Degree Of Neovascularization Research Articles

Effects of different preparation protocols of blood serum eyedrops on corneal healing after alkali burn: Clinical evaluation and α-SMA expression

Background: In cases of alkali burn, blood serum eyedrops may supply protein and growth factors. A standard preparation protocol is necessary for consistency in epitheliotropic capacity. Objective: We aim to evaluate how post-alkali-burn administration of blood serum eyedrops from different preparation protocols results in different outcomes on rabbits. Methods: This randomized experimental study used three treatments: placebo, non-diluted serum, and 25% concentration serum. We analyzed the corneal haziness, neovascularization degree, histopathological examination of neovascularization scores, and alpha-smooth muscle actin (α-SMA) expression. Results: Twelve rabbits (24 eyes) were split evenly into three treatment groups. On day 14, the group receiving the 25% concentration serum had the least corneal haziness (p = 0.005). No adverse effect was observed on the treated eyes. Histopathologically, no significant difference in neovascularization scores and α-SMA expression was observed. Conclusions: Treatment using the 25% concentration serum resulted in less corneal haziness, smaller defect size, and greater healing rate. There was also no significant difference in histopathological outcomes among the three treatment groups.

Accelerated vascularization of a novel collagen hydrogel dermal template.

Full thickness skin loss is a debilitating problem, most commonly reconstructed using split thickness skin grafts (STSG), which do not reconstitute normal skin thickness and often result in suboptimal functional and esthetic outcomes that diminish a patient's quality of life. To address the minimal dermis present in most STSG, engineered dermal templates were developed that can induce tissue ingrowth and the formation of neodermal tissue. However, clinically available dermal templates have many shortcomings including a relatively slow rate and degree of neovascularization (∼2-4weeks), resulting in multiple dressing changes, prolonged immobilization, and susceptibility to infection. Presented herein is a novel composite hydrogel scaffold that optimizes a unique scaffold microarchitecture with native hydrogel properties and mechanical cues ideal for promoting neovascularization, tissue regeneration, and wound healing. In vitro analysis demonstrated the unique combination of improved mechanical attributes with native hydrogel properties that promotes cell invasion and remodeling within the scaffold. In a novel 2-stage rat model of full thickness skin loss that closely mimics clinical practice, the composite hydrogel induced rapid cell infiltration and neovascularization, creating a healthy neodermis after only 1week onto which a skin graft could be placed. The scaffold also elicited a gradual and favorable immune response, resulting in more efficient integration into the host. We have developed a dermal scaffold that utilizes simple but unique collagen hydrogel architectural cues that rapidly induces the formation of stable, functional neodermal tissue, which holds tremendous promise for the treatment of full thickness skin loss.

Angiographic characteristics of vasculopathy in patients with idiopathic inflammatory myopathies and systemic sclerosis

To describe the peripheral angiographic features of vasculopathy in idiopathic inflammatory myopathies (IIM) and systemic sclerosis (SSc) in comparison to polyarteritis nodosa (PAN). Angiograms of 47 extremities (24 upper and 23 lower) of 11 patients with IIM (n = 5) and SSc (n = 6), and 12 patients with PAN who presented with critical limb ischemia were retrospectively analyzed with regards to the presence of stenosis, occlusion, aneurysms and delayed distal flow, and degree of neovascularization. Diffuse narrowing was more frequent (66.1 vs. 38.0%, p = 0.001), whereas multifocal stenosis (6.5% vs. 26.8%, p = 0.002), abrupt occlusion (11.3% vs. 29.6%, p = 0.010) and aneurysm formation (1.6% vs. 11.3%, p = 0.037) were less frequent in IIM/SSc than PAN. In distal arteries, tapered occlusion (95.5% vs. 76.0%, p = ns) and delayed flow (77.3% vs. 48.0%, p = 0.039) were more common in IIM/SSc than PAN. After 1 year, auto- or surgical amputation tended to be more frequent in IIM/SSc than PAN (36.4% vs. 16.7%, p = ns). In conclusion, diffuse narrowing, tapered occlusion and delayed distal flow on conventional angiograms tend to be more frequent in IIM/SSc than PAN. Further studies are needed to verify these findings in a larger prospective cohort.

