Angiogenesis Parameters Research Articles

Mitigative Effects of Topical Norfloxacin on an Imiquimod-Induced Murine Model of Psoriasis.

Psoriasis is a chronic, inflammatory dermatosis characterized by thickened, reddened, and scaly skin lesions. Norfloxacin is a fluoroquinolone antibiotic with enhanced antioxidant, anti-inflammatory, and immunomodulatory bioactivities. The aim of this study was to figure out the possible impact of topical norfloxacin on an imiquimod-induced model of psoriasis in mice. Thirty albino-type mice were split into five distinct groups of six animals each. The control group included healthy mice that had not received any treatment. The induction group was given the vehicle 2 h after the topical imiquimod, once daily for 8 days. Two hours after receiving topical imiquimod, the treatment groups including calcipotriol, norfloxacin 2.5%, and norfloxacin 5% were given topical ointments containing calcipotriol 0.005%, norfloxacin 2.5%, and norfloxacin 5%, for 8 days. Topical norfloxacin ointment significantly reduced the severity of imiquimod-exacerbated psoriatic lesions including erythema, shiny-white scaling, and acanthosis and fixed histological abnormalities. Furthermore, imiquimod-subjected mice treated with a higher concentration of norfloxacin ointment exhibited dramatically lower skin levels of inflammation-related biomarkers like IFN-γ, TNF-α, IL-6, IL-17A, IL-23, and TGF-β but higher levels of IL-10. They also demonstrated a notable decrease in angiogenesis parameters such as VEGF and IL-8, a substantial reduction in oxidative indicators like MDA and MPO, and a considerable rise in antioxidant enzymes like SOD and CAT. This study offers novel evidence that norfloxacin may assist in controlling inflammatory dermatoses like psoriasis by minimizing the severity of psoriatic plaques, correcting histological alterations, and diminishing the production of inflammatory, oxidative, and angiogenetic parameters.

Oleanolic acid rescues critical features of umbilical vein endothelial cells permanently affected by hyperglycemia.

Skin wound healing is a physiological process that involves several cell types. Among them, endothelial cells are required for inflammation resolution and neo-angiogenesis, both necessary for tissue restoration after injury. Primary human umbilical vein endothelial cells (C-HUVECs) are derived from the umbilical cord. When women develop gestational diabetes, chronic exposure to hyperglycemia induces epigenetic modifications in these cells (GD-HUVECs), leading to a permanent pro-inflammatory phenotype and impaired angiogenesis in contrast to control cells. Oleanolic acid (OA) is a bioactive triterpenoid known for its epithelial cell migration promotion stimulation and higher tensile strength of wounds. However, the potentially anti-inflammatory and pro-angiogenic properties of OA are still under investigation. We tested OA on C- and GD-HUVECs under inflammatory conditions induced by low levels of the inflammatory cytokine TNF-α. Reduced expression of adhesion molecules VCAM1, ICAM1, and SELE was obtained in OA-pre-treated C- and GD-HUVECs. Additionally, protein VCAM1 levels were also decreased by OA. Coherently, monocyte adhesion assays showed that a lower number of monocytes adhered to GD-HUVEC endothelium under OA pre-treatment when compared to untreated ones. It is noteworthy that OA improved angiogenesis parameters in both phenotypes, being especially remarkable in the case of GD-HUVECs, since OA strongly rescued their poor tube formation behavior. Moreover, endothelial cell migration was improved in C- and GD-HUVECs in scratch assays, an effect that was further confirmed by focal adhesion (FA) remodeling, revealed by paxillin staining on immunocytochemistry assays. Altogether, these results suggest that OA could be an emergent wound healing agent due to its capacity to rescue endothelial malfunction caused by hyperglycemia.

Abstract 17537: Myocardial Dysfunction is Associated With Impaired Angiogenesis in Patients With Heart Failure With Preserved Ejection Fraction

