SYNERGY OF WALL SHEAR STRESS AND CIRCUMFERENTIAL STRESS IN STRAIGHT ARTERIES

The Wall Shear Stress （WSS） generated by blood flow and Circumferential Stress （CS） driven by blood pressure have been thought to play an important role in blood flow-dependent phenomena such as angiogenesis, vascular remodeling, and atherosgenesis. The WSS and CS in straight arteries were calculated by measuring the blood pressure, center-line velocity, wall thickness, and radius of vessels. The WSS and CS in the time domain were then decomposed into the amplitude and phase in the frequency domain. The CS amplitude to the WSS amplitude ratio （referred as stress ampli tude ratio, Zs ） and the phase difference between the CS and the WSS （referred as stress phase difference, SPA） in the fre quency domain were calculated to characterize the synergy of the CS and WSS. Numerical results demonstrated that the CS is not in phase with the WSS, a time delay in the time domain or a stress phase difference in the frequency domain between the WSS and the CS exists. Theoretical analysis demonstrated that the Zs and SPA are primarily determined by the local fac tors （blood viscosity, local inertial effects, local geometry, loeal elasticity） and the input impedance of whole downstream arterial beds. Because the arterial input impedance has been shown to reflect the physiological and pathological states of whole downstream arterial beds, the stress amplitude ratio Zs and stress phase difference SPA would be thought to be the appropriate indices to reflect the effects of states of whole downstream arterial beds on the local blood flow dependent phenomena such as angiogenesis, vascular remodeling, and atherosgenesis.     

Effects of P4 Antagonist RU486 on VEGF and Its Receptors’ Signaling during the In Vivo Transition from the Preovulatory to Periovulatory Phase of Ovarian Follicles

The development of an adequate blood vessel network is crucial for the accomplishment of ovarian follicle growth and ovulation, which is necessary to support the proliferative and endocrine functions of the follicular cells. Although the Vascular Endothelial Growth Factor (VEGF) through gonadotropins guides ovarian angiogenesis, the role exerted by the switch on of Progesterone (P₄) during the periovulatory phase remains to be clarified. The present research aimed to investigate in vivo VEGF-mediated mechanisms by inducing the development of periovulatory follicles using a pharmacologically validated synchronization treatment carried out in presence or absence of P₄ receptor antagonist RU486. Spatio-temporal expression profiles of VEGF, FLT1, and FLK1 receptors and the two major MAPK/ERKs and PI3K/AKT downstream pathways were analyzed on granulosa and on theca compartment. For the first time, the results demonstrated that in vivo administration of P₄ antagonist RU486 inhibits follicular VEGF receptors’ signaling mainly acting on the theca layer by downregulating the activation of ERKs and AKTs. Under the effect of RU486, periovulatory follicles’ microarchitecture did not move towards the periovulatory stage. The present evidence provides new insights on P₄ in vivo biological effects in driving vascular and tissue remodeling during the periovulatory phase.     

原核表达融合型血管生成抑制剂Kringle 5发酵条件的优化和初步纯化

在构建融合型Kringle 5原核表达重组菌BL21(DE3)pET32a/K5的基础上,通过改变培养基的成分,确定了适宜于重组菌生长和融合Kringle 5蛋白表达的培养基.采用正变实验方法,对融合型血管生成抑制剂Kringle 5原核表达重组菌的发酵条件进行了优化,通过发酵罐中重组菌的分批培养,确定了培养液中的最佳溶氧量.采用优化后的发酵工艺,在20 L发酵罐中融合Kringle 5的表达量可达0.59 g·L-1.最后在Cu2 螯合的Sepharose Fast Flow层析介质上对融合Kringle 5进行了初步纯化,其纯度约为80%.     

Extracellular Vesicles Mediate Communication between Endothelial and Vascular Smooth Muscle Cells

The recruitment of pericytes and vascular smooth muscle cells (SMCs) that enwrap endothelial cells (ECs) is a crucial process for vascular maturation and stabilization. Communication between these two cell types is crucial during vascular development and in maintaining vessel homeostasis. Extracellular vesicles (EVs) have emerged as a new communication tool involving the exchange of microRNAs between cells. In the present study, we searched for microRNAs that could be transferred via EVs from ECs to SMCs and vice versa. Thanks to a microRNA profiling experiment, we found that two microRNAs are more exported in each cell type in coculture experiments: while miR-539 is more secreted by ECs, miR-582 is more present in EVs from SMCs. Functional assays revealed that both microRNAs can modulate both cell-type phenotypes. We further identified miR-539 and miR-582 targets, in agreement with their respective cell functions. The results obtained in vivo in the neovascularization model suggest that miR-539 and miR-582 might cooperate to trigger the process of blood vessel coverage by smooth muscle cells in a mature plexus. Taken together, these results are the first to highlight the role of miR-539 and miR-582 in angiogenesis and communication between ECs and SMCs.     

