A new fasciocutaneous flap model identifies a critical role for endothelial Notch signaling in wound healing and flap survival.

Flap surgery is a common treatment for severe wounds and a major determinant of surgical outcome. Flap survival and healing depends on adaptation of the local flap vasculature. Using a novel and defined model of fasciocutaneous flap surgery, we demonstrate that the Notch ligand Delta-like 1 (Dll1), expressed in vascular endothelial cells, regulates flap arteriogenesis, inflammation and flap survival. Utilizing the stereotyped anatomy of dorsal skin arteries, ligation of the major vascular pedicle induced strong collateral vessel development by end-to-end anastomosis in wildtype mice, which supported flap perfusion recovery over time. In mice with heterozygous deletion of Dll1, collateral vessel formation was strongly impaired, resulting in aberrant vascularization and subsequent necrosis of the tissue. Furthermore, Dll1 deficient mice showed severe inflammation in the flap dominated by monocytes and macrophages. This process is controlled by endothelial Dll1 in vivo, since the results were recapitulated in mice with endothelial-specific deletion of Dll1. Thus, our model provides a platform to study vascular adaptation to flap surgery and molecular and cellular regulators influencing flap healing and survival.

Platelet-derived concentrates influence human keratinocyte proliferation in vitro and induce wound healing in a prospective case series of chronic wounds of different entities in vivo.

Objectives: Soft tissues defects can extend into the fat layer or even deeper and can cause significant clinical disadvantages like pain, infections, and loss of function. In particular, chronic wounds are difficult to treat, as split-thickness skin grafts (STSGs) have varying success rates. To improve wound healing in chronic wounds, the authors have studied the application of platelet-mediator concentrate (PMC) in a human keratinocyte culture model in vitro and of autologous platelet concentrates (PRP) in a combination with surgical procedures in vivo as second line therapy in patients with initially failed wound closure. Methods: For in vitro testing on keratinocytes, a PMC was processed with a commercially available bedside system (ATR®, Curasan, Germany). In a clinical, nonrandomized study, five in-house patients with chronic wounds were treated using a combination of surgical debridement and autologous PRP. Time of healing as determined by epithelization as well as laser Doppler imaging to visualize blood flow was analyzed. Additionally, changes in ease of surgical wound closure were determined. Finally, the quality of life of patients was assessed using a validated questionnaire (clinicaltrials.gov # NCT03667638). Results: In vitro testing shows a significant effect of PMC on keratinocyte proliferation in cell culture. Clinical studies showed that patients treated with PRP had initiation of wound closure, higher blood flow after PRP injection, and easier wound closure as well as improved quality of life. Conclusions: The injection of platelet concentrates to treat chronic wound defects presents a favorable addition to treatment where single surgical procedures have failed and may improve current therapy options.

How to become a medical professor - a comparative analysis of academic requirements in Germany and the United States.

BACKGROUND: The acquisition of a medical professorship represents a significant step in a physician's academic career. The responsibility as well as the honor and the associated obligations are significant; however, the requirements to become a medical professor vary in Germany. OBJECTIVE: We analyzed the variable requirements for prospective medical professors in Germany, with special focus on the tenure track concept and the U.S. system. METHODS: Based on an online research, we queried German medical faculty regulations to obtain a medical professorship within Germany. RESULTS: We analyzed 35 German universities. On average, 11 publications are required after "venia legendi" to meet professorship (apl) prerequisites (median x̅ = 10, max = 24, min = 6, n = 16), whereas 6 publications with first or last authorship are required on average (x̅ = 6, max = 16, min = 4, n = 26). In most German universities, it takes an average of 4 years after gaining habilitation to apply for a professorship (x̅ = 5 years, max = 6 years, min = 2 years). Candidates for university chair positions, however, can shorten this period by an average of 38%. DISCUSSION: In the German academic system, the prerequisites to gain a professorship differ among universities. Due to different scientific cooperation and exchange programs, research and academic activities have reached an intense international exchange level. Yet there is no international or even national standardization, quality assurance, and comparability to gain a medical professorship.

