Material-driven fibronectin fibrillogenesis to engineer cell function

This thesis ventures with the extracellular matrix protein (ECM) fibronectin (FN) as an interface protein in the interaction between cells and materials to design microenvironment for future use in tissue engineering. It is studied the FN adsorption and conformations, cell behaviour to different FN conformation, cell adhesion, reorganisation and remodelling of FN at the material interface, the role of growth factors (GF) and their interactions with components of the extracellular matrix (ECM), the immunology cell response, and the stem cell fate influenced by the extrinsic signals coming from the engineered microenvironments using ECM's proteins. To investigate the FN response, in terms of adsorbed amount and conformation to different chemical properties of the material, model surfaces were used. Self assembled monolayers (SAM) with different percentages of two different chemical groups were used: CH3 and OH. FN adsorption, initial cell adhesion and signalling (focal adhesions, integrin expression and phosphorylation of FAK) is related with the reorganisation and secretion of FN and matrix degradation. It is shown that matrix degradation at the cell material interface depends on surface chemistry in metalloproteinase-dependent way. A direct relationship between FN activity at the cell-material interface and metalloproteinase 9 (MMP9) expression was found, being the product of a sequence of events that include integrin expression, focal adhesion formation, matrix reorganisation and focal adhesion kinase (FAK) phosphorylation. Two different materials with subtle variations in their chemical composition were employed as a drastically different FN conformation: from a globular conformation on PMA (poly (methyl acrylate)) to the formation of a well-interconnected FN network (similar to the FN physiological fibrillar network) triggered by PEA (poly (ethyl acrylate)). The formation of focal adhesions (vinculin), FAK expression and phosphorylation, specific integrin binding, protein and gene expression for ¿5 and ¿v was studied, seeking to correlate cell adhesion with matrix degradation. It is demonstrated that the material-driven FN fibrillogenesis on PEA triggers proteolytic activity: MMP activity is higher as a compensatory mechanism to the inability of cells to reorganise this FN network. Looking into the role of protein-material interactions and stem cell fate, and with the knowledge on PEA, we engineer different synergistic microenvironments to direct cell and stem cell fate. FN has a growth factor (GF) binding domain on its molecule (FNIII12-14) and has been demonstrated to produce a synergistic response when occurs at the same time the recognition of the cell binding domain (FNIII9-10). It is demonstrated that this domain is available on the FN coated PEA, and exploiting these interactions between PEA, FN and GF, it is developed a microenvironment to control cell behaviour and tissue repair. It is studied the BMP2 binding and presentation, the effect of BMP2 presentation on MSC proliferation and differentiation. These systems allow not only enhanced activity of GF compared to soluble administration, but also reduce GF doses, improving safety and cost effectiveness. Finally, the immunological reaction of the microenvironment developed is studied using dendritic cells, beside the conformational structure of ECM protein importance in DC integrin-based activation it is studied, helping to establish the field of adhesion-based modulation of DC as a general mechanism that has previously not been defined. The microenvironment didn't induce any maturation in DC, while different FN conformation shows differences in DC morphology and citokine level production (IL-10 and IL-12).En esta tesis se estudia la interacción de una proteina de la matriz extracelular, fibronectina (FN) como interfase en la interacción entre células y materiales, para diseñar microambientes con el propósito de ser usados en el futuro en ingeniería tisular. Se estudia la adsorción y conformación de FN y la relación con el diferente comportamiento celular: la adhesión celular, la reorganización y remodelado de la FN en la interfase célula-material, el papel que juegan los factores de crecimiento y sus interacciones con los componentes de la matriz extracelular, la respuesta immunológica y el destino celular de células madre influenciadas por las señales extrínsecas provenientes de microambientes elaborados a partir de proteínas de la matriz extracelular. Con el objetivo de investigar la respuesta a la FN en términos de conformación y cantidad absorbida a diferentes propiedades químicas del material, se usaron materiales modelo: monocapas autoensambladas (self-assembled monolayers, SAM). Las químicas estudiadas fueron CH3 and OH. La adsorption de FN, adhesion y señalización (adhesiones focales, expresión de interinas y fosforilación de quinasas de adhesiones focales (FAK)) se estudiaron en relación a la reorganización y secreción de FN y degradación de la matriz extracelular. Se demuestra que la degradación de la matriz extracelular en la interfase célula-material depende de la química de la superficie, a través de las metaloproteinasas. Se ha descubierto una relación directa entre la actividad de la FN que se encuentra en el material y la expresión de metaloproteinasa 9 (MMP9), a través de la expresión de integrinas, formación de adhesiones focales, reorganización de la matriz extracelular y fosforilación de FAK En el siguiente capítulo se emplean materiales poliméricos con una sutil diferencia en la composición química, provocando una diferencia drástica en la conformación de la FN: se pasa de una conformación globular en PMA (polimetil acrilato) a una conformación en forma de red interconectada en PEA (polietil acrilato). Con el