Induction Of Neovascularization Research Articles

Inflammation-induced subcutaneous neovascularization for the long-term survival of encapsulated islets without immunosuppression.

Cellular therapies for type-1 diabetes can leverage cell encapsulation to dispense with immunosuppression. However, encapsulated islet cells do not survive long, particularly when implanted in poorly vascularized subcutaneous sites. Here we show that the induction of neovascularization via temporary controlled inflammation through the implantation of a nylon catheter can be used to create a subcutaneous cavity that supports the transplantation and optimal function of a geometrically matching islet-encapsulation device consisting of a twisted nylon surgical thread coated with an islet-seeded alginate hydrogel. The neovascularized cavity led to the sustained reversal of diabetes, as we show in immunocompetent syngeneic, allogeneic and xenogeneic mouse models of diabetes, owing to increased oxygenation, physiological glucose responsiveness and islet survival, as indicated by a computational model of mass transport. The cavity also allowed for the in situ replacement of impaired devices, with prompt return to normoglycemia. Controlled inflammation-induced neovascularization is a scalable approach, as we show with a minipig model, and may facilitate the clinical translation of immunosuppression-free subcutaneous islet transplantation.

Biological effects of extracorporeal shockwave therapy in tendons: A systematic review.

Extracorporeal shockwave therapy was initially used for kidney stone disintegration and its application was then extended to calcific tendinitis. The therapeutic field expanded and included numerous types of tendinopathies, from shoulder to plantar fascia. The clinical benefits were documented in trials and the effects and mechanisms were studied on models including animal and human tendons. The present systematic review outlines a large spectrum of biological effects. First, an optimal dose is adapted for each species and each tendon; exceeding the optimal dose may lead to structural injury. Furthermore, the biological effects may be grouped into neovascularization induction, cellularity and extracellular matrix changes, metalloprotease and cytokine modulation, as well as lubricin production. As a result, the remodeled tendon displays improved biomechanical properties to resist stress.

Cell-Free Treatments: A New Generation of Targeted Therapies for Treatment of Ischemic Heart Disease.

Although recent progress in medicine has substantially reduced cardiovascular diseases (CVDs)-related mortalities, current therapeutics have failed miserably to be beneficial for all patients with CVDs. A wide array of evidence suggests that newly-introduced cell-free treatments (CFTs) have more reliable results in the improvement of cardiac function. The main regeneration activity of CFTs protocols is based on bypassing cells and using paracrine factors. In this article, we aim to compare various stem cell secretomes, a part of a CFTs strategy, to generalize their effective clinical outcomes for patients with CVDs. Data for this review article were collected from 70 published articles (original, review, randomized clinical trials (RCTs), and case reports/series studies done on human and animals) obtained from Cochrane, Science Direct, PubMed, Scopus, Elsevier, and Google Scholar) from 2015 to April 2020 using six keywords. Full-text/full-length articles, abstract, section of book, chapter, and conference papers in English language were included. Studies with irrelevant/insufficient/data, or undefined practical methods were excluded. CFTs approaches involved in growth factors (GFs); gene-based therapies; microRNAs (miRNAs); extracellular vesicles (EVs) [exosomes (EXs) and microvesicles (MVs)]; and conditioned media (CM). EXs and CM have shown more remarkable results than stem cell therapy (SCT). GF-based therapies have useful results as well as side effects like pathologic angiogenesis. Cell source, cell′s aging and CM affect secretomes. Genetic manipulation of stem cells can change the secretome’s components. Growing progression to end stage heart failure (HF), propounds CFTs as an advantageous method with practical and clinical values for replacement of injured myocardium, and induction of neovascularization. To elucidate the secrets behind amplifying the expansion rate of cells, increasing life-expectancy, and improving quality of life (QOL) for patients with ischemic heart diseases (IHDs), collaboration among cell biologist, basic medical scientists, and cardiologists is highly recommended.

Mesenchymal Stromal Cell-Based Therapies as Promising Treatments for Muscle Regeneration After Snakebite Envenoming.