Background: Recent evidence suggests that myocardial microvascular damage may represent a key parameter in the pathogenesis of heart failure with preserved ejection fraction (HFpEF). Aim: We sought to investigate a potential correlation between parameters of angiogenesis and myocardial dysfunction in HFpEF patients. Methods: We enrolled 30 consecutive HFpEF patients with NYHA class III, left ventricular ejection fraction (LVEF) >50%, E/e' >15, and NT-proBNP >300 pg/ml. In all patients endothelial progenitor cells (EPCs) were mobilized by granulocyte-colony stimulating factor (G-CSF; 10 mcg/kg, 5 days) and collected via apheresis. Before EPC mobilisation we performed echocardiography and measured plasma levels of biomarkers of angiogenesis. All patients underwent electroanatomical mapping for the measurement of myocardial viability; preserved viability was defined as mean unipolar voltage> 8.3 mV. Results: Myocardial viability was preserved in 11 patients (37%; Group A) and decreased in 19 patients (63%; Group B). The groups did not differ in age (63±10 years in Group A vs. 61±8 years in Group B, P=0.78), gender (male: 73% vs. 79%, P=0.70), serum creatinine (1.07±0.34 mg/dL vs. 1.04±0.40 mg/dL, P=0.43), or LVEF (60±7% vs. 57±4%, P=0.32). In Group B we found higher levels of NT-proBNP (2370±1020 pg/mL vs. 920±455 pg/mL in Group A, P=0.04) and E/e’ (23.2±3.1 vs. 17.4±1.8 pg/ in Group A, P=0.02). After G-CSF stimulation, we found significantly lower numbers of EPCs in Group B (53±42 x10 6 cells/L) than in Group A (102±52 x10 6 cells/L, P=0.001). Similarly, apheresis yielded lower numbers of EPCs in Group B (85±82 x10 6 cells vs. 200±93 x10 6 cells in Group A, P=0.003). When compared to Group A, patients in Group B displayed lower levels of pro-angiogenic biomarkers (agiopoietin-1: 7.4±2.8 ng/mL vs. 12.4±5.1 ng/mL, P=0.02; vascular endothelial growth factor: 27.5±20.1 pg/mL vs. 51.6±24.2 pg/mL, P=0.01), and higher levels of anti-angiogenic biomarkers (endostatin: 56.5±19.1 ng/mL vs. 33.2±14.3 ng/mL, P=0.01; thrombospondin-2: 16.4±4.3 ng/mL vs. 9.2±5.0 ng/mL, P=0.03). Conclusion: In patients with HFpEF, myocardial dysfunction appears to correlate with reduced mobilisation of endothelial progenitor cells and impaired angiogenesis.

Fenugreek Seed Ethanolic Extract Improves Alveolar Bone Parameters by Attenuating Inflammation in Ovariectomized Rats.

Alveolar bone residual ridge resorption remains a major challenge for dental implant placement in patients with edentulism. Fenugreek seed extracts have been reported to have potential roles in bone metabolism. This study aimed to evaluate the effects of fenugreek seed ethanolic extract (FSEE) on bone cells, inflammation, hormones, and angiogenesis parameters of alveolar bone tissue following teeth extraction in an ovariectomized (OVX) model. A total of 30 adults female Wistar rats were assigned into two major groups. Each group consisted of control, OVX, OVX+FSEE 100 mg/kg BW, OVX+FSEE 200 mg/kg BW, and OVX+FSEE 400 mg/kg BW. The FSEE treatment was applied through the intragastric route for 7 days in the first group and for 30 days in the second group of animals. The first molar tooth of the right maxilla was extracted before the FSEE treatment. The level of 17β-estradiol was measured by the ELISA method. The dissected maxilla alveolar bone processus was sectioned for histological evaluation by hematoxylin-eosin staining and an immunohistochemistry assay. This study found that FSEE reduced the blood estrogen level and increased estrogen receptor-α (ER-α) expression. FSEE administration modified the number of bone cells, angiogenesis, vascular endothelial growth factor (VEGF), sclerostin, and the osteoprotegerin/receptor activator of nuclear factor kappa-β ligand (OPG/RANKL) ratio. Alterations were seen in the inflammatory markers interleukin-6 (IL-6), transforming growth factor-β1 (TGF-β1), and the macrophage-1/macrophage-2 (M1/M2) ratio. In this study, inflammation was found to be attenuated by reductions in IL-6 and sclerostin, and an increase in TGF-β1. The maturation of bone osteocytes increased along with the increase in ER-α expression and ratio of OPG/RANKL.

In vitro, ex vivo and clinical approaches to evaluate the potential effect of Gentiana lutea extract on skin.

