumulation. It appears that FLRT2 may exist in both a membrane-bound and a shed form. Either or both of these forms may participate in cell-ECM interactions in cooperation with fibronectin or other ECM proteins.

Expression studies have implicated Fibronectin Leucine Rich Transmembrane protein 2 (FLRT2) in cranial neural crest cell migration and pre-chondrogenic cell condensation during craniofacial skeletogenesis. This aim of this study was to characterize the expression of FLRT2 and its relationship to the extracellular matrix (ECM) in ATDC5 chondroprogenitor cells. Immunofluorescence studies localized FLRT2 to the cell membrane as well as exracellularly, where it colocalized with fibronectin. FLRT2 was identified in the ATDC5-derived ECM after cell extraction. Further to its colocalization with fibronectin, FLRT2 associated with fibronectin-coated beads in cell cultures. Co-immunoprecipitation confirmed that FLRT2 and fibronectin interact, either directly or indirectly. Blocking fibronectin fibril formation in ATDC5 cell cultures demonstrated a concomitant decrease in extracellular FLRT2 accumulation. It appears that FLRT2 may exist in both a membrane-bound and a shed form. Either or both of these forms may participate in cell-ECM interactions in cooperation with fibronectin or other ECM proteins.

There is a huge unmet clinical need to treat damaged articular cartilage such as that caused by osteoarthritis (OA) with an estimated 8.75 million people in the UK having sought treatment for OA (ARUK 2013). Embryonic stem cells (ESCs) offer a promising alternative therapeutic approach, potentially providing an unlimited source of chondrocytes capable of regenerating the damaged cartilage however this is limited by the efficiency of the chondrogenic differentiation protocol. An improved understanding of the posttranscriptional regulation of chondrogenesis by microRNAs (miRNAs) may enable us to improve hESC chondrogenesis. Also the recent discovery that miRNAs are selectively packaged into exosomes which can then be transferred to and be functionally active within neighbouring cells suggests they may have a role in cell-cell communication. This project investigated the regulation of miRNA expression in relation to the transcriptome during hESCs-directed chondrogenesis and the possible role for exosomes during differentiation and in stem cell maintenance of hESCs. Small RNA-seq and whole transcriptome sequencing was performed on distinct stages of hESC-directed chondrogenesis using the Directed Differentiation Protocol (DDP) developed in our lab. Also small RNA-seq was performed on exosomes isolated from hESCs and chondroprogenitors along with the donor cells that the exosomes originated from. This revealed significant changes in the expression of several miRNAs during hESC-directed chondrogenesis including: upregulation of miRNAs transcribed from the four Hox complexes, known cartilage associated miRNAs and the downregulation of pluripotency associated miRNAs. Overall miRome and transcriptome analysis revealed the two hESC lines exhibited slightly different miRome and transcriptome profiles during chondrogenesis, with Man7 displaying larger changes in miRNA and mRNA expression as it progressed through the DDP suggesting it may be more predisposed to undergo chondrogenesis. Integration of miRomes and transcriptomes generated during hESC-directed chondrogenesis identified four key functionally related clusters of co-expressed miRNAs and protein coding genes: pluripotency associated cluster, primitive streak cluster, limb development cluster and an extracellular matrix cluster. Further investigation of these gene/miRNA clusters allowed the identification of several potential novel regulators of hESC-directed chondrogenesis. In accordance with the reported literature the exosomal miRNAs from hESCs and hESC-chondroprogenitors were enriched with a guanine rich motif. Notably, several of these were enriched with targets associated with embryonic skeletal system development suggesting they may play a role in regulating differentiation. Preliminary functional experiments examining pluripotency-associated exosomes suggests they may have a role in regulating hESC stem cell maintenance. However the molecular mechanism by which this is achieved has not been investigated. This research identified main miRome and transcriptome changes during hESC-directed chondrogenesis leading to the identification of several potential novel regulators of chondrogenesis and pluripotency which can be further investigated. This project has also highlighted the potential of exosomal miRNAs to regulate hESC stem cell maintenance and differentiation.

