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Factors down-regulated in malignant vs. benign seroma profiles. a Relative expression of cytokines down-regulated in malignant post-operative seroma fluid compared to benign seroma. b IL-16 expression in benign and malignant seroma fluid presented collectively and at specific collection time-points post surgery (group 1 = collection time point ≤8 days and group 2 = collection time-point >8 days). c IGFBP-1 expression in benign and malignant seroma fluid presented collectively and at specific collection time-points post surgery (group 1 = collection time point ≤8 days and group 2 = collection time-point >8 days). d Pearson correlation analysis showing a significant negative correlation of IL-16 expression and collection time-point of seroma post-surgery [Pearson r = −0.6082; p
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Purpose:
The accumulation of wound fluid known as seroma in the chest cavity following breast surgery is a common occurrence that can persist for many weeks. While the pro-inflammatory composition of seroma is well established, there has been remarkably little research to determine whether seroma contains pro-oncogenic factors, and whether this is...
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Traditionally, conservative breast surgery was contraindicated in centrally located breast tumors, with total mastectomy as the treatment of choice. However, restorations of the central defects by the oncoplastic volume displacement or replacement techniques have been shown to be effective. The aim of the current study was to assess the surgical ou...
Citations
... Protein extraction from blood plasma, serous fluid, and skin tissue was performed as previously described (Valeta-Magara et al., 2015;Lese et al., 2018;Lese et al., 2021). Blood and serous fluid were collected at pre-defined timepoints (PODs 14,21,28,35). ...
... Blood and serous fluid were collected at pre-defined timepoints (PODs 14,21,28,35). The samples were spun down, twice for the blood, at 1,000 rcf for 10 min, followed by 1,500 rcf for 15 min, and once for the serous fluid at 1,500 rcf for 15 min (Valeta-Magara et al., 2015). Supernatants were collected and further subjected to quantitative assessments. ...
Background: Seroma formation is a common postoperative complication. Fibrin-based glues are typically employed in an attempt to seal the cavity. Recently, the first nanoparticle (NP)-based treatment approaches have emerged. Nanoparticle dispersions can be used as tissue glues, capitalizing on a phenomenon known as ‘nanobridging’. In this process, macromolecules such as proteins physically adsorb onto the NP surface, leading to macroscopic adhesion. Although significant early seroma reduction has been shown, little is known about long-term efficacy of NPs. The aim of this study was to assess the long-term effects of NPs in reducing seroma formation, and to understand their underlying mechanism.
Methods: Seroma was surgically induced bilaterally in 20 Lewis rats. On postoperative day (POD) 7, seromas were aspirated on both sides. In 10 rats, one side was treated with NPs, while the contralateral side received only NP carrier solution. In the other 10 rats, one side was treated with fibrin glue, while the other was left untreated. Seroma fluid, blood and tissue samples were obtained at defined time points. Biochemical, histopathological and immunohistochemical assessments were made.
Results: NP-treated sides showed no macroscopically visible seroma formation after application on POD 7, in stark contrast to the fibrin-treated sides, where 60% of the rats had seromas on POD 14, and 50% on POD 21. At the endpoint (POD 42), sides treated with nanoparticles (NPs) exhibited significant macroscopic differences compared to other groups, including the absence of a cavity, and increased fibrous adhesions. Histologically, there were more macrophage groupings and collagen type 1 (COL1) deposits in the superficial capsule on NP-treated sides.
Conclusion: NPs not only significantly reduced early manifestations of seroma and demonstrated an anti-inflammatory response, but they also led to increased adhesion formation over the long term, suggesting a decreased risk of seroma recurrence. These findings highlight both the adhesive properties of NPs and their potential for clinical therapy.
... The copyright holder for this preprint (which this version posted April 28, 2023. ; https://doi.org/10.1101/2023.04.26.538467 doi: bioRxiv preprint results are consistent with events and phases of wound healing as reported in the literature [52,50]. Plots display time-dependent changes in fibroblast density, collagen density, and cytokine concentration at the cavity center as determined from simulations and histology. ...