Snakebite envenoming is a global neglected disease with an incidence of up to 2.7 million new cases every year. Although antivenoms are so-far the most effective treatment to reverse the acute systemic effects induced by snakebite envenoming, they have a limited therapeutic potential, being unable to completely neutralize the local venom effects. Local damage, such as dermonecrosis and myonecrosis, can lead to permanent sequelae with physical, social, and psychological implications. The strong inflammatory process induced by snake venoms is associated with poor tissue regeneration, in particular the lack of or reduced skeletal muscle regeneration. Mesenchymal stromal cells (MSCs)-based therapies have shown both anti-inflammatory and pro-regenerative properties. We postulate that using allogeneic MSCs or their cell-free products can induce skeletal muscle regeneration in snakebite victims, improving all the three steps of the skeletal muscle regeneration process, mainly by anti-inflammatory activity, paracrine effects, neovascularization induction, and inhibition of tissue damage, instrumental for microenvironment remodeling and regeneration. Since snakebite envenoming occurs mainly in areas with poor healthcare, we enlist the principles and potential of MSCs-based therapies and discuss regulatory issues, good manufacturing practices, transportation, storage, and related-procedures that could allow the administration of these therapies, looking forward to a safe and cost-effective treatment for a so far unsolved and neglected health problem.

Up-regulation of pro-angiogenic pathways and induction of neovascularization by an acute retinal light damage

The light damage (LD) model was mainly used to study some of the main aspects of age related macular degeneration (AMD), such as oxidative stress and photoreceptor death. Several protocols of light-induced retinal degeneration exist. Acute light damage is characterized by a brief exposure (24 hours) to high intensity light (1000 lux) and leads to focal degeneration of the retina which progresses over time. To date there are not experimental data that relate this model to neovascular events. Therefore, the purpose of this study was to characterize the retina after an acute light damage to assess whether the vascularization was affected. Functional, molecular and morphological investigations were carried out. The electroretinographic response was assessed at all recovery times (7, 60, 120 days after LD). Starting from 7 days after light damage there was a significant decrease in the functional response, which remained low up to 120 days of recovery. At 7 days after light exposure, neo-vessels invaded the photoreceptor layer and retinal neovascularization occurred. Remarkably, neoangiogenesis was associated to the up-regulation of VEGF, bFGF and their respective receptors (VEGFR2 and FGFR1) with the progression of degeneration. These important results indicate that a brief exposure to bright light induces the up-regulation of pro-angiogenic pathways with subsequent neovascularization.

Bclaf1 promotes angiogenesis by regulating HIF-1\u03b1 transcription in hepatocellular carcinoma

The development of hepatocellular carcinomas (HCC) depends on their local microenvironment and the induction of neovascularization is a decisive step in tumor progression, since the growth of solid tumors is limited by nutrient and oxygen supply. Hypoxia is the critical factor that induces transcription of the hypoxia inducible factor-1α (HIF-1α) encoding gene HIF1A and HIF-1α protein accumulation to promote angiogenesis. However, the basis for the transcriptional regulation of HIF1A expression in HCC is still unclear. Here, we show that Bclaf1 levels are highly correlated with HIF-1α levels in HCC tissues, and that knockdown of Bclaf1 in HCC cell lines significantly reduces hypoxia-induced HIF1A expression. Furthermore, we found that Bclaf1 promotes HIF1A transcription via its bZIP domain, leading subsequently to increased transcription of the HIF-1α downstream targets VEGFA, TGFB, and EPO that in turn promote HCC-associated angiogenesis and thus survival and thriving of HCC cells. Moreover, we demonstrate that HIF-1α levels and microvessel density decrease after the shRNA-mediated Bclaf1 knockdown in xenograft tumors. Finally, we found that Bclaf1 levels increase in hypoxia in a HIF-1α dependent manner. Therefore, our study identifies Bclaf1 as a novel positive regulator of HIF-1α in the hypoxic microenvironment, providing new incentives for promoting Bcalf1 as a potential therapeutic target for an anti-HCC strategy.

