Placental growth factor provides a novel local angiogenic therapy for ischemic cardiomyopathy

BACKGROUND: Heart failure occurs predominantly due to coronary artery disease and may be amenable to novel revascularization therapies. This study evaluated the effects of placental growth factor (PlGF), a potent angiogenic agent, in a rat model of ischemic cardiomyopathy. METHODS: Wistar rats underwent high proximal ligation of the left anterior descending coronary artery and direct injection of PlGF (n = 10) or saline as a control (n = 10) into the myocardium bordering the ischemic area. After 2 weeks, the following parameters were evaluated: ventricular function with an aortic flow probe and a pressure/volume conductance catheter, left ventricular (LV) geometry by histology, and angiogenesis by immunofluorescence. RESULTS: PlGF animals had increased angiogenesis compared to controls (22.8 +/- 3.5 vs. 12.4 +/- 3.2 endothelial cells/high-powered field, p < 0.03). PlGF animals had less ventricular cavity dilation (LV diameter 8.4 +/- 0.2 vs. 9.2 +/- 0.2 mm, p < 0.03) and increased border zone wall thickness (1.85 +/- 0.1 vs. 1.38 +/- 0.2 mm, p < 0.03). PlGF animals had improved cardiac function as measured by maximum LV pressure (95.7 +/- 4 vs. 73.7 +/- 2 mmHg, p = 0.001), maximum dP/dt (4206 +/- 362 vs. 2978 +/- 236 mmHg/sec, p = 0.007), and ejection fraction (25.7 +/- 2 vs. 18.6 +/- 1%, p = 0.02). CONCLUSIONS: Intramyocardial delivery of PlGF following a large myocardial infarction enhanced border zone angiogenesis, attenuated adverse ventricular remodeling, and preserved cardiac function. This therapy may be useful as an adjunct or alternative to standard revascularization techniques in patients with ischemic heart failure.     

The use of adenosine for repair of penetrating cardiac injuries: a novel method

The use of intravenous administration of adenosine to expedite cardiorrhaphy in penetrating cardiac trauma by inducing temporary asystole is described. It is quicker, more effective, and safer than the traditional methods.     

Temporal expression of the chondrogenic and angiogenic growth factor CYR61 during fracture repair.

The repair of a fractured bone is a complex biological event that essentially recapitulates embryonic development and requires the activity of a number of different cell types undergoing proliferation, migration, adhesion, and differentiation, while at the same time expressing a host of different genes. To identify such genes, we employed differential display and compared messenger RNA (mRNA) populations isolated from postfracture (PF) day 5 calluses to those of intact rat femurs. One such gene in which expression was up-regulated at PF day 5 is identified as CYR61, a member of the CCN family of secreted regulatory proteins. CYR61 is a growth factor that stimulates chondrogenesis and angiogenesis. We show that its mRNA expression during fracture repair is regulated temporally, with elevated levels seen as early as PF day 3 and day 5, rising dramatically at PF day 7 and day 10, and finally declining at PF day 14 and day 21. At the highest peak of expression (PF day 7 and day 10, which correlates with chondrogenesis), CYR61 mRNA levels are approximately 10-fold higher than those detected in intact femurs. Similarly, high protein levels are detected throughout the reparative phase of the callus, particularly in fibrous tissue and periosteum, and in proliferating chondrocytes, osteoblasts, and immature osteocytes. The secreted form of CYR61 also was detected within the newly made osteoid. No labeling was detected in hypertrophic chondrocytes or in mature cortical osteocytes. These results suggest that CYR61 plays a significant role in cartilage and bone formation and may serve as an important regulator of fracture healing.     

