Geometry–Encoded Microtrenches Stabilize Endothelium on High Shear Biomaterial Surfaces

Maintaining a confluent, antithrombotic endothelium on cardiovascular biomaterial surfaces remains a major barrier to long-term hemocompatibility, as endothelial cells (ECs) rapidly denude under supraphysiological shear in prosthetic devices. Here, we hypothesized that mesoscale surface geometry (∼100–200 µm) could reorganize near-wall hemodynamics, preserving endothelial coverage and function under extreme shear. Engineered microtrenches were introduced onto an implant biomaterial to generate spatially defined shear environments. Under supraphysiological near-wall shear (∼250 dyn/cm²), microtrenched geometries created attenuated shear and vorticity gradients. Endothelial monolayers were sustained in these flow domains for 120 hours, whereas flat controls rapidly denuded. Endothelial retention in 22.5° angled trenches increased dramatically, from an EC₅₀ of 33 to 101 dyn/cm². 45° angled trenches further increased endothelial shear resistance to an EC₅₀ of 207 dyn/cm². Endothelial monolayers demonstrated collective mechano-adaptation to ultra-high shear through VE-cadherin junction thickening and coordinated cytoskeletal and nuclear alignment. Mechanoadapted monolayers exhibited increased eNOS expression correlated with local shear and elevated nitrite production (45°: 50.4 ± 6.1 µM; 22.5°: 35.7 ± 3.3 µM; 0°: 28.4 ± 6.8 µM). In contrast, interfaces with abrupt shear transitions or elevated rotational flow exhibited reduced coverage, junctional thinning, and re-emergence of VCAM-1 and PAI-1, indicating inflammatory and pro-thrombotic activation. Structural, functional, and inflammatory readouts exhibited peak responses within a shared shear–vorticity regime. Multivariate regression identified shear–vorticity coupling as the dominant predictor of endothelial persistence, with optima clustering within a mechanical range (≈0.8–2.9 × 10⁶ dyn·cm⁻²·s⁻¹). These findings establish geometry-driven modulation of near-wall flow as a predictive, material-agnostic strategy for endothelialization and vasoprotection of high-shear cardiovascular implants.