Dark circles affect subjects of all ages and in all skin types. They can be treated by various methods, particular by topical solutions. This investigation was directed towards exploring the effect of gentiopicroside (GP) on the skin around the eyes. For this, an extract of Gentiana lutea (GIE) containing GP (65% by dry matter) was evaluated on oxidant and angiogenesis parameters using in vitro and ex-vivo studies. A clinical experimentation was also realized. The effect of GIE at different concentrations on antioxidant gene was evaluated in vitro by RT-qPCR after treatment of NHDF. The effect of 2.93 μg mL-1 GIE on the release of VEGF-A and VEGF-C by NHDF was also studied. The effect of 87.9 μg mL-1 GIE was also evaluated on pseudotube formation in a coculture system of normal dermal microvascular endothelial cells (HMVEC-d)-NHDF stimulated or not with VEGF as pro-angiogenic factor. Prior to these assays, preliminary cytotoxicity assays were performed using a standard WST-8 reduction assay. The expressions of carboxymethyl-lysine and glyoxalase-1 were quantified on skin explants topically treated with 147 μg mL-1 GIE in basal and UVA-irradiated conditions. A clinical study was conducted in 22 subjects using topical twice daily for 14 days on eye area (split-face application: cream containing 147 μg mL-1 GIE versus placebo). 3D image acquisition and skin colour measurement were performed at D0 and D14. Treatment of GIE upregulated the gene expression of NFE2L2 and downregulated the expression of CXCL8. GIE targeted AGEs pathways and reduced the formation of pseudotubes. A total of 147 μg mL-1 GIE gel cream significantly reduced significantly the average roughness and relief of the upper eyelid skin as well as the redness of dark circles after 14 days of application. By acting on the pathway of AGEs, VEGF-A and VEFG-C, GIE seems to allow a rejuvenation of the skin resulting, among others, in a decrease in redness. It now would be interesting to evaluate the efficacy of GIE on skin around eyes microbiota, antibacterial gentiopicroside property being well-established.

Data dynamics of angiogenesis visualization instrumental methods in patients with psoriasis during methotrexate therapy: Clinical and prognostic value of dermatoscopy and ultrasound dopplerography

BACKGROUND: Pathological angiogenesis is one of the important pathogenetic links of psoriasis. At the same time, the prognostic significance of instrumental methods for studying the severity of angiogenesis in psoriatic plaques remains poorly understood.
 AIM: Clinical assessment of the correlation between the dynamics of angiogenesis parameters (morphometric parameters of vascular glomeruli and Doppler blood flow in the skin and nail bed) and indicators of the methotrexate clinical efficacy during 12 months of therapy.
 MATERIALS AND METHODS: The work is based on the clinical data analysis of 82 patients with moderate and severe psoriasis vulgaris in the acute stage, who were prescribed methotrexate after initial examination. Patients underwent Doppler ultrasound examination of the skin in psoriatic plaques area and digital dermatoscopy to study the characteristics of vascular angiogenesis loci before treatment and at 1, 3 and 12 months of methotrexate therapy.
 RESULTS: As a result of comparative correlation analysis of dermatoscopy and ultrasonographic Dopplerography indicators dynamics and psoriasis clinical index values during the treatment of patients with methotrexate, moderate and strong correlations were established between the values of the average diameter of the vascular glomerulus and the degree of increased blood flow in psoriatic plaque area after 1 month of methotrexate treatment and the values of PASI at 3, 6 and 12 months from the start of treatment (r=0.540.76, p=0.0020.005), as well as between the mean vascular glomerulus diameter values and the degree of skin blood flow increase after 3 months and PASI and BSI indices after 6 and 12 months (r=0.360.74, p=0.0010.036). Disappearance of Doppler signals in psoriatic plaques skin and decrease in vascular glomeruli average diameter after 1 month from the start of therapy in 1.76 [1.38; 2.30] and 1.30 [1.38; 2.30] times increase the probability of achieving clinical improvement after three months of methotrexate treatment. The correlation coefficient between the degree of NAPSI reduction after 6 and 12 months of treatment and the dynamics of Doppler blood flow in the nail bed and resistance index of arterial vessels in the nail bed after three months from the start of treatment varied from 0.53 to 0.73.
 CONCLUSIONS: The dynamics of instrumental methods indicators for studying pathological angiogenesis correlates with the probability of achieving clinical improvement in psoriasis symptoms during systemic methotrexate therapy.

Correlation of density of microvessels and myofibroblasts with the aggressiveness of central giant cell granulomas of jaws: A preliminary study.

Central giant cell granuloma (CGCG) is a fairly common lesion involving the jaw bones. CGCG can show relatively innocuous biological behaviour or it may show clinicoradiological features suggestive of aggressive biological behaviour. To date, there are no histological parameters which can be used to predict the behaviour of these lesions. This study was conducted to assess the utility of parameters of angiogenesis, i.e., total vascular area (TVA), mean vascular area (MVA) and microvessel density (MVD), and density of myofibroblasts in aggressive and non-aggressive CGCGs. The study was undertaken as a retrospective study. A total of 20 previously diagnosed cases (10 non-aggressive and 10 aggressive) of CGCGs were included in the study. The sections were subjected to immunohistochemistry using the markers CD34 and α-SMA. For the assessment of vascular parameters, image J software was used. The density of myofibroblasts was determined in each case ranging from score-1 to 4, using the criteria given by Sridhara et al. The correlation between mean values of vascular parameters and density of myofibroblasts with aggressiveness of CGCG was assessed using Mann-Whitney U test. The result of Mann-Whitney U test suggested that the differences between the values of TVA (P < 0.001), MVA (P < 0.003) and density of myofibroblasts, i.e., SMA mean (P < 0.001) and SMA score (P < 0.001), in two groups are statistically significant. The formula for the assessment of aggressiveness was obtained using discriminant analysis. Angiogenesis and density of myofibroblasts significantly differ in aggressive and non-aggressive cases of CGCGs. The aggressiveness of CGCG case can be predicted using the obtained formula by entering the values of vascular parameters and myofibroblasts.

