Mimic Blood Vessels Research Articles

Deciphering breast cancer prognosis: a novel machine learning-driven model for vascular mimicry signature prediction.

In the ongoing battle against breast cancer, a leading cause of cancer-related mortality among women globally, the urgent need for innovative prognostic markers and therapeutic targets is undeniable. This study pioneers an advanced methodology by integrating machine learning techniques to unveil a vascular mimicry signature, offering predictive insights into breast cancer outcomes. Vascular mimicry refers to the phenomenon where cancer cells mimic blood vessel formation absent of endothelial cells, a trait associated with heightened tumor aggression and diminished response to conventional treatments. The study's comprehensive analysis spanned data from over 6,000 breast cancer patients across 12 distinct datasets, incorporating both proprietary clinical data and single-cell data from 7 patients, accounting for a total of 43,095 cells. By employing an integrative strategy that utilized 10 machine learning algorithms across 108 unique combinations, the research scrutinized 100 existing breast cancer signatures. Empirical validation was sought through immunohistochemistry assays, alongside explorations into potential immunotherapeutic and chemotherapeutic avenues. The investigation successfully identified six genes related to vascular mimicry from multi-center cohorts, laying the groundwork for a novel predictive model. This model outstripped the prognostic accuracy of traditional clinical and molecular indicators in forecasting recurrence and mortality risks. High-risk individuals identified by our model faced worse outcomes. Further validation through IHC assays in 30 patients underscored the model's extensive applicability. Notably, the model unveiled varying therapeutic responses; low-risk patients might achieve greater benefits from immunotherapy, whereas high-risk patients demonstrated a particular sensitivity to certain chemotherapies, such as ispinesib. This model marks a significant step forward in the precise evaluation of breast cancer prognosis and therapeutic responses across different patient groups. It heralds the possibility of refining patient outcomes through tailored treatment strategies, accentuating the potential of machine learning in revolutionizing cancer prognosis and management.

Development of a Novel Hierarchically Biofabricated Blood Vessel Mimic Decorated with Three Vascular Cell Populations for the Reconstruction of Small-Diameter Arteries.

The availability of grafts to replace small-diameter arteries remains an unmet clinical need. Here, the validated methodology is reported for a novel hybrid tissue-engineered vascular graft that aims to match the natural structure of small-size arteries. The blood vessel mimic (BVM) comprises an internal conduit of co-electrospun gelatin and polycaprolactone (PCL) nanofibers (corresponding to the tunica intima of an artery), reinforced by an additional layer of PCL aligned fibers (the internal elastic membrane). Endothelial cells are deposited onto the luminal surface using a rotative bioreactor. A bioprinting system extrudes two concentric cell-laden hydrogel layers containing respectively vascular smooth muscle cells and pericytes to create the tunica media and adventitia. The semi-automated cellularization process reduces the production and maturation time to 6 days. After the evaluation of mechanical properties, cellular viability, hemocompatibility, and suturability, the BVM is successfully implanted in the left pulmonary artery of swine. Here, the BVM showed good hemostatic properties, capability to withstand blood pressure, and patency at 5 weeks post-implantation. These promising data open a new avenue to developing an artery-like product for reconstructing small-diameter blood vessels.

Microtube embedded hydrogel bioprinting for vascularization of tissue-engineered scaffolds.

Vascular tissue engineering has been considered promising as one of the alternatives for viable artificial tissues and organs. Macro- and microscale hollow tubes fabricated with various techniques have been widely studied to mimic blood vessels. To date, the fabrication of biomimetic capillary vessels with sizes ranging from 1 to 10 µm is still challenging. In this paper, core-sheath microtubes were electrospun to mimic capillary vessels and were embedded in carboxymethyl cellulose/sodium alginate hydrogel for bioprinting. The results showed improved printing fidelity and promoted cell attachment. The tube concentration and tube length both had significant influences on filament size and merging area. Printed groups with higher microtube concentration showed higher microtube density, with filament/nozzle size ratio, and printed/designed grid area ratio closer to 100%. In the in vitro experiments, microtubes were not only compatible with human umbilical vein endothelial cells but also provided microtopographical cues to promote cell proliferation and morphogenesis in three-dimensional space. In summary, the microtubes fabricated by our groups have the potential for the bioprinting of vascularized soft tissue scaffolds.

Multifocal primary central nervous system angiosarcoma: illustrative case.

