Modelling stromal compartments to recapitulate the ameloblastoma tumour microenvironment

Deniz Bakkalci; Amir Zaki Abdullah Zubir; Syed Ali Khurram; Judith Pape; Kristiina Heikinheimo; Stefano Fedele; Umber Cheema

doi:10.1016/j.mbplus.2022.100125

Modelling stromal compartments to recapitulate the ameloblastoma tumour microenvironment

Matrix Biol Plus. 2022 Nov 21:16:100125. doi: 10.1016/j.mbplus.2022.100125. eCollection 2022 Dec.

Authors

Deniz Bakkalci¹, Amir Zaki Abdullah Zubir², Syed Ali Khurram², Judith Pape¹, Kristiina Heikinheimo³, Stefano Fedele⁴, Umber Cheema¹

Affiliations

¹ UCL Centre for 3D Models of Health and Disease, Division of Surgery and Interventional Science, University College London, Charles Bell House, 43-45 Foley Street, W1W 7TS London, UK.
² Unit of Oral and Maxillofacial Pathology, School of Clinical Dentistry, University of Sheffield, 19 Claremont Crescent, S10 2TA Sheffield, UK.
³ Department of Oral and Maxillofacial Surgery, Institute of Dentistry, University of Turku and Turku University Hospital, Turku, Finland.
⁴ Eastman Dental Institute, University College London, London, UK.

Abstract

Tumour development and progression is dependent upon tumour cell interaction with the tissue stroma. Bioengineering the tumour-stroma microenvironment (TME) into 3D biomimetic models is crucial to gain insight into tumour cell development and progression pathways and identify therapeutic targets. Ameloblastoma is a benign but locally aggressive epithelial odontogenic neoplasm that mainly occurs in the jawbone and can cause significant morbidity and sometimes death. The molecular mechanisms for ameloblastoma progression are poorly understood. A spatial model recapitulating the tumour and stroma was engineered to show that without a relevant stromal population, tumour invasion is quantitatively decreased. Where a relevant stroma was engineered in dense collagen populated by gingival fibroblasts, enhanced receptor activator of nuclear factor kappa-B ligand (RANKL) expression was observed and histopathological properties, including ameloblastoma tumour islands, developed and were quantified. Using human osteoblasts (bone stroma) further enhanced the biomimicry of ameloblastoma histopathological phenotypes. This work demonstrates the importance of the two key stromal populations, osteoblasts, and gingival fibroblasts, for accurate 3D biomimetic ameloblastoma modelling.

Keywords: 3D models; AM, Ameloblastoma; BPE, Bovine pituitary extract; BSA, Bovine serum albumin; CCN, Cellular communication network factor 2; DMEM, Dulbecco’s modified Eagle medium; ECM, Extracellular matrix; FBS, Foetal bovine serum; Fibroblasts; H&E, Haematoxylin and eosin; HFF-2, Human fibroblasts (HFF-2); HGF, Primary gingival fibroblasts; IF, Immunofluorescence; IHC; IHC, Immunohistochemistry; M, Molar; MEM, Minimal Essential Medium; MIQE, Minimum Information for Publication of Quantitative Real-Time PCR Experiments; MMPs, Matrix metalloproteinases; MRC5, Human lung fibroblasts; Min, Minutes; N.A, Neutralising agent; PTHLH, Parathyroid Hormone Like Hormone; RANK, The tumour necrosis factor (TNF) superfamily members receptor activator of nuclear factor kappa-B receptor; RANKL; RANKL, The tumour necrosis factor (TNF) superfamily members receptor activator of nuclear factor kappa-B ligand (TNFSF11); RT, Room temperature; SD, Standard deviation; SEM, Standard error mean; SPARC, anti-osteonectin; TGF-β, Transforming growth factor; TME, Tumour microenvironment; TNF, Tumour necrosis factor; Tumour microenvironment; dl-CGH, Double-layered collagen gel hemisphere; hOB, Human osteoblasts; α-SMA, alpha-smooth muscle actin.