Segmental flow responses in glaucomatous trabecular meshwork cells

In glaucoma, increased resistance in the conventional outflow pathway elevates intraocular pressure, yet how local flow state reshapes the mechanics of glaucomatous trabecular meshwork (TM) cells across segmental outflow regions remains unclear. Here, we treated flow as a controllable mechanical input and asked whether shear acts as a mechanical switch in TM cells isolated from tracer-defined high-flow (HF) and low-flow (LF) regions of a human glaucomatous eye. Cells were studied under no-flow→flow and flow→no-flow programs, with the switch at hour 6 and matched no-flow controls. Using time-resolved three-dimensional traction force microscopy, we quantified traction as well as strain, divergence, and curl. Pre-switch trajectories matched controls showing comparable baseline drift before the switch. Acute 6 → 8 h responses captured flow response directionality but did not separate HF from LF cells. The segmental split emerged over longer timescales. After no-flow→flow, HF cells showed a delayed traction increase (+39.2%) with parallel curl reinforcement, whereas matched controls drifted downward. After flow→no-flow, HF cells showed an acute traction decrease over 6 → 8 h (-18.5%). LF cells lacked the HF traction/curl program and instead were defined more strongly by divergence and strain, including a flow-associated LF divergence shift and suppression of strain after flow withdrawal relative to controls. Within this paired single-donor glaucomatous model, flow-state switching revealed segmental mechanophenotypes with HF cells retained traction/curl reinforcement after flow onset, whereas LF cells responded primarily through divergence and strain.