Preoperative high leukocyte count: a novel risk factor for stroke after cardiac surgery

BACKGROUND: Stroke after cardiac surgery is a devastating complication. The relationship between white blood cell count (WBC) and perioperative cerebrovascular accident (CVA) has not been investigated. An effort was made to identify how preoperative WBC may relate to CVA development during or after cardiac surgery. METHODS: Prospective data were collected from 7,483 patients who underwent coronary artery bypass grafting or valvular surgery or both. WBC was determined preoperatively and postoperatively. Differentiation of WBC was examined only preoperatively. RESULTS: There were a total of 125 CVAs (10 transient ischemic attacks [TIAs], 115 strokes). WBC was significantly higher preoperatively and directly postoperatively in patients with stroke. Qualitative changes in preoperative WBC were also found in these patients (chi2; p < 0.001). The predictive power of the stepwise logistic regression model for CVA was greater when preoperative WBC was included. The risk for perioperative CVA increased starting at preoperative WBC of 9 x 10(9)/L (p = 0.044) and progressed in higher WBC ranges. WBC had a significant impact on CVA outcome (analysis of variance, p = 0.001). CONCLUSIONS: Our studies have established the correlation between high preoperative WBC and stroke during or after cardiac surgery. Furthermore, elevated preoperative WBC was related to the clinical outcome of CVA. Preoperative measures aimed at preventing or treating conditions such as infections that may cause elevated WBC may be beneficial in the prevention of stroke during or after cardiac surgery.     

Inhibition of the cardiac angiogenic response to exogenous vascular endothelial growth factor

BACKGROUND: The angiogenic effects of vascular endothelial growth factor (VEGF) are mediated by the stimulation of endothelial nitric oxide synthase (eNOS) and nitric oxide release. Nitric oxide availability is decreased in patients with coronary disease, a possible explanation for the humble results of VEGF in clinical trials. We sought to examine the effects of exogenous VEGF in a model of endothelial dysfunction. METHODS: Miniswine fed either a regular (N = 6, group NORM) or hypercholesterolemic diet (N = 6, HICHOL) underwent ameroid placement on the circumflex artery. Three weeks later, baseline myocardial perfusion was assessed by microsphere injections, and all pigs were treated with VEGF. Four weeks later, microsphere injections were repeated and the hearts harvested. Endothelial-dependent coronary microvascular reactivity, and VEGF and eNOS expression were assessed. RESULTS: HICHOL pigs showed significant endothelial dysfunction in the ischemic territory. Post-treatment myocardial blood flow in the circumflex territory was significantly higher in the NORM compared to the HICHOL group. VEGF and eNOS levels were increased in the ischemic territory in the NORM group but decreased in the HICHOL group. CONCLUSIONS: The cardiac angiogenic response to VEGF was markedly inhibited in a hypercholesterolemia-induced porcine model of endothelial dysfunction. Coronary endothelial dysfunction could be an obstacle to the efficacy of clinical angiogenesis protocols and a putative therapeutic target.     

Basic fibroblastic growth factor as a potential meningeal angiogenic factor

Vascular supply plays a significant role in the management of skull base tumors. The diagnosis is aided by contrast-enhanced imaging and angiographic techniques, and embolization procedures are used to devascularize certain lesions. The degree of surgical technical difficulty is strongly influenced by the degree of tumor vascularity. Although the importance of this blood supply is clearly understood, the mechanism involved in developing a system of tumor-perfusing vessels is yet to be defined. The development of a vascular network, or angiogenesis, is an important event in allowing tumor proliferation to progress beyond small clusters of cells. Basic fibroblastic growth factor (bFGF) is an especially attractive candidate as an angiogenic growth factor because of its ability to stimulate processes that are characteristic of angiogenesis in vitro. Tumors that involve the meninges may have the ability to liberate normally stored bFGF, which may, in turn, induce new vessel formation for continued tumor proliferation. An immunohistochemical analysis of rodent and bovine meninges to study this phenomenon is described. The dura, arachnoid, and their associated vessels are shown clearly to contain this growth factor. Ultimately, an adjuvant therapy based on the inhibition of angiogenesis may provide a reasonable alternative to aggressive surgical approaches in skull base tumors that are incompletely resectable.     

