Transplantation of mouse embryonic stem cells into the cochlea of an auditory-neuropathy animal model: effects of timing after injury.

Application of ouabain to the round window membrane of the gerbil selectively induces the death of most spiral ganglion neurons and thus provides an excellent model for investigating the survival and differentiation of embryonic stem cells (ESCs) introduced into the inner ear. In this study, mouse ESCs were pretreated with a neural-induction protocol and transplanted into Rosenthal's canal (RC), perilymph, or endolymph of Mongolian gerbils either 1-3 days (early post-injury transplant group) or 7 days or longer (late post-injury transplant group) after ouabain injury. Overall, ESC survival in RC and perilymphatic spaces was significantly greater in the early post-injury microenvironment as compared to the later post-injury condition. Viable clusters of ESCs within RC and perilymphatic spaces appeared to be associated with neovascularization in the early post-injury group. A small number of ESCs transplanted within RC stained for mature neuronal or glial cell markers. ESCs introduced into perilymph survived in several locations, but most differentiated into glia-like cells. ESCs transplanted into endolymph survived poorly if at all. These experiments demonstrate that there is an optimal time window for engraftment and survival of ESCs that occurs in the early post-injury period.

The acellular myocardial flap: a novel extracellular matrix scaffold enriched with patent microvascular networks and biocompatible cell niches.

There is a great need for acellular, fully vascularized, and biocompatible myocardial scaffolds that provide agreeable biological, nutritional, and biomechanical niches for reseeded cells for in vitro and in vivo applications. We generated myocardial flap scaffolds comprising porcine left-anterior ventricular myocardium and its associated coronary arteries and veins and investigated the combinatorial effects of sodium dodecyl sulfate (SDS) and sodium hydroxide (NaOH) perfusion on both the myocardial extracellular matrix (ECM) and the vascular ECM. Results showed that all scaffolds displayed a fully intact and patent vasculature, with arterial burst pressures indistinguishable from native coronary arteries and perfusion to the level of capillaries. Scaffolds were free of cellular proteins and retained collagen and elastin ECM components, exhibited excellent mechanical properties, and were cytocompatible toward relevant seeded cells. SDS perfusion preserved collagen IV, laminin, and fibronectin well, but only reduced DNA content by 33%; however, this was further improved by post-SDS nuclease treatments. By comparison, NaOH was very effective in removing cells and eliminated more than 95% of tissue DNA, but also significantly reduced levels of laminin and fibronectin. Such constructs can be readily trimmed to match the size of the infarct and might be able to functionally integrate within host myocardium and be nourished by direct anastomotic connection with the host's own vasculature; they might also be useful as physiologically accurate models for in vitro studies of cardiac physiology and pathology.

Inflammation in cardiovascular tissue engineering: the challenge to a promise: a minireview.

Tissue engineering employs scaffolds, cells, and stimuli brought together in such a way as to mimic the functional architecture of the target tissue or organ. Exhilarating advances in tissue engineering and regenerative medicine allow us to envision in vitro creation or in vivo regeneration of cardiovascular tissues. Such accomplishments have the potential to revolutionize medicine and greatly improve our standard of life. However, enthusiasm has been hampered in recent years because of abnormal reactions at the implant-host interface, including cell proliferation, fibrosis, calcification and degeneration, as compared to the highly desired healing and remodeling. Animal and clinical studies have highlighted uncontrolled chronic inflammation as the main cause of these processes. In this minireview, we present three case studies highlighting the importance of inflammation in tissue engineering heart valves, vascular grafts, and myocardium and propose to focus on the endothelial barrier, the "final frontier" endowed with the natural potential and ability to regulate inflammatory signals.