Extrusion-based three-dimensional (3D) bioprinting is one of the most common methods used for tissue fabrication and is the most widely used additive manufacturing technique in all industries. In extrusion-based bioprinting, printing defects related to material deposition errors lead to a significant deviation from shape to function between the printed construct and design model. Using 3D extrusion-based bioprinter-associated optical coherence tomography (3D P-OCT), an in situ defect detection and feedback system was presented based on the accurate defect analysis and location, and a pre-built feedback mechanism. Using 3D P-OCT, multi-parameter quantification of the material deposition was carried out in real time, including the filament size, layer thickness, and layer fidelity. The material deposition errors under different paths were quantified and located specifically, including the start-stop points, straight-line path, and turnarounds. The pre-built feedback mechanism involving the control inputs, such as printing path, pressure, and velocity, provided the basis for in situ defect detection and real-time feedback control. In particular, the second printing repair can be performed after the broken filament defect is detected and located. After printing, fidelity can be quantitatively analyzed based on the point cloud registration between the 3D P-OCT result and the design model. In conclusion, 3D P-OCT enables in situ defect detection and feedback control, broken filament repair, and 3D fidelity analysis to achieve high-fidelity printing from shape to function.