propósito de relacionar la adhesión celular con la degradación de la matriz extracelular, se estudia la formación de adhesiones focales (vinculina), la expresión y fosforilación de FAK, la unión específica de integrinas y la expresión de las integrinas ¿5 and ¿v. Se demuestra que la formación de una red de FN sobre PEA induce la actividad proteolítica: la actividad de las MMPs es mayor, actuando como mecanismo compensatorio a la incapacidad de reorganización de la red de FN. Haciendo uso de la conformación de la FN sobre PEA, se estudiaron las interacciones entre la proteína-material y el destino celular de células madres. La FN posee un dominio de unión de factores de crecimiento (FNIII12-14) y se ha demostrado que se produce una respuesta sinérgica cuando el reconocimiento ocurre junto con el dominio de unión celular (FNIII9-10). En esta tesis se demuestra que el dominio de unión de factores de crecimiento está disponible en la conformación que adquiere sobre PEA y se diseñan microambientes para controlar el comportamiento celular y regeneración de tejido. Se estudia la unión y presentación de BMP2 y su efecto en la diferenciación de células madre mesenquimales. Los microambientes desarrollados, ademas de mejorar la actividad de los factores de crecimiento comparado con la administración soluble, también reduce la cantidad de factores de crecimiento que se tendría que administrar, mejorando la seguridad y efectividad. Finalmente se estudió la reacción inmunológica a los microambientes desarrollados usando células dendríticas, estudiando además la influencia de la estructura de la conformación de las proteínas en la activación de las células dendríticas a través de las integrinas. Los microambientes no indujeron ninguna maduración de células dendríticas, mientras que la conformación de la FN muestra controlEn aquesta tesi s'estudia la interacció entre una proteïna de la matriu extracel.lular, fibronectina (FN) com interfase en la interaccio entre cèl·lules i materials, per a dissenyar microambients amb el propòsit d'utilitzar-se al futur en enginyeria tissular. S'estudia l'adsorció i conformació de la FN i la relació amb el diferent comportament cel·lular: l'adhesió cel·lular, la reorganització i remodelat de la FN a la interfase cèl·lula-material, el paper que juguen els factors de creixement i les seus interaccions amb els components de la matriu extracel·lular, la resposta immunològica i el destí cel·lular de cèl·lules mare influenciades pels senyals extrínseques provinents de microambients elaborats a partir de proteïnes de la matriu extracel·lular. Amb l'objectiu d'investigar la respostar a la FN en termes de conformació i quantitat absorbida a diferents propietats químiques del material, s'utilitzaren materials model: monocapes autoacoblades (self-assembled monolayers, SAM). Les químiques estudiades van ser CH3 and OH. L'absorció de FN, adhesió i senyalització (adhesions focals, expressió d'integrines i fosforilació de quinases d'adhesions focals (FAK)) es van estudiar en relació a al reorganització i secreció de la FN i degradació de la matriu extracel·lular. Es demostra que la degradació de la matriu extracelular en la interfase cèl·lula-material depèn de la química de la superficie, a través de les metal·loproteïnases. S'ha descobert una relació directa entra l'activitat de la FN que es troba en el material i l'expressió de metaloproteinasa 9, a través de l'expressió d'integrines, formació d'adhesions focals, reorganització de la matriu extracel·lular i fosforilació de FAK. Al següent capítol es fan servir materials polimèrics amb una subtil diferència en la composició química, provocant una diferència dràstica en la conformació de la FN: es passa d'una conformació globular en PMA (polimetil acrilat) a una conformació en forma de xarxa interconnectada en PEA (polietil acrilat). Amb el propòsit de relacionar l'adhesió cel·lular amb la degradació de la matriu extracel·lular, s'estudia la formació d'adhesions focals (vinculina), l'expressió i fosforilació de FAK, la unió específica d'integrines i l'expressió de les integrines ¿5 and ¿v. Es demostra que la formació d'una xarxa de FN sobre PEA indueix l'activitat proteolítica: l'activitat de les MMPs és més gran, actuant com a mecanisme compensatori a la incapacitat de reorganització de la xarxa de FN. Fent ús de la conformació de la FN sobre PEA, es van estudiar les interaccions entre la proteïna-material i el destí cel·lular de cèl·lules mares. La FN posseeix un domini d'unió de factors de creixement (FNIII12-14) i s'ha demostrat que es produeix una resposta sinèrgica quan el reconeixement ocurreix juntament amb el domini d'unió cel·lular (FNIII9- 10). En aquesta tesi es demostra que el domini d'unió de factors de creixement està disponible a la conformació que adquireix sobre PEA i es dissenyen microambients per controlar el comportament cel·lular i regeneració de teixit. S'estudia la unió i presentació de BMP2 i el seu efecte en la diferenciació de cèl·lules mare mesenquimals. Els microambientes desenvolupats, a més de millorar l'activitat dels factors de creixement comparat amb l'administració soluble, també redueix la quantitat de factors de creixement que s'hauria d'administrar, millorant la seguretat i efectivitat. Finalment es va estudiar la reacció immunològica als microambients desenvolupats usant cèl·lules dendrítiques, estudiant a més la influència de l'estructura de la conformació de les proteïnes en l'activació de les cèl·lules dendrítiques a través de les integrines. Els microambients no van induir cap maduració de cèl·lules dendrítiques, mentre que la conformació de la FN mostra controlar la morfologia de les cèl·lules dendrítiques iLlopis Hernández, V. (2017). Material-driven fibronectin fibrillogenesis to engineer cell function [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/90412TESI

