Evaluation of recombinant human Vascular Endothelial Growth Factor VEGF121-loaded poly-l-lactide microparticles as a controlled release delivery system

Abraham, Sunil; Rangaswamy, Srinivasa Prasad; Amutha, C.

doi:10.3906/biy-1908-32

Cited by 3 publications

(8 citation statements)

References 44 publications

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“…HESC (5000 cells/well) were cultivated in 96-well plates (Corning, NY, USA) consecutively, in the specified medium with growth factors, in 5% CO 2 at 37 °C incubator for the cells to adhere to the wells. The medium was replaced with fresh serum and growth factor reduced medium with rVEGF 121 (200 and 500 ng) and VEGF-loaded PLA microparticles (days 1, 20, and 30) [ 12 ] and determined its effects on proliferation. MTT assay was performed as per the manufacturer’s protocol (HiMedia, Mumbai, India).…”

Section: Methodsmentioning

confidence: 99%

“…The monolayer was scratched with a sterile tip and the cellular debris was washed with PBS. The cells were thereafter incubated with VEGF (study group) and without rVEGF 121 (200 and 500 ng) (control group) and VEGF-loaded PLA microparticles (days 1, 20, and 30) [ 12 ] in serum-free DMEM for at least 48 h for the migration of the cells to the scratched area. The plates were imaged in a camera connected with an inverted microscope (Leica).…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…VEGF 121 , the shortest isoform of VEGF-A, is abundantly expressed in the mid-late proliferative phase of human endometrium stromal cells [ 12 ]. Recombinant VEGF 121 and its microparticles were already characterized in our earlier studies [ 12 ]. This 16 kDa protein is more angiogenic than other isoforms, mainly because it can freely diffuse from the cells producing it [ 13 ].…”

Section: Introductionmentioning

confidence: 99%

“…Thus, there is a need for a controlled delivery mechanism that will release the bioactive molecules in a sustained mode to the delivery site. Analysis and characterization of rVEGF 121 and PLA-VEGF were already reported in our earlier publication [ 12 ]. Here we have studied the role of VEGF 121 - and PLA-VEGF-mediated signaling for its angiogenesis effect in vitro especially in endometrial stromal cells.…”

Section: Introductionmentioning

confidence: 99%

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Encapsulated VEGF121-PLA microparticles promote angiogenesis in human endometrium stromal cells

Abraham

Sanjay

Majiyd³

et al. 2021

Journal of Genetic Engineering and Biotechnology

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Background In this study, Vascular Endothelial Growth Factor 121 expressed abundantly in endometrial stromal cells is encapsulated with poly-l-lactide and characterized the properties for endometrial angiogenesis. We studied the migration, proliferation and the protein levels of human immortalized endometrium stromal cells after treating the cells with recombinant Vascular Endothelial Growth Factor (200 and 500 nanogram), and poly-l-lactide loaded Vascular Endothelial Growth Factor 121 (day 1, 20 and 30). The present study explains endometrium angiogenesis because endometrium plays an important role in pregnancy. Results Migration and proliferation studies in endometrium cells proved the efficiency of Vascular Endothelial Growth Factor and poly-l-lactide loaded Vascular Endothelial Growth Factor 121. This proliferated and increased the migration of the cells in vitro and also activated the Protein kinase B, Phosphatidylinositol-4, 5-Bisphosphate 3-Kinase Catalytic Subunit Beta, α-Smooth muscle actin and vascular endothelial growth factor receptor 2 pathways. Western blot analysis showed the increased expression levels of kinases, smooth muscle actin and vascular endothelial growth factor receptor 2 after the treatment with Vascular Endothelial Growth Factor and poly-l-lactide loaded Vascular Endothelial Growth Factor 121 particles in comparison to the control group. The elevated levels of α-Smooth muscle actin in endometrium cells with Vascular Endothelial Growth Factor prove the regulation of angiogenesis in vitro. Conclusion Endometrium thickness is one of the important factors during implantation of embryo and pregnancy. Slow release of VEGF from PLA encapsulated microparticle further controls the endothelial cell proliferation and migration and helps in the promotion of angiogenesis. The combined effect studied in vitro could be used as a pro-angiogenic drug on further in vivo confirmation.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Encapsulated VEGF121-PLA microparticles promote angiogenesis in human endometrium stromal cells

Abraham

Sanjay

Majiyd³

et al. 2021

Journal of Genetic Engineering and Biotechnology

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VEGF-A and FGF4 Engineered C2C12 Myoblasts and Angiogenesis in the Chick Chorioallantoic Membrane

2022

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Angiogenesis is the formation of new blood vessels from pre-existing vessels. Adequate oxygen transport and waste removal are necessary for tissue homeostasis. Restrictions in blood supply can lead to ischaemia which can contribute to disease pathology. Vascular endothelial growth factor (VEGF) is essential in angiogenesis and myogenesis, making it an ideal candidate for angiogenic and myogenic stimulation in muscle. We established C2C12 mouse myoblast cell lines which stably express elevated levels of (i) human VEGF-A and (ii) dual human FGF4-VEGF-A. Both stably transfected cells secreted increased amounts of human VEGF-A compared to non-transfected cells, with the latter greater than the former. In vitro, conditioned media from engineered cells resulted in a significant increase in endothelial cell proliferation, migration, and tube formation. In vivo, this conditioned media produced a 1.5-fold increase in angiogenesis in the chick chorioallantoic membrane (CAM) assay. Delivery of the engineered myoblasts on Matrigel demonstrated continued biological activity by eliciting an almost 2-fold increase in angiogenic response when applied directly to the CAM assay. These studies qualify the use of genetically modified myoblasts in therapeutic angiogenesis for the treatment of muscle diseases associated with vascular defects.

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Microvascular Experimentation in the Chick Chorioallantoic Membrane as a Model for Screening Angiogenic Agents including from Gene-Modified Cells

Kennedy

Coen

Wheatley

et al. 2021

IJMS

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The chick chorioallantoic membrane (CAM) assay model of angiogenesis has been highlighted as a relatively quick, low cost and effective model for the study of pro-angiogenic and anti-angiogenic factors. The chick CAM is a highly vascularised extraembryonic membrane which functions for gas exchange, nutrient exchange and waste removal for the growing chick embryo. It is beneficial as it can function as a treatment screening tool, which bridges the gap between cell based in vitro studies and in vivo animal experimentation. In this review, we explore the benefits and drawbacks of the CAM assay to study microcirculation, by the investigation of each distinct stage of the CAM assay procedure, including cultivation techniques, treatment applications and methods of determining an angiogenic response using this assay. We detail the angiogenic effect of treatments, including drugs, metabolites, genes and cells used in conjunction with the CAM assay, while also highlighting the testing of genetically modified cells. We also present a detailed exploration of the advantages and limitations of different CAM analysis techniques, including visual assessment, histological and molecular analysis along with vascular casting methods and live blood flow observations.

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Evaluation of recombinant human Vascular Endothelial Growth Factor VEGF121-loaded poly-l-lactide microparticles as a controlled release delivery system

Abstract: Vascular endothelial growth

Cited by 3 publications

References 44 publications

Encapsulated VEGF121-PLA microparticles promote angiogenesis in human endometrium stromal cells

Encapsulated VEGF121-PLA microparticles promote angiogenesis in human endometrium stromal cells

VEGF-A and FGF4 Engineered C2C12 Myoblasts and Angiogenesis in the Chick Chorioallantoic Membrane

Microvascular Experimentation in the Chick Chorioallantoic Membrane as a Model for Screening Angiogenic Agents including from Gene-Modified Cells

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