AB035. Vasohibin-1 as a potential therapeutic target for erectile dysfunction

Induction of neovascularization is a promising strategy to treat erectile dysfunction (ED). Here we for the first time report the unexpected role of vasohibin-1, which is previously defined as an anti-angiogenic factor, in the restoration of erectile function by enhancing cavernous (penile) angiogenesis in an animal model of diabetic ED. The cavernous expression of vasohibin-1 was down-regulated in diabetic mice and in patients with diabetic ED; vasohibin-1 was mainly expressed in endothelial cells. Vasohibin-1 knockout mice revealed decrease in cavernous endothelial and pericyte content compared with wild type mice, which resulted in deterioration of erectile function. Local delivery of vasohibin-1 peptide into the corpus cavernosum of diabetic mice induced significant restoration of erectile function, which reached up to 85% of control values at the concentration of 4 µg/20 µL. Vasohibin-1 significantly increased cavernous endothelial cell content and induced eNOS phosphorylation (Ser1177). Vasohibin-1 decreased extravasation of oxidized-LDL by restoring pericytes and endothelial cell-cell junction proteins in the diabetic mice. By using proteome profiler array kit, we found that the induction of angiogenic factors, including angiopoietin-1, vascular endothelial growth factor, and basic fibroblasts growth factor, mainly in fibroblasts is a major molecular mechanism responsible for vasohibin-1-mediated cavernous angiogenesis and subsequent restoration of erectile function. Our findings suggest that vasohibin-1 is proangiogenic in diabetic penis and is a promising therapeutic target for ED from vascular causes.

ADSC-sheet Transplantation to Prevent Stricture after Extended Esophageal Endoscopic Submucosal Dissection

In past years, the cell-sheet construct has spurred wide interest in regenerative medicine, especially for reconstructive surgery procedures. The development of diversified technologies combining adipose tissue-derived stromal cells (ADSCs) with various biomaterials has led to the construction of numerous types of tissue-engineered substitutes, such as bone, cartilage, and adipose tissues from rodent, porcine, or human ADSCs. Extended esophageal endoscopic submucosal dissection (ESD) is responsible for esophageal stricture formation. Stricture prevention remains challenging, with no efficient treatments available. Previous studies reported the effectiveness of mucosal cell-sheet transplantation in a canine model and in humans. ADSCs are attributed anti-inflammatory properties, local immune modulating effects, neovascularization induction, and differentiation abilities into mesenchymal and non-mesenchymal lineages. This original study describes the endoscopic transplantation of an ADSC tissue-engineered construct to prevent esophageal stricture in a swine model. The ADSC construct was composed of two allogenic ADSC sheets layered upon each other on a paper support membrane. The ADSCs were labeled with the PKH67 fluorophore to allow probe-based confocal laser endomicroscopy (pCLE) monitoring. On the day of transplantation, a 5-cm and hemi-circumferential ESD known to induce esophageal stricture was performed. Animals were immediately endoscopically transplanted with 4 ADSC constructs. The complete adhesion of the ADSC constructs was obtained after 10 min of gentle application. Animals were sacrificed on day 28. All animals were successfully transplanted. Transplantation was confirmed on day 3 with a positive pCLE evaluation. Compared to transplanted animals, control animals developed severe strictures, with major fibrotic tissue development, more frequent alimentary trouble, and reduced weight gain. In our model, the transplantation of allogenic ADSCs, organized in double cell sheets, after extended ESD was successful and strongly associated with a lower esophageal stricture rate.

ADSC-sheet Transplantation to Prevent Stricture after Extended Esophageal Endoscopic Submucosal Dissection.

In past years, the cell-sheet construct has spurred wide interest in regenerative medicine, especially for reconstructive surgery procedures. The development of diversified technologies combining adipose tissue-derived stromal cells (ADSCs) with various biomaterials has led to the construction of numerous types of tissue-engineered substitutes, such as bone, cartilage, and adipose tissues from rodent, porcine, or human ADSCs. Extended esophageal endoscopic submucosal dissection (ESD) is responsible for esophageal stricture formation. Stricture prevention remains challenging, with no efficient treatments available. Previous studies reported the effectiveness of mucosal cell-sheet transplantation in a canine model and in humans. ADSCs are attributed anti-inflammatory properties, local immune modulating effects, neovascularization induction, and differentiation abilities into mesenchymal and non-mesenchymal lineages. This original study describes the endoscopic transplantation of an ADSC tissue-engineered construct to prevent esophageal stricture in a swine model. The ADSC construct was composed of two allogenic ADSC sheets layered upon each other on a paper support membrane. The ADSCs were labeled with the PKH67 fluorophore to allow probe-based confocal laser endomicroscopy (pCLE) monitoring. On the day of transplantation, a 5-cm and hemi-circumferential ESD known to induce esophageal stricture was performed. Animals were immediately endoscopically transplanted with 4 ADSC constructs. The complete adhesion of the ADSC constructs was obtained after 10 min of gentle application. Animals were sacrificed on day 28. All animals were successfully transplanted. Transplantation was confirmed on day 3 with a positive pCLE evaluation. Compared to transplanted animals, control animals developed severe strictures, with major fibrotic tissue development, more frequent alimentary trouble, and reduced weight gain. In our model, the transplantation of allogenic ADSCs, organized in double cell sheets, after extended ESD was successful and strongly associated with a lower esophageal stricture rate.