Breast cancer is the most commonly diagnosed cancer type worldwide. Given high survivorship, increased focus has been placed on long-term treatment outcomes and patient quality of life. While breast-conserving surgery (BCS) is the preferred treatment strategy for early-stage breast cancer, anticipated healing and breast deformation (cosmetic) outcomes weigh heavily on surgeon and patient selection between BCS and more aggressive mastectomy procedures. Unfortunately, surgical outcomes following BCS are difficult to predict, owing to the complexity of the tissue repair process and significant patient-to-patient variability. To overcome this challenge, we developed a predictive computational mechanobiological model that simulates breast healing and deformation following BCS. The coupled biochemical-biomechanical model incorporates multi-scale cell and tissue mechanics, including collagen deposition and remodeling, collagen-dependent cell migration and contractility, and tissue plastic deformation. Available human clinical data evaluating cavity contraction and histopathological data from an experimental porcine lumpectomy study were used for model calibration. The computational model was successfully fit to data by optimizing biochemical and mechanobiological parameters through the Gaussian Process. The calibrated model was then applied to define key mechanobiological parameters and relationships influencing healing and breast deformation outcomes. Variability in patient characteristics including cavity-to-breast volume percentage and breast composition were further evaluated to determine effects on cavity contraction and breast cosmetic outcomes, with simulation outcomes aligning well with previously reported human studies. The proposed model has the potential to assist surgeons and their patients in developing and discussing individualized treatment plans that lead to more satisfying post-surgical outcomes and improved quality of life.
... The differential expression of pro-oncogenic growth factors and cytokines is secreted from malignant to benign lesions in the post-surgical seroma in breast tumors. Valeta-Magara and These results showed the preconditioning effect of normal surrounding tissue on the tumor and provided a pro-oncogenic environment that remains after the removal of the tumor by surgery (108). In a recent study, Agresti and collaborators detected 34 cytokines, growth factors, and chemokines in seroma of 27 BC patients that promote the initiation and development of cancer. ...
Intraoperative radiotherapy (IORT) has become a growing therapy for early-stage breast cancer (BC). Some studies claim that wound fluid (seroma), a common consequence of surgical excision in the tumor cavity, can reflect the effects of IORT on cancer inhibition. However, further research by our team and other researchers, such as analysis of seroma composition, affected cell lines, and primary tissues in two-dimensional (2D) and three-dimensional (3D) culture systems, clarified that seroma could not address the questions about IORT effectiveness in the surgical site. In this review, we mention the factors involved in tumor recurrence, direct or indirect effects of IORT on BC, and all the studies associated with BC seroma to attain more information about the impact of IORT-induced seroma to make a better decision to remove or remain after surgery and IORT. Finally, we suggest that seroma studies cannot decipher the mechanisms underlying the effectiveness of IORT in BC patients. The question of whether IORT-seroma has a beneficial effect can only be answered in a trial with a clinical endpoint, which is not even ongoing.
... Seroma contains several growths and inflammatory factors due to physiologic responses to the operation and wound healing process (5). Some studies suggest that seroma induces an increase in cell proliferation, migration, and invasion in the BC cell line (6)(7)(8). However, the proteomics analysis of IORT-treated seroma has shown inhibition of cell proliferation and invasion in the BC cell line (9,10). ...
... It has been shown that seroma can cause the migration and invasion of the cancer cell lines (6)(7)(8). Our results indicate that IORT-treated seroma causes an inhibition in the migration and invasion of the cancer cell lines. We have analyzed the expression level of MMP-9, which is an invasion biomarker. ...
Background: Intraoperative radiation therapy (IORT) is a novel approach to breast cancer (BC) treatment. Objectives: In this study, we compared the cellular and molecular effects of IORT-treated post-lumpectomy wound fluid (seroma) at the point of IOeRT versus IOxRT on the BC cell line. Methods: Immortalized human BC cell lines: MCF-7, MDA-MB-231, and MCF10 were incubated with seroma from 3 groups of patients (as a pilot study). The first group received Intraoperative electron radiation therapy (IOeRT, Boost dose=12Gy), the second one received IOeRT (Radical dose=21Gy), and the third group was prescribed Intraoperative x-ray radiation therapy (IOxRT, X-ray=20Gy). Cellular and molecular tests were used to investigate how cells are influenced by the IORT-treated seroma. Results: We evaluated the effects of dose-time and source-dependent IORT-treated seroma on BC cell lines. In this study, we observed that IOxRT-treated seroma has the most significant effects on the reduction of proliferation, induced cell cycle arrest, and apoptosis. Furthermore, inhibited migration and invasion of BC cell lines were compared to IOeRT -treated seroma. Conclusions: Although this is a pilot study, we suggest that at 24 h, the IORT (specifically IOxRT)-treated seroma may play an important protective role in the breast tumor bed, which is followed by local recurrence decreases.
