Pape Judith, Bakkalci Deniz, Hosni Rawiya Al, Simpson Benjamin S, Heikinheimo Kristiina, Fedele Stefano, Cheema Umber
UCL Centre for 3D Models of Health and Disease, Division of Surgery and Interventional Science, University College London, London, UK.
Research Department of Targeted Intervention, Division of Surgery and Interventional Science, University College London, London, UK.
J Tissue Eng. 2022 Dec 24;13:20417314221140500. doi: 10.1177/20417314221140500. eCollection 2022 Jan-Dec.
Ameloblastoma is a benign, locally invasive epithelial odontogenic neoplasm of the jaw. Treatment of choice is jaw resection, often resulting in significant morbidity. The aim of this study was to recapitulate ameloblastoma in a completely humanised 3D disease model containing ameloblastoma cells, osteoblasts and activated osteoclasts to investigate the RANKL pathway within the ameloblastoma stromal environment and its response to the RANKL antibody denosumab. In vitro bone was engineered by culturing human osteoblasts (hOB) in a biomimetic, dense collagen type I matrix, resulting in extensive mineral deposits by day 21 forming alizarin red positive bone like nodules throughout the 3D model. Activated TRAP + human osteoclasts were confirmed through the differentiation of human CD14+ monocytes after 10 days within the model. Lastly, the ameloblastoma cell lines AM-1 and AM-3 were incorporated into the 3D model. RANKL release was validated through TACE/ADAM17 activation chemically or through hOB co-culture. Denosumab treatment resulted in decreased osteoclast activation in the presence of hOB and ameloblastoma cells. These findings stress the importance of accurately modelling tumour and stromal populations as a preclinical testing platform.
成釉细胞瘤是一种颌骨的良性、局部侵袭性上皮性牙源性肿瘤。首选治疗方法是颌骨切除，这通常会导致显著的发病率。本研究的目的是在一个完全人源化的三维疾病模型中重现成釉细胞瘤，该模型包含成釉细胞瘤细胞、成骨细胞和活化的破骨细胞，以研究成釉细胞瘤基质环境中的RANKL通路及其对RANKL抗体地诺单抗的反应。通过在仿生的致密I型胶原基质中培养人成骨细胞（hOB）来构建体外骨，到第21天时形成大量矿化沉积，在整个三维模型中形成茜素红阳性的骨样结节。在模型中10天后，通过人CD14+单核细胞的分化确认了活化的TRAP+人破骨细胞。最后，将成釉细胞瘤细胞系AM-1和AM-3纳入三维模型。通过化学激活TACE/ADAM17或通过与hOB共培养来验证RANKL的释放。地诺单抗治疗导致在存在hOB和成釉细胞瘤细胞的情况下破骨细胞活化减少。这些发现强调了准确模拟肿瘤和基质群体作为临床前测试平台的重要性。