The Impact of Tumor-associated Macrophages on ChemoresistanceviaAngiogenesis in Colorectal Cancer

The tumor microenvironment plays an important role in tumor progression. Tumor-associated macrophages (TAMs) have been reported to promote proliferation, invasion, metastasis, angiogenesis, and immunosuppression. Furthermore, angiogenesis has been reported to induce chemoresistance due to the inefficient distribution of drugs to cancer cells. However, the impact of TAMs on chemoresistance via angiogenesis in colorectal cancer (CRC) remains unclear. The aim of the study was to evaluate the impact of TAMs on the chemotherapeutic outcome in CRC. We enrolled 54 patients who underwent chemotherapy for unresectable metastatic CRC after resection of the primary tumor. We evaluated the density of TAMs and the degree of angiogenesis by immunohistochemistry and then explored the correlation between the density of TAMs and chemotherapeutic outcome. Furthermore, we assessed any correlation between the density of TAMs and that of neovascularity. The high-TAMs group had a significantly worse progression-free survival (p=0.0006) and a poorer response rate (p=0.0274) than the low-TAMs group. In addition, a positive correlation was observed between the density of TAMs and the degree of neovascularity (r=0.665, p=0.0004). TAMs were shown to promote chemoresistance via angiogenesis in CRC.

Anti-thrombotic and anti-inflammatory activity of sulodexide compared to aspirin in the rat model.

Although the number of vascular surgeries performed is increasing, the incidence of complications associated with this surgery has not improved and re-operations are frequently required. Thrombosis in a vessel is the most hazardous postoperative complication. The aim of this study was to evaluate the anti-thrombotic and anti-inflammatory effects of sulodexide compared to aspirin in a rat model. We divided the animals into three groups (sham (saline), aspirin, and sulodexide). The abdominal aorta was surgically opened and closed, primarily with 8/0 Prolene sutures. Postoperatively, saline, aspirin, or sulodexide was administered by oral gavage for 14 days to the rats. The degree of neovascularization, thrombus, calcification, inflammatory infiltrates, and fibrosis were analyzed histopathologically by hematoxylin and eosin staining. There was no significant difference in the incidence of postoperative thrombogenesis, but less calcification and inflammatory infiltrates were observed in the sulodexide group compared to the aspirin group. Histopathologic score revealed less infiltration of inflammatory cells and mild calcification for the sulodexide group (0.17±0.41 and 1.33±0.52, respectively) compared to the aspirin group (0.67±0.52 and 1.67±0.52, respectively) at days 14. This study offers the possibility that sulodexide could be used as an aspirin substitute for the postoperative management of vascular patients, with low gastrointestinal discomfort. In addition, it may also offer reduced postoperative calcification and inflammation.

Endoscopic suturing promotes healing of mucosal defects after gastric endoscopic submucosal dissection: endoscopic and histologic analyses in in vivo porcine models (with video)

Endoscopic suturing of mucosal defects after endoscopic submucosal dissection (ESD) is expected to prevent postoperative adverse events. We aimed to endoscopically and histologically evaluate the healing process of post-ESD mucosal defects closed with endoscopic hand suturing (EHS) in invivo porcine models. Twelve mucosal defects (2 cm in size) were created in 2 pigs (6 defects per pig). Initially, 2 defects were created: one was closed with EHS (sutured group) and the other was kept open (control group). On postoperative days (PODs) 7 and 14, 2 additional defects were created in each session, and they were treated in the same manner as in the initial procedure. On POD 21, the entire stomach, with the 6 lesion sites, was extracted for histologic evaluation after endoscopic observation. Endoscopically, all sutured sites remained closed in all sessions (PODs 7, 14, and 21). Histologically, on POD 14, the epithelium and muscularis mucosae were appropriately connected. The mucosae were covered with the epithelium without inversion of the mucosal edge in the sutured group, whereas the ulcer bed was exposed in the control group. Furthermore, the degree of neovascularity and fibroblasts in the submucosa was smaller in the sutured group than that in the control group. Our findings suggest that endoscopic suturing promotes healing of post-ESD mucosal defects histologically in invivo porcine models. Thus, endoscopic mucosal closure after ESD might be clinically useful for the prevention of delayed perforation/bleeding if secure suturing is performed endoscopically.