Comparison of postoperative corneal changes between dry eye and non-dry eye in a murine cataract surgery model.

To compare the effects of the surgical insult of cataract surgery on corneal inflammatory infiltration, neovascularization (NV) and lymphangiogenesis (LY) between the dry eye and non-dry eye in murine cataract surgery models. We established two groups of animals, one with normal eyes (non-dry eye) and the second with induced dry eyes. In both groups, we used surgical insults to mimic human cataract surgery, which consisted of lens extraction, corneal incision and suture. After harvesting of corneas on the 9(th) postoperative day and immunohistochemical staining, we compared NV, LY and CD11b+ cell infiltration in the corneas. Dry eye group had significantly more inflammatory infiltration (21.75%±7.17% vs 3.65%±1.49%; P=0.049). The dry eye group showed significantly more NV (48.21%±4.02% vs 26.24%±6.01%; P=0.016) and greater levels of LY (9.27%±0.48% vs 4.84%±1.15%; P=0.007). In corneas on which no surgery was performed, there was no induction of NV in both the dry and non-dry group, but dry eye group demonstrated more CD11b+ cells infiltration than the non-dry eye group (0.360%±0.160% vs 0.023%±0.006%; P=0.068). Dry eye group showed more NV than non-dry eye group in both topical PBS application and subconjunctival PBS injection (P=0.020 and 0.000, respectively). In a murine cataract surgery model, preexisting dry eye can induce more postoperative NV, LY, and inflammation in corneal tissue.

Multimodality noninvasive imaging for assessing therapeutic effects of exogenously transplanted cell aggregates capable of angiogenesis on acute myocardial infarction

Although the induction of neovascularization by cell-based approaches has demonstrated substantial potential in treating myocardial infarction (MI), the process of cell-mediated angiogenesis and its correlation with therapeutic mechanisms of cardiac repair remain elusive. In this work, three-dimensional (3D) aggregates of human umbilical vein endothelial cells (HUVECs) and cord-blood mesenchymal stem cells (cbMSCs) are constructed using a methylcellulose hydrogel system. By maximizing cell–cell and cell–ECM communications and establishing a hypoxic microenvironment in their inner cores, these cell aggregates are capable of forming widespread tubular networks together with the angiogenic marker αvβ3 integrin; they secret multiple pro-angiogenic, pro-survival, and mobilizing factors when grown on Matrigel. The aggregates of HUVECs/cbMSCs are exogenously engrafted into the peri-infarct zones of rats with MI via direct local injection. Multimodality noninvasive imaging techniques, including positron emission tomography, single photon emission computed tomography, and echocardiography, are employed to monitor serially the beneficial effects of cell therapy on angiogenesis, blood perfusion, and global/regional ventricular function, respectively. The myocardial perfusion is correlated with ventricular contractility, demonstrating that the recovery of blood perfusion helps to restore regional cardiac function, leading to the improvement in global ventricular performance. These experimental data reveal the efficacy of the exogenous transplantation of 3D cell aggregates after MI and elucidate the mechanism of cell-mediated therapeutic angiogenesis for cardiac repair.

In vivo angiogenic activity induction by collagen- soaked Poly-L-lactic acid scaffolds

