Integrating bioprinted oral epithelium with millifluidics for fluorouracil perfusion and Fusobacterium infection to bioengineer oral mucositis-on-a-chip

• Magnetic 3D bioassembly or bioprinting can promptly produce physiologically relevant multi-layered oral epithelial sheets from human oral keratinocytes. • Our dynamic millifluidic system can generate an oral mucositis (OM) disease model within an organ-on-a-chip from bioprinted oral epithelium after 5-fluorouracil (5FU) chemotherapy drug perfusion. • This millifluidic system was meaningful to confirm the effects of our in-house plant-produced EGF towards the promotion of epithelial migration in wound healling, mitigation of oxidative stress and reduction of DNA damage on the bioengineerined OM-on-a-chip in the presence of Fusobacterium nucleatum (Fn), an OM-associated pathogen . • P-EGF may constitute a promising therapeutic modality to quickly promote oral re-epithelialization and wound healing. Oral mucositis (OM) remains a painful complication of anticancer chemotherapy (CT), tending to progress in severity in the presence of Fusobacterium nucleatum ( Fn ). Yet, no effective therapy exists to suppress OM since in vitro models mimicking CT-induced OM are lacking, halting the discovery of new drugs. Here, we developed an integrated millifluidic in vitro tissue culture system for OM disease modeling. This bioengineered system integrates magnetically bioassembled oral epithelium sheets with millifluidics for CT-based 5-fluorouracil perfusion and Fn infection to model CT-induced OM. After modeling OM with all pro-inflammatory hallmarks, we were able to suppress OM with our in-house plant-produced epidermal growth factor (P-EGF), a well-known re-epithelialization cue. Thus, this the first instance where a milifluidic system enabled OM modeling in the presence of CT drug perfusion and Fn infection. This bioengineered system is a novel tool for drug discovery as it propelled P-EGF as a promising therapy for OM.