Fibrosarcoma in the hock joint of an ostrich (Struthio camelus) - A case report

A one-year old female ostrich (Struthio camelus) was presented for surgical excision with the history of growth near left hock joint (ankle) and progressively increasing in size. The tumorous mass was surgically excised after sedation and local infiltrative anesthesia. Histopathological examination of the mass revealed a large number of fibrocytes and fibroblasts arranged in an irregular manner accompanied with high degree of neovascularisation. Few areas also showed infiltration of inflammatory cells, degenerative fibroblast cells, high mitotic figures and extensive local invasion to surrounding soft tissues. These features were indicative of clear malignancy and condition was diagnosed as a case of fibrosarcoma.

P3394Carotid plaque neovascularization correlates to increased intraplaque blood-like tissue in acute coronary syndrome

Abstract Background The assessment of carotid plaque composition has attracted great interest as a tool to predict plaque vulnerability and subsequent risk of cardiovascular events. These vulnerable plaque lesions are characterized by the presence of intraplaque neovascularization (IPN). Previously, grayscale pixel ranges of ultrasound images have been correlated to various human tissue types. Thus, using these ranges, it is possible to assess the composition of atherosclerotic plaque non-invasively. Purpose This study sought to determine the relationship between carotid plaque composition and the degree of neovascularization in patients presenting with acute coronary syndrome (ACS), when compared to patients with stable angina. Methods A carotid focused vascular ultrasound was performed in 92 participants who had recently undergone coronary angiography due to stable angina or ACS. In 81 participants found to have atherosclerotic plaque in at least one carotid artery, plaque composition was assessed for tissue-like types by pixel distribution analysis: grayscale ranges 0–4 (blood), 8–26 (fat), 41–76 (muscle), 112–196 (fibrous), and 211–255 (calcium) (Figure 1). Participants also received microbubble contrast to assess IPN. IPN was graded based on the presence and location of microbubbles within each plaque (0, not visible; 1, peri-adventitial; 2, plaque core) and averaged to obtain an overall score. An independent t-test was used to compare continuous variables. Pearson pairwise correlations were used to assess associations between IPN and tissue composition. Results In the overall sample population, increased average IPN score correlated with increased plaque % blood tissue type (rho=0.28; p=0.01). Patients presenting with ACS (n=32) had increased neovascularization (IPN score of 1.47 vs 1.09; p=0.4), compared to patients with stable angina. In addition, ACS patients with an IPN score ≥1.25 had higher levels of plaque % blood (1.97% vs 0.75%; p=0.02).In ACS patients only, % blood increased with IPN score (r=0.34; p=0.06) and % fibrous tissue decreased with IPN score (r=−0.42; p=0.02). Figure 1 Conclusion The proportion of blood-like tissue in carotid plaque is associated with neovascularization and is increased in ACS patients. This may indicate hemorrhage within the plaque leading to rupture. Increased fibrous tissue may have a protective role, stabilizing the plaque when present. This data suggests that carotid plaque composition may be used as an imaging biomarker for cardiovascular risk, and its incorporation into routine screening practices may improve patient risk stratification. Acknowledgement/Funding CIHR CGS-Doctoral Award, Heart and Stroke Foundation of Canada, Canada Foundation for Innovation

Assessment of Laparoscopically Harvested Omental Flap Used in Immediate Reconstruction in Breast Cancer Cases Eligible for Breast Conservative Surgery

AbstractBackground and Purpose: Breast conservative surgery is considered a standard surgical modality for treatment of early breast cancer. Omental pedicled flap is used for volume replacement after breast conservative surgery.Aim of Study: The aim of this study was to assess the fate of laparoscopically harvested omental flap used in immediate reconstruction in breast cancer cases eligible for breast con-servative surgery.Patients and Methods: Ten female patients were included in this retrospective study which was conducted at The Surgical Oncology Unit, Department of General Surgery, Tanta Uni-versity Hospitals, from June 2016 to July 2018. Every patient was subjected to mammography with complementary ultra-sonography, Doppler ultrasound and Tru-cut needle biopsy from the omental flap and histopathology with different stains.Results: Nodules were detected in 40% of cases which were proved to be benign in nature by Histopathology. Doppler revealed that two cases had partial infarction and degeneration of the omental flap. Histopathology revealed minimal inflam-matory reaction, extensive fat necrosis and dense fibrosis in 50% of the studied cases. Vasculitis was detected in 30% of the studied cases. No recurrence of malignancy could be detected over the follow-up period. With CD31, various degrees of neovascularization were detected in the omental flap, mild in 20% of cases, moderate in 40% of cases and severe in 40% of cases. With CD8, T-Killer cells were found mild in 50% of cases and moderate in 40% of cases. CD44 detected presence of stem cells in 20% of cases.Conclusion: The laparoscopically harvested omental flap had a promising oncological safety during the follow up. In spite of the fibrosis and nodulation found, no change was observed in the shape or contour of the breast. The inflamma-tory reaction and enhanced local immunity by the omentum is not studied as a possible preventive factor for recurrence. Vasculitis, angiogenesis and the presence of stem cells are not studied as a possible potentiating factor for recurrence of breast cancer. So, long term follow-up is essential to see whether the omental flap could be a barrier against recurrence or it may enhance it.