Angiogenesis is essential in tissue integration and it is involved in the biological response to biomaterials. Poly-L-lactic acid (PLLA), a synthetic polymer, is utilized as scaffolding to regenerate new tissues. This study investigated the short and long term degradation and the induction of neovascularization of both native PLLA (n-PLLA) and collagen type I soaked PLLA (c-PLLA) porous scaffolds, implanted subcutane - ously in balb/c mice. The comparative analysis by phase contrast, optical, and scanning electron micros- copy (SEM) of scaffolds 7 and 21 days after implantation showed a mild inflam- matory response at the implant site of c-PLLA scaffold. No significant difference in systemic immune response was detected by hematology analyzer, and by histologi- cal evaluation of lymph node and spleen features. On the contrary, immune reaction was moderate in n-PLLA. Pores of both PLLA networks laying on the muscle fibers were partially infiltrated by appositional collagen/elastin tissue, phagocytic cells, and fibroblast, with respect to the inner side. The presence of numerous and large blood vessels into pores of c-PLLA scaffolds showed an enhancing vascularization rate. These characteristics appeared to be less conspicuous in n-PLLA. At longer time points (42 and 84 days), there was low difference in inflammato- ry cell presence into scaffold pores and the number of cells infiltrating each implant was significantly decreased. In fact, we did not observe difference in the migration of inflammatory cells into PLLA scaffolds. Polymer degradation was detected in both PLLA networks, but there are no considerable differences, as confirmed by the SEM analysis. Our results suggest that tissue integration of PLLA is enhanced when it is soaked with collagen, as well as the angiogenic activity on c-PLLA. Furthermore, the colla- gen soaking makes PLLA polymer more suitable for supporting cell attachment, pro- liferation, and function by mimicking the natural extra cellular matrix.

The corneal micropocket assay: a model of angiogenesis in the mouse eye.

The mouse corneal micropocket assay is a robust and quantitative in vivo assay for evaluating angiogenesis. By using standardized slow-release pellets containing specific growth factors that trigger blood vessel growth throughout the naturally avascular cornea, angiogenesis can be measured and quantified. In this assay the angiogenic response is generated over the course of several days, depending on the type and dose of growth factor used. The induction of neovascularization is commonly triggered by either basic fibroblast growth factor (bFGF) or vascular endothelial growth factor (VEGF). By combining these growth factors with sucralfate and hydron (poly-HEMA (poly(2-hydroxyethyl methacrylate))) and casting the mixture into pellets, they can be surgically implanted in the mouse eye. These uniform pellets slowly-release the growth factors over five or six days (bFGF or VEGF respectively) enabling sufficient angiogenic response required for vessel area quantification using a slit lamp. This assay can be used for different applications, including the evaluation of angiogenic modulator drugs or treatments as well as comparison between different genetic backgrounds affecting angiogenesis. A skilled investigator after practicing this assay can implant a pellet in less than 5 min per eye.

Cancer cells release anaphylatoxin C5a from C5 by serine protease to enhance invasiveness.

Anaphylatoxin C5a indirectly fosters cancer cells through recruitment of myeloid-derived suppressor cells (MDS) for inhibiting antitumor CD8+ T cells and induction of neovascularization. We recently found activation of cancer cells by C5a directly via the C5a-receptor (C5aR; CD88) to enhance invasiveness. Thus, C5a possibly contributes to cancer progression rather than elimination. C5a generation in cancer tissues has been reported; however, the mechanism is not fully elucidated. Cancer cell expression of complement regulatory molecules suggests inefficient C5a generation through activation of the complement system in response to cancer cells. To explore another C5a generation mechanism in cancer tissues, we examined cancer cells for C5a-releasing activity from C5. C5a was present in C5-supplemented culture media of cancer cells including C5aR-expressing cells, and the media enhanced C5aR-expressing cancer cell invasion, which was abolished by anti-C5a antibody. The C5a-releasing activity was absent in the supernatants of the media and was inhibited by aprotinin, a serine protease inhibitor, and decanoyl-Arg-Val-Lys-Arg-chloromethylketone but not by inhibitors specific for cysteine, acid, or metal proteases. These results indicated C5a release from C5 by a cancer cell membrane-bound serine protease that can cleave peptide bonds at the carboxy-terminal site of paired basic amino acid residues. Cancer cell C5a release from the complement-immobilized plasma supported feasibility of this cancer cell protease-dependent C5a generation in cancer tissues. The new mechanism of C5a generation suggests self-activation of C5aR-expressing cancer cells to enhance invasiveness and induction of MDS recruitment and neovascularization to create a microenvironment favorable for cancer progression.

Cardiomyocyte-specific Estrogen Receptor Alpha Increases Angiogenesis, Lymphangiogenesis and Reduces Fibrosis in the Female Mouse Heart Post-Myocardial Infarction.

