MUMBAI, India, Aug. 17 -- Intellectual Property India has published a patent application (202641096827 A) filed by Swathi Priyadarshinini; Dr. D H Gopalan; Dr. M. Vani; Dr. J. Chanemougavally; and D. Savitha on August 11, 2026, for A Method For Assessing Dose-Dependent Modulation Of Angiogenesis By Chlorpyrifos (cpf) And Its Metabolite 3,5,6-Trichloro-2-Pyridinol (tcpy) Using Chick Chorioallantoic Membrane (cam) Assay With Gene Expression Profiling Of Vegf, Vegfr2, And N-Cadherin.
Inventors include Swathi Priyadarshinini; Dr. D H Gopalan; Dr. M. Vani; Dr. J. Chanemougavally; and D. Savitha.
The application for the patent was published on August 14, 2026, under issue no. 33/2026.
Abstract: The present invention discloses a novel method for assessing the dose-dependent, bi-phasic modulation of angiogenesis by the organophosphate insecticide chlorpyrifos (CPF) and its primary metabolite 3,5,6-trichloro-2-pyridinol (TCPy) using an ex vivo chick chorioallantoic membrane (CAM) assay model. Nine-day-old fertilised country eggs (Gallus gallus) are inoculated via air-sac injection with CPF or TCPy at doses of 0.001, 0.005, 0.01, and 0.05 mg per egg (n=6 per group), along with untreated and vehicle controls. On embryonic day 11, the retrieved CAM is subjected to three-parameter analysis: (i) quantitative image analysis of blood vessel density and branching pattern using AngioSys 2.0 software; (ii) histomorphometric measurement of vessel wall thickness and lumen diameter from haematoxylin and eosin- stained paraffin sections using ImageJ software; and (iii) semiquantitative RT-PCR gene expression analysis of VEGF, VEGFR2, and N-cadherin using specific validated primer sequences. The method reveals that at low doses (0.001-0.005 mg/egg), CPF and TCPy are pro-angiogenic, increasing vessel density (up to 28.34 branches), wall thickness (9-9.8 microns), and lumen diameter (~30 microns), with upregulation of all three angiogenic genes. At high doses (0.01-0.05 mg/egg), CPF and TCPy are anti-angiogenic, reducing vessel density (down to 12 branches), wall thickness (3.4-5.2 microns), and lumen diameter (~9 microns), with specific downregulation of N-cadherin while VEGF and VEGFR2 remain upregulated. The pro-angiogenic effect is attributed to estrogenic mimicry and oxidative stress- mediated VEGF/VEGFR2 activation; the anti-angiogenic effect at high doses is attributed to apoptotic mechanisms and N-cadherin-mediated pericyte-endothelial disruption. This method provides a valuable toolkit for developmental toxicology, environmental risk assessment of organophosphate pesticides, and mechanistic studies of pesticide effects on embryonic vascular development, tumour angiogenesis, wound healing, and diabetic vascular complications.
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