The effectiveness and safety of topical 0.1% tacrolimus after high-risk penetrating keratoplasty

Objective: To observe the effectiveness and safety of topical 0.1% tacrolimus(FK506) as immunosuppressant in high-risk penetrating corneal transplantation to prevent the immune rejection and to compare the outcomes with topical 1% Cyclosporin A (CsA). Methods: The study consists of 49 patients (50 eyes), who were fitted with the high-risk corneal transplantation standard and undergone the penetrating keratoplasty(PKP) or combined operation in Beijing Tongren hospital between March 2015 to September. With the time sequence, the patients were divided into observation group (FK506 group) and the control group (CsA group). The observation group included 9 females and 16 males with an average age of 57.8±14.8. Twenty-four patients were in the control group (25 eyes), including 10 females and 14 males, with an average age of (45.1±16.2). Observation group was treated with topical 0.1% tacrolimus, and the control group treated with topical 1%CsA. Both groups' treatment combined glucocorticoid as well. Two groups had 1 year follow-up observation. The incidence of rejection was compared by statistical methods of Cox regression. The adverse reactions were graded and compared using Mann-Whitney U test. Results: After one year, 22 cases of the observation group and 23 cases of the control group were accomplished all observations. The rejection rate was 4.54% in observation group and 27.23% in control group. The difference between the groups was statistically significant (χ(2)=4.291, P=0.038). Control group had high rejection rate. Besides, there was no severe side effects happened in both groups. After 1 month after surgery, 36.4% of the observation group showed mild corneal edema. The ratio of mild to moderate corneal edema in the control group was 26.1% and 8.7%. Three months after surgery, 4.5% of the observation group showed mild corneal edema, while 13.0% and 13.0% of the control group was found mild to moderate corneal edema. Six months after surgery, 4.5% of the observation group showed moderate corneal edema. The ratio of mild, moderate to severe corneal edema in the control group was 17.4%, 17.4% and 8.7%. The degree of corneal edema in the control group was more serious in three monthes(Z=-2.770, -2.018, -2.941, P<0.05). The differences in both monthes were statistically significant. Mild neovascularization occurred in the 13.6% of observation group. Mild to severe neovascularization occurred in the 13.0%, 4.3%, and 4.3% control groups. The degree of neovascularization in the control group was higher than that in the observation group(Z=-3.221, P=0.001). The differences in both months were statistically significant. Mild to moderate neovascularization occurred in the 18.2% and 9.1% of observation group. Mild to extremely severe neovascularization occurred in the 17.4%, 26.1%, 4.3% and 4.3% control groups. The degree of neovascularization in the control group was higher than that in the observation group(Z=-1.988, P=0.047).The differences in both monthes were statistically significant. Conclusions: Both 0.1% tacrolimus and 1% cyclosporine A are safe and effective in reducing the rejection after high-risk corneal transplantation. (Chin J Ophthalmol, 2019, 55: 419-427).

Using Pressure Massage for Achilles Tendinopathy: A Single-Blind, Randomized Controlled Trial Comparing a Novel Treatment Versus an Eccentric Exercise Protocol

