Figure 9

Proposed schematic showing direct and bystander effects of WF of IORT. After surgery, tumor spheroids were established to study the tumor behavior in 3D culturing in tumor on-chip. Tumor spheroids contain many cell types, cancer cells, cancer-associated fibroblasts (CAFs), and Tumor-associated macrophages (TAMs). Negative margin exposure to IORT shows the direct effect of radiation that inhibits several signaling pathways that its effect could target the residual cancer cells in the negative margin. Wound fluid (WF) produced in the surgical cavity contains several cytokines, growth factors, and free radicals after IORT, which have a bystander effect. After 24 h of IORT, WF was collected and studied on 3D spheroid and 2D cell culturing that show several biological processes, including arrested cell cycle through free radicals and DNA fragmentation, downregulation of Hsp-90 that stabilized TP-53 in cancer cells, upregulation P21 and P16 that promote cell cycle arrest and finally senescence. Senescence-associated secretory phenotype (SASP) factors also activated pathways to inhibit the cell cycle and activated several biological processes such as apoptosis and senescence. Growth factors were increased to promote the wound healing process in the surgical cavity after IORT and increase proliferation, motility, and migration of some spheroids and cell lines. The figure was created using Biorender (https://biorender.com).