... Studies on primary breast cancer cells indicate that wound fluid promotes cell chemoresistance 86 . Further, the expression level of pro-oncogenic cytokines and growth factors in surgical-induced wound fluid differ significantly between benign and malignant lesions 87 . Several reports have indicated that the accumulation of surgery-induced wound fluid in the surgical cavity after lumpectomy stimulates wound healing processes, which likely contribute to the increased risk of local recurrences in patients with breast cancer [88][89][90] . ...
Intraoperative radiotherapy (IORT) could abrogate cancer recurrences, but the underlying mechanisms are unclear. To clarify the effects of IORT-induced wound fluid on tumor progression, we treated breast cancer cell lines and human-derived tumor spheroids in 2D and microfluidic cell culture systems, respectively. The viability, migration, and invasion of the cells under treatment of IORT-induced wound fluid (WF-RT) and the cells under surgery-induced wound fluid (WF) were compared. Our findings showed that cell viability was increased in spheroids under both WF treatments, whereas viability of the cell lines depended on the type of cells and incubation times. Both WFs significantly increased sub-G1 and arrested the cells in G0/G1 phases associated with increased P16 and P21 expression levels. The expression level of Caspase 3 in both cell culture systems and for both WF-treated groups was significantly increased. Furthermore, our results revealed that although the migration was increased in both systems of WF-treated cells compared to cell culture media-treated cells, E-cadherin expression was significantly increased only in the WF-RT group. In conclusion, WF-RT could not effectively inhibit tumor progression in an ex vivo tumor-on-chip model. Moreover, our data suggest that a microfluidic system could be a suitable 3D system to mimic in vivo tumor conditions than 2D cell culture.
... Cytokines and growth factors secreted by tumour cells and surrounding cells are affected by radiation therapy and by invasive procedures, such as surgery [21,22]. IFN, TGF-β, IL-1β, IL-6, IL-7, and granulocytemacrophage colony-stimulating factor (GMCF) are known to regulate pro-and anti-immune responses [23]. ...
Background
Pancreatic cancer has highly aggressive features, such as local recurrence that leads to significantly high morbidity and mortality and recurrence after successful tumour resection. Intraoperative radiation therapy (IORT), which delivers targeted radiation to a tumour bed, is known to reduce local recurrence by directly killing tumour cells and modifying the tumour microenvironment.
Methods
Among 30 patients diagnosed with pancreatic cancer, 17 patients received IORT immediately after surgical resection. We investigated changes in the immune response induced by IORT by analysing the peritoneal fluid (PF) and blood of patients with and without IORT treatment after pancreatic cancer surgery. Further, we treated three pancreatic cell lines with PF to observe proliferation and activity changes.
Results
Levels of cytokines involved in the PI3K/SMAD pathway were increased in the PF of IORT-treated patients. Moreover, IORT-treated PF inhibited the growth, migration, and invasiveness of pancreatic cancer cells. Changes in lymphocyte populations in the blood of IORT-treated patients indicated an increased immune response.
Conclusions
Based on the characterisation and quantification of immune cells in the blood and cytokine levels in the PF, we conclude that IORT induced an anti-tumour effect by activating the immune response, which may prevent pancreatic cancer recurrence.
Clinical trial registration
NCT03273374.
... Cytokines and growth factors secreted by tumour cells and surrounding cells are affected by radiation therapy and by invasive procedures, such as surgery. [20,21] IFN, TGF-β, IL-1β, IL-6, IL-7, and granulocytemacrophage colony-stimulating factor (GMCF) are known to regulate pro-and anti-immune responses. [22] In the current study, we identi ed cytokines secreted at elevated levels in the PF of IORT-treated pancreatic cancer patients compared with those in the PF of patients who underwent surgical resection alone. ...
Background: Pancreatic cancer has highly aggressive features, such as local recurrence that leads to significantly high morbidity and mortality and recurrence after successful tumour resection. Intraoperative radiation therapy (IORT), which delivers targeted radiation to a tumour bed, is known to reduce local recurrence by directly killing tumour cells and modifying the tumour microenvironment.
Methods: Among 30 patients diagnosed with pancreatic cancer, 17 patients received IORT immediately after surgical resection. We investigated changes in the immune response induced by IORT by analysing the peritoneal fluid (PF) and blood of patients with and without IORT treatment after pancreatic cancer surgery. Further, we treated three pancreatic cell lines with PF to observe proliferation and activity changes.
Results: Levels of cytokines involved in the PI3K/SMAD pathway were increased in the PF of IORT-treated patients. Moreover, IORT-treated PF inhibited the growth, migration, and invasiveness of pancreatic cancer cells. Changes in lymphocyte phenotype populations in the blood of IORT-treated patients indicated an increased immune response.
