Role of Post-Intraoperative Radiation Therapy Wound Fluids in Interaction with White Blood Cells on Cancer Cell Growth
Authors: Delshad Belal , Abadijoo Hamed , Simaee Hossein , Khayamian Mohammad Ali , Ghaderinia Mohammadreza , Yazdanparast Seyed Mojtaba , Beheshti Jalil , Shamsi Khosro , Avatefi Afkham Maryam , Mansouri Sepideh , Akbari Mohammad Esmaeil , Abdolahad Mohammad 
Published: January 2024

Background: Intraoperative radiation therapy (IORT), is a motivating method which has been widely applied in breast cancer lumpectomy to exclude or reduce the requirement of external beam radiotherapy. Although its effect on destructing remaining vital cancer cells in tumor bed was approved, maintaining or draining post intraoperative radiation therapy wound fluids (PIWFs) is challenging. Moreover, the roles of immune cells in interaction with PIWFs (either in favor or against of therapeutic approaches) haven’t been studied before which is the main investigation of this paper.
Methods:  Surgical wound fluids were collected from 24 IDC patients one day after lumpectomy. Patients were divided to control and IORT groups. The IORT group patients were subjected to an IORT boost while the control group patients did not receive radiotherapy. Collected wound fluids were centrifuged for 20 minutes at 2000 rpm. Subsequential imaging in time evolution was applied to monitor immune cells and cancer cells interactions with each other. The concentration of tumor-associated cytokines and inflammasomes were recorded using the immunoassays. Also we interacted the PIWFs, wound fluids (WFs) and serum of breast cancer patients undergoing BCS (in one group) and BCS + IORT (in another group) with White blood cells (WBCs) and  MDA-MB-231 breast cancer cells to investigate the co=effect of the fluids on both cancer and immune cells.
Results: Surgical wound fluids were collected from 24 IDC patients one day after lumpectomy. Patients were divided to control and IORT groups. The IORT group patients were subjected to an IORT boost while the control group patients did not receive radiotherapy. Collected wound fluids were centrifuged for 20 minutes at 2000 rpm. Subsequential imaging in time evolution was applied to monitor immune cells and cancer cells interactions with each other. The concentration of tumor-associated cytokines and inflammasomes were recorded using the immunoassays. Also we interacted the PIWFs, wound fluids (WFs) and serum of breast cancer patients undergoing BCS (in one group) and BCS + IORT (in another group) with White blood cells (WBCs) and  MDA-MB-231 breast cancer cells to investigate the co=effect of the fluids on both cancer and immune cells.
Conclusion: In summary, post IORT wound fluids (PIWFs) not only stimulate the residue of cancer cells in cavity sides in favor of disease progression but also can not stimulate the immuno cells against cancer cells. We recently showed the effect of PIWFs in increasing the growth and mitosis of tumor cell residues in breast cancer patients. Here we have focused on the effect of PIWFs on the proactivation or deactivation of white blood cells (WBCs) in the tumor bed environment. By sequential imaging in time-transient intervals from the interaction between WBCs and cancer cells, we showed that PIWFs have no additive proactivating effect on immune cells of tumor bed while they stimulated the aggressive functions of cancer cells. Also, cytokine profiling of patients' PIWFs showed no significant immune cell-activating trend with respect to non-IORT cases. In contrast, tumor-associated cytokines were overexpressed in IORT-treated cases. Due to the over-expression of tumor-associated cytokines inside the PIWF of the IORT-treated cases, drainage of PIWFs may be mandatory.