Angiogenesis, or the formation of new blood vessels from the preexisting vasculature, is a key component in numerous physiologic and pathologic responses and has broad impact in many medical and surgical specialties. In this review, we discuss the key cellular steps which lead to the neovascularization of tissues, and highlight the main molecular mechanisms and mediators in this process. We include discussions on proteolytic enzymes, cell/matrix interactions, pertinent cell signaling pathways, and end with a survey of the mechanisms which lead to the stabilization and maturation of neovasculatures.

Blood stream infection (BSI) is a serious complication of hematopoietic stem cell transplantation (HSCT). The aim of this retrospective cohort analysis was to describe BSI after HSCT, and to assess the predictors and outcomes of BSI after HSCT using multivariable modeling. Of the 243 subjects transplanted, 56% received allogeneic HSCT and 106 (43.6%) developed BSI. Of the 185 isolates, 68% were Gram-positive cocci, 21% were Gram-negative bacilli (GNR) and 11% were fungi. Type of allogeneic HSCT was an independent risk factor for BSI (hazard ratio (HR) 3.26, 95% confidence interval (CI) 1.50, 7.07, P ¼ 0.01), as was the degree of HLA matching (HR 1.84, 95% CI 1.00, 3.37, P ¼ 0.05). BSI was a significant independent predictor of mortality after HSCT (HR 1.79, 95% CI 1.18, 2.73, P ¼ 0.007), after adjusting for acute graft-versus-host disease (GVHD) and allogeneic HSCT (both predicting death p3 months after HSCT). In contrast to the effects of acute GVHD and allogeneic HSCT, the effect of BSI was evident throughout the post-HSCT period. GNR BSI and vancomycin-resistant enterococcal BSI also were significantly associated with death. We concluded that BSI is a common complication of HSCT associated with increased mortality throughout the post-HSCT period.

Neovascularization can be categorized into two general processes: vasculogenesis and angiogenesis. Angiogenesis is the formation of new capillaries from pre-existing vessels, requiring growth factor driven recruitment, migration, proliferation, and differentiation of endothelial cells (ECs). Complex cell-cell and cell-extracellular matrix (ECM) interactions contribute to this process, leading finally to a network of tube-like formations of endothelial cells supported by surrounding mural cells. The study of angiogenesis has broad clinical implications in the fields of peripheral and coronary vascular disease, oncology, hematology, wound healing, dermatology, and ophthalmology, among others. As such, novel, clinically relevant models of angiogenesis in vitro are crucial to the understanding of angiogenic processes. We highlight some of the advances made in the development of these models, and discuss the importance of incorporating the three-dimensional cell-matrix and EC-mural cell interactions into these in vitro assays of angiogenesis. This review also discusses our own 3-D angiogenesis assay and some of the in vitro results from our lab as they relate to therapeutic neovascularization and tissue engineering of vascular grafts.

Cardiovascular disease continues to be the leading cause of death worldwide, and the prevalence of cardiovascular disease has reached epidemic proportions worldwide. Not surprisingly this has led to an increasing number of vascular procedures annually. Unfortunately, the success of these procedures over time continues to limit their long-term effects. Biomedical engineering approaches to improve upon current prosthetic grafts, developing new prosthetic grafts, and creating tissue engineered blood vessels for clinical application offer hope of improving the durability of vascular interventions and improving patients' treatment for cardiovascular disease.

Smooth muscle cells (SMCs) and collagen scaffolds are widely used in vascular tissue engineering but their interactions in remodeling at the microscale level remained unclear. We characterized microscale morphologic alterations of collagen remodeled by SMCs in six dimensions: three spatial, time, multi-channel and multi-position dimensions. In live imaging assays, computer-assisted cell tracking showed locomotion characteristics of SMCs; reflection and fluorescent confocal microscopy and spatial reconstruction images of each time point showed detailed morphologic changes of collagen fibers and spatial collagen-SMC interactions. The density of the collagen around the SMCs was changed dynamically by the leading edges of the cells. The density of the collagen following 24 h of cell-induced remodeling increased 51.61 ± 9.73% compared to unremodeled collagen containing cells for 1 h (P < 0.0001, n = 40) (NS vs. collagen without cells). Fast Fourier transform analysis showed that the collagen fibers' orientation changed from random (alignment index = 0.047 ± 0.029, n = 40) after 1 h into concordant with that of the SMCs (alignment index = 0.379 ± 0.098, P < 0.0001, n = 40) after 24 h. Mosaic imaging extended the visual field from a single cell to a group of cells in one image without loss of optical resolution. Direct visualization of alignment of actin fibers and collagen fibers showed the molecular machinery of the process of scaffold remodeling. This is a new approach to better understanding the mechanism of scaffold remodeling and our techniques represent effective tools to investigate the interactions between cells and scaffold in detail at the microscale level.

The delivery of growth factors to cellularize biocompatible scaffolds like fibrin is a commonly used strategy in tissue engineering. We characterized SMC proliferation and chemotaxis in response to PDGF-BB and FGF-2, alone and in combination, in 2-D culture and in 3-D fibrin hydrogels. While both growth factors induced an equipotent mitogenic response in 2-D culture, only FGF-2 was significantly mitogenic for SMCs in 3-D culture. Only PDGF-BB was significantly chemotactic in a modified Boyden chamber assay. In a 3-D assay of matrix invasion, both growth factors induced an invasive response into the fibrin hydrogel in both proliferating and non-proliferating, mitomycin C (MMC) treated cells. The invasive response was less attenuated by the inhibition of proliferation in PDGF-BB stimulated cells compared with FGF-2 stimulated cells. We conclude that SMCs cultured in fibrin hydrogels have a more robust chemotactic response to PDGF-BB compared with FGF-2, and that the response to FGF-2 is more dependent on cell proliferation. Delivery of both growth factors together potentiates the chemotactic, but not mitogenic response to either growth factor alone.

We investigated the delivery of R136K-CBD (a collagen binding mutuat chimera of fibroblast growth factor-1) with a type I collagen scaffold as the delivery vehicle to smooth muscle cells (SMCs) for vascular tissue engineering. The binding affinity of R136K-CBD to 3-D collagen scaffolds was investigated both in the presence and absence of cells and/or salts. 2-D and 3-D visualization of delivery of R136K-CBD into SMCs was accomplished by combined fluorescent and reflection confocal microscopy. The mitogenic effect of collagen-immobilized R136K-CBD on SMCs in 3-D collagen was studied by Cyquant assay at different time intervals. In the group devoid of salt and cells, no detectable release of R136K-CBD into overlying culture media was found, compared with burst-and-continuous release of R136K and FGF-1 over a 14-day period in all other groups. The release rate of R136K-CBD was 1.7 and 1.6 fold less than R-136K and FGF-1 when media was supplemented with 2M salt (P<0.0001), and 2.6 and 2.5 fold less in cell-populated collagen hydrogels (P<0.0001), respectively. R136K-CBD showed essentially uniform binding to collagen and its distribution was dependent on that of the collagen scaffold. Internalization of R136K-CBD into SMCs was documented by confocal microscopy. 3-D local delivery of collagen-immobilized R136K-CBD increased the proliferation of SMCs in the collagen matrix to significantly greater levels and for a significantly greater duration than R136K or FGF-1, with 2.0 and 2.1 fold more mitogenicity than R136K and FGF-1 respectively (P<0.0001) at day 7. The results suggest that our collagen binding fusion protein is an effective strategy for growth factor delivery for vascular tissue engineering.

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