Angiosarcoma is a malignant mesenchymal tumor derived from vascular endothelial cells in which a primary intracranial origin is extremely rare. Most previous reports of primary central nervous system (CNS) angiosarcoma have been solitary cases. The authors report a case of primary CNS angiosarcoma that caused the development of multiple disseminated cerebral hemorrhagic lesions within a short period of time. This rapid progression of symptoms resulted in the death of the patient. During surgery, several nodules suggestive of a tumor were removed from just below the surface of the brain, mixed into the hematoma. A pathological examination revealed atypical cells mimicking blood vessels in the subarachnoid space that were positive for specific vascular endothelial markers. In this case, multifocal angiosarcoma occurred on the brain surface and ventricles, suggesting cerebrospinal fluid dissemination. If multiple cerebral hemorrhages are found on the brain surface, multifocal angiosarcoma should also be considered.

Engineering a dynamic three-dimensional cell culturing microenvironment using a ‘sandwich’ structure-liked microfluidic device with 3D printing scaffold

Most of in vivo tissue cells reside in 3D extracellular matrix (ECM) with fluid flow. To better study cell physiology and pathophysiology, there has been an increasing need in the development of methods for culturing cells in in vivo like microenvironments with a number of strategies currently being investigated including hydrogels, spheroids, tissue scaffolds and very promising microfluidic systems. In this paper, a ‘sandwich’ structure-liked microfluidic device integrated with a 3D printing scaffold is proposed for three-dimensional and dynamic cell culture. The device consists of three layers, i.e. upper layer, scaffold layer and bottom layer. The upper layer is used for introducing cells and fixing scaffold, the scaffold layer mimicking ECM is used for providing 3D attachment areas, and the bottom layer mimicking blood vessels is used for supplying dynamic medium for cells. Thermally assisted electrohydrodynamic jet (TAEJ) printing technology and microfabrication technology are combined to fabricate the device. The flow field in the chamber of device is evaluated by numerical simulation and particle tracking technology to investigate the effects of scaffold on fluid microenvironment. The cell culturing processes are presented by the flow behaviors of inks with different colors. The densities and viabilities of HeLa cells are evaluated and compared after 72 h of culturing in the microfluidic devices and 48-well plate. The dose-dependent cell responses to doxorubicin hydrochloride (DOX) are observed after 24 h treatment at different concentrations. These experimental results, including the evaluation of cell proliferation and in vitro cytotoxicity assessment of DOX in the devices and plate, demonstrate that the presented microfluidic device has good biocompatibility and feasibility, which have great potential in providing native microenvironments for in vitro cell studies, tissue engineering and drug screening for tumor therapy.

Network topology-directed design of molecular CPU for cell-like dynamic information processing.

Natural cells (NCs) can automatically and continuously respond to fluctuant external information and distinguish meaningful stimuli from weak noise depending on their powerful genetic and protein networks. We herein report a network topology–directed design of dynamic molecular processing system (DMPS) as a molecular central processing unit that powers an artificial cell (AC) able to process fluctuant information in its immediate environment similar to NCs. By constructing a mixed cell community, ACs and NCs have synchronous response to fluctuant extracellular stimuli under physiological condition and in a blood vessel–mimic circulation system. We also show that fluctuant bioinformation released by NCs can be received and processed by ACs. The molecular design of DMPS-powered AC is expected to allow a profound understanding of biological systems, advance the construction of intelligent molecular systems, and promote more elegant bioengineering applications.

High-fidelity deconvolution for acoustic-resolution photoacoustic microscopy enabled by convolutional neural networks

Acoustic-resolution photoacoustic microscopy (AR-PAM) image resolution is determined by the point spread function (PSF) of the imaging system. Previous algorithms, including Richardson–Lucy (R–L) deconvolution and model-based (MB) deconvolution, improve spatial resolution by taking advantage of the PSF as prior knowledge. However, these methods encounter the problems of inaccurate deconvolution, meaning the deconvolved feature size and the original one are not consistent (e.g., the former can be smaller than the latter). We present a novel deep convolution neural network (CNN)-based algorithm featuring high-fidelity recovery of multiscale feature size to improve lateral resolution of AR-PAM. The CNN is trained with simulated image pairs of line patterns, which is to mimic blood vessels. To investigate the suitable CNN model structure and elaborate on the effectiveness of CNN methods compared with non-learning methods, we select five different CNN models, while R–L and directional MB methods are also applied for comparison. Besides simulated data, experimental data including tungsten wires, leaf veins, and in vivo blood vessels are also evaluated. A custom-defined metric of relative size error (RSE) is used to quantify the multiscale feature recovery ability of different methods. Compared to other methods, enhanced deep super resolution (EDSR) network and residual in residual dense block network (RRDBNet) model show better recovery in terms of RSE for tungsten wires with diameters ranging from 30 μm to 120 μm. Moreover, AR-PAM images of leaf veins are tested to demonstrate the effectiveness of the optimized CNN methods (by EDSR and RRDBNet) for complex patterns. Finally, in vivo images of mouse ear blood vessels and rat ear blood vessels are acquired and then deconvolved, and the results show that the proposed CNN method (notably RRDBNet) enables accurate deconvolution of multiscale feature size and thus good fidelity.