[A specialized university outpatient clinic of the department of Plastic and Aesthetic Surgery]. / Spezialisierte Hochschulambulanz mit Querschnittscharakter: Die Poliklinik für Plastische und Ästhetische Chirurgie.

University outpatient clinics are essential for medical research, teaching and treatment of complex and special diseases based on current knowledge and guidelines. Furthermore new therapeutic approaches are developed for complicated and rare diseases. The diversity of disease entities and treatment decisions provided by a specialized university outpatient clinic for plastic and aesthetic surgery is so far unknown and subject to our research. The study results highlight the value of the specialized university outpatient clinic for plastic and aesthetic surgery for medical training purposes, research and advanced health care. METHOD: In a retrospective study, outpatients consulting the clinic of the Department of Plastic and Aesthetic Surgery of Hannover Medical School university hospital between 2013 and 2014 were analyzed. The epidemiology and distribution of disease entities previously treated in the different medical specialties such as surgery, internal medicine, dermatology, neurology and psychiatry, dentistry and ophthalmology were investigated. RESULTS: During a two-year period (2013/2014) 9272 patients with 821 different ICD-coded diagnoses were seen. 57 % of the primary and secondary diagnoses were related to surgery and 36 % to internal medicine. 7 % of the diagnoses were derived from other clinical specialties such as dermatology, neurology, psychiatry, dentistry and ophthalmology. Geriatric surgery comprised 22 %, post oncological 7 %, postoperative complication management 6 %, sequelae of conservative (non-surgical) treatment 3 % and congenital malformations 3 %. In 29 % of patients, surgical treatment was recommended. CONCLUSION: The analysis shows that patients with complex and a wide range of disease entities presented at our outpatient clinic of plastic- and aesthetic surgery. The variety of diagnoses requires not only extensive specialized expertise, but also broad medical knowledge in order to reach adequate treatment strategies. The setting of complex as well as a broad spectrum of diseases of the university outpatient clinic is thus particularly suitable for teaching of differential diagnoses, diagnostic algorithms and development of treatment strategies during medical training and student education.

Retinal myeloid cells regulate tip cell selection and vascular branching morphogenesis via Notch ligand Delta-like 1.

During angiogenesis, single endothelial cells (EC) specialize into tip cells that guide vessel sprouting towards growth factor gradients and instruct the adjacent vessel stalk. The balance between tip and stalk cells is regulated by endothelial Notch signalling through the expression of Notch ligand Delta-like 4 (Dll4) in tip cells, which suppresses a tip cell fate in adjacent stalk cells. Here we show, using genetic reporter and conditional deletion strategies, that myeloid cells regulate tip cell numbers and Dll4 expression via the Notch ligand Dll1 during vascular development in the retina. Dll1 is selectively expressed by a subpopulation of retinal myeloid cells, which progressively localizes to the sprouting vascular network. Conditional, myeloid-specific deletion of Dll1 impairs endothelial Dll4 tip-stalk gradient resulting in an increase of endothelial tip cells and EC filopodia, accompanied by an increase in vascular density and branching. In vitro, co-culture of human EC with monocyte-derived macrophages induced Dll1 upregulation in macrophages and Dll4 upregulation and an endothelial tip cell signature in EC. Furthermore, culturing human EC on recombinant DLL1 induced endothelial Dll4 expression and a tip cell program, indicating that changes are Dll1-dependent. Thus, myeloid cells regulate tip cell fate and angiogenesis through expression of Notch ligand Dll1.

[Imaging of bacteria in burn wounds treated with split-thicknessgrafts in MEEK/MESH technique: a pilot study with first experiences in clinical wound evaluation with autofluorescence]. / Visualisierung von Bakterien auf Verbrennungswunden und Spalthauttransplantaten nach der MEEK/MESH-Technik ­ Eine Pilotstudie mit ersten Erfahrungen der klinischen Wundbeurteilung durch Autofluoreszenz.