Pro-Angiogenetic Effects of Purified Extracts from Helix aspersa during Zebrafish Development

Helix aspersa is a species of land snail belonging to the Helicidae family, widespread in the Mediterranean and continental area up to Northern Europe. In some areas it is appreciated as a food, but is mostly considered a parasite of gardens and cultivated fields. The mucus of Helix aspersa has found multiple applications in the cosmetic and health fields. In the present study, we investigated for the first time the angiogenetic properties of purified extracts from Helix aspersa using a transgenic zebrafish line Tg (kdrl:EGFP). The angiogenesis induced by purified snail extracts was demonstrated by their capability to increase the three well-established parameters of angiogenesis: generation of intersegmental vessels, modeling of caudal venous plexus, and formation of sub-intestinal venous plexus. The effects appeared to be mediated by the vascular endothelial growth factor (VEGF) pathway, being prevented by pretreatment of embryos with the selective VEGF receptor antagonist SU5416, and supported by the increased VEGF mRNA levels found in snail-extract-treated embryos. Insufficient vascular supply is underlined by low VEGF signaling, primarily because of its indispensable role in preventing capillary loss. Our findings might have a pharmacological impact by counteracting VEGF hypofunction and promoting angiogenesis to maintain adequate microvascular and vascular density in normal and suffering tissues and organs.

Pro-angiogenic potential of a functionalized hydrogel scaffold as a secretome delivery platform: An innovative strategy for cell homing-based dental pulp tissue engineering.

Angiogenesis is a key process that provides a suitable environment for successful tissue engineering and is even more crucial in regenerative endodontic procedures, since the root canal anatomy limits the development of a vascular network supply. Thus, sustainable and accelerated vascularization of tissue-engineered dental pulp constructs remains a major challenge in cell homing approaches. This study aimed to functionalize a chitosan hydrogel scaffold (CS) as a platform loaded with secretomes of stem cells from human exfoliated deciduous teeth (SHEDs) and evaluate its bioactive function and pro-angiogenic properties. Initially, the CS was loaded with SHED secretomes (CS-S), and the release kinetics of several trophic factors were assessed. Proliferation and chemotaxis assays were performed to analyze the effect of functionalized scaffold on stem cells from apical papilla (SCAPs) and the angiogenic potential was analyzed through the Matrigel tube formation assay with co-cultured of human umbilical vein endothelial cells and SCAPs. SHEDs and SCAPs expressed typical levels of mesenchymal stem cell surface markers. CS-S was able to release the trophic factors in a sustained manner, but each factor has its own release kinetics. The CS-S group showed a significantly higher proliferation rate, accelerated the chemotaxis, and higher capacity to form vascular-like structures. CS-S provided a sustained and controlled release of trophic factors, which, in turn, improved proliferation, chemotaxis and all angiogenesis parameters in the co-culture. Thus, the functionalization of chitosan scaffolds loaded with secretomes is a promising platform for cell homing-based tissue engineering.

Extracorporeal Shock Wave Therapy Salvages Critical Limb Ischemia in B6 Mice through Upregulating Cell Proliferation Signaling and Angiogenesis.

(1) This study tests hypothesis whether extracorporeal shock wave (ECSW) therapy effectively salvages mouse critical limb ischemia (CLI). In vitro result demonstrated that the angiogenesis parameters (i.e., tubular length/cluster/network formation) and protein expressions of EGFR/VEGFR2/RAS/c-Raf/MEK/ERK/VEGF/p-PI3K/p-Akt/p-m-TOR were significantly and progressively increased with stepwise augmentation of ECSW energy (0.1/0.14/0.20 mJ/mm2/140 impulses). On the other hand, they were suppressed by administration of Avastin (20 μM). Adult male B6 mice (n = 24) were equally categorized into group 1 (sham-operated control), group 2 (CLI), group 3 [CLI + ECSW (0.12 mJ/mm2/120 impulses/at days 1/3/7 after CLI induction)] and group 4 [CLI + ECSW (0.12 mJ/mm2/120 impulses) + Avastin (1 mg/intramuscular-injection)] at days 1/3/7 after CLI induction] and quadriceps were harvested by day 14. The laser Doppler result showed that the ratio of left (ischemia) to right (normal) limb blood flow was highest in group 1, lowest in group 2, and significantly higher in group 3 than in group 4 by days 7/14 after the CLI procedure (p < 0.0001). The protein expressions of cell proliferation/migration/angiogenesis receptors (EGFR/VEGFR2), angiogenesis biomarkers (VEGF/CXCR4/SDF-1) and cell proliferation/growth/survival (Ras/c-Raf/MEK/ERK)/(PI3K/Akt/m-TOR) and cell motility/proliferation (p-FAK/p-Scr) signaling biomarkers were significantly higher in group 3 than in groups 1/2/4, and significantly lower in group 1 than in groups 2/4, but they did not show a difference between groups 2 and 4 (all p < 0.001). The small vessel density and cellular levels of endothelial cell surface marker (CD31+) exhibited an identical pattern of blood flow, whereas the angiogenesis (CXCR4+/VEGF+) displayed an identical pattern of VEGFR2 among the groups (all p < 0.0001). The in vitro and in vivo studies found ECSW salvaged the CLI mainly through upregulating Ras-Raf-MEK/ERK/cell motility, cell proliferation/growth pathways and angiogenesis.