Strategic behaviour in Schelling dynamics: theory and experimental evidence

In this paper we experimentally test Schelling’s (1971) segregation model and obtain the striking result of full segregation in most of the cases. In addition, we extend Schelling’s model theoretically by adding strategic behaviour and moving costs. We obtain a unique subgame perfect equilibrium in which rational agents facing moving costs may find it optimal not to move (anticipating other participants’ movements). This equilibrium is far from full segregation. We run experiments for this extended Schelling model, and find that the percentage of full segregated societies is notably reduced when the cost of moving is high, but it is not affected when it is low, relative to the baseline with costless moving. We also find that the degree of segregation depends on the distribution of strategic subjects relative of a baseline model where moving is costless

Strategic behaviour in Schelling dynamics: Theory and experimental evidence

In this paper we experimentally test Schelling's (1971) segregation model and confirm the striking result of segregation. In addition, we extend Schelling's model theoretically by adding strategic behavior and moving costs. We obtain a unique subgame perfect equilibrium in which rational agents facing moving costs may find it optimal not to move (anticipating other participants' movements). This equilibrium is far from full segregation. We run experiments for this extended Schelling model, and find that the percentage of strategic players dramatically increases with the cost of moving and that the degree of segregation depends on the distribution of strategic subjects

Decision making under uncertainty in environmental projects using mathematical simulation modeling