Background:Eccentric exercises are the only conservative treatment that has shown good clinical results in studies of Achilles tendinopathy (AT), but success rates vary, indicating the need for alternative treatments. Soft tissue treatments are widely used for AT, but strong scientific evidence is lacking to support those treatments.Purpose/Hypotheses:This study aimed to determine whether pressure massage to the calf muscles is a useful treatment for AT and to compare this treatment versus an eccentric exercise protocol. Our first hypothesis was that pressure massage treatment is equivalent or superior to eccentric exercises with regard to pain reduction time (ie, pain would be reduced more quickly with pressure massage). The second hypothesis was that pressure massage is equivalent or superior to eccentric exercises with regard to function of the calf muscles.Study Design:Randomized controlled trial; Level of evidence, 1.Methods:A total of 60 patients with AT were randomized into 3 groups: group 1 underwent an eccentric exercise protocol, group 2 underwent pressure massage, and group 3 underwent pressure massage and the eccentric exercise protocol. Patients were evaluated with the Icelandic version of the Victorian Institute of Sports Assessment–Achilles questionnaire (VISA-A-IS), an algometer to test the pressure pain threshold (PPT) of the Achilles tendon, tests for ankle range of motion (ROM), and real-time ultrasonographic (US) scanning of tendon thickness and degree of neovascularization. Measurements for VISA-A-IS, PPT, and ROM were taken at 0, 4, 8, 12, and 24 weeks. US scan measurements were taken at 0, 12, and 24 weeks. Mixed-model analysis of variance was used for statistical analysis.Results:All groups improved when evaluated with VISA-A-IS scores (P < .0001). The pressure massage group improved significantly more than the eccentric exercise group at week 4, which was the only between-group difference. Ankle ROM increased significantly over time (ROM bent knee P = .006 and ROM straight knee P = .034), but no significant difference was found between groups. No significant difference was found in evaluations of PPT or US scan measurements.Conclusion:Pressure massage is a useful treatment for Achilles tendinopathy. Compared with eccentric exercise treatment, pressure massage gives similar results. Combining the treatments did not improve the outcome.

FA05.05: PROLONGED LAPAROSCOPIC GASTRIC ISCHEMIC CONDITIONING PRIOR TO ESOPHAGECTOMY IN PATIENTS OF ESOPHAGEAL CANCER: FEASIBILITY AND CLINICAL SIGNIFICANCE

Abstract Background The studies on gastric ischemic preconditioning in improving blood flow to the distal gastric conduit and reducing the incidence of anastomotic complications have yielded variable results. Lack of adequate time interval between gastric ischemic preconditioning and esophagectomy has been postulated as one of the reasons for lack of clinical benefit. Hence the present study was done to evaluate the feasibility and clinical outcome of prolonged gastric ischemic conditioning prior to esophagectomy. Methods Patients with locally advanced squamous cell carcinoma of the esophagus planned for neoadjuvant chemoradiotherapy followed by esophagectomy and gastric pull through were included in the prospective study. Prior to initiation of neoadjuvant therapy laparoscopic feeding jejunostomy with ischemic preconditioning by the ligation of left and the short gastric vessel was performed. All patients underwent thoracolaparoscopic esophagectomy with two field esophagectomy and cervical esophagogastric anastomosis using modified Orringer technique. Radiological assessment of the caliber of the right gastroepiploic artery (RGEA) and Immunohistologic analysis of Hypoxia-inducible factor – 1 alpha (HIF-1alpha) expression in the fundus to determine the degree of neovascularization was performed after ischemic preconditioning. The feasibility of laparoscopic gastric mobilization post ischemic preconditioning and cervical anastomotic leak rate was documented and compared with patients who did not undergo ischemic preconditioning. Results Fifteen patients underwent thoracolaparoscopic esophagectomy after a median time of 124 days post laparoscopic gastric ischemic preconditioning. Minor adhesions present in the gastrosplenic region did not interfere with laparoscopic gastric mobilization. The mean pre-ischemic conditioning diameter of RGEA increased from 2.18 mm to 2.25 mm post ischemic conditioning (P = 0.21). The median HIF-1 alpha index post ischemic conditioning was 0.6. The cervical anastomotic leak rate was less in patients who underwent ischemic preconditioning (1/14, 7.1%) compared to the patients who did not undergo ischemic preconditioning (6/16, 37.5%), although the difference was not statistically significant (P = 0.24) Conclusion Prolonged gastric ischemic preconditioning prior to esophagectomy is safe and does not interfere with laparoscopic gastric mobilization. It can potentially reduce cervical anastomotic complications post neoadjuvant chemoradiotherapy. Disclosure All authors have declared no conflicts of interest.

