RESUMO
Developing, homeostatic, and regenerating tissues are full of various gradients, including mechanical, chemical, porosity and growth-factor gradients. However, it remains challenging to replicate these gradients using common tissue engineering approaches. Here, we use electrospinning to create scaffolds with in-depth gradients. We created a fiber diameter gradient and pore size gradient throughout the depth of electrospun (ESP) scaffolds by a continuous gradient of polymer concentration. As an alternative to this established method, we developed a novel method to create fiber diameter gradients by changing the voltage on both needle and collector, keeping the total voltage constant. In this way, fiber diameter could be changed in a gradient matter by focusing the electrospinning spot. Using this method, we created a fiber diameter and pore size gradient, while keeping all other parameters constant. Lastly, we developed a novel method to create functional group gradients, which can potentially be used in a wide variety of polymer solutions to couple peptides and proteins to ESP scaffolds. A scaffold with an in-depth gradient of functional groups was created by adding functionalized poly(ethylene glycol) additives to the polymer solution, a novel method with potentially wide applications. The techniques demonstrated here could be applied to a wide variety of polymers and applications and can aid in developing physiologically relevant gradient scaffolds.
Assuntos
Materiais Biocompatíveis/química , Eletroquímica/métodos , Poliésteres/química , Polímeros/química , Proteínas/química , Engenharia Tecidual/métodos , Alicerces Teciduais , Humanos , Teste de Materiais , Polietilenoglicóis/química , Porosidade , RegeneraçãoRESUMO
Both biological and mechanical signals are known to influence cell proliferation. However, biological signals are mostly studied in two-dimensions (2D) and the interplay between these different pathways is largely unstudied. Here, we investigated the influence of the cell culture environment on the response to bFGF, a widely studied and important proliferation growth factor. We observed that human mesenchymal stromal cells (hMSCs), but not fibroblasts, lose the ability to respond to soluble or covalently bound bFGF when cultured on microfibrillar substrates. This behavior correlated with a downregulation of FGF receptor 1 (FGFR1) expression of hMSCs on microfibrillar substrates. Inhibition of actomyosin or the MRTF/SRF pathway decreased FGFR1 expression in hMSCs, fibroblasts and MG63 cells. To our knowledge, this is the first time FGFR1 expression is shown to be regulated through a mechanosensitive pathway in hMSCs. These results add to the sparse literature on FGFR1 regulation and potentially aid designing tissue engineering constructs that better control cell proliferation.