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Oral Health

A High Throughput Microfluidic Model of Human Gingival Tissue

/ by Lifeline® Cell Technology

The Link Between Oral Health and Overall Health

Oral health is closely connected to overall health and well-being. The mouth is home to a complex microbial environment, and when oral hygiene is compromised, bacteria and inflammation can result in common conditions such as tooth decay and gum disease. In fact, nearly half of American adults suffer from some form of gum disease, ranging from mild gingival inflammation (gingivitis) to more severe disease states like periodontitis.

Periodontitis is a persistent bacterial infection that allows inflammatory mediators and oral pathogens to enter the bloodstream. This systemic exposure has been associated with an increased risk of multiple chronic conditions, including cardiovascular disease, stroke, diabetes, respiratory disease, certain cancers, and Alzheimer’s disease. This emphasizes oral health as an important and often underrecognized contributor to systemic health.

In our latest publication feature, researchers describe the development of a high throughput microphysiological triculture system that models the human gum tissue in vitro, to facilitate the investigation of gum disease progression and support therapeutic development.

Modeling Inflammation in Gingival Tissues

Gard and colleagues describe the Microfluidic model of Oral physiology for Understanding Tissue Health (MOUTH) model, which is a multi-layered, multi-phenotypic gingival tissue established in a 96-device microfluidic membrane bilayer platform, known as PREDICT96. MOUTH is a triculture model consists of human gingival fibroblasts (hGFs) cultured on the top surface of the membrane, human dermal microvascular endothelial cells (hMVECs) on the bottom surface, and human oral keratinocytes (hOKs) layered above the hGFs within the microfluidic device. Both hGFs and hOKs were sourced from Lifeline Cell Technology.

Baseline Characterization of the MOUTH Model

The MOUTH model maintained a stratified gingival tissue morphology consisting of five to eight keratinocyte layers, with a thickness of up to 200 µm. While many in vitro gingival tissue models preserve this architecture for only up to seven days, MOUTH sustained tissue structure for as long as 28 days. A multi-week experimental window of approximately one month was established, during which stable barrier formation was maintained, as measured by transepithelial electrical resistance (TEER).

Baseline tissue health and inflammatory status were assessed by profiling 18 inflammatory biomarkers throughout the culture period. None of the biomarkers showed a significant increase over time, indicating sustained barrier integrity and low basal inflammation. These consistent trends over 30 days demonstrate the reproducibility and durability of the long-term gingival tissue culture within a microfluidic device.

Inflammation was subsequently induced using a clinically relevant, pathophysiological cytokine cocktail of 300 ng/mL TNF-α and 300 ng/mL IL-1β applied for 24 hours. TEER was measured daily, and inflammatory biomarkers were analyzed over a 48-hour post-stimulation window. Following cytokine exposure, TEER values in stimulated devices decreased by approximately 14% at 48 hours, whereas vehicle-treated tissues (0.1% BSA) remained largely unchanged.

Secreted prostaglandin E2 (PGE2) levels measured in the bottom channel increased more than sixfold relative to controls within 24 hours and remained elevated at greater than fivefold above vehicle levels for at least 48 hours. Additionally, the inflammatory cytokines MIP-3α, IL-10, and IFN-γ were significantly upregulated at both 24- and 48-hour time points compared with control conditions. Together, these findings demonstrate a robust, multi-factor inflammatory response in the MOUTH model following cytokine challenge.

Treating inflammation in MOUTH

A small-molecule inhibitor, SB203580, was used in combination with inflammatory stimulation to assess its potential preventative effect on gingival tissue in response to IL-1β and TNF-α. Both barrier function and PGE2 release, the main metrics of inflammation, were assessed after dosing MOUTH with SB203580.

Pre-treatment with SB203580 prior to IL-1β and TNF-α stimulation significantly reduced the inflammatory response in the MOUTH model compared with stimulated tissues that did not receive the inhibitor, as quantified by reduced PGE2 secretion. These results demonstrate the model’s sensitivity to pharmacological modulation of inflammation.

Importantly, the long-term culture stability and high-throughput format of the MOUTH platform enable evaluation of a broad therapeutic parameter space, including different dosing regimens and early/late treatment. In this study, MOUTH tissues responded to a second dose of SB203580, illustrating the ability to perform repeated dosing and extended, consecutive studies within the same tissue constructs.

Overall, the MOUTH model maintains physiologically relevant tissue architecture, mucosal barrier integrity, and stable protein biomarker expression and secretion over several weeks. By enabling reproducible induction of inflammation, the model provides a human-relevant platform for evaluating novel therapeutic targets and supporting preclinical drug efficacy and safety studies in gum disease.

Lifeline Cell Technology Oral Cells and Media

High quality primary cells are essential to establishing complex microphysiological models, such as the one described in our publication highlight. Lifeline Cell Technology offers a comprehensive portfolio of primary oral cells directly isolated from source tissue, along with specialized media formulations optimized to support the development of advanced human-relevant models for preclinical research.

Explore our blog to see how our cells and culture media are advancing biomedical research worldwide. If you have used our products in your publication, we’d love to feature your work here!

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