The efficacy of moxifloxacin in patients with bacterial keratitis

Background/Aim. Keratitis is a sociomedical problem of moderately developed countries. including Serbia, too. The incidence of bacterial keratitis in the world is about 20% in relation to all keratitis, but its frequency in Serbia is still not known. Bearing in mind the complications in the front segment of the eye after bacterial keratitis (ulcer, neovascularization, fibrosis) and decline in visual acuity, it was necessary to assess the efficacy of local therapy by moxifloxacin which will shorten the healing time and reduce complications. The aim of the study was to analyze the efficiency of shortening the treatment of bacterial keratitis with moxifloxacin. Methods. The study was designed as prospective, randomized, double-blind study. The study included 30 patients with diagnosed keratitis and locally applied antibiotic moxifloxacin, and 60 patients in a control group, with locally applied artificial tears. All participants were subjected to complete clinical ophthalmologic analysis (2015/16), for a period of 1?15 days after the application of therapies (healing time of corneal pathology). The following was determined in all patients: degree of hyperemia, degree of epithelial defect, level of corneal sensitivity, level of tear secretion and tear quality, degree of conjunctival secretion, degree of neovascularisation and corneal scarring, degree of visual acuity, score of subjective symptomatology and correlation of ophthalmological findings and subjective symptoms. Results. There was a statistically significant difference in times of therapeutic efficacy/clinical response between the study group patients who received moxifloxacin and the control group patients who received arteficial tears. Conclusion. Local application of moxifloxacin had therapeutic effect (total benefit) both in terms of the effective shortening of the healing time and also the reduction of the complications of bacterial keratitis, without unwanted effects.

Abstract W P396: Positive Correlation Between Neovascularization Degree of Carotid Atherosclerosis Determined by Contrast-enhanced Ultrasound and Level of Serum C-reactive Protein

Objective: To correlate plaque thickness, artery stenosis and levels of serum C-reactive protein (CRP) with the degree of intraplaque enhancement determined by contrast-enhanced ultrasound (CEUS). Methods: CEUS was performed on 72 carotid atherosclerotic plaques in 48 patients. Contrast enhancement within the plaque was categorized as grade 1, 2 or 3. Maximum plaque thickness was measured in short-axis view. Carotid artery stenosis was categorized as mild, moderate or severe. Serum levels of CRP were detected in the fasting state during the same period. Results: Patients were divided into three groups according to the degree of contrast enhancement. Plaque contrast enhancement was not associated with the degree of artery stenosis (P&gt;0.05) or with plaque thickness (P&gt;0.05). Serum CRP levels were positively correlated with the number of new vessels in the plaque. CRP levels increased in the three groups(Grade 1: 3.72±1.79mg/L; Grade 2: 7.88±4.24 mg/L; Grade 3: 11.02±3.52 mg/L), with significant differences among them (F=10.14, P&lt;0.01), and significant differences between each two groups (P&lt;0.05). Spearman’s rank correlation analysis showed that serum CRP levels were positively correlated with the degree of carotid plaque enhancement (rs =0.69, P&lt;0.01). Conclusions: The degree of plaque enhancement on CEUS was not significantly correlated with plaque thickness or vascular stenosis. However, CEUS could semi-quantitatively evaluate new vessels in plaques, and there was a positive correlation between the degree of intraplaque neovascularization and serum CRP levels. The combination of serum CRP levels and intraplaque neovascularization detected by CEUS may allow more accurate evaluation of plaque stability.

Efecto de la adición de fracción vasculoestromal de grasa a la sutura de lesiones meniscales crónicas en zona avascular del menisco de cerdo

ObjectiveThe addition of mesenchymal stem cells may increase the chance of healing of meniscal lesions in the avascular zone. The objective of this study is to find out if the postoperative application of the stromal vascular fraction (SVF) from adipose tissue can have an effect on vascularization and cell proliferation in the sutured meniscus. MethodsFour pigs (8 knees) underwent a longitudinal injury of 10mm in the avascular area of the medial meniscus in both knees. Two months later the injury was repaired on both knees with a simple suture. At the same time 50g of abdominal fat was removed and 2cm3 of SVF was purified (in which it was found to contain approximately 1-5×106mesenchymal stem cells). This was injected into one of the knees of each pig 4h after the surgery (SVF group), using the other knee as the control group. The animals were sacrificed after 15 days. The following histological parameters with hematoxylin-eosin staining were evaluated: degree of healing, neo-vascularization of tissue and the proliferation of fibro-chondrocytes around the lesion and the presence of new chondrocytes. ResultsThe healing rate (unhealed / partial healing / healed) for the SVF group was 1/1/2, and was 2-2-0 for control group (non-significant differences). The degree of neo-vascularization was higher in the SVF group (3.25 vs 2). The degree of fibro-chondrocytes proliferation was greater in the SVF group (2.5 vs 1.75). The formation of isogenic chondrocytes was observed in 2 menisci of the SVF group and none in the control group. ConclusionsThe postoperative intra-articular injection of SVF might increase the neovascularization, the proliferation of fibro-chondrocytes and the repopulation of chondrocytes 15 days after the repair of meniscal injury in pigs. Evidence levelExperimental study. Clinical relevanceA system for the use of stem cells is presented that might be easier to implement in humans that cell cultures.