TiO2 Nanocrystals and Annona crassiflora Polyphenols Used Alone or Mixed Impact Differently on Wound Repair.

Wounds treated with TiO2 nanoparticles (TiO2-NPs) show an improvement in healing time. However, little is known about the parameters that can contribute to this result. On the other hand, the treatment of wounds with polyphenols is widely known. These compounds are found in the peel of Annona crassiflora fruit and have antioxidant, analgesic and anti-inflammatory properties. In this study, we evaluated the healing effect of TiO2 nanocrystals (TiO2-NCs), polyphenolic fractions obtained from ethanolic extract of A. crassiflora fruit peel (PFAC) and mix (PFAC + TiO2-NCs) on the parameters of wound closure, inflammation, collagen deposition, metalloproteinase activity (MMPs) and angiogenesis. TiO2-NCs and PFAC have activity for wound healing, showed anti-inflammatory action and a shorter wound closure time. These treatments also contributed to increased collagen deposition, while only treatment with TiO2-NCs increased MMP-2 activity, parameters essential for the migration of keratinocytes and for complete restoration of the injured tissue. The combination of PFAC + TiO2-NCs reduced the effectiveness of individual treatments by intensifying the inflammatory process, in addition to delaying wound closure. We conclude that the interaction between the hydroxyl groups of PFAC polyphenols with TiO2-NCs may have contributed to difference in the healing activity of skin wounds.

Significance of HIF-1α and CD105 in establishing oral squamous cell carcinoma associated with oral submucous fibrosis a distinct clinicopathological entity.

Oral squamous cell carcinoma associated with oral submucous fibrosis (OSCC with OSMF) is clinicopathologically a distinct entity. However, scientific proof in view of assessment of biomarkers of hypoxia and neoangiogenesis to differentiate them are lacking. The expression of hypoxia-inducible factor 1-α (HIF-1α) and CD105 in OSCC with and without OSMF possibly will be explicated along these lines. This study aims to evaluate the molecular basis of hypoxia and neoangiogenesis in terms of immunohistochemical expression of HIF-1α and CD105 in OSCC with and without OSMF cases. A retrospective cohort. The study comprise of 203 histopathologically diagnosed surgically operated cases of OSCC retrieved from the departmental archives. The OSCC cases were subgrouped into two, OSCC with OSMF (Group I) and OSCC without OSMF (Group II). The evaluation of hypoxia and angiogenesis was carried out by immunohistochemical markers, HIF-1α and CD105. MVD is the parameter of angiogenesis expressed by CD105. Differences in CD105, and HIF-1α immunoreactivity between study groups were done using descriptive statistics using "Kruskal-Wallis test," "Mann-Whitney test." Statistical significance was set at P < 0.05. On comparison of MVD in Group I and II, statistically significant difference was found in MVD (8.88 ± 3.41, 16.13 ± 5.86, P = 0.0001). The HIF1-α expression was less in Group I (6.85 ± 2.62) as compare to Group II (7.22 ± 3.08) but the difference was statistically nonsignificant (P = 0.35). The OSCC with OSMF is not only clinicopathologically distinct entity of OSCC but also diverse in its molecular pathogenesis as explicited by distinct expression of HIF-1 α and CD105.

