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，导致喉癌功能的严重恶化。喉部分切除术仍然是最安全的根治性治疗（使器官得以保存），但由于软骨丢失，患者经常遭受误吸和永久性气管切开术。因此，迫切需要一种具有精确形状和良好生物相容性的支架进行重建。我们之前的研究表明，高静水压力治疗在保留支撑组织的结构和生物力学的同时，特异性地使细胞失活。在这里，我们介绍用高静水压力使肿瘤浸润软骨失活，以促进自体和原位再植。我们将在控制细胞毒性试验中确认所有癌细胞的失活，同时通过光镜和电子显微镜验证软骨基质的保存。坏死细胞和生长因子残留在基质中，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癌细胞增殖的增加与失活肿瘤共培养。此外，我们将检测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，并分别暴露于压力处理的癌细胞中。最后，我们将用高静水压力治疗侵犯兔耳廓软骨的VX2癌。皮下再植入术后，存活的肿瘤将被排除。本研究通过对压力处理的自体软骨移植的深入研究，向真正的解剖喉部重建迈出了一步。因此，未来的应用可能会改善喉部分切除术后患者气道保护和发声。我们期望我们的项目成为进一步研究的起点，利用高流体静压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.