Annular Fiber Probe for Interstitial Illumination in Photoacoustic Guidance of Radiofrequency Ablation.

Unresectable liver tumors are commonly treated with percutaneous radiofrequency ablation (RFA). However, this technique is associated with high recurrence rates due to incomplete tumor ablation. Accurate image guidance of the RFA procedure contributes to successful ablation, but currently used imaging modalities have shortcomings in device guidance and treatment monitoring. We explore the potential of using photoacoustic (PA) imaging combined with conventional ultrasound (US) imaging for real-time RFA guidance. To overcome the low penetration depth of light in tissue, we have developed an annular fiber probe (AFP), which can be inserted into tissue enabling interstitial illumination of tissue. The AFP is a cannula with 72 optical fibers that allows an RFA device to slide through its lumen, thereby enabling PA imaging for RFA device guidance and ablation monitoring. We show that the PA signal from interstitial illumination is not affected by absorber-to-surface depth compared to extracorporeal illumination. We also demonstrate successful imaging of the RFA electrodes, a blood vessel mimic, a tumor-mimicking phantom, and ablated liver tissue boundaries in ex vivo chicken and bovine liver samples. PA-assisted needle guidance revealed clear needle tip visualization, a notable improvement to current US needle guidance. Our probe shows potential for RFA device guidance and ablation detection, which potentially aids in real-time monitoring.

A biomimetic orthogonal-bilayer tubular scaffold for the co-culture of endothelial cells and smooth muscle cells.

In blood vessels, endothelial cells (ECs) grow along the direction of blood flow, while smooth muscle cells (SMCs) grow circumferentially along the vessel wall. To mimic this structure, a polycaprolactone (PCL) tubular scaffold with orthogonally oriented bilayer nanofibers was prepared via electrospinning and winding. ECs were cultured on the inner layer of the scaffold with axial nanofibers and SMCs were cultured on the outer layer of the scaffold with circumferential nanofibers. Fluorescence images of the F-actin distribution of ECs and SMCs indicated that cells adhered, stretched, and proliferated in an oriented manner on the scaffold. Moreover, layers of ECs and SMCs formed on the scaffold after one month of incubation. The expression levels of platelet-endothelial cell adhesion molecule 1 (PECAM-1) and a contractile SMC phenotype marker in the EC/SMC co-culture system were much higher than those in individual culture systems, thus demonstrating that the proposed biomimetic scaffold promoted the intercellular junction of ECs and preserved the contractile phenotype of SMCs.

Моделирование пульсирующих потоков крови для задач оптической когерентной томографии в офтальмологии.

Optical coherence tomography (OCT) is a method of non-invasive medical diagnostics based on scanning a biological object with near-infrared radiation, followed by registration and analysis of the back-reflected and back-scattered photons. The most of OCT systems are used in ophthalmology for diagnostics conditions of the retina, optic nerve and anterior segment of the eye. The purpose of this research is to increase the reliability of controlled experiments of the retina using phantoms by taking into account the partial clamping and vibrations of the walls of blood vessels that occur in real living objects. Methods of a retinal phantom molding containing special cavities (mimicking blood vessels) suitable for pumping blood-imitating fluid are described. The retina is shaped as a multilayered structure. Two-component transparent liquid silicone is used as to mold each layer. Indian ink with known optical properties is used to mimic absorption. Titanium dioxide particles are used as scatterers. The layers are formed sequentially from the bottom (choroid) to the top (inner boundary membrane). Mass fractions of the additives, as well as the thickness of each layer, are chosen on an individual basis. All blood vessels are represented as elongated and connected cavities. The formation of the base and complex network of the molded blood vessels is performed using a 3D printer. A device for making the palatial blood flow in phantoms of the retina, containing imitators of blood vessels and a pump with an adjustable flow velocity for pumping blood-imitating fluid, electric motors and a vibration motor for adjustable deformations and vibrations of the flexible tube walls is presented. When the characteristics of the pump and vibration motor are changed, turbulence is created after the formed laminar flow in the microtubes, which increases quality and blood flow reliability of the phantoms. Series of experiments applying the developed technique and the vessel phantoms with pulsating flow have been carried out using the method of optical coherence tomography. The developed phantoms and the controller can be used to test ophthalmic ultrasound and OCT systems for biomedical studies as well as to develop and adjust new modifications of the OCT imaging.

