Current Status of Angiogenic Cell Therapy and Related Strategies Applied in Critical Limb Ischemia

Critical limb ischemia (CLI) constitutes the most severe form of peripheral arterial disease (PAD), it is characterized by progressive blockade of arterial vessels, commonly correlated to atherosclerosis. Currently, revascularization strategies (bypass grafting, angioplasty) remain the first option for CLI patients, although less than 45% of them are eligible for surgical intervention mainly due to associated comorbidities. Moreover, patients usually require amputation in the short-term. Angiogenic cell therapy has arisen as a promising alternative for these “no-option” patients, with many studies demonstrating the potential of stem cells to enhance revascularization by promoting vessel formation and blood flow recovery in ischemic tissues. Herein, we provide an overview of studies focused on the use of angiogenic cell therapies in CLI in the last years, from approaches testing different cell types in animal/pre-clinical models of CLI, to the clinical trials currently under evaluation. Furthermore, recent alternatives related to stem cell therapies such as the use of secretomes, exosomes, or even microRNA, will be also described.     

Defining Dry Eye from a Clinical Perspective

Over the past decades, the number of patients with dry eye disease (DED) has increased dramatically. The incidence of DED is higher in Asia than in Europe and North America, suggesting the involvement of cultural or racial factors in DED etiology. Although many definitions of DED have been used, discrepancies exist between the various definitions of dry eye disease (DED) used across the globe. This article presents a clinical consensus on the definition of DED, as formulated in four meetings with global DED experts. The proposed new definition is as follows: “Dry eye is a multifactorial disease characterized by a persistently unstable and/or deficient tear film (TF) causing discomfort and/or visual impairment, accompanied by variable degrees of ocular surface epitheliopathy, inflammation and neurosensory abnormalities.” The key criteria for the diagnosis of DED are unstable TF, inflammation, ocular discomfort and visual impairment. This definition also recommends the assessment of ocular surface epitheliopathy and neurosensory abnormalities in each patient with suspected DED. It is easily applicable in clinical practice and should help practitioners diagnose DED consistently. This consensus definition of DED should also help to guide research and clinical trials that, to date, have been hampered by the lack of an established surrogate endpoint.     

Extracellular Matrix Deposition and Remodeling after Corneal Alkali Burn in Mice

Corneal transparency relies on the precise arrangement and orientation of collagen fibrils, made of mostly Type I and V collagen fibrils and proteoglycans (PGs). PGs are essential for correct collagen fibrillogenesis and maintaining corneal homeostasis. We investigated the spatial and temporal distribution of glycosaminoglycans (GAGs) and PGs after a chemical injury. The chemical composition of chondroitin sulfate (CS)/dermatan sulfate (DS) and heparan sulfate (HS) were characterized in mouse corneas 5 and 14 days after alkali burn (AB), and compared to uninjured corneas. The expression profile and corneal distribution of CS/DSPGs and keratan sulfate (KS) PGs were also analyzed. We found a significant overall increase in CS after AB, with an increase in sulfated forms of CS and a decrease in lesser sulfated forms of CS. Expression of the CSPGs biglycan and versican was increased after AB, while decorin expression was decreased. We also found an increase in KS expression 14 days after AB, with an increase in lumican and mimecan expression, and a decrease in keratocan expression. No significant changes in HS composition were noted after AB. Taken together, our study reveals significant changes in the composition of the extracellular matrix following a corneal chemical injury.     

基于MLP神经网络的圆锥角膜辅助诊断北大核心CSCD

圆锥角膜在病变过程中会导致角膜中央部位向前凸出,使角膜呈现出圆锥形,而且会导致高度不规则近视和散光,对视力造成不同程度损害。疾病一般发生于青少年时期,为了能及时治疗避免病变严重,筛查区分圆锥角膜具有十分重要的意义。而且临床上对于圆锥角膜诊断通常是采用角膜地形图的方法,可以得到角膜形态学的改变,但是有一定的误诊率。目前研究发现,角膜力学特性改变先于形态学,所以本文从角膜生物力学角度出发,提出一种基于多层感知机(multi-layer perceptron,MLP)神经网络区分圆锥角膜的模型。首先,利用可视化生物力学分析仪(corneal visualization scheimpflug technology,Corvis-ST)测得角膜的生物力学视频,进行处理计算得到角膜生物力学参数作为数据集,其中包含正常角膜和圆锥角膜2种类别;然后,针对角膜生物力学参数数据集构建MLP神经网络模型,将70%数据集作为训练集,30%数据集作为测试集。在数据集上训练及测试的结果表明,该模型区分圆锥角膜的准确率为97.6%。     

Design and implementation of optical system for Placido-disc topography

Corneal topography provides powerful support in the diagnosis and treatment of corneal disease by displaying the corneal surface topography in data or image format. To realize the precise detection of corneal surface topography, an optical system for the corneal topography that is based on a Placido disc is designed, which includes a ring distribution on a Placido disc, an imaging system and a collimating illumination system. First, a mathematical model that is based on the corneal topography working principles is established with MATLAB to determine the distribution of white-and-black rings on the Placido disc, in which the ellipsoid facial rings-target of the Placido disc is utilized. Second, the imaging lens structure is designed and optimized by Zemax software. Last, the collimating illumination lens structure is designed by paraxial ray trace equations. The quality of the corneal topography, which is based on our designed optical system, is evaluated. The high-contrast image of uniformly distributed white-and-black rings is observed through the CCD camera. Our optical system for the corneal topography has high precision, with a measuring region of the cornea with a diameter of approximately 10 mm. Therefore, the creation of this optical system offers guidance for designing and improving the optical system of Placido-disc topography.     