The final publication is available at Springer via http://dx.doi.org/10.1007/s12665-016-6135-yIn decision-making processes, reliability and risk aversion play a decisive role. The aim of this study is to perform an uncertainty assessment of the effects of future scenarios of sustainable groundwater pumping strategies on the quantitative and chemical status of an aquifer. The good status of the aquifer is defined according to the terms established by the EU Water Framework Directive (WFD). A decision support systems (DSS) is presented, which makes use of a stochastic inverse model (GC method) and geostatistical approaches to calibrate equally likely realizations of hydraulic conductivity (K) fields for a particular case study. These K fields are conditional to available field data, including hard and soft information. Then, different future scenarios of groundwater pumping strategies are generated, based on historical information and WFD standards, and simulated for each one of the equally likely K fields. The future scenarios lead to different environmental impacts and levels of socioeconomic development of the region and, hence, to a different degree of acceptance among stakeholders. We have identified the different stakeholders implied in the decision-making process, the objectives pursued and the alternative actions that should be considered by stakeholders in a public participation project (PPP). The MonteCarlo simulation provides a highly effective way for uncertainty assessment and allows presenting the results in a simple and understandable way even for non-experts stakeholders. The methodology has been successfully applied to a real case study and lays the foundations to performa PPP and stakeholders' involvement in a decisionmaking process as required by the WFD. The results of the methodology can help the decision-making process to come up with the best policies and regulations for a groundwater system under uncertainty in groundwater parameters and management strategies and involving stakeholders with conflicting interests.Llopis Albert, C.; Palacios Marqués, D.; Merigó -Lindahl, JM. (2016). Decision making under uncertainty in environmental projects using mathematical simulation modeling. Environmental Earth Sciences. 75(19):1-11. doi:10.1007/s12665-016-6135-yS1117519Arhonditsis GB, Perhar G, Zhang W, Massos E, Shi M, Das A (2008) Addressing equifinality and uncertainty in eutrophication models. Water Resour Res 44:W01420. doi: 10.1029/2007WR005862Capilla JE, Llopis-Albert C (2009) Gradual conditioning of non-gaussian transmissivity fields to flow and mass transport data. J Hydrol 371:66–74. doi: 10.1016/j.jhydrol.2009.03.015CHJ (Júcar Water Agency) (2016) Júcar river basin authority. http://www.chj.es/CHS (Segura Water Agency) (2016) Segura river basin authority. http://www.chsegura.es/Custodio E (2002) Aquifer overexploitation: what does it mean? Hydrogeol J 10:254–277EC (2000). Directive 2000/60/EC of the European Parliament and of the Council of October 23 2000, establishing a framework for community action in the field of water policy. Official Journal of the European Communities L327/1eL327/72. 22.12.2000EC (2006) Directive 2006/118/EC of the European Parliament and of the Council of 12 December 2006 on the protection of groundwater against pollution and deteriorationGómez-Hernández JJ, Srivastava RM (1990) ISIM3D: an ANSI-C three dimensional multiple indicator conditional simulation program. Comput Geosci 16(4):395–440Harbaugh AW, Banta ER, Hill MC and McDonald MG (2000) MODFLOW- 2000, The US geological survey modular groundwater model-user guide to modularization concepts and the groundwater flow process. US Geol. Surv. Open-File Rep 00–92, 12Hu LY (2000) Gradual deformation and iterative calibration of Gaussian related stochastic models. Math Geol 32(1):87–108Jagelke J, Barthel R (2005) Conceptualization and implementation of a regional groundwater model for the Neckar catchment in