Conclusions: Based on the characterisation and quantification of immune cells in the blood and cytokine levels in the PF, IORT induced an anti-tumour effect by activating the immune response, which may prevent pancreatic cancer recurrence.
Clinical Trial Registration: NCT03273374
... Stimulatory effects of post-lumpectomy seroma on cell proliferation via activation of stat-3 was reported [10]. In addition, a research reported pro-oncogenic cytokines expression in post-surgical wound fluid [13]. It is not clear if seroma remained about a certain time after surgery or if it can have positive impact on changing the margin composition; consequently, a reducing local recurrence rate would take place. ...
Introduction: Post-lumpectomy wound fluid (seroma) contains many proteins from tumor bed due to physiologic answer to operation and wound healing process. Some cellular tests had been performed on different types of breast cancer (BC) cell lines and normal cell line while treated with seroma. Materials and Methods: The wound fluid samples were collected from BC patients. The human BC cell lines included MCF-7, MDA-MB-231 as well as normal non-tumorigenic epithelial cell line (MCF-10). Results: Seroma could inhibit various cancer cells proliferation pattern in comparison with the normal cell. Concerning the cell death, aggressive MDA-MB-231 cells were put into the apoptosis process. Besides, seroma could decrease colony count and size and changed the clone morphology from holoclone to paraclone. Regarding the invasion assay, seroma significantly inhibited cell motility. Conclusion: By remaining in tumor bed, seroma can induce inhibitory pattern of proliferation, and change the morphology of cancer cell colony and cell motility, consequently leading to positive impact on patients who suffer from cancer.
... Although researchers have shown great interest in a single high-dose clinical approaches for different cancers, limited research has examined the biological and molecular basis of single high-dose effects, especially after IORT [24]. Studies have described that Post-surgery wound fluids (WF) induces an increase in the cell proliferation, migration in addition invasion in BC cell line [25][26][27]. However, the proteomics analysis of IORT-treated WF has shown an inhibition of cell proliferation and invasion in the BC cell line [28,29]. ...
Background Radiotherapy (RT) is recommended to all patients undergoing Breast Conserving Surgery (BCS). Two strategies can be applied to irradiation, External Beam RT (EBRT) in addition, Intraoperative Radiation Therapy (IORT). The aim of this study was to introduce a protein biomarker panel related to molecular function under IORT.
Methods Six Breast Cancer (BC) patients as a pilot study were treated by 12 Gy (Boost dose) and 21 Gy (Radical dose). Samples tissue included Margin before IORT (MB), and Margin 24 hours After IORT (MA24 h). Isobaric Tag for Relative and Absolute Quantitation (iTRAQ) was performed to study proteomic of IORT-treated tumor bed.
Results We classified 110 differentially expressed proteins (DEPs) as a protein biomarker panel by mapping the annotated coding region sequences to the reference canonical pathways in the KEGG database.
Conclusion Our findings indicate that the DEPs may be key proteins in IORT-treated tumor bed and may serve as potential Effective biomarkers under IORT.
... Furthermore, cells surrounding an incipient tumor, the cellular and molecular interaction between the tumor and its surroundings, and the type of treatment plus how to use them [17]. In fact, seroma induces an increase in the cell proliferation; migration and invasion in BC cell line [18][19][20]. ...
Aim:
Intraoperative electron Radiotherapy, herein referred to, as IOeRT is a novel approach in breast cancer (BC) treatment. This study designed to investigate short-term molecular effects of 12Gy as Boost versus 21Gy as Radical dose of IOeRT using high throughput approaches.
Materials and methods:
Six BC patients as a pilot study were treated with IOeRT following two separate strategies, including Boost and Radical doses. Approximately 100 mg of tumor bed tissue retrieved from each patient (before IOeRT,immediately, 24 h post-treatment). mRNA sequencing also Isobaric tag for relative and absolute quantitation (iTRAQ) were performed to study the transcriptome and proteome profile of IOeRT-treated tumor bed.
Results:
Using NGS, ~6 Giga base (GB) clean data per individual samples were generated. Moreover, by iTRAQ for proteome quantification, in total, 1,045,410 spectrums were generated, likewise 5860 proteins were identified (FDR <0.01).
Conclusion:
Functional annotation and gene ontology (GO) indicated that significant enrichment in molecular pathways on BC treatment is somehow single high dose-independent. This means that, key molecular pathways in radiotherapy (RT) are equally enriched by both Boost and Radical doses. Generally, by modification of the Radical dose, with the same effectiveness, it is possible to reduce single high dose irradiation in BC.