Experimental studies showed that 17β-estradiol (E2) and activated Estrogen Receptors (ER) protect the heart from ischemic injury. However, the underlying molecular mechanisms are not well understood. To investigate the role of ER-alpha (ERα) in cardiomyocytes in the setting of myocardial ischemia, we generated transgenic mice with cardiomyocyte-specific overexpression of ERα (ERα-OE) and subjected them to Myocardial Infarction (MI). At the basal level, female and male ERα-OE mice showed increased Left Ventricular (LV) mass, LV volume and cardiomyocyte length. Two weeks after MI, LV volume was significantly increased and LV wall thickness decreased in female and male WT-mice and male ERα-OE, but not in female ERα-OE mice. ERα-OE enhanced expression of angiogenesis and lymphangiogenesis markers (Vegf, Lyve-1), and neovascularization in the peri-infarct area in both sexes. However, attenuated level of fibrosis and higher phosphorylation of JNK signaling pathway could be detected only in female ERα-OE after MI. In conclusion, our study indicates that ERα protects female mouse cardiomyocytes from the sequelae of ischemia through induction of neovascularization in a paracrine fashion and impaired fibrosis, which together may contribute to the attenuation of cardiac remodelling.

Extracorporeal shock wave therapy in periodontics: A new paradigm.

The quest for exploring new frontiers in the field of medical science for efficient and improved treatment modalities has always been on a rise. Extracorporeal shock wave therapy (ESWT) has been enormously used in medical practice, principally, for the management of urolithiasis, cholelithiasis and also in various orthopedic and musculoskeletal disorders. The efficacy of ESWT in the stimulation of osteoblasts, fibroblasts, induction of neovascularization and increased expression of bone morphogenic proteins has been well documented in the literature. However, dentistry is no exception to this trend. The present article enlightens the various applications of ESWT in the field of dentistry and explores its prospective applications in the field of periodontics, and the possibility of incorporating the beneficial properties of shock waves in improving the treatment outcome.

Echo-guided extracorporeal shock wave therapy for refractory angina improves regional myocardial blood flow as assessed by PET imaging

Purpose: Low-intensity extracorporeal shock wave (SW) therapy (SWT) has been shown to improve symptoms and exercise tolerance in patients (pts.) with coronary artery disease (CAD) not suitable for conventional revascularization strategies. Induction of neovascularization and improvement of myocardial perfusion are mechanisms hypothetized to be involved. Methods: 43 pts. with advanced CAD (mean age 67±10 years) not suitable for catheter-based or surgical revascularization were suffering from severe stable angina pectoris (CCS class III or IV) refractory to individually optimized medical treatment underwent a series of 9 echocardiography-targeted SW applications (3 SW applications/week during weeks 1, 5, and 9). The antero-septal wall (LAD territory) was targeted in 19, the lateral wall (RCX territory) in 18, and the inferior wall (RCA territory) in 6 pts. A series of 300-500 shock waves was applied per session. Anti-anginal medication (combination of 2 or 3 drugs) was kept unchanged. Regional myocardial blood flow (MBF) was measured quantitatively by NH3-PET at baseline and 4-6 weeks after completion of SWT. Results: Complications of SWT did not occur, markers of myocardial cell damage were all negative during SWT. At follow-up, 30 pts. (64%) reported improvement of angina to a tolerable level. CCS angina class decreased from 3.1±0.6 to 2.5±0.6 (p<0.0001). Maximum ergometric workload increased from 78±53 to 90±46 watts (p=0.04). MBF in the LV region targeted by SWT improved from 119+42 mL/min/100g at baseline to 129±48 mL/min/100g at follow up (p=0.047), while there was no change in the opposite wall (136±52 vs. 137±53 ml/min/100g; p=0.9). Conclusions: SWT improves symptoms in a sizeable number of pts. with chronic refractory angina. Regional improvement of MBF in the region targeted by SWT was also documented by PET imaging. Additional studies are warranted to clarify the role of SWT in the armamentarium for this challenging patient group.

Efficacy of a poly glycolic acid (PGA)/collagen composite nanofibre scaffold on cell migration and neovascularisation in vivo skin defect model

Application of tissue engineering currently provides promising therapeutic options in the fields of plastic surgery and wound management. The ability of scaffold material for cell proliferation and differentiation is the key for tissue engineering. This study has developed a novel nanofibre composed of poly glycolic acid (PGA) and collagen, both of which have their own respective beneficial properties. This study aimed to estimate the in vivo efficiency of the PGA/collagen nanofibre on granulation histology and its ability to induce neovascularisation. The electrospinning technique produced the PGA/collagen nanofiber with a diameter of 500 nm and weight mixing ratio of 40%. The skin defects on the mouse model were covered with PGA/collagen or a commercially available collagen matrix (n = 9). The PGA/collagen group histologically showed significantly higher cell density and a fine microstructure with greater number of migrating cells as compared to collagen matrix. Then, both materials were applied to the microcirculatory angiogenesis model. The PGA/collagen group (n = 8) revealed significantly higher functional capillary density on days 5 and 7 after application. The findings substantiated the fact that our material had a superior ability regarding cellular migration and induction of neovascularisation compared with the elementary collagen matrix product. This better result might be attributed to the nano-size effect of fine structure and the incorporation of PGA, which has been associated with enhanced angiogenesis.