the framework of an integrated regional model. Adv Geosci 5:105–111Llopis-Albert C (2008) Stochastic inverse modeling conditional to flow, mass transport and secondary information. Universitat Politècnica de València, València. ISBN 978-84-691-9796-7Llopis-Albert C, Capilla JE (2009a) Gradual conditioning of non-gaussian transmissivity fields to flow and mass transport data. Demonstration on a synthetic aquifer. J Hydrol 371:53–55. doi: 10.1016/j.jhydrol.2009.03.014Llopis-Albert C, Capilla JE (2009b) Gradual conditioning of non-gaussian transmissivity fields to flow and mass transport data. Application to the macrodispersion experiment (MADE-2) site, on Columbus air force base in Mississippi (USA). J Hydrol 371:75–84. doi: 10.1016/j.jhydrol.2009.03.016Llopis-Albert C, Capilla JE (2010a) Stochastic simulation of non-gaussian 3D conductivity fields in a fractured medium with multiple statistical populations: a case study. J Hydrol Eng 15(7):554–566. doi: 10.1061/(ASCE)HE.1943-5584.0000214Llopis-Albert C, Capilla JE (2010b) Stochastic inverse modeling of hydraulic conductivity fields taking into account independent stochastic structures: a 3D case study. J Hydrol 391:277–288. doi: 10.1016/j.jhydrol.2010.07.028Llopis-Albert C, Pulido-Velazquez D (2014) Discussion about the validity of sharp-interface models to deal with seawater intrusion in coastal aquifers. Hydrol Process 28(10):3642–3654Llopis-Albert C, Pulido-Velazquez D (2015) Using MODFLOW code to approach transient hydraulic head with a sharp-interface solution. Hydrol Process 29(8):2052–2064. doi: 10.1002/hyp.10354Llopis-Albert C, Palacios-Marqués D, Merigó JM (2014) A coupled stochastic inverse-management framework for dealing with nonpoint agriculture pollution under groundwater parameter uncertainty. J Hydrol 511:10–16. doi: 10.1016/j.jhydrol.2014.01.021Llopis-Albert C, Merigó JM, Palacios-Marqués D (2015) Structure adaptation in stochastic inverse methods for integrating information. Water Resour Manage 29(1):95–107. doi: 10.1007/s11269-014-0829-2Llopis-Albert C, Merigó JM, Xu Y (2016) A coupled stochastic inverse/sharp interface seawater intrusion approach for coastal aquifers under groundwater parameter uncertainty. J Hydrol 540:774–783. doi: 10.1016/j.jhydrol.2016.06.065McDonald MG and Harbaugh AW (1988) A modular three-dimensional finite-difference groundwater flow model. US geological survey technical manual of water resources investigation, Book 6, US geological survey, Reston, Virginia, 586Molina JL, Pulido-Velazquez M, Llopis-Albert C, Peña-Haro S (2013) Stochastic hydro-economic model for groundwater quality management using Bayesian networks. Water Sci Technol 67(3):579–586. doi: 10.2166/wst.2012.598Peña-Haro S, Llopis-Albert C, Pulido-Velazquez M (2010) Fertilizer standards for controlling groundwater nitrate pollution from agriculture: El Salobral-Los Llanos case study, Spain. J Hydrol 392:174–187. doi: 10.1016/j.jhydrol.2010.08.006Peña-Haro S, Pulido-Velazquez M, Llopis-Albert C (2011) Stochastic hydro-economic modeling for optimal management of agricultural groundwater nitrate pollution under hydraulic conductivity uncertainty. Environ Model Softw 26(8):999–1008. doi: 10.1016/j.envsoft.2011.02.010Pulido-Velazquez D, Llopis-Albert C, Peña-Haro S, Pulido-Velazquez M (2011) Efficient conceptual model for simulating the effect of aquifer heterogeneity on natural groundwater discharge to rivers. Adv Water Resour 34(11):1377–1389. doi: 10.1016/j.advwatres.2011.07.010Reichert P, Borsuk M, Hostmann M, Schweizer S, Spörri C, Tockner K, Truffer B (2005) Concepts of decision support for river rehabilitation. Environ Model Softw 22:188–201Wright SAL, Fritsch O (2011) Operationalising active involvement in the EU water framework directive: why, when and how? Ecol Econ 70(12):2268–2274Zhou H, Gómez-Hernández JJ, Li L (2014) Inverse methods in hydrogeology: evolution and recent trends. Adv Water Resour 63:22–37. doi: 10.1016/j.advwatres.2013.10.01

