Conditioned Medium Derived from the Human Amniotic Membrane Prevents Brain Damage against Cerebral Ischemia/Reperfusion in Three Phases, Subacute, Acute, and Chronic in a Stroke Model in Rats

Abstract

Introduction: Stem cells isolated from amniotic membrane can produce and freedom substances that have the ability to regenerate damaged tissues and contain proteins and other factors that apply via numerous major and minor mechanisms leading to increasing angiogenesis and tissue survival. The research was conducted to prove the defensive characteristics of the secretome in the face of temporary focal cerebral ischemia in mouse stroke models. Methods: Cerebral ischemia protocol in a specific area was implemented in rats with middle cerebral artery occlusion for 60 minutes and then reperfusion was given for 6, 20 and 30 minutes. Within 30 minutes after the start of reperfusion, conditioned medium derived from human amniotic membrane (AMSC-CM) was poured into the right ventricle (ICV) with a dose of 0.5 microliters. Finally, the volume of the injury, cerebral tissue water, sensorimotor activity and the strength of the blood-brain barrier integrity were evaluated 24 hours after drug injection. Results: ICV injection of conditioned medium at the start of reperfusion phase considerably decrease the volume of the injury in 6, 20, and 30 hours after reperfusion compared MCAO operated group (p < 0.01), cerebral tissue water in the treatment group decreased considerably after intervention in comparison with the MCAO group in core and penumbral area not in the subcortical area (p < 0.05), Also, the Evans Blue penetration rate in all times in the core and penumbral area in AMSC-CM group considerably decreased paralleled with the MCAO group (p < 0.05). Conclusion: The results show that treatment with AMSC-CM during 6-30 h after ischemia-reperfusion insult exerts some beneficial effects against ischemia-reperfusion injury. These findings provide an important vision for more complementary research and treatment of stroke.