Trks are novel oncogenes involved in the induction of neovascularization, tumor progression, and nodal metastasis in oral squamous cell carcinoma

The function of tropomyosin receptor kinase (Trk) family including TrkA, TrkB, and TrkC in cancer remains unknown. The role of Trks in oral squamous cell carcinoma (OSCC) was examined. Knockdown of Trks provided inhibition of growth or invasion and decrease of apoptosis in OSCC cells, which expressed Trks at high levels. VEGF expression was associated with TrkA and TrkB expression; a decrease of VEGF-C and VEGF-D was observed in OSCC cells with TrkB knockdown. TrkC did not affect the expression of VEGF family. An immunohistochemical analysis of 102 OSCCs showed that TrkB expression was related to microvessel density (MVD), lymph vessel density (LVD), and poor prognosis. TrkC expression was correlated with clinical stage, lymph node metastasis, MVD, LVD, and poor prognosis. TrkA expression was associated with VEGF expression, whereas TrkB expression was associated with the expressions of VEGF, VEGF-C and VEGF-D. No significant association was found between the expression of TrkC and genes of the VEGF family. Expression of Trks was not associated with RUNX3 silencing by methylation in OSCC cells. Trks expression was inversely correlated with RUNX3 expression in the OSCC cases. These results suggested that Trks enhances progression of OSCC through angiogenesis and lymphangiogenesis.

Abstract 515: Increased Adventitial Inflammation Occurs in Regions of Low Endothelial Shear Stress in a Swine Model of Coronary Atherosclerosis

Introduction: Plaque inflammation is a critical step in the initiation and progression of atherosclerosis. Such inflammation is thought to originate from the luminal surface of plaque and infiltrate the intima-media in advanced lesions. Low endothelial shear stress (ESS) is known to induce intima-media inflammation and plaque growth. In this study we investigated in-vivo the hypothesis that low ESS induces also adventitial inflammation. Methods: We studied 11 swine at 23 (baseline) and 30 (followup) weeks after the induction of diabetes and hyperlipidemia. Using angiography and intravascular ultrasound data, we performed 3D coronary reconstruction of coronary arteries and calculated the ESS with computational fluid dynamics. In 56 segments, we assessed the adventitial inflammatory (CD45) and antigen-presenting (MHC-II) cell content at followup with immunohistochemistry. Segments were classified as low (≤ 1 Pa) or higher (&gt;1 Pa) ESS. Results: MHC-II content in the adventitia (1.3±0.3%) was higher than in the media (0.3±0.1%, p&lt;0.001) but lower than in the intima (9.3±2.3%, p&lt;0.001). Low-ESS regions had increased adventitial MHC-II content compared to high-ESS regions (1.6±0.4 vs. 0.9±0.3%, p&lt;0.05). Adventitial MHC-II content was not influenced by plaque size (large plaques 1.7±0.5 vs. intermediate plaques 0.8±0.3%, p=0.24), internal elastic lamina fragmentation (IEL) grade (minimal/absent 0.6±0.3 vs. moderate/large 1.4±0.3, p=0.9), or remodeling pattern (inadequate 1.7±0.6 vs. compensatory 1.2±0.4 vs. expansive 0.6±0.2%, p=0.6). Adventitial CD45 content was not significantly different between low- and high-ESS segments, and also not different in large vs. intermediate plaques, in segments with various IEL fragmentation grades or remodeling patterns. Conclusion: Although total inflammation is not dependent on ESS, low ESS induces higher adventitial activated inflammatory cell content, as assessed by MHC-II immunostaining. This, in conjunction with the higher MHC-II content in the adventitia than in the media and the presence of an intact IEL suggests an additional source of inflammation in low-ESS plaque regions, originating from the vessel outer wall. The induction of neovascularization possibly accounts for this phenomenon.