Strategic Endothelial Cell Tube Formation Assay: Comparing Extracellular Matrix and Growth Factor Reduced Extracellular Matrix

Xie, Daniel; Ju, Donghong; Speyer, Cecilia L.; Gorski, David; Kosir, Mary A.

doi:10.3791/54074-v

Cited by 3 publications

(2 citation statements)

References 2 publications

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“…Tube formation assays were conducted, which, as a more sophisticated in vitro model, could simulate all the steps involved in the angiogenic process in vivo: adhesion, migration, protease activity, alignment, and tube formation. [33] Consistent with the wound closure results, a similar trend was observed in the tube formation assays (Figure 2E-G). Relative to the DMEM group, the MR-T plus GM6001 and MR n -T groups showed little change in tube formation.…”

Section: Mt1-mmp-activated Proangiogenic Ability Of Mr-t-pd In Vitrosupporting

confidence: 88%

Enzyme‐Activated Prodrug‐Based Smart Liposomes Specifically Enhance Tumor Hemoperfusion with Efficient Drug Delivery to Pancreatic Cancer Cells and Stellate Cells

Duan

Song

et al. 2021

Adv Funct Materials

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Tumor‐specific enhanced delivery of chemotherapeutics and modulators to tumor cells and activated pancreatic stellate cells (aPSCs), respectively, represents safer and more effective therapy for pancreatic cancer. Herein, a membrane type 1‐matrix metalloproteinase (MT1‐MMP)‐cleavable spacer is used to assemble low‐density cRGDfK onto thermosensitive liposomes loaded with phosphorylated calcipotriol (PCAL) and doxorubicin (DOX), yielding MR‐T‐PD. The liposome‐linked cRGDfK prodrug on MR‐T‐PD surface is first activated by MT1‐MMP, which is selectively expressed on tumor endothelial cells, to release cRGDfK. The free cRGDfK specifically promotes tumor angiogenesis, leading to 3.4‐fold higher accumulation and a wider distribution of MR‐T‐PD in tumors. Furthermore, MR‐T‐PD rapidly releases PCAL and DOX into the interstitium under heat treatment. The released DOX enters tumor cells to induce apoptosis, whereas the PCAL prodrug is converted to CAL by alkaline phosphatase on the surface of aPSCs; CAL can then enter aPSCs to induce quiescence and promote the antitumor effect of DOX. Finally, by enhancing the exposure of DOX and CAL to tumor cells and aPSCs, respectively, in a tumor‐specific manner, MR‐T‐PD exerts superior efficacy (a 5.9‐fold decrease in tumor weight) without causing additional side effects. Overall, this prodrug‐based smart liposome system represents a promising paradigm for pancreatic cancer therapy.

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Section: Mt1-mmp-activated Proangiogenic Ability Of Mr-t-pd In Vitrosupporting

confidence: 88%

Enzyme‐Activated Prodrug‐Based Smart Liposomes Specifically Enhance Tumor Hemoperfusion with Efficient Drug Delivery to Pancreatic Cancer Cells and Stellate Cells

Duan

Song

et al. 2021

Adv Funct Materials

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“…The EC networks shown in Figure were analyzed via image quantification to determine the overall interconnected network volumes, , identify the overall features of the network, and measure the average diameter of the tube-like structures (Figure ). The total volume of an EC network was defined as the sum of all volumes between two branch points or between a branch point and a terminal point.…”

Section: Resultsmentioning

confidence: 99%

VEGF-Encoding, Gene-Activated Collagen-Based Matrices Promote Blood Vessel Formation and Improved Wound Repair