Effects of remote ischemic preconditioning (RIPC) and chronic remote ischemic preconditioning (cRIPC) on levels of plasma cytokines, cell surface characteristics of monocytes and in-vitro angiogenesis: a pilot study

Remote ischemic preconditioning (RIPC) protects the heart against myocardial ischemia/reperfusion (I/R) injury and recent work also suggested chronic remote ischemic conditioning (cRIPC) for cardiovascular protection. Based on current knowledge that systemic immunomodulatory effects of RIPC and the anti-inflammatory capacity of monocytes might be involved in cardiovascular protection, the aim of our study was to evaluate whether RIPC/cRIPC blood plasma is able to induce in-vitro angiogenesis, identify responsible factors and evaluate the effects of RIPC/cRIPC on cell surface characteristics of circulating monocytes. Eleven healthy volunteers were subjected to RIPC/cRIPC using a blood pressure cuff inflated to > 200 mmHg for 3 × 5 min on the upper arm. Plasma and peripheral blood monocytes were isolated before RIPC (Control), after 1 × RIPC (RIPC) and at the end of 1 week of daily RIPC (cRIPC) treatment. Plasma concentrations of potentially pro-angiogenic humoral factors (CXCL5, Growth hormone, IGFBP3, IL-1α, IL-6, Angiopoietin 2, VEGF, PECAM-1, sTie-2, IL-8, MCSF) were measured using custom made multiplex ELISA systems. Tube formation assays for evaluation of in-vitro angiogenesis were performed with donor plasma, monocyte conditioned culture media as well as IL-1α, CXCL5 and Growth hormone. The presence of CD14, CD16, Tie-2 and CCR2 was analyzed on monocytes by flow cytometry. Employing in-vitro tube formation assays, several parameters of angiogenesis were significantly increased by cRIPC plasma (number of nodes, P < 0.05; number of master junctions, P < 0.05; number of segments, P < 0.05) but were not influenced by culture medium from RIPC/cRIPC treated monocytes. While RIPC/cRIPC treatment did not lead to significant changes of the median plasma concentrations of any of the selected potentially pro-angiogenic humoral factors, in-depth analysis of the individual subjects revealed differences in plasma levels of IL-1α, CXCL5 and Growth hormone after RIPC/cRIPC treatment in some of the volunteers. Nevertheless, the positive effects of RIPC/cRIPC plasma on in-vitro angiogenesis could not be mimicked by the addition of the respective humoral factors alone or in combination. While monocyte conditioned culture media did not affect in-vitro tube formation, flow cytometry analyses of circulating monocytes revealed a significant increase in the number of Tie-2 positive and a decrease of CCR2 positive monocytes after RIPC/cRIPC (Tie-2: cRIPC, P < 0.05; CCR2: RIPC P < 0.01). Cardiovascular protection may be mediated by RIPC and cRIPC via a regulation of plasma cytokines as well as changes in cell surface characteristics of monocytes (e.g. Tie-2). Our results suggest that a combination of humoral and cellular factors could be responsible for the RIPC/cRIPC mediated effects and that interindividual variations seem to play a considerable part in the RIPC/cRIPC associated mechanisms.

The combined effect of pomegranate extract and tangeretin on the DMBA-induced breast cancer model

The aim of this study was to investigate the protective effects of pomegranate extract and tangeretin alone or in combination in DMBA-induced rat breast cancer model.A total of 68 female rats were randomly divided into 8 groups. The first 4 groups were designed as controls for cancer and treatment groups, and the control groups were composed of only control (C), Pomegranate (P), Tangeretin (T), and Pomegranate+Tangeretin (P+T) groups. The other four groups were designed as cancer and treatment groups and were composed of DMBA (D) and DMBA+Pomegranate (D+P), DMBA+Tangeretin (D+T), DMBA+Pomegranate+Tangeretin (D+P+T) groups. Tumor markers and angiogenesis parameters were studied from plasma samples obtained from rats. Histopathological, immunohistochemical, and TUNEL analyses and expressions of proteins affecting apoptosis and cell cycle were determined in breast tissue samples.In the DMBA group, plasma CA15-3, CEA, VEGF, MMP-9, and NF-κB levels were significantly increased compared to the controls, but significant decreases were observed in these parameters except MMP-9 in the treatment groups. It was observed that p53 and Bax expressions significantly increased in both D+P and D+P+T groups compared to the DMBA group, and these findings were supported by Tunel and immunohistochemical findings. Cyclin D1 expressions were found to be significantly decreased only in the D+T group and supported by TUNEL and immunohistochemical findings. Immunohistochemical ER-α and Ki-67 immune reactivities were significantly decreased in all treatment groups compared to the DMBA group.Our results showed that combined application of pomegranate extract and tangeretin may be more beneficial in preventing breast cancer development.

The Dependence between Urinary Levels of Angiogenesis Factors, 8-Iso-prostaglandin F2α, ɣ-Synuclein, and Interleukin-13 in Patients with Bladder Cancer: A Pilot Study.

