Enhanced potency of cell-based therapy for ischemic tissue repair using an injectable bioactive epitope presenting nanofiber support matrix.

The translation of cell-based therapies for ischemic tissue repair remains limited by several factors, including poor cell survival and limited target site retention. Advances in nanotechnology enable the development of specifically designed delivery matrices to address these limitations and thereby improve the efficacy of cell-based therapies. Given the relevance of integrin signaling for cellular homeostasis, we developed an injectable, bioactive peptide-based nanofiber matrix that presents an integrin-binding epitope derived from fibronectin, and evaluated its feasibility as a supportive artificial matrix for bone marrow-derived pro-angiogenic cells (BMPACs) used as a therapy in ischemic tissue repair. Incubation of BMPACs with these peptide nanofibers in vitro significantly attenuated apoptosis while enhancing proliferation and adhesion. Pro-angiogenic function was enhanced, as cells readily formed tubes. These effects were, in part, mediated via p38, and p44/p42 MAP kinases, which are downstream pathways of focal adhesion kinase. In a murine model of hind limb ischemia, an intramuscular injection of BMPACs within this bioactive peptide nanofiber matrix resulted in greater retention of cells, enhanced capillary density, increased limb perfusion, reduced necrosis/amputation, and preserved function of the ischemic limb compared to treatment with cells alone. This self-assembling, bioactive peptide nanofiber matrix presenting an integrin-binding domain of fibronectin improves regenerative efficacy of cell-based strategies in ischemic tissue by enhancing cell survival, retention, and reparative functions.

Supramolecular nanostructures that mimic VEGF as a strategy for ischemic tissue repair.

There is great demand for the development of novel therapies for ischemic cardiovascular disease, a leading cause of morbidity and mortality worldwide. We report here on the development of a completely synthetic cell-free therapy based on peptide amphiphile nanostructures designed to mimic the activity of VEGF, one of the most potent angiogenic signaling proteins. Following self-assembly of peptide amphiphiles, nanoscale filaments form that display on their surfaces a VEGF-mimetic peptide at high density. The VEGF-mimetic filaments were found to induce phosphorylation of VEGF receptors and promote proangiogenic behavior in endothelial cells, indicated by an enhancement in proliferation, survival, and migration in vitro. In a chicken embryo assay, these nanostructures elicited an angiogenic response in the host vasculature. When evaluated in a mouse hind-limb ischemia model, the nanofibers increased tissue perfusion, functional recovery, limb salvage, and treadmill endurance compared to controls, which included the VEGF-mimetic peptide alone. Immunohistological evidence also demonstrated an increase in the density of microcirculation in the ischemic hind limb, suggesting the mechanism of efficacy of this promising potential therapy is linked to the enhanced microcirculatory angiogenesis that results from treatment with these polyvalent VEGF-mimetic nanofibers.

Human factors affect the quality of cardiopulmonary resuscitation in simulated cardiac arrests.

AIM: Cardiopulmonary resuscitation is a team endeavour. There are only limited data on whether team performance during cardiopulmonary resuscitation is influenced by behavioural issues. The aim of the study was to determine whether and how human factors affect the quality of cardiopulmonary resuscitation. METHODS: 16 teams, each consisting of three health-care workers, were studied in a patient simulator. A scenario of witnessed cardiac arrest due to ventricular fibrillation was used. Ventricular fibrillation could be converted into sinus rhythm by two countershocks administered during the first 2 min or by two countershocks administered during the first 5 min provided that uninterrupted basic life support was started in under 60 s. Teams were rated to be successful if ventricular fibrillation was converted into sinus rhythm. Behavioural rating included leadership, task distribution, information transfer, and conflicts. RESULTS: Only six out of 16 teams were successful. Compared with successful teams, teams that failed exhibited significantly less leadership behaviour (P=0.033) and explicit task distribution (P=0.035). All teams shared among them sufficient theoretical knowledge to successfully treat the simulated cardiac arrest. CONCLUSIONS: In a scenario of simulated witnessed cardiac arrest almost two thirds of teams composed of qualified health-care workers failed to provide basic life support and/or defibrillation within an appropriate time window. Absence of leadership behaviour and absence of explicit task distribution were associated with poor team performance. Failure to translate theoretical knowledge into effective team activity appears to be a major problem.