Facing contrast-enhancing gliomas: perfusion MRI in grade III and grade IV gliomas according to tumor area.

Tumoral neoangiogenesis characterizes high grade gliomas. Relative Cerebral Blood Volume (rCBV), calculated with Dynamic Susceptibility Contrast (DSC) Perfusion-Weighted Imaging (PWI), allows for the estimation of vascular density over the tumor bed. The aim of the study was to characterize putative tumoral neoangiogenesis via the study of maximal rCBV with a Region of Interest (ROI) approach in three tumor areas—the contrast-enhancing area, the nonenhancing tumor, and the high perfusion area on CBV map—in patients affected by contrast-enhancing glioma (grades III and IV). Twenty-one patients were included: 15 were affected by grade IV and 6 by grade III glioma. Maximal rCBV values for each patient were averaged according to glioma grade. Although rCBV from contrast-enhancement and from nonenhancing tumor areas was higher in grade IV glioma than in grade III (5.58 and 2.68; 3.01 and 2.2, resp.), the differences were not significant. Instead, rCBV recorded in the high perfusion area on CBV map, independently of tumor compartment, was significantly higher in grade IV glioma than in grade III (7.51 versus 3.78, P = 0.036). In conclusion, neoangiogenesis encompasses different tumor compartments and CBV maps appear capable of best characterizing the degree of neovascularization. Facing contrast-enhancing brain tumors, areas of high perfusion on CBV maps should be considered as the reference areas to be targeted for glioma grading.

Adipose tissue‐derived stem cells (ADSCs) transplantation promotes regeneration of expanded skin using a tissue expansion model

Transplantation of adipose tissue-derived stem cells (ADSCs) is a promising method that has been used in regenerative medicine because it has shown the capacity to accelerate wound healing. However, roles of ADSCs transplantation in expanded-skin regeneration have remained unknown. To clarify the roles, a tissue expansion model was used in this study. The study comprised three groups of 13 rats in each group: the ADSCs group, the fibroblast (FB) group, and the control group. The skin regeneration in the ADSCs group was enhanced, as evidenced by increased cell proliferation and a higher hydroxyproline content and degree of neovascularization, all with p < 0.05, when compared with both the FB group and the control group. Consistent with enhanced cell proliferation and neovascularization, the regenerated skin in the ADSCs group was much thicker, which further reduced the retraction ratio of the expanded skin. Four weeks after operation, 5'-bromo-2'-deoxyuridine-labeled ADSCs appeared in subcutaneous tissue, vascular vessels, and hair follicles. The up-regulation of protein expression, such as epidermal growth factor and vascular endothelial growth factor, primarily emerged in the ADSC group, with the up-regulated basic fibroblast growth factor appearing in the FB group. Collectively, these results suggest that the transplantation of ADSCs could enhance the regeneration of expanded skin by participating in skin structures and up-regulating the secretion of epidermal growth factor and vascular endothelial growth factor.

Effect of Glucocorticoid (Triamcinolone Acetonide) Pretreatment in a Murine Penetrating Keratoplasty and Suture Model