Orientador: Ibsen Bellini Coimbra / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-21T06:48:06Z (GMT). No. of bitstreams: 1 Bombini_MarianaFreschi_M.pdf: 1816531 bytes, checksum: 7a3edba503588d6825d034f24c884991 (MD5) Previous issue date: 2012 / Resumo: A utilização de células-tronco mesenquimais (CTM) para a reconstrução da cartilagem articular é uma promissora alternativa terapêutica, devido à vulnerabilidade do tecido a lesões e processo degenerativo irreversível. O objetivo deste estudo foi investigar o potencial condrogênico de CTM de líquido amniótico humano (CTM-LA) em sistema de cultura de micromass (alta densidade celular) com TGF-?3 por 21 dias. Métodos: 53 líquido amnióticos (LA) foram obtidos de mulheres submetidas à amniocentese durante o segundo trimestre de gravidez. A indicação da amniocentese foi feita pela obstetrícia, conforme protocolo específico do serviço de medicina fetal da UNICAMP. Foram selecionadas células-tronco mesenquimais, caracterizadas por citometria de fluxo. Estas células foram expandidas para obter um número populacional para o desenvolvimento do micromass. O micromass foi realizado em placa de cultura de 96 poços com fundo em "v", em cada poço foram pipetados 10?l contendo 5x105 CTM-LA e meio para diferenciação condrogênica contendo TGF-?3. Esta condição se manteve por 21 dias, e então, o potencial condrogênico foi avaliado pela presença da proteína do colágeno II pela técnica de western blotting, também foi avaliada a expressão gênica do Sox-9, colágeno II e agrecano pela técnica da PCR em tempo real (qRT-PCR). Comparamos CTM-LA em monocamada a CTM-LA submetidas ao sistema de cultura de micromass e como controle positivo utilizamos a cartilagem adulta humana. Resultados: Confirmamos o potencial condrogênico pela diferenciação das CTM-LA em condrócitos através da expressão dos genes SOX-9, colágeno tipo II e agrecano, bem como a proteína do colágeno II. A expressão de SOX-9 em micromass foi significativamente maior do que na cartilagem adulta. Conclusão: A condrogênese foi desenvolvida a partir da combinação de uma fonte de célula tronco recém descrita proveniente do líquido amniótico humano com o sistema de cultura de micromass. Esta fonte apresentou alto potencial condrogênico e dessa forma, fortes evidências para aplicações clínicas. Os resultados são promissores e sugerem a possibilidade de investimentos em bancos de líquido amniótico / Abstract: Introduction: The use of mesenchymal stem cells (MSC) for reconstruction of articular cartilage, leads to a promising therapeutic alternative, due to the tissue vulnerability to injuries and irreversible degenerative process. The aim of this study was to investigate the chondrogenic potential of MSC from human amniotic fluid in Micromass system (high-density cell culture) with TGF-?3 for 21 days. Methodology: The amniotic fluid was obtained from 53 pregnant women. The micromass was performed using MSC that was cultured in monolayer and chondrocytes from adult human normal cartilage as control groups. After 21 days, the chondrogenic potential was determined by metabolic products released from the cell, such as SOX-9, type II collagen and aggrecan. This study was approved by the ethics committee. Results: The proteic production of type II collagen was observed by Western Blotting. The genetic expression of SOX-9 was analyzed by PCR in real time, and this was found to be significantly higher than in adult cartilage. The same procedure was used to determinate the genetic expression of aggrecan and type II collagen, verifying positive result for both. Conclusion: Chondrogenesis was developed from the unique combination of the newly discovered source of mesenchymal stem cells from human amniotic fluid with micromass, and it demonstrated a satisfactory expression. Thus, this source is extremely viable for clinical applications, and the results suggest the possibility of investments in human amniotic fluid banks / Mestrado / Clinica Medica / Mestra em Clínica Médica