Tissue-engineered cardiac constructs for cardiac repair

Several recent basic research studies have described surgical methods for cardiac repair using tissue cardiomyoplasty. This review summarizes recent advances in cardiac repair using bioengineered tissue from the viewpoint of the cardiac surgeon. We conclude that the results of many basic and preclinical studies indicate that bioengineered tissue can be adapted to conventional surgical techniques. However, no clinical studies have yet proved bioengineered tissue is effective as a treatment for human heart failure. Today's cardiac surgeons can look forward to the advent of new techniques to benefit patients who respond poorly to existing treatment for heart failure.     

Comparison of developmental endothelial locus-1 angiogenic factor with vascular endothelial growth factor in a porcine model of cardiac ischemia

BACKGROUND: This study compared the angiogenic effects of developmental endothelial locus-1 (DEL-1), vascular endothelial growth factor (VEGF), as well as the negative control, beta-galactosidase (beta-gal), in a porcine model of cardiac ischemia. METHODS: Twenty pigs underwent left circumflex artery occlusions. After 3 weeks, the animals received myocardial injections of adenovirus expressing beta-gal (n=6), DEL-1 (n=7), or VEGF (n=7). At 7 weeks, animals were assessed for both function and coronary flow and compared with baseline measurements. RESULTS: Regional wall motion index and global ejection fraction showed deterioration in function in the beta-gal group and no change in the VEGF and DEL-1 groups between the treatment and harvest time points. Preload recruitable stroke work suggested functional improvement in the VEGF group (35.8 +/- 8.6 vs 56.4 +/- 17.8, p = 0.033). The increase in the DEL-1 group was not statistically significant (27.3 +/- 9.8 vs, 40.2 +/- 19.4, p = 0.067). The beta-gal group exhibited minimal change (30.7 +/- 14.8 vs 35.9 +/- 12.1, p = 0.96). Regional blood flow as assessed by fluorescent microspheres was improved under stress conditions in the VEGF group (1.00 +/- 0.15 vs 1.15 +/-0.22, p = 0.03). CONCLUSIONS: Treatment with VEGF led to a modest improvement in regional blood flow and cardiac function in previously ischemic myocardial tissue.     

Interleukin-17, a regulator of angiogenic factor release by synovial fibroblasts

OBJECTIVE: Angiogenesis is a process stimulated in inflamed synovium of patients with osteoarthritis (OA), and contributes to the progression of the disease. Synovial fibroblasts secrete angiogenic factors, such as vascular endothelial growth factor (VEGF), an up-regulator of angiogenesis, and this ability is increased by interleukin (IL)-1beta. The purpose of this study was to verify whether IL-17 contributes and/or synergizes with IL-1beta and tumor necrosis factor (TNF)-alpha in vessel development in articular tissues by stimulating the secretion of proangiogenic factors by synovial fibroblasts. DESIGN: We stimulated in vitro synovial fibroblasts isolated from OA, rheumatoid arthritis (RA) and fractured patients (FP) with IL-17 and IL-1beta and from OA patients with IL-17, IL-1beta and TNF-alpha. In the supernatants from the cultures, we assayed the amount of VEGF by immunoassay and other angiogenic factors (keratinocyte growth factor, KGF; hepatocyte growth factor, HGF; heparin-binding endothelial growth factor, HB-EGF; angiopoietin-2, Ang-2; platelet-derived growth factor B, PDGF-BB; thrombopoietin, TPO) by chemiluminescence; semiquantitative RT-PCR was used to state mRNA expression of nonreleased angiogenic factors (Ang-2 and PDGF-BB) and tissue inhibitors of metalloproteinase (TIMP)-1. RESULTS: IL-17, TNF-alpha and IL-1beta increased VEGF secretion by synovial fibroblasts from OA patients. IL-17 and IL-1beta also increased VEGF secretion in RA and FP. Besides, IL-17 increased KGF and HGF secretions in OA, RA and FP; in OA and RA, IL-17 also increased the HB-EGF secretion and the expression of TIMP-1 as protein and mRNA. In OA patients IL-17 had an additive effect on TNF-alpha-stimulated VEGF secretion. CONCLUSIONS: These results suggest that IL-17 is an in vitro stimulator of angiogenic factor release, both by its own action and by cooperating with TNF-alpha.     