Microfluidic Devices and Three Dimensional-Printing Strategies for in vitro Models of Bone.

Bone is a complex and highly dynamic tissue, which has been worldwide studied, from fundamental biology to tissue engineering fields. Even so, current in vitro models do not truly replicate the native bone tissue environment. For so, new and improved in vitro tissue models are necessary to obtain more reliable data, not only in a development point of view, but also to fasten the translation of new drugs into the clinics. In this reasoning, tissue-engineering strategies were applied to develop mimetic and three-dimensional (3D) microenvironments, which were associated with microfluidic devices for the development of more complex and realistic systems. Such devices mimic blood vessels that are present in the native tissue, thus enabling the study of complex biological mechanism as such as bone angiogenesis. More recently, 3D printing has been pursued to produce more intricate microfluidic devices and engineered tissues in a single step. The ability to print spatially controlled structures composed of different biomaterials, growth factors and cells caught the attention of scientists for the development of more efficient in vitro models. Additionally, it allows obtaining microfluidic devices and/or engineered tissues with the desired architecture within a small amount of time and with reduced costs. Recently, the use of high-resolution scanning boosted the production of patient-specific implants. Despite the difficulties associated with 3D printed structures that still need to be overcome, it has been proven to be a valuable tool to accomplish a new generation of 3D bioprinted bone-on-a-chip platforms.

Effective Delivery of Anti-Cancer Drug Molecules with Shape Transforming Liquid Metal Particles.

Liquid metals are being studied intensively because of their potential as a drug delivery system. Eutectic gallium–indium (EGaIn) alloy liquid metals have a low melting point, low toxicity, and excellent tissue permeability. These properties may enable them to be vascular embolic agents that can be deformed by light or heat. In this study, we developed EGaIn particles that can deliver anticancer drugs to tumor cells in vitro and change their shapes in response to external stimuli. These particles were prepared by sonicating a solution containing EGaIn and amphiphilic lipids. The liquid metal (LM)/amphiphilic lipid (DSPC, 1,2-distearoyl-sn-glycero-3-phosphocholin) particles formed a vehicle for doxorubicin, an anticancer drug, which was released (up to 50%) when the shape of the particles was deformed by light or heat treatment. LM/DSPC particles are non-toxic and LM/DSPC/doxorubicin particles have anticancer effects (resulting in a cell viability of less than 50%). LM/DSPC/doxorubicin particles were also able to mimic blood vessel embolisms by modifying their shape using precisely controlled light and heat in engineered microchannels. The purpose of this study was to examine the potential of EGaIn materials to treat tumor tissues that cannot be removed by surgery.

Shear-resistant hydrogels to control permeability of porous tubular scaffolds in vascular tissue engineering.

Aiming to perfuse porous tubular scaffolds for vascular tissue engineering (VTE) with controlled flow rate, prevention of leakage through the scaffold lumen is required. A gel coating made of 8% w/v alginate and 6% w/v gelatin functionalized with fibronectin was produced using a custom-made bioreactor-based method. Different volumetric proportions of alginate and gelatin were tested (50/50, 70/30, and 90/10). Gel swelling and stability, and rheological, and uniaxial tensile tests reveal superior resistance to the aggressive biochemical microenvironment, and their ability to withstand physiological deformations (~10%) and wall shear stresses (5-20 dyne/cm2). These are prerequisites to maintain the physiologic phenotypes of vascular smooth muscle cells and endothelial cells (ECs), mimicking blood vessels microenvironment. Gels can induce ECs proliferation and colonization, especially in the presence of fibronectin and higher percentages of gelatin. The custom-designed bioreactor enables the development of reproducible and homogeneous tubular gel coating. The permeability tests show the effectiveness of tubular scaffolds coated with 70/30 alginate/gelatin gel to occlude wadding pores, and therefore prevent leakages. The synthesized double-layered tubular scaffolds coated with alginate/gelatin gel and fibronectin represent both promising substrate for ECs and effective leakproof scaffolds, when subjected to pulsatile perfusion, for VTE applications.

