Analyzing Structure and Function of Vascularization in Engineered Bone Tissue by Video-rate Intravital Microscopy and 3D Image Processing (original) (raw)

2015, Tissue engineering. Part C, Methods

Vascularization is a key challenge in tissue engineering. 3D structure and microcirculation are two fundamental parameters for evaluating vascularization. Microscopic techniques with cellular level resolution, fast continuous observation and robust 3D post image processing are essential for evaluation but have not been applied previously because of technical difficulties. Here we report novel video rate confocal microscopy and 3D post image processing techniques to accomplish this goal. In an immune-deficient mouse model, vascularized bone tissue was successfully engineered using human bone marrow mesenchymal stem cells (hMSCs) and human umbilical vein endothelial cells (HUVECs) in a poly (DL-lactide-co-glycolide) (PLGA) scaffold. Video rate (30 FPS) intravital confocal microscopy was applied in vitro and in vivo to visualize the vascular structure in the engineered bone and the micro circulation of the blood cells. Post image processing was applied to perform 3D image reconstructio...

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