Mesenchymale Stammzellen bieten eine interessante Option in der regenerativen Medizin, da sie praktisch unlimitiert verfügbar sind. Um das Verhalten von humanen MSC zu studieren, werden Untersuchungen momentan an immundefizienten Mäusen durchgeführt, deren Verwendung kostenintensiv und aufwendig ist. Fra-gestellung war, ob durch Immunisolation (Alginat, Dialyseschlauch, Diffusionskammer) die Knorpel erhaltenden -, bzw. bildenden Eigenschaften von MSC-Konstrukten ebenso gut in immunkompetenten Mäusen untersucht werden können. Gleichzeitig sollte geprüft werden, ob die mit einer Immunabschirmung einhergehende Reduktion der Zellversorgung und damit die Annäherung an die Gelenksituation ihre Mineralisierung vermindern kann und ob Mauszellen für eine Veränderung der vordifferenzierten Knorpelpellets verantwortlich sind. Hierzu wurden hBMSC chondrogen differenziert. Die Zellpellets wurden mit Alginat, dem Dialyseschlauch oder der Diffusionskammer verkapselt und parallel zu unver-kapselten Kontrollpellets subkutan in immundefiziente SCID-Mäuse sowie in immunkompetente BDF1-Mäuse implantiert. Die Explantate wurden mit Alzianblau-, Alizarinrot-, Kollagen Typ II-Färbungen, sowie einer ALU in-situ Hybridisierung mar-kiert und mittels Histologiescore doppelt blind bewertet (MannWhitneyU). Überra-schenderweise zeigten die unverkapselten Kontrollen in den BDF1-Mäusen weder Zeichen von Inflammation noch von Destruktion und 4/5 der Pellets waren auf Kol-lagen Typ-II und Alzianblau positiv. Gleichzeitig war der Grad der Mineralisierung in den BDF1-Mäusen gegenüber SCID-Mäusen reduziert (p = 0,03). Durch Alginat wurde die Mineralisierung in den BDF1 Mäusen (0/8) völlig verhindert, während in den SCID-Mäusen noch 7/8 der Pellets Kalzifizierung zeigten (p = 0,001). Die Verkapselung mit Alginat verglichen mit der Kontrolle führte in beiden Mausstämmen zu höheren Scores für Kollagen Typ II (SCID: p = 0,013, BDF1: p = 0,042) und zeigte gleichzeitig eine Reduktion der Mineralisierung (SCID: p = 0,018, BDF1: p = 0,031). In SCID-Mäusen war außerdem der Alzianblau-Wert gegenüber den Kontrollen erhöht (p = 0,003). Die Diffusionskammer erwies sich als ungeeignet, da die Pellets ihre knorpeligen Eigenschaften verloren. Durch die Verwendung des Dialyseschlauches konnte lediglich in der SCID-Maus eine Erhöhung der Kollagen Typ II (p = 0,03) und eine Reduktion der Kalzifizierung (p = 0,004) erreicht werden. Sowohl im Alginatbead in der BDF1-Maus (1/3 Spendern), als auch im Dialyseschlauch mit Kollagenmembran (2/3 Spendern) konnte eine erfolgreiche in vivo Chondrogenese durchgeführt werden. Zur Untersuchung der in vivo Stabilität knorpeliger MSC-basierter Konstrukte stellt die BDF1-Maus eine attraktive, kostengünstige Alternative mit einer gegenüber der SCID-Maus verringerten Mineralisierungsrate dar. Die in vitro gebildete knorpelige Extrazellulärmatrix erzeugt dabei bereits eine Immunisolation, welche die Transplantatdestruktion verhindert. Ob ein intaktes lymphozytäres System die Knorpelstabilität gegenüber defizienten Immunsystemen begünstigt, muss durch die Untersuchung weiterer Ansätze belegt werden. Im Gegensatz zur Diffusionskammer bietet Alginat das richtige Maß an Versorgungsreduktion, um die Stabilisierung des Knorpelphänotyps der Konstrukte zu ermöglichen.

