Page 179 - AWSAR 2.0

lining provide a fertile environment. And, the estrogen hormone is vital in keeping the endometrium fertile. However, if the thickness of the endometrium is below 7 mm then it is considered a thin endometrium. In that case, the embryo does not withstand and results in miscarriage and the menstrual cycle begins again.
Other reasons may include improper blood flow/poor angiogenesis, abnormal periods, D&C or pelvic surgeries, intrauterine adhesions, fibroids, too much usage of Clomid and long-term use of birth control pills.
Keeping this preview in mind, we started working on Endometrium AngiogenesisVEGF121, the shortest isoform which is abundantly expressed in endometrial stromal cells and was amplified using specific primers; cloned in pET vector and expressed in BL21 (DE3) pLysS (E.coli) expression system. Expressing proteins in E.coli are advantageous which includes,
easy scale-up, high expression
level at low cost. VEGF121,
one of the splice variants of
VEGF A, is an important factor
in angiogenesis. This 16 kDa
protein is more angiogenic than
other isoforms; it is because it
can freely diffuse from the cells
producing it.
On appropriate administration, VEGF can
promote angiogenesis and
signal vascular endothelial
cells, which help in cell
proliferation and migration but the therapeutic effects of this protein can be achieved at high doses but, it may also lead to the development of a vascular tumour.
Thus, we planned to encapsulate rVEGF121 with Poly-l-lactide (PLA), which is an FDA-approved biodegradable polymer that undergoes hydrolysis and breaks down
Mr. Sunil Abraham || 155
to its monomers when injected into the body and becomes biologically compatible material because of their polyester nature. Tricarboxylic acid cycle (TCA) helps in removal of the PLA compound from the body.
In our study, we examined the usage of poly-l-lactide (PLA) as a delivery system for rVEGF121 and characterized the chemical properties of PLA encapsulated rVEGF121 microparticle for endometrial angiogenesis. To study the angiogenic potential of rVEGF121, we purified the recombinant VEGF121 using His-tagged column and then encapsulated it with PLA. We analyzed the particles using Scanning Electron Microscopy (SEM), which provided us information about how the particles had formed and their size. And, if the PLA-VEGF121 compound is injected into a specific site then how can it supply the protein totheaffectedarea?Tounderstandthesame, the encapsulated compound was subjected to
release kinetics. The compound was mixed in media and was incubated in a shaker, every 24 hours the sample was collected up to 30 days and tested using VEGF ELISA. Released VEGF121 performed better thus the release was found daily till day 30. This suggested that PLA degrades and releases VEGF in a sustained motion which on injection does the same function.
If PLA releases the encapsulated VEGF121 then how does one confirm that this encapsulated compound is performing angiogenesis? To answer this question we analyzed the PLA compound for its bioactivity. We studied the migration and proliferation effect of PLA- VEGF121 on Human Umbilical Vein Endothelial Cells (HUVEC). As a result, the cells were able to migrate in the presence of rVEGF and
Endometrium lining is very important for a female uterus because it supports implantation. If there is no healthy endometrium lining there is no way for the embryos to implant, which results in menstruation.