BACKGROUND: Partial and full thickness burns require surgical treatment, such as early débridement and skin transplantation in MEEK/MESH technique or further reconstructive surgery. Infections of burns or transplanted areas limit surgical success and increase patient mortality. For split-thickness grafts in MEEK technique a superficial silk is applied as a protective on-top dressing, whereas in MESH technique fatty gauze and foam are used as standard protective covers over five to seven days. However, wound occlusion by both materials provides the soil for growth of microorganisms. The timely identification of impending infections is necessary to initiate early removal in order to safe and preserve skin grafts. Early identification of infections and removal of foreign material should therefore be attempted. MATERIAL AND METHODS: Burn wounds treated with split-thickness skin grafts processed by MEEK/MESH technique and covered with silk or foam overlayers were analyzed for signs of bacterial infection using the MolecuLight i:X™ device. In addition, swaps for microbiological analysis where taken from fluorescent areas and correlated with florescent image results. RESULTS: We examined burn wounds (n = 14) of three different intensive care patients. The MolecuLight i:X™ camera showed a strong colonization of the transplanted areas and foreign materials, that were in line with microbiological analysis findings. The representation of the excitation load showed high values in the foreign materials. The take rate of MEEK-transplants was 90 % compared to MESH-transplanted with about 60 %. The positive predictive value was 81.8 % for detection of a wound infection with autofluorescence. The negative predictive value was 90.3 % with a sensitivity of 86.7 % and a specificity of 87.5 %. CONCLUSION: The representation of the fluorescence exciter load shows high concentrations of pathogens both in the MEEK silk layer as well as in foam linkers. Overall split-thickness grafts according to the MEEK technique showed a higher healing rate compared to MESH technique. Screening of burns wounds with autofluorescence imaging can be helpful for an additive wound assessment. Split-thickness graft covers should be applied only for a minimum time period required to ensure stable grafting.

Multimodal and Multiscale Analysis Reveals Distinct Vascular, Metabolic and Inflammatory Components of the Tissue Response to Limb Ischemia.

Ischemia triggers a complex tissue response involving vascular, metabolic and inflammatory changes. METHODS: We combined hybrid SPECT/CT or PET/CT nuclear imaging studies of perfusion, metabolism and inflammation with multicolor flow cytometry-based cell population analysis to comprehensively analyze the ischemic tissue response and to elucidate the cellular substrate of noninvasive molecular imaging techniques in a mouse model of hind limb ischemia. RESULTS: Comparative analysis of tissue perfusion with [99mTc]-Sestamibi and arterial influx with [99mTc]-labeled albumin microspheres by SPECT/CT revealed a distinct pattern of response to vascular occlusion: an early ischemic period of matched suppression of tissue perfusion and arterial influx, a subacute ischemic period of normalized arterial influx but impaired tissue perfusion, and a protracted post-ischemic period of hyperdynamic arterial and normalized tissue perfusion, indicating coordination of macrovascular and microvascular responses. In addition, the subacute period showed increased glucose uptake by [18F]-FDG PET/CT scanning as the metabolic response of viable tissue to hypoperfusion. This was associated with robust macrophage infiltration by flow cytometry, and glucose uptake studies identified macrophages as major contributors to glucose utilization in ischemic tissue. Furthermore, imaging with the TSPO ligand [18F]-GE180 showed a peaked response during the subacute phase due to preferential labeling of monocytes and macrophages, while imaging with [68Ga]-RGD, an integrin ligand, showed prolonged post-ischemic upregulation, which was attributed to labeling of macrophages and endothelial cells by flow cytometry. CONCLUSION: Combined nuclear imaging and cell population analysis reveals distinct components of the ischemic tissue response and associated cell subsets, which could be targeted for therapeutic interventions.