Les maladies cardiovasculaires sont la première cause de mortalité dans le monde. Une meilleure compréhension des mécanismes physiopathologiques est nécessaire. Dans ce travail de thèse nous nous sommes intéressés à deux pathologies cardiovasculaires : l’athérosclérose et l’insuffisance cardiaque. Récemment, nous avons identifié le récepteur LRP1 et la protéine adaptatrice ShcA comme étant deux protéines impliquées dans deux de ces pathologies cardiovasculaires. Nousavons montré que ShcA joue un rôle protecteur dans l’insuffisance cardiaque. Chez les souris déficientes en ShcA au niveau cardiaque, nous observons une cardiomyopathie caractérisée par une dilatation du ventricule gauche associée à une perte de la contractilité. Nous avons montré que ShcA est essentiel à l’organisation des sarcomères et ceci très tôt durant l’embryogenèse. Dans une deuxième partie nous avons montré qu’en l’absence de PPARgamma, LRP1 était nécessaire à la calcification vasculaire en activant la voie prochondrogénique de Wnt5a. Nous avons montré que PPARgamma protège de la calcification vasculaire en induisant l’expression de Sfrp2 qui agit comme un antagoniste de Wnt5a. / Cardiovascular disease is the number one cause of death worldwide. A better understanding of the pathophysiological mechanisms is necessary. In this thesis we are focused on two cardiovascular diseases: atherosclerosis and heart failure. Recently, we identified the LRP1 receptor and the adapter protein ShcA as two proteins involved in two of these cardiovascular diseases. We have shown that ShcA exerts a protective role against heart failure. Mutant mice lacking ShcA in the heart exhibit a dilated cardiomyopathy with reduced cardiac contractility. Myocyte ultrastructure analysis shows that Shc A is essential to maintain sarcomeric intégrity in early embryonic heart development. in last part we have shown vascular calcification in the absence of PPARgamma requires expression of LRP1 in vascular smooth muscle cells. LRP1 promotes a Wnt5a-dependent prochondrogenic pathway. We show that PPARgamma protects against vascular calcification by inducing the expression of secreted frizzled-related protein-2 (Sfrp2, wich functions as a Wnt5a antagonist.

[EN] Articular cartilage is a tissue with low capacity for self-restoration due to its avascularity and low cell population. It is located on the surface of the subchondral bone covering the diarthrodial joints. Degeneration of articular cartilage can appear in athletes, in people with genetic degenerative processes (osteoarthritis or rheumatoid arthritis) or due to a trauma; what produces pain, difficulties in mobility and progressive degeneration that finally leads to joint failure. Self-restoration is only produced when the defect reaches the subchondral bone and bone marrow mesenchymal stem cells (MSCs) invade the defect. However, this new formed tissue is a fibrocartilaginous type cartilage and no a hyaline cartilage, which finally leads to degeneration. Transplantation of autologous chondrocytes has been proposed to regenerate articular cartilage but this therapy fails mainly to the absence of a material support (scaffold) for the adequate stimulation of cells. Matrix-induced autologous chondrocyte implantation uses a collagen hydrogel as scaffold for chondrocytes; however, it does not have the adequate mechanical properties, does not provide the biological cues for cells and regenerated tissue is not articular cartilage but fibrocartilage. Different approaches have been done until now in order to obtain a scaffold that mimics better articular cartilage properties and composition. Hydrogels are a good option as they retain high amounts of water, in a similar way to the natural tissue, and can closely mimic the composition of natural tissue by the combination of natural derived hydrogels. Their three-dimensionality plays a critical role in articular cartilage tissue engineering to maintain chondrocyte function, since monolayer culture of chondrocytes makes them dedifferentiate towards a fibroblast-like phenotype secreting fibrocartilage. Recently, injectable hydrogels have attracted attention for the tissue engineering of articular cartilage due to their ability to encapsulate cells, injectability in the injury with minimal invasive surgeries and adaptability to the shape of the defect. Following this new approach we aimed at synthesizing two new families of injectable hydrogels based on the natural protein gelatin for the tissue engineering of articular cartilage. The first series of materials consisted on the combination of injectable gelatin with loose reinforcing polymeric microfibers to obtain injectable composites with improved mechanical properties. Our