Hwang

Kiick

Sullivan

2023

ACS Appl. Mater. Interfaces

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Disruption in vascularization during wound repair can severely impair healing. Proangiogenic growth factor therapies have shown great healing potential; however, controlling growth factor activity and cellular behavior over desired healing time scales remains challenging. In this study, we evaluated collagen-mimetic peptide (CMP) tethers for their capacity to control growth factor gene transfer and growth factor activity using our recently developed gene-activated hyaluronic acid-collagen matrix (GAHCM). GAHCM was comprised of DNA/polyethyleneimine (PEI) polyplexes that were retained on hyaluronic acid (HA)-collagen hydrogels using CMPs. We hypothesized that using CMP-collagen tethers to control vascular endothelial growth factor-A (VEGF-A) gene delivery in fibroblasts would provide a powerful strategy to modulate the proangiogenic behaviors of endothelial cells (ECs) for blood vessel formation, resulting in enhanced wound repair. In co-culture experiments, we observed that CMP-modified GAHCM induced tunable gene delivery in fibroblasts as predicted, and correspondingly, VEGF-A produced by the fibroblasts led to increased growth and persistent migration of ECs for at least 7 days, as compared to non-CMP-modified GAHCM. Moreover, when ECs were exposed to fibroblast-containing VEGF-GAHCM with higher levels of CMP modification (50% CMP-PEI, or 50 CP), high CD31 expression was stimulated, resulting in the formation of an interconnected EC network with a significantly higher network volume and a larger diameter network structure than controls. Application of VEGF-GAHCM with 50 CP in murine splinted excisional wounds facilitated prolonged prohealing and proangiogenic responses resulting in increased blood vessel formation, improved granulation tissue formation, faster re-epithelialization, and overall enhanced repair. These findings suggest the benefits of CMP-collagen tethers as useful tools to control gene transfer and growth factor activity for improved treatment of wounds.

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Effects of FTMT Expression by Retinal Pigment Epithelial Cells on Features of Angiogenesis

Buyandelger

Walker

Yanagisawa

et al. 2020

IJMS

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Aberrant angiogenesis is a pathological feature of a number of diseases and arises from the uncoordinated expression of angiogenic factors as response to different cellular stresses. Age-related macular degeneration (AMD), a leading cause of vision loss, can result from pathological angiogenesis. As a mutation in the mitochondrial ferritin (FTMT) gene has been associated with AMD, its possible role in modulating angiogenic factors and angiogenesis was investigated. FTMT is an iron-sequestering protein primarily expressed in metabolically active cells and tissues with high oxygen demand, including retina. In this study, we utilized the human retinal pigment epithelial cell line ARPE-19, both as undifferentiated and differentiated cells. The effects of proinflammatory cytokines, FTMT knockdown, and transient and stable overexpression of FTMT were investigated on expression of pro-angiogenic vascular endothelial growth factor (VEGF) and anti-angiogenic pigment epithelial-derived factor (PEDF). Proinflammatory cytokines induced FTMT and VEGF expression, while NF-κB inhibition significantly reduced FTMT expression. VEGF protein and mRNA expression were significantly increased in FTMT-silenced ARPE-19 cells. Using an in vitro angiogenesis assay with endothelial cells, we showed that conditioned media from FTMT-overexpressing cells had significant antiangiogenic effects. Collectively, our findings indicate that increased levels of FTMT inhibit angiogenesis, possibly by reducing levels of VEGF and increasing PEDF expression. The cellular models developed can be used to investigate if increased FTMT may be protective in angiogenic diseases, such as AMD.

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Strategic Endothelial Cell Tube Formation Assay: Comparing Extracellular Matrix and Growth Factor Reduced Extracellular Matrix

Cited by 3 publications

References 2 publications

Enzyme‐Activated Prodrug‐Based Smart Liposomes Specifically Enhance Tumor Hemoperfusion with Efficient Drug Delivery to Pancreatic Cancer Cells and Stellate Cells

Enzyme‐Activated Prodrug‐Based Smart Liposomes Specifically Enhance Tumor Hemoperfusion with Efficient Drug Delivery to Pancreatic Cancer Cells and Stellate Cells

VEGF-Encoding, Gene-Activated Collagen-Based Matrices Promote Blood Vessel Formation and Improved Wound Repair

Effects of FTMT Expression by Retinal Pigment Epithelial Cells on Features of Angiogenesis

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