Microenvironment-Controlled Micropatterned Microfluidic Model (MMMM) for Biomimetic In Situ Studies

Attachment of trophozoites to the intestine is an indispensable step for Giardia's survival and pathogenicity in almost 280 million infections worldwide each year. However, the analysis of the attachment mechanism is difficult due to the lack of methods that can create a favorable microaerobic atmosphere. Herein, we developed an osmotic-pressure, pH, excretion, nutrition, gas, ionic-strength, flow-rate, and temperature microenvironment-controlled micropatterned microfluidic model to simulate the in vivo microenvironment to study in situ the stress applied to Giardia in the intestinal tract. We designed three nonbiological surfaces with stagger arrangement manners and integrated them with a resistance microfluidic network to split Giardia-attaching forces ingeniously and developed the term "attaching contribution rate" (ACR) to describe their corresponding contributions. Our study shows that the total attaching force measured is 49.58 Pa, with three components being 22.66 Pa (suction force), 12.52 Pa (clutching force), and 14.4 Pa (combined electrostatic and van der Waals force), respectively, with ACRs being 46%, 25%, and 29%, respectively. By decomposing the attaching force and analyzing each force component and their structure and composition basis, whole profiles of the attachment mechanisms were revealed. Our method enables the analysis of the surface attachment mechanisms and their ACRs for Giardia.