[The Humboldtian model in Plastic and Reconstructive Surgery - a student survey on teaching and venia legendi]. / Das Humboldt'sche Bildungsideal in der Plastischen Chirurgie ­ Studentenumfrage zur Lehre und Venia Legendi.

BACKGROUND: The workload of university hospitals and hospitals with university association includes clinical patient care as well as teaching and research in particular. The current development with focus on financial issues leads to a reduction of teaching and research capacities. Economic focus in university medicine changes priorities of academic surgery. METHODS: An online survey questioned medical students with regard to subjective assessment of quality of the academic body of university hospitals and current teaching quality. Students evaluated the current quality of teaching of postdoctoral lecturers in relation to their career stage and made suggestions for quality of teaching improvement. RESULTS: A total of 166 students participated in the survey. Of 123 students, about 78 % stated that the reputation of postdoctoral lecturers increases with the habilitation but about 85 % stated that professional expectations also rise. About 43 % of the students aim to achieve a postdoctoral lecture qualification. DISCUSSION: Among students academic career is still attractive, but restructuring and modernization of established working models is an essential prerequisite.

The chemokine receptor CX3CR1 coordinates monocyte recruitment and endothelial regeneration after arterial injury.

Regeneration of arterial endothelium after injury is critical for the maintenance of normal blood flow, cell trafficking, and vascular function. Using mouse models of carotid injury, we show that the transition from a static to a dynamic phase of endothelial regeneration is marked by a strong increase in endothelial proliferation, which is accompanied by induction of the chemokine CX3CL1 in endothelial cells near the wound edge, leading to progressive recruitment of Ly6Clo monocytes expressing high levels of the cognate CX3CR1 chemokine receptor. In Cx3cr1-deficient mice recruitment of Ly6Clo monocytes, endothelial proliferation and regeneration of the endothelial monolayer after carotid injury are impaired, which is rescued by acute transfer of normal Ly6Clo monocytes. Furthermore, human non-classical monocytes induce proliferation of endothelial cells in co-culture experiments in a VEGFA-dependent manner, and monocyte transfer following carotid injury promotes endothelial wound closure in a hybrid mouse model in vivo Thus, CX3CR1 coordinates recruitment of specific monocyte subsets to sites of endothelial regeneration, which promote endothelial proliferation and arterial regeneration.

Blood vessel control of macrophage maturation promotes arteriogenesis in ischemia.

Ischemia causes an inflammatory response that is intended to restore perfusion and homeostasis yet often aggravates damage. Here we show, using conditional genetic deletion strategies together with adoptive cell transfer experiments in a mouse model of hind limb ischemia, that blood vessels control macrophage differentiation and maturation from recruited monocytes via Notch signaling, which in turn promotes arteriogenesis and tissue repair. Macrophage maturation is controlled by Notch ligand Dll1 expressed in vascular endothelial cells of arteries and requires macrophage canonical Notch signaling via Rbpj, which simultaneously suppresses an inflammatory macrophage fate. Conversely, conditional mutant mice lacking Dll1 or Rbpj show proliferation and transient accumulation of inflammatory macrophages, which antagonizes arteriogenesis and tissue repair. Furthermore, the effects of Notch are sufficient to generate mature macrophages from monocytes ex vivo that display a stable anti-inflammatory phenotype when challenged with pro-inflammatory stimuli. Thus, angiocrine Notch signaling fosters macrophage maturation during ischemia.Molecular mechanisms of macrophage-mediated regulation of artery growth in response to ischemia are poorly understood. Here the authors show that vascular endothelium controls macrophage maturation and differentiation via Notch signaling, which in turn promotes arteriogenesis and ischemic tissue recovery.

[Careers in plastic and aesthetic surgery: a review of habilitation and professorship of members of the DGPRAEC]. / Karriere in der Plastischen und Ästhetischen Chirurgie ­ Untersuchung zur Habilitation und Professur der Mitglieder der DGPRÄC.

