Biocompatibility of decellularized spinach leaves

Previously, we demonstrated the potential of decellularized spinach leaves to act as prevascularized scaffolds for tissue engineering. In this follow-up study, we aim to further probe this technology's efficacy through investigation of leaf scaffold biocompatibility. Decellularization protocols were modified to limit possible toxicity by lowering the time and concentration of detergents used. Decellularized leaves that were processed with this modified technique were found to have limited effects on cell viability in a leaching assay. To evaluate biocompatibility, decellularized leaves were implanted subcutaneously in Sprague Dawley rats. Leaves maintained a limited immunological response 4 weeks post-implantation. A separate group of decellularized leaves were pre-functionalized with an RGD-dopamine peptide and subsequently implanted. These functionalized leaf implants integrated into the host tissue within 1-week, with visible rat collagen deposition found within the leaf scaffold. These findings further demonstrate the suitability of leaves as scaffolding for tissue engineering and suggest exploration for clinical use.