Decellularized Human Amnion-Chorion Membrane as a Novel Scaffold for Expansion of Human Umbilical Cord Tissue- Derived Mesenchymal Stem Cells and Production of Chlorogenic Acid-Loaded Extracellular Vesicles Against Glioblastoma

Abstract Background: Brain cancer named glioblastoma multiforme often called GBM is hard to manage. Survival tends to stretch just 14 to 16 months, despite aggressive therapies. Research now turns toward tiny bubbles released by mesenchymal stem cells, known as extracellular vesicles. These particles slip through the blood-brain barrier without trouble. Meanwhile, attention grows around caffeic acid, a substance found in plants. Early findings hint it may slow tumor growth in the brain. Yet issues remain. Poor absorption into tissues marks one limitation, while access to the brain proves especially difficult. A new approach emerges here. In this study, a biological layer decellularized human amnion-chorion membrane, known as dHACM was tested to cultivate stem cells from umbilical cords, labeled hUCT-MSCs. From these, extracellular vesicles form, capable of transporting CGA against aggressive brain tumors like glioblastoma multiforme. From dHACM tissue, MSCs were isolated through a targeted technique, afterward verifying cell identity with precision. Flow cytometry joined karyotyping in confirming traits, ensuring accuracy without reliance on assumptions. Extracellular vesicles emerged from these cultured cells, captured for further study. Nanoparticle tracking analysis, along with transmission electron microscopy, revealed their physical details clearly. Microwave-assisted loading was applied to insert CGA into the vesicles successfully. Each phase unfolded systematically, maintaining alignment with initial design goals. Moreover, network pharmacology was applied alongside molecular docking to explore how CGA behaves. It turned out EGFR was a key target of CGA activity. When examining U87 glioblastoma multiforme cells, both EVs and CGA showed measurable effects. Health status of the hUCT-MSCs remained strong throughout testing. Abundant levels of CD90 and CD105 appeared in the sample, yet CD31 remained low. Viability was confirmed through standard measures. With particles falling within expected dimensions, binding between CGA and EGFR occurred exactly as intended. Exposure to U87 cells revealed growth inhibition by CGA alone; effects intensified once EVs entered the mix. Findings highlight freeze-thawed dHACM supports MSC expansion, enabling production of EVs capable of delivering CGA against glioblastoma multiforme. Natural compounds like this may one day play a role in targeting aggressive brain tumors. A single advance, but meaningful.