Background Over the last few decades plastic and aesthetic surgery careers aimed at holding a chair as head of the department or clinical director. The current career trend shows a drain of academic teaching staff to peripheral hospitals with sole clinical focus. The achievement of a doctorate in German university medicine or obtaining the venia legendi appears to be the termination of academic careers. This brain drain with loss of expertise and scientific output imposes a problem to future progress in clinical and scientific plastic and reconstructive surgery. The causative role of our present work profile, workload and financial compensation will be discussed in this paper. Methods In order to understand this brain drain, the scientific and clinical developments of all habilitands, Assistant Professors and University Directors enlisted in our specialist society (DGPRAEC) were analyzed. The evaluation included the duration of the residency, the time span from being a specialist physician to habilitation, as well as gaining a leadership position after habilitation. Finally, the current activity of the members at university and non-university institutions was evaluated. Results A total of 1238 members were analyzed. Among these, 177 (14.3 %) members had completed the habilitation. In total, 114 (9.21 %) were included based on full available CVs. Of the listed members, 80 members (6,5 %) had an APL professorship/university professorship in April 2017. 88 CVs showed an average time span of 4.2 years from specialization to habilitation. 80 CVs revealed a 5 year time span to achieve an APL professorship/university professorship. After an average of 4.2 years, leadership positions were held. Of the analyzed habilitations, 60 % were active in peripheral hospitals at the time April 2017. Discussion The loss of scientific and clinical expertise should be prevented in order to preserve academic plastic surgery with focus on patient care, academic education and research. This could be achieved by creating more attractive working conditions.

Blood flow controls bone vascular function and osteogenesis.

While blood vessels play important roles in bone homeostasis and repair, fundamental aspects of vascular function in the skeletal system remain poorly understood. Here we show that the long bone vasculature generates a peculiar flow pattern, which is important for proper angiogenesis. Intravital imaging reveals that vessel growth in murine long bone involves the extension and anastomotic fusion of endothelial buds. Impaired blood flow leads to defective angiogenesis and osteogenesis, and downregulation of Notch signalling in endothelial cells. In aged mice, skeletal blood flow and endothelial Notch activity are also reduced leading to decreased angiogenesis and osteogenesis, which is reverted by genetic reactivation of Notch. Blood flow and angiogenesis in aged mice are also enhanced on administration of bisphosphonate, a class of drugs frequently used for the treatment of osteoporosis. We propose that blood flow and endothelial Notch signalling are key factors controlling ageing processes in the skeletal system.

Regulation of monocyte cell fate by blood vessels mediated by Notch signalling.

A population of monocytes, known as Ly6C(lo) monocytes, patrol blood vessels by crawling along the vascular endothelium. Here we show that endothelial cells control their origin through Notch signalling. Using combinations of conditional genetic deletion strategies and cell-fate tracking experiments we show that Notch2 regulates conversion of Ly6C(hi) monocytes into Ly6C(lo) monocytes in vivo and in vitro, thereby regulating monocyte cell fate under steady-state conditions. This process is controlled by Notch ligand delta-like 1 (Dll1) expressed by a population of endothelial cells that constitute distinct vascular niches in the bone marrow and spleen in vivo, while culture on recombinant DLL1 induces monocyte conversion in vitro. Thus, blood vessels regulate monocyte conversion, a form of committed myeloid cell fate regulation.

MAP-Kinase Activated Protein Kinase 2 Links Endothelial Activation and Monocyte/macrophage Recruitment in Arteriogenesis.

