In situ monitoring of perfusable vascular network formation and development in an electrochemical microfluidic chip.

The formation of perfusable vascular networks is essential for constructing physiologically relevant in vitro tissue models, yet quantitative approaches for evaluating functional perfusion during vascular network development remain limited. Here, we developed a triparallel microfluidic vascular chip integrated with an electrochemical detection system to dynamically monitor molecular transport associated with vascular functional development. Through dynamic electrochemical monitoring of molecular transport, we quantitatively characterized functional perfusion during vascular network formation and maturation under static and flow-stimulated conditions. We characterized vascular development as a stage-dependent process and found that under static conditions, functional perfusion was established after morphological network formation, revealing a temporal lag between vascular structure and transport function. Following perfusion onset, continued vessel fusion and enlargement further enhanced tracer transport, which was captured by increased electrochemical responses. Under flow stimulation, functional perfusion emerged concurrently with morphological network formation, eliminating the delayed transition observed under static culture. These results demonstrate that flow not only accelerates vascular development but also regulates the temporal coupling between structural assembly and functional maturation. Overall, this electrochemical microfluidic platform provides a quantitative approach for evaluating vascular functional development beyond morphology-based analysis and offers a tool for studying perfusion maturation in engineered vascularized models.