抗FGF-2纳米抗体抑制碱烧伤诱导大鼠角膜血管新生

目的：研究抗成纤维细胞生长因子（FGF-2）纳米抗体对碱烧伤诱导的大鼠角膜血管生成的治疗作用。方法：将SD大鼠分为：假手术组（Sham），模型组（Model，直径为3 mm的浸有1 mol/L NaOH溶液圆形滤纸贴于大鼠眼角膜中央处30 s，制备大鼠碱烧伤血管生成模型）和治疗组（Treatment，术后7天至21天用3 mg/mL的抗FGF-2纳米抗体溶液滴眼，每日3次，每次10 μL，共14天）。通过体视显微镜和CD31免疫组织化学染色计算大鼠角膜血管生成情况。实时荧光定量PCR、酶联免疫吸附测定和免疫组织化学染色3种方法检测抗血管内皮生长因子（VEGF）和FGF-2的mRNA和蛋白表达水平。结果：（1）血管：治疗组较模型组的面积显著减少，血管管腔较窄（P<0.05），在药物干预14天后，差异最为显著；（2）FGF-2的mRNA和蛋白表达水平：模型组与治疗组的结果相近（P>0.05）；（3）VEGF的mRNA和蛋白表达水平：治疗组显著高于模型组（P<0.05）。此外，假手术组的持续给药也使得VEGF表达显著增加（P<0.05）。 结论：抗FGF-2纳米抗体可抑制由碱烧伤诱导的角膜血管新生，但也使得正常大鼠角膜或病理大鼠角膜的VEGF表达水平代偿性升高。     

Mucoadhesive liposomes as ocular delivery system: physical,microbiological, and in vivo assessment

Background: Mucoadhesive drug delivery is a promising strategy to overcome ocular biopharmaceutical constraints. Objective and methods: Ciprofloxacin HCl-loaded reverse phase evaporation liposomes were coated with different concentrations and molecular weights of mucoadhesive biocompatible chitosan polymer to form chitosomes. This colloidal mucoadhesive system was evaluated in vitro and in vivo with respect to deliver the antibiotic to ocular surface. Results and conclusion: The results obtained pointed out that liposome coating process resulted in entrapment efficiency reduction and higher chitosan concentration, and molecular weight showed a more pronounced effect. No morphological differences between coated and uncoated liposomes were observed. Diffusion was the drug release mechanism from chitosomes. Concerning rheological behavior, pseudoplastic flow was characteristic to the prepared chitosomal dispersions. In addition, chitosan coating improved the ocular permeation of ciprofloxacin HCl. Microbiologically; this formulated system enhanced antimicrobial activity of ciprofloxacin HCl against both Gram-positive and Gram-negative bacteria. Moreover, this mucoadhesive system was able to inhibit the growth of Pseudomonas aeruginosa in rabbits' eyes for 24 hours when compared to the marketed preparation. In vivo bacterial conjunctivitis model elucidated that symptoms were controlled by the prolonged release formulation such as that done by the marketed product.     

Image processing techniques in a preliminary morphometric characterization of corneal epithelium by scanning electron microscopy

Several processing techniques of digital images allowed us to quantify the percentage of cell surface covered by microprojections (microvilli or microplicae) (SCM), the adhesion between epithelial cells by the parameter intercellular junctions (IJ), the size (cell area), shape (cell shape) and shade (cell shade) of cells on the corneal epithelium of nine rabbits. The data were analyzed and the epithelial cells were classified into three groups by cluster analysis. Assuming the representativeness of the sample, our findings suggest that for a normal corneal epithelium, 80% of the cells could show SCM >41%, and IJ >0.98 (being one a cell to cell junction without disruptions). Standard deviations of cell shade lower than 21 gray levels could indicate a tendency to lose the cell shade mosaic. Normal corneas could show a majority of cells (54–69%) included in group 2 with smaller mean size (80% of cells with cell area <242 μm²), higher SCM (80% of cells with SCM >44.83%), polygonal mean shape and brighter shade. Group 1 (15–30% of cells) could show a larger mean size (80% of cells with cell area >494 μm²), lower SCM (although 80% of cells with SCM >32.61%), circular mean shape and darker electron reflex. Group 3 could display a medium mean size, higher SCM (similar to group 2), circular mean shape (similar to group 1), and brighter shade. These analyses could possibly be used to further assess the corneal response to ocular drugs, contact lens, and surgical procedures or to discriminate between pathology stages. Microsc. Res. Tech. 73:1059–1066, 2010. © 2010 Wiley‐Liss, Inc.     

Tumor Vessels Fuel the Fire in Glioblastoma

Glioblastoma, a subset of aggressive brain tumors, deploy several means to increase blood vessel supply dedicated to the tumor mass. This includes typical program borrowed from embryonic development, such as vasculogenesis and sprouting angiogenesis, as well as unconventional processes, including co-option, vascular mimicry, and transdifferentiation, in which tumor cells are pro-actively engaged. However, these neo-generated vascular networks are morphologically and functionally abnormal, suggesting that the vascularization processes are rather inefficient in the tumor ecosystem. In this review, we reiterate the specificities of each neovascularization modality in glioblastoma, and, how they can be hampered mechanistically in the perspective of anti-cancer therapies.