Sinus augmentation with bovine hydroxyapatite/synthetic peptide in a sodium hyaluronate carrier (PepGen P-15 Putty): a clinical investigation of different healing times.

PURPOSE: Augmentation of the sinus floor with autogenous bone often requires an extra donor site, which is associated with a risk of morbidity, and current grafting protocols involve healing times of up to 9 months. In this prospective in vivo study, the time-dependent efficacy of PepGen P-15 Putty, a combination of bovine hydroxyapatite and synthetic peptide in a sodium hyaluronate carrier, was evaluated in sinus grafting. MATERIALS AND METHODS: Twenty-four edentulous patients received bilateral sinus augmentations with PepGen P-15 Putty, which mimics the cell-binding domain of type I collagen responsible for cell migration, differentiation, and proliferation. The patients were randomly divided into four groups of six patients each, corresponding to 2, 4, 6, and 9 months of healing postaugmentation. After these time intervals, bone biopsy specimens were retrieved through the alveolar bone crest into the augmented sinus, and subsequently oral implants were placed. The specimens were immersed in buffered formalin, scanned with a desktop microcomputed tomography machine, and processed for histologic and histomorphometric evaluation. A mixed model was used for statistical analysis. RESULTS: Three-dimensional microcomputed tomography depicted the distinct structure of trabecular bone encompassing remnants of PepGen, and histologic evaluation revealed osteoblasts and osteoid with osteocytes in the vicinity of the PepGen particles at all healing stages. Histomorphometric results indicated an increase in the newly formed bone fraction in the specimens as follows: 21.3% (± 2.33) at 2 months, 21.9% (± 8.9) at 4 months, 28.5% (± 6.9) at 6 months, and 29.8% (± 11.8) at 9 months. The differences were statistically insignificant. All implants placed in the augmented sites integrated and were restored prosthetically. CONCLUSIONS: PepGen can be used successfully for maxillary sinus augmentation. These data provide evidence that implant placement, even after only 2 months of healing, may be possible.

Sequential expression of adhesion and costimulatory molecules in graft-versus-host disease target organs after murine bone marrow transplantation across minor histocompatibility antigen barriers.

Graft-versus-host disease (GVHD) is a potentially fatal complication after allogeneic bone marrow transplantation. However, few data exist thus far on the molecular signals governing leukocyte trafficking during the disease. We therefore investigated the sequential pattern of distinct adhesion, costimulatory, and apoptosis-related molecules in GVHD organs (ileum, colon, skin, and liver) after transplantation across minor histocompatibility barriers (B10.D2 --> BALB/c, both H-2d). To distinguish changes induced by the conditioning regimen from effects achieved by allogeneic cell transfer, syngeneic transplant recipients (BALB/c --> BALB/c) and irradiated nontransplanted mice were added as controls. Irradiation upregulated the expression of vascular cell adhesion molecule (VCAM)-1, intercellular adhesion molecule (ICAM)-l, and B7-2 in ileum, as well as VCAM-1 and B7-2 in colon, on day 3 in all animals. Whereas in syngeneic mice these effects were reversed from day 9 on, allogeneic recipients showed further upregulation of VCAM-1, ICAM-1, B7-1, and B7-2 in these organs on day 22, when GVHD became clinically evident. Infiltration of CD4+ and CD8+ donor T cells was noted on day 9 in skin and liver and on day 22 in ileum and colon. Surprisingly, the expression of several other adhesion molecules, such as ICAM-2, platelet-endothelial cell adhesion molecule 1, E-selectin, and mucosal addressin cell adhesion molecule 1, did not change. Proapoptotic and antiapoptotic markers were balanced in GVHD organs with the exception of spleen, in which a preferential expression of the proapoptotic Bax could be noted. Our results indicate that irradiation-induced upregulation of VCAM-1, ICAM-1, and B7-2 provides early costimulatory signals to incoming donor T cells in the intestine, followed by a cascade of proinflammatory signals in other organs once the alloresponse is established.