A study of digital image analysis on the acrylamide derivative monomers induced apoptosis in rat cerebrum

Nowadays, apoptosis is mostly evaluated visually in histological studies. By using the quantitative digital image analysis, this study aimed to investigate the effect of acrylamide-based monomers (acrylamide [AAm], methacrylamide [MAAm], N-isopropylacrylamide [NIPAm]) on the cerebrum tissues in rats, which are the most common water-soluble monomers in the production of polymeric hydrogels used as biomaterials. The Wistar albino rats weighing ~220-240 g were divided into control and three test groups. The control group received 1 mL of saline, and the test groups received 1 mL of aqueous 50 mg/kg/day intramuscular injection of AAm, MAAm, and NIPAm, respectively. At the end of the experiments, brain tissues of all rats euthanized by intramuscular injection of sodium pentobarbital were removed. Terminal deoxynucleotide transferase dUTP nick and labeling (TUNEL) method was applied to brain tissue sections. The monomers have been shown to cause apoptosis due to oxidative stress in cerebrum tissue. Based on apoptosis by tunneling method, quantitative digital image analysis of cell fragments was performed with Olympus cellSens Dimension 1.15 software, and the number, total count area, selected area, average area, and ROI% values of the fragments were found. In addition, the total area and ROI% values of the fragments increased linearly with increasing the molar mass of monomers from the digital image analysis data. Quantitative digital image analysis can facilitate the monitoring of apoptosis caused by the oxidative stress of monomers used in the production of the biomaterials.