During the last decade, a significant increase in the incidence of bladder cancer (BC) has been observed. Angiogenesis plays a key role in the process of tumor growth and metastasis. Additionally, the participation of oxidative stress and chronic inflammation in BC pathogenesis is indicated. The aim of the study was to evaluate the urinary levels of parameters of angiogenesis, stimulating angiogenin (ANG) and inhibiting angiostatin (ANGST), 8-iso-prostaglandin F2α (8-iso-PGF2α) as a marker of oxidative stress, ɣ-synuclein (SNCG) as a cancer progression parameter, and interleukin-13 (IL-13) as an anti-inflammatory immunomodulator. The levels of ANG, ANGST, 8-iso-PGF2α, SNCG, and IL-13 in the urine of BC patients and healthy controls were measured by the enzyme-linked immunosorbent assay. These parameters were examined in the whole group of BC patients and in subgroups depending on the clinical stage: nonmuscle-invasive bladder cancer (NMIBC) and muscle-invasive bladder cancer (MIBC); histopathologic malignancy: low grade (LG) and high grade (HG) and in primary and recurrent BC. Significantly, higher urinary parameters were found in BC patients in comparison to controls. Levels of all parameters increased with the development of cancer, with the exception of 8-iso-prostaglandin F2α, in which the level was higher in the early stages of the disease, but these differences were not statistically significant. Some correlations have been demonstrated between parameters in BC patients. Based on the receiver operating characteristic curves, ANG and ANGST had the best diagnostic value for BC. The obtained results indicate the important role of the examined parameters of angiogenesis, oxidative stress, and inflammation in the pathogenesis and development of BC. It is reasonable to continue research in order to thoroughly assess the impact of various associated processes on the course of BC. It is also important to carry out similar tests in patients with other urological diseases.

Evaluation of the Healing Effects of Hypericum perforatum and Curcumin on Burn Wounds in Rats.

Background For centuries, medicinal plants have been extensively used in wound healing of burn injuries. The aim of this study is to analyze comparatively the effects of curcumin and Hypericum perforatum (HP) on second-degree burn wounds in rats. Materials and Methods. This experimental study was conducted on 24 male Sprague-Dawley rats with second-degree burns. The animals were randomly divided into three groups. The burns were treated with curcumin (Group B) and Hypericum perforatum (HP) on second-degree burn wounds in rats. Results All histological parameters of the control group showed statistically significant difference than the other groups (p < 0.05). There was no statistically significant difference between Groups B and C in terms of reepithelization and inflammation (p < 0.05). There was no statistically significant difference between Groups B and C in terms of reepithelization and inflammation (p < 0.05). There was no statistically significant difference between Groups B and C in terms of reepithelization and inflammation (p < 0.05). There was no statistically significant difference between Groups B and C in terms of reepithelization and inflammation (Conclusion Both curcumin and Hypericum perforatum oil are effective in burn wound healing. Our findings showed a better quality of healing in curcumin-treated rats.Hypericum perforatum (HP) on second-degree burn wounds in rats.

In Vivo Anticancer Activity of Cleome viscose Linn. alcoholic extract and its fractions against Ehrlich’s Ascites Carcinoma (EAC) Cell Line

The investigation aimed to evaluate the anti-cancer properties of alcoholic extracts and the fractions Cleome viscose Linn. against Ehrlich Ascites Carcinoma (EAC) cell lines in Swiss albino mice. In the case of EAC tumor, 24 hours after tumor inoculation, the extract and fractions are administered every day within 14 days. On the fifteenth day the mice were sacrificed to monitor antitumor activity. The effect of alcoholic extracts and their fraction in terms of mean survival time, percentage increase in life span, spleen weight, ascitic fluid volume, ascitic fluid volume and angiogenesis of EAC-bearing mice and simultaneous alteration in hematological profile were estimated. Alcoholic extracts and their fractures exhibited an impact on mean survival time, percentage increase in life span, spleen weight, ascitic fluid volume, ascitic fluid volume, angiogenesis and haematological parameters in EAC tumor bearing mice. Hematological profile was reverted to normal level in the extracts and their fractions treated mice. From the current study of alcoholic extracts and its fraction of Cleome viscose Linn. exhibited antitumor action in a dose dependent manner comparable to that of standard drug. So, the current research provides a scientific basis for the curative use of Cleome viscose Linn. Which are largely attributable to the improver or synergies effect of their constituents.