Tissue-engineered blood vessel mimics in complex geometries for intravascular device testing.

ObjectiveIntravascular stents are commonly used to treat occluded arteries during coronary heart disease. After coronary stent implantation, endothelial cells grow over the stent, which is referred to as re-endothelialization. Re-endothelialization prevents blood from clotting on the stent surface and is a good predictor of stent success. Blood vessel mimics (BVMs) are in vitro tissue-engineered models of human blood vessels that may be used to preclinically test stents for re-endothelialization. BVMs have been developed in straight geometries. However, the United States Food and Drug Administration recommends that devices intended to treat coronary occlusions be preclinically tested in bent and bifurcated vessels due to the complex geometries of native coronary arteries. The main objectives of this study were to develop and characterize BVMs in complex geometries.DesignBioreactors were designed and constructed so that BVMs could be cultivated in bent (>45°) and bifurcated geometries. Human umbilical vein endothelial cells were sodded onto complex-shaped scaffolds, and the resulting BVMs were characterized for cell deposition. For a final proof of concept, a coronary stent was deployed in a severely angulated BVM.ResultsThe new bioreactors were easy to use and mounting scaffolds in complex geometries in the bioreactors was successful. After sodding scaffolds with cells, there were no statistically significant differences between the cell densities along the length of the BVMs, on the top and bottom halves of the BVMs, or on the inner and outer halves of the BVMs. This suggests cells deposited evenly throughout the scaffolds, resulting in consistent complex-geometry BVMs. Also, a coronary stent was successfully deployed in a severely angulated BVM.ConclusionsBioreactors can be constructed for housing complex-shaped vessels. BVMs can be developed in the complex geometries observed in native coronary arteries with endothelial cells evenly dispersed throughout BVM lumens.

LED-Based Light Source Combined with Quantum Dot for Spectral Imaging

Quantum dot (QD) is a kind of semiconductor nanoparticle and shows photoluminescence, in which the light with narrow spectral band is emitted from the QD after the absorption of the excitation light. In this study, it is demonstrated that the LED-based light source combined with a QD film can be applied to spectral imaging. The polymer film with QD was prepared by photopolymerization. Through measurement of the emission spectrum, it was confirmed that the optical intensity at the peak wavelength of the spectral bands of 410 and 540 nm could be controlled by changing the thickness of the film. In the study using a home-made phantom mimicking blood vessels, the patterns’ contrast in the phantom images could be enhanced by using LED combined with QD as compared with the white light image. The LED light combined with the QD film provides a useful solution for illumination for spectral imaging.

Tissue-engineered aneurysm models for in vitro assessment of neurovascular devices.

Preclinical testing of neurovascular devices is crucial for successful device design and is commonly performed using in vivo organisms such as the rabbit elastase-induced aneurysm model; however, simple in vitro models may help further refine this testing paradigm. The purpose of the current work was to evaluate, and further develop, tissue-engineered blood vessel mimics (BVMs) as simple, early-stage models to assess neurovascular devices in vitro prior to animal or clinical use. The first part of this work used standard straight-vessel BVMs to evaluate flow diverters at 1, 3, and 5 days post-deployment. The second part developed custom aneurysm-shaped scaffolds to create aneurysm BVMs. Aneurysm scaffolds were characterized based on overall dimensions and microstructural features and then used for cell deposition and vessel cultivation. It was feasible to deploy flow diverters within standard BVMs and cellular linings could withstand and respond to implanted devices, with increasing cell coverage over time. This provided the motivation and foundation for the second phase of work, where methods were successfully developed to create saccular, fusiform, and blister aneurysm scaffolds using a wax molding process. Results demonstrated appropriate fiber morphology within different aneurysm shapes, consistent cell deposition, and successful cultivation of aneurysm BVMs. It is feasible to use tissue-engineered BVMs for assessing cellular responses to flow diverters, and to create custom aneurysm BVMs. This supports future use of these models for simple, early-stage in vitro testing of flow diverters and other neurovascular devices.

Optimal wavelengths for subdiffuse scanning laser oximetry of the human retina.

