High hydrostatic pressure devitalization of head and neck cancer with cartilage invasion: Oncological safety and immune response after reimplantation

喉癌的软骨侵犯会降低微创手术、化疗和放疗的疗效，导致功能严重恶化。部分喉切除术仍然是最安全的治愈性治疗（能够保留器官），患者经常因软骨损失而遭受吸入和永久性气管造口术。因此，迫切需要一种形状精确、生物相容性好的支架材料。我们之前的研究表明，高静水压力治疗可以在保持支持组织结构和生物力学的同时，特异性地使细胞失活。在这里，我们介绍了通过高静水压力使肿瘤浸润软骨失活，以促进自体和原位再植。我们将在受控细胞毒性试验中确认所有癌细胞的失活，同时通过光学和电子显微镜验证软骨基质的保存。失活细胞和生长因子的残留物保留在基质中，并可能影响使组织恢复活力的细胞。我们将排除干细胞的转化和与失活肿瘤共培养中癌细胞增殖的增加。此外，我们将检查对失活组织的免疫反应：从患者血液中分离巨噬细胞和淋巴细胞，并暴露于各自的压力处理的癌细胞。最后，我们将用高静水压治疗侵袭兔耳廓软骨的VX 2癌。皮下再植入后，将排除存活癌症的生长。本研究通过对压力处理的自体软骨移植物的深入研究，向真实的解剖喉重建迈进了一步。因此，未来的应用可能会改善部分喉切除术后患者的气道保护和发声。我们预计我们的项目将成为进一步研究的起点，使用高静水压力来对抗不同的侵袭性癌症实体，并开发更有效的癌症免疫疗法。

Cartilage invasion in laryngeal cancer leads to serious deterioration of the functional outcome by diminishing the efficacy of minimal-invasive surgery, chemo- and radiation therapy. Following partial laryngectomy, which remains the safest curative treatment (enabling organ preservation), patients frequently suffer aspiration and permanent tracheostomy due to loss of cartilage. Therefore, a scaffold of precise shape and superb biocompatibility for the reconstruction is urgently needed. Our previous research showed that high hydrostatic pressure treatment specifically devitalizes cells while preserving the structure and the biomechanics of supporting tissue.Here, we introduce the devitalization of tumor-infiltrated cartilage by means of high hydrostatic pressure to facilitate autologous and orthotopic reimplantation. We will confirm the devitalization of all cancer cells in controlled cytotoxicity assays, while the preservation of the cartilage matrix is verified by light and electron microscopy. Remnants of devitalized cells and growth factors remain in the matrix and may affect cells that revitalize the tissue. We will exclude the transformation of stem cells and the increase of cancer cell proliferation in co-culture with the devitalized tumor. Furthermore, we will examine the immune response to the devitalized tissue: Macrophages and lymphocytes are isolated from patients’ blood and exposed to the respective pressure-treated cancer cells. Finally, we will treat VX2 cancer invading the rabbit auricle cartilage with high hydrostatic pressure. After subcutaneous reimplantation outgrowth of viable cancer will be ruled out. This study represents a step towards real anatomic laryngeal reconstruction by the profound investigation of pressure-treated cartilage autografts. Hence, a future application may improve airway protection and voicing for patients after partial laryngectomy. We anticipate our project to be a starting point for further studies using high hydrostatic pressure to fight different aggressive cancer entities and to develop more efficient cancer immunotherapy.