hiPSC-Derived Gingival Epithelium-On-Chip for Modeling Inflammation and Immune Response

The gingival epithelium plays a central role in coordinating epithelial-immune interactions during oral inflammation. However, many existing in vitro models are constrained by limited scalability, donor variability, or restricted inflammatory signal amplitude, reducing sensitivity for comparative and screening-oriented studies. We present a gingival epithelium-on-chip (GEoC) platform based on human induced pluripotent stem cell (hiPSC)-derived oral keratinocytes cultured in a dual-channel microfluidic device with a spatial calcium gradient, enabling controlled, calcium-dependent epithelial differentiation within a three-dimensional (3D) microenvironment. The differentiated keratinocytes exhibit oral lineage commitment and resemble primary gingival keratinocytes in phenotypic profiles. Upon lipopolysaccharide (LPS) stimulation, the GEoC model exhibits markedly amplified pro-inflammatory cytokine responses relative to parallel two-dimensional (2D) cultures, yielding a substantially expanded dynamic range of inflammatory output. This enhanced sensitivity enables clear discrimination of immunomodulatory effects that are muted in 2D systems, as demonstrated by differential cytokine attenuation following rapamycin and stannous fluoride treatment. Rather than recapitulating fully stratified or barrier-mature gingival tissue, the GEoC is designed as a scalable and controllable immunocompetent epithelial-immune inflammation platform that enhances inflammatory signal detection and, supports reproducible and comparative evaluation of anti-inflammatory interventions using a renewable hiPSC-derived epithelial source and standardized microfluidic hardware.