To evaluate the effect of glucocorticoid (triamcinolone acetonide injectable suspension) pretreatment on corneal neovascularization, lymphangiogenesis, and inflammation in a murine penetrating keratoplasty (PK) and corneal suture model. For the PK model, BALB/c mice were used as recipients and C57BL/6 mice were used as donors. A group pretreated with subconjunctival glucocorticoid and a combination of post-subconjunctival and topical glucocorticoids (group I) was compared with two groups that did not receive glucocorticoid pretreatment [one group received a combination of subconjunctival and topical glucocorticoids postoperatively (group II) and the other group received only topical glucocorticoid treatment postoperatively (group III)]. All groups were treated with subconjunctival glucocorticoid on the day of surgery. For the corneal suture model, BALB/c mice were used. A group receiving only pre-suture glucocorticoid treatment (group A) and a group receiving only post-suture glucocorticoid treatment (group C) were compared with a control group that did not receive glucocorticoid therapy (group B). The degree of neovascularization, lymphangiogenesis, and inflammatory infiltration was compared in each of these models. In the PK model, the group receiving glucocorticoid pretreatment (group I) showed less neovascularization compared with the posttreatment-only groups (group II, P=0.043; group III, P=0.020) and less lymphangiogenesis compared with group III (P=0.005). In the corneal suture model, the glucocorticoid pretreatment group showed a similar level of neovascularization, lymphangiogenesis, and inflammatory infiltration as the posttreatment-only groups (P>0.05). Glucocorticoid pretreatment before PK decreases neovascularization and lymphangiogenesis compared with posttransplant glucocorticoid treatment alone.

Vitreous TIMP-1 levels associate with neovascularization and TGF-β2 levels but not with fibrosis in the clinical course of proliferative diabetic retinopathy

In proliferative diabetic retinopathy (PDR), vascular endothelial growth factor (VEGF) and CCN2 (connective tissue growth factor; CTGF) cause blindness by neovascularization and subsequent fibrosis. This angio-fibrotic switch is associated with a shift in the balance between vitreous levels of CCN2 and VEGF in the eye. Here, we investigated the possible involvement of other important mediators of fibrosis, tissue inhibitor of metalloproteinases (TIMP)-1 and transforming growth factor (TGF)-β2, and of the matrix metalloproteinases (MMP)-2 and MMP-9, in the natural course of PDR. TIMP-1, activated TGF-β2, CCN2 and VEGF levels were measured by ELISA in 78 vitreous samples of patients with PDR (n = 28), diabetic patients without PDR (n = 24), and patients with the diabetes-unrelated retinal conditions macular hole (n = 10) or macular pucker (n = 16), and were related to MMP-2 and MMP-9 activity on zymograms and to clinical data, including degree of intra-ocular neovascularization and fibrosis. TIMP-1, CCN2 and VEGF levels, but not activated TGF-β2 levels, were significantly increased in the vitreous of diabetic patients, with the highest levels in PDR patients. CCN2 and the CCN2/VEGF ratio were the strongest predictors of degree of fibrosis. In diabetic patients with or without PDR, activated TGF-β2 levels correlated with TIMP-1 levels, whereas in PDR patients, TIMP-1 levels, MMP-2 and proMMP-9 were associated with degree of neovascularization, like VEGF levels, but not with fibrosis. We confirm here our previous findings that retinal fibrosis in PDR patients is significantly correlated with vitreous CCN2 levels and the CCN2/VEGF ratio. In contrast, TIMP-1, MMP-2 and MMP-9 appear to have a role in the angiogenic phase rather than in the fibrotic phase of PDR.

Association between carotid plaque enhancement shown by multidetector CT angiography and histologically validated microvessel density

Carotid plaques analysed by MDCTA can show contrast enhancement. The purpose of this study was to explore the association between carotid plaque enhancement (CPE) and microvessel density. We obtained IRB approval. Twenty-nine consecutive (male, 20; median age, 63) symptomatic patients studied with 16-detector CT were prospectively analysed. Examinations were performed before and after intravenous contrast medium administration, and analysis of plaque enhancement was performed. Patients underwent "en bloc" carotid endarterectomy; histological sections were prepared and the presence of microvessels quantified. Logistic regression analysis as well as ROC curve and area under the curve was calculated. A statistically significant association between the degree of CPE and microvessel density (P = 0.009; rho = 0.553) was observed. The ROC curve analysis confirmed this association with an area under the curve of 0.906, 0.735, 0.644 and 0.546 for CPE of 10 HU, 15 HU, 20 HU and 25 HU respectively. There was a statistically significant difference between the CPE and the degree of neovascularisation (P = 0.0003). Results of this preliminary study suggest that CPE might be associated with the microvessel density. Histological analysis seems to demonstrate that the degree of intra-plaque neo-vascularisation is statistically associated with CPE. Carotid artery plaque enhancement at CT is associated with microvessel density. The degree of intra-plaque neo-vascularisation is statistically associated with carotid plaque enhancement. Plaque enhancement at CT should be considered when assessing vulnerable plaques.