Arteriogenesis, the growth of natural bypass arteries, is triggered by hemodynamic forces within vessels and requires a balanced inflammatory response, involving induction of the chemokine MCP-1 and recruitment of leukocytes. However, little is known how these processes are coordinated. The MAP-kinase-activated-proteinkinase-2 (MK2) is a critical regulator of inflammatory processes and might represent an important link between cytokine production and cell recruitment during postnatal arteriogenesis. Therefore, the present study investigated the functional role of MK2 during postnatal arteriogenesis. In a mouse model of hindlimb ischemia (HLI) MK2-deficiency (MK2KO) significantly impaired ischemic blood flow recovery and growth of collateral arteries as well as perivascular recruitment of mononuclear cells and macrophages. This was accompanied by induction of endothelial MCP-1 expression in wildtype (WT) but not in MK2KO collateral arteries. Following HLI, MK2 activation rapidly occured in the endothelium of growing WT arteries in vivo. In vitro, inflammatory cytokines and cyclic stretch activated MK2 in endothelial cells, which was required for stretch- and cytokine-induced release of MCP-1. In addition, a monocyte cell autonomous function of MK2 was uncovered potentially regulating MCP-1-dependent monocyte recruitment to vessels: MCP-1 stimulation of WT monocytes induced MK2 activation and monocyte migration in vitro. The latter was reduced in MK2KO monocytes, while in vivo MK2 was activated in monocytes recruited to collateral arteries. In conclusion, MK2 regulates postnatal arteriogenesis by controlling vascular recruitment of monocytes/macrophages in a dual manner: regulation of endothelial MCP-1 expression in response to hemodynamic and inflammatory forces as well as MCP-1 dependent monocyte migration.

Genetic reporter analysis reveals an expandable reservoir of OCT4+ cells in adult skin.

The transcription factor Oct4 (Pou5f1) is a critical regulator of pluripotency in embryonic and induced pluripotent stem cells. Therefore, Oct4 expression might identify somatic stem cell populations with inherent multipotent potential or a propensity for facilitated reprogramming. However, analysis of Oct4 expression is confounded by Oct4 pseudogenes or non-pluripotency-related isoforms. Systematic analysis of a transgenic Oct4-EGFP reporter mouse identified testis and skin as two principle sources of Oct4 (+) cells in postnatal mice. While the prevalence of GFP(+) cells in testis rapidly declined with age, the skin-resident GFP(+) population expanded in a cyclical fashion. These cells were identified as epidermal stem cells dwelling in the stem cell niche of the hair follicle, which endogenously expressed all principle reprogramming factors at low levels. Interestingly, skin wounding or non-traumatic hair removal robustly expanded the GFP(+) epidermal cell pool not only locally, but also in uninjured skin areas, demonstrating the existence of a systemic response. Thus, the epithelial stem cell niche of the hair follicle harbors an expandable pool of Oct4+ stem cells, which might be useful for therapeutic cell transfer or facilitated reprogramming.

Urokinase receptor associates with myocardin to control vascular smooth muscle cells phenotype in vascular disease.

OBJECTIVE: The urokinase-type plasminogen activator (uPA) and its specific receptor (uPAR) are a potent multifunctional system involved in vascular remodeling. The goal of the study was to unravel the mechanisms of uPA/uPAR-directed vascular smooth muscle cell (VSMC) differentiation. METHODS AND RESULTS: Using cultured human primary VSMCs, we identified a new molecular mechanism controlling phenotypic modulation in vitro and in vivo. We found that the urokinase-type plasminogen activator receptor (uPAR) acts together with the transcriptional coactivator myocardin to regulate the VSMC phenotype. uPAR, a glycosylphosphatidylinositol-anchored cell-surface receptor family member, undergoes ligand-induced internalization and nuclear transport in VSMCs. Platelet-derived growth factor receptor ß and SUMOylated RanGAP1 mediate this trafficking. Nuclear uPAR associates with myocardin, which is then recruited from the promoters of serum response factor target genes and undergoes proteasomal degradation. This chain of events initiates the synthetic VSMC phenotype. Using mouse carotid artery ligation model, we show that this mechanism contributes to adverse vascular remodeling after injury in vivo. We then cultured cells on a microstructured biomaterial and found that substrate topography induced uPAR-mediated VSMC differentiation. CONCLUSIONS: These findings reveal the transcriptional activity of uPAR, controlling the differentiation of VSMCs in a vascular disease model. They also suggest a new role for uPAR as a therapeutic target and as a marker for VSMC phenotyping on prosthetic biomaterials.

