New strategies for tissue regeneration

My PhD project is divided in two parts, focusing on the development of new strategies for orthopedic tissue regeneration. In particular, the first part is about cartilage regeneration using human lipoaspirate as autologous injectable active scaffold for one-step repair of cartilage defects, the second part is about bone regeneration, through an injectable medicated graft substitute active on bone tissue regeneration. I. Cartilage regeneration Research on mesenchymal stem cells from adipose tissue (ASC) shows promising results for cell-based therapy in cartilage lesions: in these studies cells have been isolated, expanded, and differentiated in vitro, before transplantation into the damaged cartilage, or onto materials used as scaffolds to deliver cells to the impaired area. The present study employed in vitro assays to investigate the potential of intra-articular injection of micro-fragmented lipoaspirate, as a one-step repair strategy; it aimed to determine whether adipose tissue can act as a scaffold for cells naturally present at their anatomical site. Cultured clusters of lipoaspirate showed a spontaneous outgrowth of cells with mesenchymal phenotype and with multilineage differentiation potential. Transduction of lipoaspirate clusters by lentiviral vectors expressing GFP underlined the propensity of the outgrown cells to repopulate fragments of damaged cartilage. On the basis of the results, which showed an induction of proliferation and extracellular matrix (ECM) production of human primary chondrocytes, it was hypothesized that lipoaspirate may play a paracrine role. Moreover, the structure of a floating culture of lipoaspirate, treated for three weeks with chondrogenic growth factors, changed: tissue with a high fat component was replaced by a tissue with a lower fat component and connective tissue rich in glycosaminoglycan (GAG) and in collagen type I, increasing the mechanical strength of the tissue. From these promising in vitro results, it may be speculated that an injectable autologous biologically-active scaffold (lipoaspirate), employed intra-articularly, may: 1) become a fibrous tissue that provides mechanical support for the load on the damaged cartilage; 2) induce host chondrocytes to proliferate and produce ECM; 3) provide cells at the site of injury, which could regenerate or repair the damaged or missing cartilage. II. Bone regeneration With the aim to obtain an injectable medicated scaffold, which speeds bone formation in sinus lift augmentation, in bony void and in fracture repair, we have developed a three-dimensional (3D) jelly collagen containing Lysophosphatidic acid (LPA) and 1a,25-Dihydroxyvitamin D3 (1,25D3) using soluble native collagen prepared from rat tail tendons. We have demonstrated with an in vitro 3D culture model of bone fracture an osteoblasts\u2019Rho-kinase mediated contraction of the collagen that causes an approach of human bone trabecular fragments with the formation of new union tissue within 3 weeks of organ culture. The contraction was faster in LPA medicated collagen while 1,25D3 enhanced the mineralization of the new formed tissue that showed also increased tensile strength. LPA was shown to modulate gel contraction rate not only mechanically, working in cytoskeleton reorganization, but also osteoconductively evidencing activity on proliferation, differentiation and migration of human primary osteoblasts (hOB). When LPA was used in combination with 1,25D3 a synergism on hOB\u2019s activity in term of alkaline phosphatase and mineralization was seen. On the basis of these data, collagen can be considered as an injectable natural scaffold that allows the migration of cells from the side of bone defect and its enrichment with LPA and 1,25D3 could be used in vivo to accelerate bone growth and fracture healing

Canadá más allá del TLCAN

La renegociación del TLCAN pone nuevamente en evidencia la fuerte dependencia del comercio canadiense con EEUU, pero si bien puede entenderse como una amenaza a la situa-ción comercial actual de Canadá, se presenta como una chance de efectuar cambios significa-tivos en las relaciones económicas con el resto del mundo. Ante este escenario y la incerti-dumbre del resultado de dichas negociaciones, la posibilidad de diversificar geográficamente las exportaciones y extender sus acuerdos con otros socios comerciales se presenta como una alternativa cada vez más necesaria.Departamento de América del NorteInstituto de Relaciones Internacionale

Canadá más allá del TLCAN

La renegociación del TLCAN pone nuevamente en evidencia la fuerte dependencia del comercio canadiense con EEUU, pero si bien puede entenderse como una amenaza a la situa-ción comercial actual de Canadá, se presenta como una chance de efectuar cambios significa-tivos en las relaciones económicas con el resto del mundo. Ante este escenario y la incerti-dumbre del resultado de dichas negociaciones, la posibilidad de diversificar geográficamente las exportaciones y extender sus acuerdos con otros socios comerciales se presenta como una alternativa cada vez más necesaria.Departamento de América del NorteInstituto de Relaciones Internacionale

Canadá más allá del TLCAN

La renegociación del TLCAN pone nuevamente en evidencia la fuerte dependencia del comercio canadiense con EEUU, pero si bien puede entenderse como una amenaza a la situa-ción comercial actual de Canadá, se presenta como una chance de efectuar cambios significa-tivos en las relaciones económicas con el resto del mundo. Ante este escenario y la incerti-dumbre del resultado de dichas negociaciones, la posibilidad de diversificar geográficamente las exportaciones y extender sus acuerdos con otros socios comerciales se presenta como una alternativa cada vez más necesaria.Departamento de América del NorteInstituto de Relaciones Internacionale

