Endothelialization of chitosan porous conduits via immobilization of a recombinant fibronectin fragment (rhFNIII7–10)

The present study aimed to develop a pre-endothelialized chitosan (CH) porous hollowed scaffold for application in spinal cord regenerative therapies. CH conduits with different degrees of acetylation (DA; 4% and 15%) were prepared, characterized (microstructure, porosity and water uptake) and functionalized with a recombinant fragment of human fibronectin (rhFNIII7–10). Immobilized rhFNIII7–10 was characterized in terms of amount (125I-radiolabelling), exposure of cell-binding domains (immunofluorescence) and ability to mediate endothelial cell (EC) adhesion and cytoskeletal rearrangement. Functionalized conduits revealed a linear increase in immobilized rhFNIII7–10 with rhFNIII7–10 concentration, and, for the same concentration, higher amounts of rhFNIII7–10 on DA 4% compared with DA 15%. Moreover, rhFNIII7–10 concentrations as low as 5 and 20 lgml 1 in the coupling reaction were shown to provide DA 4% and 15% scaffolds, respectively, with levels of exposed cell-binding domains exceeding those observed on the control (DA 4% scaffolds incubated in a 20 lgml 1 human fibronectin solution). These grafting conditions proved to be effective in mediating EC adhesion/cytoskeletal organization on CH with DA 4% and 15%, without affecting the endothelial angiogenic potential. rhFNIII7–10 grafting to CH could be a strategy of particular interest in tissue engineering applications requiring the use of endothelialized porous matrices with tunable degradation rates.

Habitação modular evolutiva

Da necessidade de criação de habitações que se adaptem ao constante dinamismo do ser humano, surgiu a ideia da concepção de um edifício evolutivo com base na construção modular. A construção modular poderá ser entendida como sendo a principal resposta ao combate da construção de estruturas físicas e imóveis, com características inalteráveis ao longo do tempo. Com este trabalho idealizou-se uma habitação evolutiva, baseado nos conceitos da Coordenação Modular, identificando-se as suas especificações, exigências de desempenho e características funcionais. Foi considerada como medida modular base 0,10m. Os módulos foram pensados com o intuito de que o seu desempenho prático seja satisfatório. A sua base estrutural foi executada a partir do rearranjo de contentores marítimos. Previu-se ainda, o uso de coberturas verdes por serem uma solução comprovada de minorar notoriamente o efeito pernicioso que a construção tem sobre o meio ambiente, ao mesmo tempo que apresentam vantagens térmicas e acústicas. A questão do incentivo do uso da pré-fabricação em Portugal a uma maior escala, não poderia ser descurada. A apresentação de eventuais desenvolvimentos futuros é também relevante, dada a importância que é inerente ao âmbito da industrialização da construção. Do estudo realizado ficaram ainda, bem patentes, os benefícios da racionalização e da industrialização, tanto em termos económicos como em termos ambientais. É possível executar um projecto com níveis de perdas mais baixos e com índices de qualidade superiores a um menor preço.

Acetyl-L-Carnitine Prevents Methamphetamine-Induced Structural Damage on Endothelial Cells via ILK-Related MMP-9 Activity

Methamphetamine (METH) is a potent psychostimulant highly used worldwide. Recent studies evidenced the involvement of METH in the breakdown of the blood-brain-barrier (BBB) integrity leading to compromised function. The involvement of the matrix metalloproteinases (MMPs) in the degradation of the neurovascular matrix components and tight junctions (TJs) is one of the most recent findings in METH-induced toxicity. As BBB dysfunction is a pathological feature of many neurological conditions, unveiling new protective agents in this field is of major relevance. AcetylL-carnitine (ALC) has been described to protect the BBB function in different paradigms, but the mechanisms underling its action remain mostly unknown. Here, the immortalized bEnd.3 cell line was used to evaluate the neuroprotective features of ALC in METH-induced damage. Cells were exposed to ranging concentrations of METH, and the protective effect of ALC 1 mM was assessed 24 h after treatment. F-actin rearrangement, TJ expression and distribution, and MMPs activity were evaluated. Integrin-linked kinase (ILK) knockdown cells were used to assess role of ALC in ILK mediated METHtriggered MMPs’ activity. Our results show that METH led to disruption of the actin filaments concomitant with claudin-5 translocation to the cytoplasm. These events were mediated by MMP-9 activation in association with ILK overexpression. Pretreatment with ALC prevented METH-induced activation of MMP-9, preserving claudin-5 location and the structural arrangement of the actin filaments. The present results support the potential of ALC in preserving BBB integrity, highlighting ILK as a new target for the ALC therapeutic use.