Immunomodulatory effects of conditioned medium from high hydrostatic pressure-treated head and neck squamous cell carcinoma cells regarding macrophage polarization
5 affiliations
- 1Department of Orthopaedics, Biomechanics and Implant Technology Research Laboratory, Rostock University Medical Center
- 2Department of Otorhinolaryngology, Head and Neck Surgery “Otto Körner”, Rostock University Medical Center
- 3Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital RWTH Aachen
- 4Center for Integrated Oncology (CIO), University Hospital Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen
- 5Department of Oto-Rhino-Laryngology, Plastic, Aesthetic and Reconstructive Head and Neck Surgery, University of Würzburg
Abstract
Surgical resection of cartilage-infiltrating head and neck squamous cell carcinoma (HNSCC) frequently results in significant tissue defects, which require reconstructive interventions. Orthotopic reimplantation of autologous laryngeal cartilage tissue could be a promising approach due to its anatomical conformity and excellent biocompatibility. High hydrostatic pressure (HHP) at 300 MPa effectively eliminates residual malignant cells while preserving the extracellular matrix and biomechanical properties of hyaline cartilage. However, HHP alters the release of tumor-derived cytokines and damage-associated molecular patterns. Given the critical role of balanced inflammatory and immunomodulatory responses in tissue integration, this in vitro study investigated whether factors released after HHP treatment influence macrophage polarization. Conditioned medium (CM) from HHP-treated or untreated HNSCC cell lines was used to stimulate adherent mononuclear cells for 72 hours. Unstimulated cells, M1 (GM-CSF + LPS), and M2 (M-CSF + IL-4) macrophage controls served as controls. Macrophage morphology, gene expression, and protein secretion were analyzed for M1 markers (TNF-α, IL-1β, IL-6, IL-8, IL-12 p70, IL-23, CXCL9, CXCL10) and M2 markers (IL-10, CCL17, CCL18, MMP-9). Surface markers HLA-DR, CD80, CD86, CD192 (M1-like) and CD206, CX3CR1 (M2-like) were assessed. CM from HHP-treated cells induced a heterogeneous macrophage morphology, including spindle-shaped and rounded cells. Gene expression revealed significant upregulation of IL-1β and IL-8, a trend for IL-6, and no increase in TNF-α or IL-10. At the protein level, IL-1β and IL-8 were significantly elevated, with a trend toward increased IL-6 and CXCL9, while IL-12 p70, and IL-23 remained unchanged. IL-10 levels stayed low, and no induction of CCL17, CCL18, or CCL22 was observed. Phenotypically, most cells expressed CD86 and HLA-DR, with a high proportion positive for CD192, but only a minority expressed CD80. A notable fraction retained CD206 and CX3CR1 expression regardless of stimulation. Overall, CM derived from HHP-treated HNSCC cells induced a heterogeneous, partially activated macrophage state with predominantly M1-like features rather than a distinct M1 or M2 phenotype. These findings suggest that CM may influence the inflammatory microenvironment relevant to cartilage reimplantation without promoting a strongly immunosuppressive macrophage response. Further studies using tissue models and in vivo validation are required to establish its relevance for tissue integration.
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BibTeX
@article{grote2026immunomodulatory,
title = {{Immunomodulatory effects of conditioned medium from high hydrostatic pressure-treated head and neck squamous cell carcinoma cells regarding macrophage polarization}},
author = {Vivica Freiin Grote and Laura Schaffran and Daniel Strüder and Julia Kristin Brach and Agmal Scherzad and Stephan Hackenberg and Rainer Bader and Anika Jonitz-Heincke},
journal = {Frontiers in Immunology},
year = {2026},
month = oct,
doi = {10.3389/fimmu.2026.1885288},
url = {https://www.frontiersin.org/articles/10.3389/fimmu.2026.1885288/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Immunomodulatory effects of conditioned medium from high hydrostatic pressure-treated head and neck squamous cell carcinoma cells regarding macrophage polarization
AU - Vivica Freiin Grote
AU - Laura Schaffran
AU - Daniel Strüder
AU - Julia Kristin Brach
AU - Agmal Scherzad
AU - Stephan Hackenberg
AU - Rainer Bader
AU - Anika Jonitz-Heincke
PY - 2026
DA - 2026/10/19
JO - Frontiers in Immunology
DO - 10.3389/fimmu.2026.1885288
UR - https://www.frontiersin.org/articles/10.3389/fimmu.2026.1885288/full
AB - Surgical resection of cartilage-infiltrating head and neck squamous cell carcinoma (HNSCC) frequently results in significant tissue defects, which require reconstructive interventions. Orthotopic reimplantation of autologous laryngeal cartilage tissue could be a promising approach due to its anatomical conformity and excellent biocompatibility. High hydrostatic pressure (HHP) at 300 MPa effectively eliminates residual malignant cells while preserving the extracellular matrix and biomechanical properties of hyaline cartilage. However, HHP alters the release of tumor-derived cytokines and damage-associated molecular patterns. Given the critical role of balanced inflammatory and immunomodulatory responses in tissue integration, this in vitro study investigated whether factors released after HHP treatment influence macrophage polarization. Conditioned medium (CM) from HHP-treated or untreated HNSCC cell lines was used to stimulate adherent mononuclear cells for 72 hours. Unstimulated cells, M1 (GM-CSF + LPS), and M2 (M-CSF + IL-4) macrophage controls served as controls. Macrophage morphology, gene expression, and protein secretion were analyzed for M1 markers (TNF-α, IL-1β, IL-6, IL-8, IL-12 p70, IL-23, CXCL9, CXCL10) and M2 markers (IL-10, CCL17, CCL18, MMP-9). Surface markers HLA-DR, CD80, CD86, CD192 (M1-like) and CD206, CX3CR1 (M2-like) were assessed. CM from HHP-treated cells induced a heterogeneous macrophage morphology, including spindle-shaped and rounded cells. Gene expression revealed significant upregulation of IL-1β and IL-8, a trend for IL-6, and no increase in TNF-α or IL-10. At the protein level, IL-1β and IL-8 were significantly elevated, with a trend toward increased IL-6 and CXCL9, while IL-12 p70, and IL-23 remained unchanged. IL-10 levels stayed low, and no induction of CCL17, CCL18, or CCL22 was observed. Phenotypically, most cells expressed CD86 and HLA-DR, with a high proportion positive for CD192, but only a minority expressed CD80. A notable fraction retained CD206 and CX3CR1 expression regardless of stimulation. Overall, CM derived from HHP-treated HNSCC cells induced a heterogeneous, partially activated macrophage state with predominantly M1-like features rather than a distinct M1 or M2 phenotype. These findings suggest that CM may influence the inflammatory microenvironment relevant to cartilage reimplantation without promoting a strongly immunosuppressive macrophage response. Further studies using tissue models and in vivo validation are required to establish its relevance for tissue integration.
SN - 1664-3224
ER -