Rat igf 1

To determine the production level of growth factors and inflammatory cytokines, blood samples were drawn under anesthesia and collected into sterile blood collection tubes from the eyes of controls rats, untreated rats, only TBO–chit–Au–AgNPs treated rats and TBO–chit–Au–AgNPs+ laser light treated rats. Thereafter, all rats were immediately sacrificed as per the guidelines. The samples were centrifuged at 5000 rpm for 5 min. Thereafter, the serum supernatant was aliquoted in microcentrifuge tubes and stored at − 80 °C for further analysis⁷⁴ .The commercially available enzyme-linked immunosorbent assay (ELISA) kits were used according to the manufacturers’ protocol: Rat TNF-α (Tumour Necrosis Factor-alpha) ELISA Kit (RAY BIOTECH, CAT: ELR-TNFΑ), Rat IL-6(Interleukin-6) ELISA Kit (RAY BIOTECH, CAT: ELR-IL6), Rat TGF-β1 (Transforming growth factor beta-1) ELISA Kit (ELABSCIENCE, CAT: E-EL-0162), Rat EGF (Epidermal growth factor) ELISA Kit (ELABSCIENCE, CAT: E-EL-R0369), Rat VEGF-A (Vascular endothelial growth factor-A) ELISA Kit (ELABSCIENCE, CAT: E-EL-R2603), Rat IGF-1 (Insulin-like growth factor-1) ELISA Kit (ELABSCIENCE, CAT: E-EL-R3001). The levels of TNF-α, IL-6, TGF-β1, EGF, VEGF-A and IGF-1 were measured by ELISA reader (BIORAD MICROPLATE READER, INDIA).