Electrospun fiber - Hydrogel composites for nucleus pulposus tissue engineering

New materials are needed to replace degenerated intervertebral disc tissue and to provide longer-term solutions for chronic back-pain. Replacement tissue potentially could be engineered by seeding cells into a scaffold that mimics the architecture of natural tissue. Many natural tissues, including the nucleus pulposus (the central region of the intervertebral disc) consist of collagen nanofibers embedded in a gel-like matrix. Recently it was shown that electrospun micro- or nano-fiber structures of considerable thickness can be produced by collecting fibers in an ethanol bath. Here, randomly aligned polycaprolactone electrospun fiber structures up to 50 mm thick are backfilled with alginate hydrogels to form novel composite materials that mimic the fiber-reinforced structure of the nucleus pulposus. The composites are characterized using both indentation and tensile testing. The composites are mechanically robust, exhibiting substantial strain-to-failure. The method presented here provides a way to create large biomimetic scaffolds that more closely mimic the composite structure of natural tissue. © 2012 Materials Research Society.

Development of an ex vivo organ culture model using human gastro-intestinal tissue and Campylobacter jejuni.

Campylobacter jejuni is an important food-borne pathogen. However, relatively little is understood regarding its pathogenesis, and research is hampered by the lack of a suitable model. Recently, a number of groups have developed assays to study the pathogenic mechanisms of C. jejuni using cell culture models. Here, we report the development of an ex vivo organ culture model, allowing for the maintenance of intestinal mucosal tissue, to permit more complex host-bacterium interactions to be studied. Ex vivo organ culture highlights the propensity for C. jejuni to adhere to mucosal tissue via the flagellum, either as discrete colonies or as multicellular units.

Microfabricated tissue gauges to measure and manipulate forces from 3D microtissues

Physical forces generated by cells drive morphologic changes during development and can feedback to regulate cellular phenotypes. Because these phenomena typically occur within a 3-dimensional (3D) matrix in vivo, we used microelectromechanical systems (MEMS) technology to generate arrays of microtissues consisting of cells encapsulated within 3D micropatterned matrices. Microcantilevers were used to simultaneously constrain the remodeling of a collagen gel and to report forces generated during this process. By concurrently measuring forces and observing matrix remodeling at cellular length scales, we report an initial correlation and later decoupling between cellular contractile forces and changes in tissue morphology. Independently varying the mechanical stiffness of the cantilevers and collagen matrix revealed that cellular forces increased with boundary or matrix rigidity whereas levels of cytoskeletal and extracellular matrix (ECM) proteins correlated with levels of mechanical stress. By mapping these relationships between cellular and matrix mechanics, cellular forces, and protein expression onto a bio-chemo-mechanical model of microtissue contractility, we demonstrate how intratissue gradients of mechanical stress can emerge from collective cellular contractility and finally, how such gradients can be used to engineer protein composition and organization within a 3D tissue. Together, these findings highlight a complex and dynamic relationship between cellular forces, ECM remodeling, and cellular phenotype and describe a system to study and apply this relationship within engineered 3D microtissues.