Canadá-Mercosur: negociaciones hacia el TLC

Entre el 22 y 29 de febrero de 2019, se llevó a cabo la V Ronda de Negociaciones para el establecimiento de un Tratado de Libre Comercio entre Canadá y el MERCOSUR. Aproximadamente un año después del inicio formal de las negociaciones, resulta relevante repasar el estado de las mismas y destacar los puntos desfavorables que cada una de las partes puede afrontar eventualmente.Centro de Estudios CanadiensesInstituto de Relaciones Internacionale

Canadá-Mercosur: negociaciones hacia el TLC

Entre el 22 y 29 de febrero de 2019, se llevó a cabo la V Ronda de Negociaciones para el establecimiento de un Tratado de Libre Comercio entre Canadá y el MERCOSUR. Aproximadamente un año después del inicio formal de las negociaciones, resulta relevante repasar el estado de las mismas y destacar los puntos desfavorables que cada una de las partes puede afrontar eventualmente.Centro de Estudios CanadiensesInstituto de Relaciones Internacionale

Human lipoaspirate as autologous injectable active scaffold for one-step repair of cartilage defects

Research on mesenchymal stem cells from adipose tissue shows promising results for cell-based therapy in cartilage lesions. In these studies, cells have been isolated, expanded, and differentiated in vitro before transplantation into the damaged cartilage or onto materials used as scaffolds to deliver cells to the impaired area. The present study employed in vitro assays to investigate the potential of intra-articular injection of microfragmented lipoaspirate as a one-step repair strategy; it aimed to determine whether adipose tissue can act as a scaffold for cells naturally present at their anatomical site. Cultured clusters of lipoaspirate showed a spontaneous outgrowth of cells with a mesenchymal phenotype and with multilineage differentiation potential. Transduction of lipoaspirate clusters by lentiviral vectors expressing GFP evidenced the propensity of the outgrown cells to repopulate fragments of damaged cartilage. On the basis of the results, which showed an induction of proliferation and ECM production of human primary chondrocytes, it was hypothesized that lipoaspirate may play a paracrine role. Moreover, the structure of a floating culture of lipoaspirate, treated for 3 weeks with chondrogenic growth factors, changed: tissue with a high fat component was replaced by a tissue with a lower fat component and connective tissue rich in GAG and in collagen type I, increasing the mechanical strength of the tissue. From these promising in vitro results, it may be speculated that an injectable autologous biologically active scaffold (lipoaspirate), employed intra-articularly, may 1) become a fibrous tissue that provides mechanical support for the load on the damaged cartilage; 2) induce host chondrocytes to proliferate and produce ECM; and 3) provide cells at the site of injury, which could regenerate or repair the damaged or missing cartilage

Evaluación de la meta 17.2 de los Objetivos de Desarrollo Sostenible para los países del Comité de Ayuda al Desarrollo

El logro de los Objetivos de Desarrollo Sostenible requiere de un gran compromiso y esfuerzo de los distintos actores públicos y privados a nivel internacional, pero en particular precisa de un volumen de ayuda financiera considerable. La meta 17.2 permite monitorear estos aspectos de la cooperación, focalizándose en aquellas naciones más desarrolladas. Pese a los progresos observados, principalmente en las últimas dos décadas, resta aún mucho camino por recorrer y quizás sea necesario un replanteo a nivel global de las responsabilidades, procurando completar la Agenda 2030 sin “que nadie se quede atrás”.Departamento de Cooperación InternacionalInstituto de Relaciones Internacionale

Objetivos de Desarrollo Sustentable, comercio de bienes y los Países Menos Avanzados: mismas reglas, ¿distintos resultados?

La concreción de las metas de los Objetivos de Desarrollo Sustentable se encuentra fuertemente condicionada por el marco normativo. Asociar cuestiones de desarrollo a reglas que son criticadas en este aspecto, resulta contradictorio y más aún, cuando se esperan resultados significativos que no han sido alcanzados en el pasado. El Trato Especial y Diferenciado entra en esta categoría de reglas y sus logros pueden ser cuestionables, evidenciando la necesidad de cambios que permitan adecuar estos instrumentos a las verdaderas necesidades de los Países Menos Avanzados.Instituto de Relaciones Internacionale

Lipoaspirate Shows In Vitro Potential for Wound Healing

Mesenchymal stem cells (MSCs) are a promising therapy in wound healing, although extensive time and manipulation are necessary for their use. In our previous study on cartilage regeneration, we demonstrated that lipoaspirate acts as a natural scaffold for MSCs and gives rise to their spontaneous outgrowth, together with a paracrine effect on resident cells that overcome the limitations connected to MSC use. In this study, we aimed to investigate in vitro whether the microfragmented adipose tissue (lipoaspirate), obtained with Lipogems® technology, could promote and accelerate wound healing. We showed the ability of resident cells to outgrow from the clusters of lipoaspirate encapsulated in a 3D collagen substrate as capability of repopulating a culture of human skin. Moreover, we demonstrated that the in vitro lipoaspirate paracrine effect on fibroblasts and keratinocytes proliferation, migration, and contraction rate is mediated by the release of trophic/reparative proteins. Finally, an analysis of the paracrine antibacterial effect of lipoaspirate proved its ability to secrete antibacterial factors and its ability to modulate their secretion in culture media based on a bacterial stimulus. The results suggest that lipoaspirate may be a promising approach in wound healing showing in vitro regenerative and antibacterial activities that could improve current therapeutic strategies