Modelling Ameloblastoma Behaviour With Bone-Like Co-Culture Scaffolds

Objectives: Ameloblastoma (AM) is a benign yet destructive tumour of the jawbones and surrounding soft tissues with a high recurrence potential. Therapy of AM is typically surgical, involving large resection of the jawbone and surrounding soft tissues. Disease recurrence is caused by the presence of cell islands surrounding the main tumour, which can result in incomplete tumour removal. Little is known about the cellular mechanisms of tissue invasion of this tumour.
We have developed an in vitro bone-like co-culture scaffold to model the initial AM tumour growth, invasion and examine changes in gene expression.
Methods: The co-culture models were assembled from two parts: first, a cellularised ‘hard’ tissue-like construct was made using plastic compressed collagen and bone cells. These constructs were placed next to collagen gels with cells from the AM-1 cell line to create co-culture models. Confocal microscopy, quantitative real-time PCR, scanning electron microscopy and cell proliferation assays were used to characterise the behaviour of the cells in the scaffolds.
Results: The in vitro model described here was shown to closely mimic the native ameloblastoma growth environment. AM cells were shown to interact with the bone cells, upregulate bone resorptive genes, and express fibroblast-like markers with increasing culture time.
This model enables close characterisation of AM cells in the model using fluorescence microscopy, as well as gene and protein expression assays. Furthermore, using this model we have been able to easily assess the effects of potential therapeutic agents on AM growth in the model prior to clinical studies.
Conclusions: Our novel model can be used to determine the underlying molecular events in AM growth and to efficiently test drugs in an in vitro organotypic setting.
Furthermore, with minor refinement, this scaffold may be used to model other oral tumours.