results demonstrate that there is an influence of the shape and distribution of the fibers in the mechanical properties of the composite. More importantly bad fiber-matrix interaction is not able to reinforce the hydrogel. Due to this, our composites were optimized by improving matrix-fiber interaction through a hydrophilic grafting onto the microfibers, with very successful results. The second series of materials were inspired in the extracellular matrix of articular cartilage and consisted of injectable mixtures of gelatin and hyaluronic acid. Gelatin molecules in the mixtures provided integrin adhesion sites to cells, and hyaluronic acid increased the mechanical properties of gelatin. This combination demonstrated ability for the differentiation of MSCs towards the chondrocytic lineage and makes these materials very good candidates for the regeneration of articular cartilage. The last part of this thesis is dedicated to the synthesis of a non-biodegradable material with mechanical properties, swelling and permeability similar to cartilage. This material intends to be used as a platform in a bioreactor in which the typical loads of the joint are simulated, so that the hydrogels or scaffolds would fit in the recesses in the platform. The function of the platform is to simulate the effect of the surrounding tissue on the scaffold after implantation and could reduce animal experimentation by simulating in vivo conditions. / [ES] El cartílago articular es un tejido con baja capacidad de auto-reparación debida a su avascularidad y baja población celular. Se encuentra en la superficie del hueso subcondral cubriendo las articulaciones. La degeneración del cartílago articular puede aparecer en atletas, en personas con procesos genéticos degenerativos o debido a un trauma; lo que produce dolor, dificultades en la movilidad y degeneración progresiva que lleva al fallo de la articulación. La auto-reparación sólo se produce cuando el defecto alcanza el hueso subcondral y las células madre (MSCs) de la médula ósea invaden el defecto. Sin embargo, este nuevo tejido es un cartílago de tipo fibrocartilaginoso y no un cartílago hialino, el cual finalmente lleva a la degeneración. El trasplante de condrocitos autólogos ha sido propuesto para regenerar el cartílago articular pero esta terapia falla principalmente por la ausencia de un material soporte (scaffold) que estimule adecuadamente a las células. El implante de condrocitos autólogos mediante un hidrogel de colágeno no tiene las propiedades mecánicas apropiadas, no proporciona las señales biológicas a las células y el tejido regenerado no es cartílago articular sino fibrocartílago. Se han realizado diferentes enfoques para obtener un scaffold que mimetice mejor las propiedades y la composición del cartílago articular. Los hidrogeles son una buena opción ya que retienen elevadas cantidades de agua, de forma similar al tejido natural, y pueden imitar de cerca la composición del tejido natural mediante la combinación de derivados de hidrogeles naturales. Su tridimensionalidad juega un papel crítico para mantener la función de los condrocitos, ya que el cultivo en monocapa de los condrocitos hace que desdiferencien hacia un fenotipo similar al fibroblasto secretando fibrocartílago. Los hidrogeles inyectables han acaparado la atención en la ingeniería tisular de cartílago articular debido a su capacidad para encapsular células, su inyectabilidad en el daño con cirugías mínimamente invasivas y su adaptabilidad a la forma del defecto. Siguiendo este nuevo enfoque hemos sintetizado dos nuevas familias de hidrogeles inyectables basados en la proteína natural gelatina para la ingeniería tisular del cartílago articular. La primera serie de materiales combina una gelatina inyectable con microfibras poliméricas sueltas de refuerzo para obtener composites inyectables con propiedades mecánicas mejoradas. Nuestros resultados demuestran que hay una influencia de la forma y la distribución de las fibras en las propiedades mecánicas del composite. Además, la mala interacción entre las fibras y la matriz no es capaz de reforzar el hidrogel. Debido a esto, nuestros composites han sido optimizados mediante la mejora de la interacción fibra-matriz a través de un injerto hidrófilo sobre las microfibras, con resultados muy exitosos. La segunda serie de materiales se ha inspirado en la matriz extracelular del cartílago articular y ha consistido en mezclas inyectables de gelatina y ácido hialurónico. Las moléculas de gelatina proporcionan los dominios de adhesión mediante integrinas a las células, y el ácido hialurónico aumenta las propiedades mecánicas de la gelatina. Esta combinación ha demostrado la habilidad para la diferenciación