EFFECTIVENESS OF HUMAN FLUORESCENT ANTIBODIES IN EVALUATING THE REMODELING OF TISSUE-ENGINEERED CONSTRUCTS HAVING UNDERGONE PRECLINICAL TESTING IN AN ANIMAL MODEL

Highlights

• Multiplex immunofluorescence imaging panels have been developed for monitoring the remodeling of tissue‑engineered vascular prostheses in baboons using human-specific antibodies.
• Scanning laser microscopy has confirmed that antibodies can detect key elements of newly formed vascular tissue 6 months after implantation of prostheses into the femoral arteries of baboons. The identified elements include endothelial cells, extracellular matrix, smooth muscle–like and fibroblast-like cells.
• The remodeled tissue formed on the basis of TSP is similar in many respects to the native vascular wall. This confirms the effectiveness of the developed panels for preclinical testing and expands the possibilities of detecting remodeling processes in baboon models.

Aim. To develop immunofluorescence assay panels based on human-specific antibodies to enable the detection of tissue-engineered construct remodeling during preclinical trials in baboon models.

Methods. A comparative assessment of the sensitivity of immunofluorescent antibodies was performed using samples of intact human internal mammary artery and explanted tissue-engineered vascular prostheses implanted into the femoral arteries of baboon for six months. The cross-reactivity and targeting capability of human antibodies against CD31, vWF, collagen IV, α-smooth muscle actin, and vimentin were demonstrated in relation to baboon tissues. Immunofluorescence visualization panels for assessing the remodeling of tissue-engineered vascular prostheses were developed. Fluorescence intensity was evaluated using scanning laser microscopy. Fluorescence intensity was assessed using scanning laser microscopy. Statistical data processing was performed using GraphPad Prism 8.

Results. It was demonstrated that the use of human antibodies against CD31, vWF, collagen IV, α-smooth muscle actin, and vimentin enabled the detection of key components of newly formed vascular tissue six months after implantation of tissue-engineered vascular prostheses into the femoral arteries of baboons, including endothelial cells, extracellular matrix, smooth muscle–like cells, and fibroblast-like cells.

Conclusion. The developed multiplex immunofluorescence imaging panels reflected the main features of tissue-engineered construct remodeling in primates, which is similar to the native vascular wall in many parameters, confirming their effectiveness for monitoring the formation of newly formed vascular tissue.

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