Comparative Evaluation of the Cytotoxic and Angiogenic Effects of Minocycline and Clindamycin: An In Vitro Study

IntroductionThis study aimed to compare the cytocompatibility and angiogenic potential of 2 antibiotics (clindamycin [CLIN] and minocycline [MINO]) at distinct concentrations on dental pulp stem cells (DPSCs) and human umbilical vein endothelial cells (HUVECs). MethodsDPSCs and HUVECs were exposed to cell culture media modified with CLIN or MINO at concentrations ranging from 30 μg/mL–1000 μg/mL. Cell toxicity and proliferation were investigated using the lactate dehydrogenase and tetrazolium reduction assays, respectively. A capillarylike tube formation in vitro assay was conducted to determine the angiogenic potential associated with each antibiotic. Additionally, selected morphometric angiogenesis parameters were determined using dedicated software (WimTube; Onimagin Technologies SCA, Córdoba, Spain). All statistical analyses were performed using 1-way analysis of variance and the Tukey post hoc test (α= .05). ResultsThe collected data showed that compared with the control (cell culture media, alpha-minimum essential medium Eagle) increasing the antibiotic concentration significantly decreased cell viability and proliferation of both DPSCs and HUVECs. In terms of angiogenic potential, when tested at 30 μg/mL and 50 μg/mL, CLIN significantly amplified tube formation when compared with MINO with angiogenesis parameters (ie, tube length and tube number) similar to the effect promoted by exogenous vascular endothelial growth factor (50 ng/mL). ConclusionsCLIN was less cytotoxic when compared with MINO at higher concentrations. Of note, CLIN did not hinder the proangiogenic activity induced by vascular endothelial growth factor to the same extent as MINO, suggesting that the replacement of MINO by CLIN might translate into positive implications in the overall regenerative outcome.

Maternal cigarette smoking and expression of selected micro-RNAS in the plasma collected after spontaneous vaginal delivery

Several studies have shown a gradual increase in the extent of bone marrow angiogenesis in various stages of proliferative plasma cell disorders, from monoclonal gammopathy of undetermined significance (MGUS) to active multiple myeloma (MM). The main aim of this study was to evaluate tumor angiogenesis parameters in detail and to correlate them with the expression of osteopontin (OPN) and vascular endothelial growth factor (VEGF) in the bone marrow of patients with MGUS and MM. In addition, we wanted to determine their prognostic significance in active MM.Ninety-five patients were enrolled in the study: 14 diagnosed with MGUS, 13 with asymptomatic myeloma (AMM) and 68 with active MM. Computer assisted image analysis was used to determine the angiogenesis parameters, the quantity of microvessels per 1 mm2 (MVD), the area occupied by microvessels per 1 mm2 and the percentage of microvessel area in total section area (TVA). Double immunohistochemical methods CD138 + VEGF and CD138 + OPN were used to evaluate expression of these proteins in plasma cells, and OPN was also analyzed for its interstitial expression (iOPN).A significant positive correlation was determined between VEGF and iOPN with angiogenic parameters in the MGUS stage of the disease. In advanced stages of the disease, a significant negative correlation was recorded between OPN and iOPN with parameters of angiogenesis. Overall survival was significantly shorter for patients with negative iOPN (p = 0.002) and higher angiogenic parameters, MVD (p = 0.009), TVA (p = 0.008) and area of microvessels per 1 mm2 (p = 0.02). Positive VEGF expression in our model predicted a better three-year survival of patients with active MM (OR: 5.25, p = 0.03; HR: 0.44, p = 0.04).The results of our study suggested a possible key role of VEGF and OPN in the induction of angiogenesis in early-stage disease.

The relationship between the rheological behavior of RBCs and angiogenesis in the morbidly obese.

The aim of this study was to investigate the relationship between red blood cell (RBC) aggregation and deformability and angiogenesis parameters in obese patients. We studied 35 obese subjects and 20 non-obese people as a control group. Angiogenesis was detected using Bio-Plex Pro Human Angiogenesis Multiplex Assays. The RBC aggregation and deformability of the red blood cell aggregation were performed by the Laser-assisted Optical Rotational Cell Analyser - LORCA. The aggregation index and the syllectogram's amplitude were significantly higher in the obese patients, whereas the aggregation half-time (t1/2) was lower compared with the control group. The deformability of RBC expressed as EI was significantly lower in the obese group than it was in the control group. All angiogenesis parameters were higher in obese individuals than they were in the control group. Significant differences were observed in angiopoietin 2 (p = 0.048), folistin (p = 0.0017), G-CSF (p = 0.042), HGF (p = 0.016), and PECAM-1 (p = 0.014). The VEGF tended to be higher in the obese patients than in the control group (p = 0.09); nevertheless, the concentration of PDGF-BB was similar in both groups. EI at shear stresses of 18.49 Pa and 30.2 Pa was strongly correlated with all angiogenesis parameters. No correlations were found between the studied RBC aggregation indices and angiogenesis parameters. Multivariate analyses indicated that only HGF was an independent predictor of RBC deformability at 18.49 Pa (β-0.83, P < 0.000005) and at 30.2 Pa (β-0.83, P < 0.00005). The study found that there are relationships between enhanced RBC rigidity and angiogenesis status in obese subjects. Because this correlation between angiogenesis and RBC deformability is presented for the first time, the physiological importance of the relationship requires further research.