de MSCs hacia el linaje condrocítico y convierte a estos materiales en buenos candidatos para la regeneración del cartílago articular. La última parte de esta tesis se dedica a la síntesis de un material no biodegradable con propiedades mecánicas, hinchado y permeabilidad similar al cartílago. Este material pretende ser empleado como plataforma en un biorreactor en el que se simulan las cargas típicas de las articulaciones, de forma que los scaffolds encajarían en los huecos de la plataforma. Su función es simular el efecto del tejido circundante en el scaffold después de su implantación y podría reducir la experimentación anim / [CA] El cartílag articular es un teixit amb baixa capacitat d'auto-reparació deguda a la seua avascularitat i baixa població cel·lular. Es troba en la superfície de l'ós subcondral cobrint les articulacions. La degeneració del cartílag articular pot aparèixer en atletes, en persones amb processos genètics degeneratius o degut a un trauma; produeix dolor, dificultats a la mobilitat i degeneració progressiva que finalment porta a la fallida de l'articulació. L'auto-reparació es produeix quan el defecte arriba fins a l'ós subcondral i les cèl·lules mare (MSCs) de la medul·la òssia envaeixen el defecte. No obstant això, aquest nou teixit format es un cartílag de tipus fibrocartilaginós i no un cartílag hialí, el qual finalment porta a la degeneració. El transplantament de condròcits autòlegs ha sigut proposat per a regenerar el cartílag articular però aquesta teràpia falla principalment per la absència d'un material de suport (scaffold) que estimuli adequadament a les cèl·lules. L'implant de condròcits autòlegs en un hidrogel de col·lagen per als condròcits no té les propietats mecàniques apropiades, no proporciona les senyals biològiques a les cèl·lules i el teixit regenerat no és cartílag articular sinó fibrocartílag. Diferents enfocs han sigut realitzats fins ara per a obtenir un scaffold que mimetitzi millor les propietats i la composició del cartílag articular. Els hidrogels son una bona opció ja que retenen elevades quantitats d'aigua, de forma similar al teixit natural, i poden imitar acuradament la composició del teixit natural mitjançant la combinació d'hidrogels naturals. La seua tridimensionalitat juga un paper crític per a mantenir la funció dels condròcits, ja que el cultiu en monocapa dels condròcits fa que aquests desdiferencien cap a un fenotip similar al fibroblàstic secretant fibrocartílag. Recentment, els hidrogels injectables han acaparat l'atenció en l' enginyeria tissular de cartílag articular degut a la seua capacitat per a encapsular cèl·lules, la seua injectabilitat en el dany amb cirurgies mínimament invasives i la seua adaptabilitat a la forma del defecte. Seguint aquesta nova aproximació hem sintetitzat dues noves famílies d'hidrogels injectables basats en la proteïna natural gelatina per a l'enginyeria tissular del cartílag articular. La primera sèrie de materials combina una gelatina injectable amb microfibres polimèriques soltes de reforç per a obtenir compòsits injectables amb propietats mecàniques millorades. Els nostres resultats demostren que hi ha una influència de la forma i la distribució de les fibres en les propietats mecàniques del compòsit. Més importantment, la mala interacció entre les fibres i la matriu no és capaç de reforçar l'hidrogel. Degut a això, els nostres compòsits han segut optimitzats mitjançant la millora de la interacció fibra-matriu a traves d'un empelt hidròfil sobre les fibres, amb resultats molt exitosos. La segona sèrie de materials està inspirada en la matriu extracel·lular del cartílag articular i ha consistit en mescles injectables de gelatina i àcid hialurònic. Les molècules de gelatina proporcionen els dominis d'adhesió mitjançant integrines a les cèl·lules, i l'àcid hialurònic augmenta les propietats mecàniques de la gelatina. Esta combinació ha demostrat l'habilitat per a la diferenciació de MSCs cap al llinatge condrocític i converteix a aquests materials en bons candidats per a la regeneració del cartílag articular. L'última part d'aquesta tesi és dedicada a la síntesi d'un material no biodegradable amb propietats mecàniques, inflat i permeabilitat similar al cartílag. Aquest material pretén ser utilitzat com a plataforma a un bioreactor que simula les cargues típiques de les articulacions, de manera que els hidrogels o scaffolds encaixarien als buits de la plataforma. La seua funció es simular l'efecte del teixit circumdant al scaffold després d / Poveda Reyes, S. (2016). Protein-based injectable hydrogels towards the regeneration of articular cartilage [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/61392 / Premios Extraordinarios de tesis doctorales