RESUMEN
In vivo, cells interact with the extracellular matrix (ECM), which provides a multitude of biophysical and biochemical signals that modulate cellular behavior. Inspired by this, we explored a new methodology to develop a more physiomimetic polysaccharide-based matrix for 3D cell culture. Maleimide-modified alginate (AlgM) derivatives were successfully synthesized using DMTMM to activate carboxylic groups. Thiol-terminated cell-adhesion peptides were tethered to the hydrogel network to promote integrin binding. Rapid and efficient in situ hydrogel formation was promoted by thiol-Michael addition "click" chemistry via maleimide reaction with thiol-flanked protease-sensitive peptides. Alginate derivatives were further ionically crosslinked by divalent ions present in the medium, which led to greater stability and allowed longer cell culture periods. By tailoring alginate's biofunctionality we improved cell-cell and cell-matrix interactions, providing an ECM-like 3D microenvironment. We were able to systematically and independently vary biochemical and biophysical parameters to elicit specific cell responses, creating custom-made 3D matrices. DMTMM-mediated maleimide incorporation is a promising approach to synthesizing AlgM derivatives that can be leveraged to produce ECM-like matrices for a broad range of applications, from in vitro tissue modeling to tissue regeneration.
Asunto(s)
Alginatos , Química Clic , Matriz Extracelular , Hidrogeles , Maleimidas , Compuestos de Sulfhidrilo , Humanos , Alginatos/química , Adhesión Celular/efectos de los fármacos , Reactivos de Enlaces Cruzados/química , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Hidrogeles/química , Hidrogeles/síntesis química , Maleimidas/química , Compuestos de Sulfhidrilo/químicaRESUMEN
Campylobacter is the leading cause of foodborne bacterial gastroenteritis in humans worldwide, often associated with the consumption of undercooked poultry. In Jordan, the majority of broiler chicken production occurs in semi-commercial farms, where poor housing conditions and low bio-security are likely to promote campylobacter colonisation. While several studies provided estimates of the key parameters describing the within-flock transmission dynamics of campylobacter in typical high-income countries settings, these data are not available for Jordan and Middle-East in general. A Bayesian model framework was applied to a longitudinal dataset on Campylobacter jejuni infection in a Jordan flock to quantify the transmission rate of C. jejuni in broilers within the farm, the day when the flock first became infected, and the within-flock prevalence (WFP) at clearance. Infection with C. jejuni is most likely to have occurred during the first 8 days of the production cycle, followed by a transmission rate value of 0.13 new infections caused by one infected bird/day (95% CI 0.11-0.17), and a WFP at clearance of 34% (95% CI 0.24-0.47). Our results differ from published studies conducted in intensive poultry production systems in high-income countries but are well aligned with the expectations obtained by means of structured questionnaires submitted to academics with expertise on campylobacter in Jordan. This study provides for the first time the most likely estimates and credible intervals of key epidemiological parameters driving the dynamics of C. jejuni infection in broiler production systems commonly found in Jordan and the Middle-East and could be used to inform Quantitative Microbial Risk Assessment models aimed to assess the risk of human exposure/infection to campylobacter through consumption of poultry meat.