Retinal blood vessel oxygenation is considered to be an important marker for numerous eye diseases. Oxygenation is typically assessed by imaging the retinal vessels at different wavelengths using multispectral imaging techniques, where the choice of wavelengths will affect the achievable measurement accuracy. Here, we present a detailed analysis of the error propagation of measurement noise in retinal oximetry, to identify optimal wavelengths that will yield the lowest uncertainty in saturation estimation for a given measurement noise level. In our analysis, we also investigate the effect of hemoglobin packing in discrete blood vessels (pigment packaging), which may result in a nonnegligible bias in saturation estimation if unaccounted for under specific geometrical conditions, such as subdiffuse sampling of smaller blood vessels located deeper within the retina. Our analyses show that using 470, 506, and 592nm, a fairly accurate estimation of the whole oxygen saturation regime [0 1] can be realized, even in the presence of the pigment packing effect. To validate the analysis, we developed a scanning laser ophthalmoscope to produce high contrast images with a maximum pixel rate of 60kHz and a maximum 30-deg imaging field of view. Confocal reflectance measurements were then conducted on a tissue-mimicking scattering phantom with optical properties similar to retinal tissue including narrow channels filled with absorbing dyes to mimic blood vessels. By imaging at three optimal wavelengths, the saturation of the dye combination was calculated. The experimental values show good agreement with our theoretical derivations.

Circular-shaped microfluidic device to study the effect of shear stress on cellular orientation.

Understanding the effects of shear stress on mammalian cells is a crucial factor for understanding a number of biological processes and diseases. Here, we show the development of a circular-shaped microfluidic device for the facile generation of shear stress gradients. With this microfluidic device, the effect of shear stress on orientation of human umbilical vein endothelial cells was studied. This microfluidic device, which enables to control the alignment of human umbilical vein endothelial cells within a microchannel, can be a valuable tool to mimic blood vessels.

Directional migration of mesenchymal stem cells under an SDF-1α gradient on a microfluidic device.

Homing of peripheral stem cells is regulated by one of the most representative homing factors, stromal cell-derived factor 1 alpha (SDF-1α), which specifically binds to the plasma membrane receptor CXCR4 of mesenchymal stem cells (MSCs) in order to initiate the signaling pathways that lead to directional migration and homing of stem cells. This complex homing process and directional migration of stem cells have been mimicked on a microfluidic device that is capable of generating a chemokine gradient within the collagen matrix and embedding endothelial cell (EC) monolayers to mimic blood vessels. On the microfluidic device, stem cells showed directional migration toward the higher concentration of SDF-1α, whereas treatment with the CXCR4 antagonist AMD3100 caused loss of directionality of stem cells. Furthermore, inhibition of stem cell’s main migratory signaling pathways, Rho-ROCK and Rac pathways, caused blockage of actomyosin and lamellipodia formation, decreasing the migration distance but maintaining directionality. Stem cell homing regulated by SDF-1α caused directional migration of stem cells, while the migratory ability was affected by the activation of migration-related signaling pathways.

Antiangiogenic Activity of rTRAIL is Potentiated by Sflt-1, a VEGF Trap

Objective: Anti-angiogenesis and pro-apoptosis are the two processes which are strategically used to target tumors. The soluble form of VEGF receptor 1 (Flt-1) is a modulator of VEGF activity and could be useful as a trap to intrinsically sequester VEGF in tumor cells. In the present study, we have investigated the role of rTRAIL in inhibiting VEGF-mediated angiogenesis and its synergistic potentiation by sFlt-1. Methods: We have expressed human recombinant TRAIL, to study its role in anti-angiogenesis and a 2-domain active variant of sFlt-1 by bacterial expression to be used as a trap for VEGF. 3[H] thymidine incorporation assay was used to confirm the inhibitory activity of sFlt-1 and/or rTRAIL on cell proliferation. The synergistic anti-migratory and anti-angiogenic activity of sFlt-1 and rTRAIL were assessed using endothelial cell wound healing and tube formation assay in-vitro and by rat corneal micropocket assay in-vivo. Results: Inhibition of cell proliferation was evident in sFlt-1 and rTRAIL treated cells in a dose -dependent manner with more than 60% reduction in proliferation rate as compared to the tumor cells treated with sFlt-1 and rTRAIL alone. Furthermore, the anti-metastatic and anti-angiogenic activity of sFlt-1 and rTRAIL in combination was evident in endothelial wound healing and tube formation with a significant reduction in the number of cells migrated into the wounded area and the number of honey comb-like structures with the length of the tubes formed to mimic blood vessel formation in-vitro respectively. The in-vivo corneal micropocket assay confirmed the anti-angiogenic effect of sFlt-1 and rTRAIL in combination. Conclusion: The synergistic role of sFlt-1 and rTRAIL as the potential inhibitors of VEGF- mediated angiogenesis could be therapeutically exploited.