ADAM8: a novel osteoclast stimulating factor.

We used polymerase chain reaction (PCR)-selective complementary DNA (cDNA) subtraction hybridization with an immortalized murine osteoclast (OCL) precursor cell line to identify genes that are highly expressed in OCLs compared with OCL precursors and which may be involved in the OCL differentiation process. ADAM8 was one of the 50 genes identified. ADAM (a disintegrin and metalloproteinase) peptides are membrane-bound proteins that can act as cell-to-cell and cell-to-matrix adhesion molecules, degrade the extracellular matrix, and play a role in tissue morphogenesis. Addition of antisense (AS) S-oligonucleotides for ADAM8 (1-10 nM) to mouse bone marrow cultures treated with 10(-9) M 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] significantly inhibited OCL formation compared with treatment with the control S-oligonucleotide. Furthermore, conditioned media from 293 cells transiently transfected with a secretable form of the ADAM8 cDNA increased OCL formation in a dose-dependent manner. In addition, treatment of OCLs with soluble ADAM8 conditioned media significantly increased pit formation per dentin slice compared with control OCLs. Time course studies indicated that ADAM8 increased OCL formation only when it was present during days 4-7 of the 7-day culture period. Structural analysis, using truncated constructs of ADAM8, showed that the cysteine-rich/disintegrin domain was responsible for its OCL stimulatory activity. Western blot analysis confirmed that the soluble form of ADAM8 is present in normal marrow cultures. These data suggest that ADAM8 plays an important role in OCL formation and acts primarily at the later stages of OCL differentiation.     

Evaluation of a novel biomaterial for intrasubstance muscle laceration repair.

The authors compare the effects of small intestinal submucosa (SIS) treatment to suture repair with respect to histologic and functional outcomes for complete muscle lacerations in a rabbit model. The authors hypothesized that SIS treatment of full-thickness muscle belly lacerations would significantly improve muscle function, strength, and regeneration compared to the current standard-of-care treatment. Muscle belly lacerations were created in the extensor digitorum longus (EDL) of both hind limbs of each rabbit. After randomization, lacerations were left unrepaired (n = 48) or repaired using a 4-0 Prolene modified Kessler stitch (n = 48). A flap of SIS graft was sutured into half (n = 24 each) of the repaired and unrepaired muscles forming four study groups. Suture repair with SIS augmentation of complete muscle lacerations resulted in healed tissue that most closely resembled normal muscle in terms of morphology and function when compared to current standard-of-care treatments. Active force production in this group reached 79% of uninjured controls 12 weeks after surgery. SIS may have important clinical advantages over suture repair alone and warrants further clinical study.     

Transforming growth factor type e is a novel mediator of wound repair

Transforming growth factor type e is a potent mitogen for epithelial and fibroblastic cells in vitro. It is primarily produced by epithelial tissues and cells, including normal skin keratinocytes. In this study we examined the effects of transforming growth factor type e in vivo and in vitro. On application of transforming growth factor type e to the chorioallantoic membrane of 8-day-old chick embryos, we noted a prominent thickening of the membrane caused primarily by fibroblastic and epithelial proliferation. In a subsequent study, transforming growth factor type e suspended in Matrigel was injected subcutaneously into the abdominal wall of Swiss National Institutes of Health mice. Significant influx of first neutrophils and then macrophages into the gel of transforming growth factor type e-treated mice, but not of control mice, was observed. Transforming growth factor type e also exhibited a potent chemoattractive effect on the J774 macrophage cell line in vitro. In transcutaneous mouse wounds, topical administration of transforming growth factor type e led first to an influx of neutrophils and macrophages, followed by an increased formation of granulation tissue in the transforming growth factor type e-treated wounds. These results suggest that transforming growth factor type e plays an important role in wound healing as a chemoattractant for neutrophils and macrophages and as a mitogen for epithelial and fibroblastic cells.