ASC and SVF Cells Synergistically Induce Neovascularization in Ischemic Hindlimb Following Cotransplantation

Previously, we reported the angio-vasculogenic properties of human stromal vascular fraction (SVF) and adipose tissue-derived mesenchymal stem cells (ASCs). In this study, we investigated whether the combination of ASCs and SVF cells exhibited synergistic angiogenic properties. We conducted quantitative (q)RT-PCR, Matrigel plug, tube formation assays, and in vivo therapeutic assays using an ischemic hind limb mouse model. Immunohistochemical analysis was also conducted. qRT-PCR results revealed that FGF-2 was highly upregulated in ASCs compared with SVF, while PDGF-b and VEGF-A were highly upregulated in SVF. Conditioned medium from mixed cultures of ASCs and SVF (A+S) cells showed higher Matrigel tube formation and endothelial cell proliferation in vitro. A+S cell transplantation into ischemic mouse hind limbs strongly prevented limb loss and augmented blood perfusion compared with SVF cell transplantation. Transplanted A+S cells also showed high capillary density, cell proliferation, angiogenic cytokines, and anti-apoptotic potential in vivo compared with transplanted SVF. Our data indicate that A+S cell transplantation results in synergistic angiogenic therapeutic effects. Accordingly, A+S cell injection could be an alternative therapeutic strategy for treating ischemic diseases.     

Modulation of Mesenchymal Stem Cells for Enhanced Therapeutic Utility in Ischemic Vascular Diseases

Mesenchymal stem cells are multipotent stem cells isolated from various tissue sources, including but not limited to bone marrow, adipose, umbilical cord, and Wharton Jelly. Although cell-mediated mechanisms have been reported, the therapeutic effect of MSCs is now recognized to be primarily mediated via paracrine effects through the secretion of bioactive molecules, known as the “secretome”. The regenerative benefit of the secretome has been attributed to trophic factors and cytokines that play neuroprotective, anti-angiogenic/pro-angiogenic, anti-inflammatory, and immune-modulatory roles. The advancement of autologous MSCs therapy can be hindered when introduced back into a hostile/disease environment. Barriers include impaired endogenous MSCs function, limited post-transplantation cell viability, and altered immune-modulatory efficiency. Although secretome-based therapeutics have gained popularity, many translational hurdles, including the heterogeneity of MSCs, limited proliferation potential, and the complex nature of the secretome, have impeded the progress. This review will discuss the experimental and clinical impact of restoring the functional capabilities of MSCs prior to transplantation and the progress in secretome therapies involving extracellular vesicles. Modulation and utilization of MSCs–secretome are most likely to serve as an effective strategy for promoting their ultimate success as therapeutic modulators.     

The Role of the VEGF Family in Atherosclerosis Development and Its Potential as Treatment Targets

The vascular endothelial growth factor (VEGF) family, the crucial regulator of angiogenesis, lymphangiogenesis, lipid metabolism and inflammation, is involved in the development of atherosclerosis and further CVDs (cardiovascular diseases). This review discusses the general regulation and functions of VEGFs, their role in lipid metabolism and atherosclerosis development and progression. These functions present the great potential of applying the VEGF family as a target in the treatment of atherosclerosis and related CVDs. In addition, we discuss several modern anti-atherosclerosis VEGFs-targeted experimental procedures, drugs and natural compounds, which could significantly improve the efficiency of atherosclerosis and related CVDs’ treatment.     

Systemic Delivery of a STING Agonist-Loaded Positively Charged Liposome Selectively Targets Tumor Immune Microenvironment and Suppresses Tumor Angiogenesis

Although stimulator of interferon genes (STING) agonists has shown great promise in preclinical studies, the clinical development of STING agonist therapy is challenged by its limited systemic delivery. Here, positively charged fusogenic liposomes loaded with a STING agonist (PoSTING) are designed for systemic delivery and to preferentially target the tumor microenvironment. When PoSTING is administered intravenously, it selectively targets not only tumor cells but also immune and tumor endothelial cells (ECs). In particular, delivery of STING agonists to tumor ECs normalizes abnormal tumor vasculatures, induces intratumoral STING activation, and elicits robust anti-tumor T cell immunity within the tumor microenvironment. Therefore, PoSTING can be used as a systemic delivery platform to overcome the limitations of using STING agonists in clinical trials.     

Cuttlebone-Derived Organic Matrix: A Facile Periosteum Substitute for Bone Regeneration

Artificial periosteum has become a novel strategy for bone healing. Several efforts are made to develop novel periosteum substitutes with excellent osteogenesis and angiogenesis performance. The problem, however, is that many inconveniences, such as unattainable materials and complex preparation processes, limit its clinical application. In this study, cuttlebone-derived organic matrix (CDOM) film is developed as a facile obtained artificial periosteum with good bioactivity, biodegradability, and biosafety. In this study, CDOM films are prepared by etching calcium carbonate of cuttlebone with hydrochloric acid (HCl), phosphoric acid (H₃PO₄), and ethylenediaminetetraacetic acid (EDTA), respectively. CDOM films possess “S” oriented grooves on one surface, providing suitable space for survival, adhesion, and proliferation of rat mesenchymal stem cells and human umbilical vein endothelial cells. Among three CDOM films, CDOM-EDTA is the most efficient at enhancing bone regeneration in vivo, with an increase in neovascularization and new bone formation. Meanwhile, CDOM-EDTA is degradable in two months and of extremely low toxicity. There is a great deal of promise in the future for this novel periosteum substitute to be used for bone defect healing in clinical settings.