InmunoProteómica de la respuesta a la infección sistémica en ratón con una cepa Sccharomyces cerevisiae procedente de suplementos dietéticos

Comunicaciones a congreso

Integration of ICT into the higher education process: The case of Colombia

This paper has as its aim to establish a pedagogical strategy aimed at supporting the integration of ICTs (Information and Communication Technologies) into the higher education process. A survey was carried out aiming focus on the analysis of teachers’ practices so that competences in ICT use at university education in Colombia could be enlarged. 81 universities answered the survey questionnaire: 51 private and 30 public, and a total of 423 lecturers from different academic disciplines. Our findings show the degree of attention paid by teachers to strengthening ICT skills in their training tasks. This research is complemented with Strengths, Weaknesses, Opportunities, and Threats (SWOT) analysis performed with 50 professionals experienced in ICT use in higher education

Haplotypes of "Candidatus Liberibacter solanacearum" identified in Umbeliferous crops in Spain

[EN] 'Candidatus Liberibacter solanacearum' is a phloem-limited Gram-negative bacterium that causes serious damage to different crops of the botanical families Solanaceae and Apiaceae. Five haplotypes have been described: LsoA and LsoB are present in solanaceous crops in America and vectored by the tomato/potato psyllid Bactericera cockerelli; LsoC affects carrots from Northern and Central Europe, and is transmitted by the carrot psyllid Trioza apicalis; haplotypes LsoD and LsoE are present in Southern Europe and Morocco in carrot and celery, and are associated with the psyllid Bactericera trigonica. Thirty-four 'Ca. L. solanacearum' isolates were collected in six different regions of Spain from distinct Apiaceae hosts (carrot, celery, parsley and parsnip) in eight consecutive years and were analysed. Their haplotypes were determined by a sequence analysis of 16S ribosomal RNA, the 16S-26S ribosomal RNA intergenic spacer, and the 23S ribosomal RNA and rplJ and rplL genes. Both haplotypes LsoD and LsoE were found across Spain, and no host specificity appeared between these two haplotypes. This is the first report of 'Ca. L. solanacearum' associated with parsley and parsnip.This work has been supported by grant INIA (RTA2011-00142). This paper is dedicated to the memory of F.J. Villaescusa (1981-2011). The technical support of S. Sanjuan and J.C. Ferrandiz from Agricola Villena Coop. V. is also acknowledged.Alfaro Fernández, AO.; Hernández-Llopis, D.; Font San Ambrosio, MI. (2017). Haplotypes of "Candidatus Liberibacter solanacearum" identified in Umbeliferous crops in Spain. European Journal of Plant Pathology. 149(1):127-131. https://doi.org/10.1007/s10658-017-1172-21271311491Alfaro-Fernández A., Cebrián M.C., Villaescusa F.J., Hermoso de Mendoza A., Ferrándiz J.C., Sanjuán S., Font M.I. (2012). ‘Candidatus Liberibacter solanacearum’ associated with Bactericera trigonica affected carrots in the Canary Islands. Plant Disease 96, 581.Bertolini, E., Teresani, G. R., Loiseau, M., Tanaka, F. A. O., Barbé, S., Martínez, C., Gentit, P., López, M. M., & Cambra, M. (2014). Transmission of ‘Candidatus Liberibacter solanacearum’ in carrot seeds. Plant Pathology, 64, 276–285.EPPO. (2013). Data sheets on pests recommended for regulation. Candidatus Liberibacter solanacearum. EPPO Bulletin, 43, 197–201.Green, M. J., Thompson, D. A., & Mackenzie, D. J. (1999). Easy and efficient DNA extraction from woody plants for the detection of phytoplasmas by polymerase chain reaction. Plant Disease, 83, 482–485.Hansen, A. K., Trumble, J. T., Stouthamer, R., & Paine, T. D. (2008). A new huanglongbing species, “Candidatus Liberibacter psyllaurous,” found to infect tomato and potato, is vectored by the psyllid Bactericera cockerelli (Sulc). Applied Environmental Microbiology, 74, 5862–5865.Larkin, M. A., Blackshields, G., Brown, N. P., Chenna, R., McGettigan, P. A., McWilliam, H., Valentin, F., Wallace, I. M., Wilm, A., Lopez, R., Thompson, J. D., Gibson, T. J., & Higgins, D. G. (2007). Clustal W and Clustal X version 2.0. Bioinformatics, 23, 2947–2948.Loiseau, M., Garnier, S., Boirin, V., Merieau, M., Leguay, A., Renaudin, I., Renvoisé, J.-P., & Gentit, P. (2014). First report of ‘Candidatus Liberibacter solanacearum’ in carrot in France. Plant Disease, 98, 839.Munyaneza, J. E. (2012). Zebra chip disease of potato: biology, epidemiology and management. American Journal of Potato Research, 89, 329–350.Munyaneza, J., Buchman, J., Upton, J., Goolsby, J., Crosslin, J., & Bester, G. (2008). Impact of different potato psyllid populations on zebra chip disease incidence, severity, and potato yield. Subtropical Plant Science, 60, 27–37.Munyaneza, J., Sengoda, V., Crosslin, J., de la Rosa-Lorenzo, G., & Sanchez, A. (2009). First report of ‘Candidatus Liberibacter psyllaurous’ in potato tubers with zebra Chip disease in Mexico. Plant Disease, 93, 552.Munyaneza, J. E., Fisher, T. W., Sengoda, V. G., & Garczynski, S. F. (2010a). First report of ‘Candidatus Liberibacter solanacearum’ associated with psyllid-affected carrots in Europe. Plant Disease, 94, 639.Munyaneza, J. E., Fisher, T. W., Sengoda, V. G., Garczynski, S. F., Nissinen, A., & Lemmetty, A. (2010b). Association of ‘Candidatus Liberibacter solanacearum’ with the psyllid Trioza apicalis (hemiptera: Triozidae). Journal of Economic Entomology, 103, 1060–1070.Munyaneza, J. E., Swisher, K. D., Hommes, M., Willhauck, A., Buck, H., & Meadow, R. (2015). First report of ‘Candidatus Liberibacter solanacearum’ associated with psyllid-infested carrots in Germany. Plant Disease, 99, 1269.Nelson, W. R., Fisher, T. W., & Munyaneza, J. E. (2011). Haplotypes of “Candidatus Liberibacter solanacearum” suggest long-standing separation. European Journal of Plant Patholology, 130, 5–12.Nelson, W. R., Sengoda, V. G., Alfaro-Fernández, A., Font, M. I., Crosslin, J. M., & Munyaneza, J. E. (2013). A new haplotype of ‘Candidatus Liberibacter solanacearum’ identified in the Mediterranean region. European Journal of Plant Pathology, 135, 633–639.Tahzima, R., Maes, M., Achbani, E. H., Swisher, K. D., Munyaneza, J. E., & De Jonghe, K. (2014). First report of ‘Candidatus Liberibacter solanacearum’ on carrot in Africa. Plant Disease, 98, 1426.Teresani, G. R., Bertolini, E., Alfaro-Fernández, A., Martínez, C., Tanaka, F. A. O., Kitajima, E. W., Roselló, M., Sanjuán, S., Ferrándiz, J. C., López, M. M., Cambra, M., & Font, M. I. (2014). Association of ‘Candidatus Liberibacter solanacearum’ with a vegetative disorder of celery in Spain and development of a real-time PCR method for its detection. Phytopathology, 104, 804–811.Teresani, G., Hernández, E., Bertolini, E., Siverio, F., Marroquín, C., Molina, J., de Hermoso Mendoza, A., & Cambra, M. (2015). Search for potential vectors of ‘Candidatus Liberibacter solanacearum’: population dynamics in host crops. Spanish Journal of Agricultural Research, 13, e10–002