HSP70 enhances immunosuppressive function of CD4(+)CD25(+)FoxP3(+) T regulatory cells and cytotoxicity in CD4(+)CD25(-) T cells.

Human CD4(+)CD25(+)FoxP3(+) T regulatory cells (Tregs) control effector T cells and play a central role in peripheral tolerance and immune homeostasis. Heat shock protein 70 (HSP70) is a major immunomodulatory molecule, but its effect on the functions of Tregs is not well understood. To investigate target-dependent and -independent Treg functions, we studied cytokine expression, regulation of proliferation and cytotoxicity after exposure of Tregs to HSP70. HSP70-treated Tregs significantly inhibited proliferation of CD4(+)CD25(-) target cells and downregulated the secretion of the proinflammatory cytokines IFN-γ and TNF-α. By contrast, HSP70 increased the secretion of Treg suppressor cytokines IL-10 and TGF-ß. Treatment with HSP70 enhanced the cytotoxic properties of Tregs only to a minor extent (4-fold), but led to stronger responses in CD4(+)CD25(-) cells (42-fold). HSP70-induced modulation of T-cell responses was further enhanced by combined treatment with HSP70 plus IL-2. Treatment of Tregs with HSP70 led to phosphorylation of PI3K/AKT and the MAPKs JNK and p38, but not that of ERK1/2. Exposure of Tregs to specific inhibitors of PI3K/AKT and the MAPKs JNK and p38 reduced the immunosuppressive function of HSP70-treated Tregs as indicated by the modified secretion of specific target cell (IFN-γ, TNF-α) and suppressor cytokines (IL-10, TGF-ß). Taken together, the data show that HSP70 enhances the suppressive capacity of Tregs to neutralize target immune cells. Thus HSP70-enhanced suppression of Tregs may prevent exaggerated immune responses and may play a major role in maintaining immune homeostasis.

Overexpression of integrin beta 5 enhances the paracrine properties of circulating angiogenic cells via Src kinase-mediated activation of STAT3.

OBJECTIVE: To determine the intracellular mechanisms mediating the angiogenic effects of integrin alpha v beta 5 overexpression in circulating angiogenic cells (CACs). METHODS AND RESULTS: Integrin alpha v beta 5 is expressed on angiogenic endothelial cells, and integrin alpha v beta 5 activation was shown to improve the reparative functions of endothelial progenitors within the cardiovascular system. CACs were transiently transfected with the full-length cDNA of human integrin beta 5 (CAC-ITGB5) or control-vector (CAC-vector). Integrin beta 5 overexpression was confirmed using flow cytometry, Western blot, and PCR analysis; it enhanced the angiogenic capacities of CACs in vitro (spheroid and Matrigel angiogenesis assay) and stimulated new vessel formation in vivo (murine hind limb ischemia model). Overexpression of ITGB5 resulted in integrin alpha v beta 5 phosphorylation and activation of Src kinase and signal transducer and activator of transcription (STAT) 3. Furthermore, elevated mRNA and protein expression of the CXC chemokine CXCL8 and the CC chemokine CCL2 was detected in CAC-ITGB5, and conditioned medium from CAC-ITGB5 enhanced the sprouting of coincubated human endothelial cells in a STAT3-, CXCL8-, and CCL2-dependent manner. CONCLUSIONS: Src kinase-mediated activation of STAT3 and subsequent angiogenic gene expression mediate the effects of integrin alpha v beta 5 and may be exploited to enhance the paracrine activities of CACs.