Autofluorescence Quenching in Decellularized Plant Scaffolds for Tissue Engineering.

Area of Science:

• Biomaterials Science
• Tissue Engineering
• Optical Imaging

Background:

• Plant-derived scaffolds offer a sustainable alternative for tissue engineering.
• Autofluorescence in these scaffolds hinders cellular visualization using common fluorescent markers.
• Overlapping autofluorescence limits detailed analysis of cell behavior and scaffold integration.

Purpose of the Study:

• To evaluate copper sulfate, ammonium chloride, and sodium borohydride as autofluorescence quenching agents.
• To determine if these agents compromise scaffold mechanical integrity or cell viability.
• To improve fluorescence imaging in decellularized plant scaffolds for tissue engineering.

Main Methods:

• Decellularized scaffolds from leatherleaf viburnum, spinach, and parsley were treated with quenching agents.
• Spectral scans characterized baseline autofluorescence.
• Scaffolds were imaged using Hoechst and FITC channels; autofluorescence intensity, quenching stability, mechanical properties, and cell viability were assessed.

Main Results:

• Copper sulfate demonstrated superior autofluorescence reduction and improved nuclear visualization across scaffold types.
• Scaffold mechanics (tensile strength, elastic modulus) remained unaffected by treatments.
• Endothelial cell viability decreased with copper sulfate in leatherleaf and parsley scaffolds but was preserved in spinach scaffolds.

Conclusions:

• Copper sulfate effectively quenches autofluorescence in plant scaffolds, enhancing imaging post-fixation.
• Scaffold-specific toxicity of copper sulfate limits its application in live-cell studies.
• Ammonium chloride and sodium borohydride are viable alternatives when cell viability is paramount, with no compromise to scaffold mechanics.