Material-driven fibronectin assembly for high-efficiency presentation of growth factors

Growth factors (GFs) are powerful signaling molecules with the potential to drive regenerative strategies, including bone repair and vascularization. However, GFs are typically delivered in soluble format at supraphysiological doses because of rapid clearance and limited therapeutic impact. These high doses have serious side effects and are expensive. Although it is well established that GF interactions with extracellular matrix proteins such as fibronectin control GF presentation and activity, a translation-ready approach to unlocking GF potential has not been realized. We demonstrate a simple, robust, and controlled material-based approach to enhance the activity of GFs during tissue healing. The underlying mechanism is based on spontaneous fibrillar organization of fibronectin driven by adsorption onto the polymer poly(ethyl acrylate). Fibrillar fibronectin on this polymer, but not a globular conformation obtained on control polymers, promotes synergistic presentation of integrin-binding sites and bound bone morphogenetic protein 2 (BMP-2), which enhances mesenchymal stem cell osteogenesis in vitro and drives full regeneration of a nonhealing bone defect in vivo at low GF concentrations. This simple and translatable technology could unlock the full regenerative potential of GF therapies while improving safety and cost-effectiveness

Role of Surface Chemistry in Protein Remodeling at the Cell-Material Interface

Background: The cell-material interaction is a complex bi-directional and dynamic process that mimics to a certain extent the natural interactions of cells with the extracellular matrix. Cells tend to adhere and rearrange adsorbed extracellular matrix (ECM) proteins on the material surface in a fibril-like pattern. Afterwards, the ECM undergoes proteolytic degradation, which is a mechanism for the removal of the excess ECM usually approximated with remodeling. ECM remodeling is a dynamic process that consists of two opposite events: assembly and degradation. Methodology/Principal Findings: This work investigates matrix protein dynamics on mixed self-assembled monolayers (SAMs) of –OH and –CH3 terminated alkanethiols. SAMs assembled on gold are highly ordered organic surfaces able to provide different chemical functionalities and well-controlled surface properties. Fibronectin (FN) was adsorbed on the different surfaces and quantified in terms of the adsorbed surface density, distribution and conformation. Initial cell adhesion and signaling on FN-coated SAMs were characterized via the formation of focal adhesions, integrin expression and phosphorylation of FAKs. Afterwards, the reorganization and secretion of FN was assessed. Finally, matrix degradation was followed via the expression of matrix metalloproteinases MMP2 and MMP9 and correlated with Runx2 levels. We show that matrix degradation at the cell material interface depends on surface chemistry in MMP-dependent way. Conclusions/Significance: This work provides a broad overview of matrix remodeling at the cell-material interface, establishing correlations between surface chemistry, FN adsorption, cell adhesion and signaling, matrix reorganization and degradation. The reported findings improve our understanding of the role of surface chemistry as a key parameter in the design of new biomaterials. It demonstrates the ability of surface chemistry to direct proteolytic routes at the cell-material interface, which gains a distinct bioengineering interest as a new tool to trigger matrix degradation in different biomedical applications