From leaf to cellulose scaffold: Decellularization and multi-scale characterization of <i>Neolamarckia cadamba</i> leaf for biomedical applications

Exploring biological "orthogonality" between plant and animal kingdoms provides a unique opportunity to mimic biological features of both kingdoms. This novel approach can provide solutions to problems that animal kingdom faces. In this study, we aimed to develop and characterize the Neolamarckia cadamba leaf-derived cellulose scaffolds (NCCS). Fresh Neolamarckia cadamba leaves were decutinized using an n-hexanes-diethyl ether and decellularized using 5% sodium dodecyl sulfate (SDS) for 120h and 2% sodium deoxycholate (SDC) solution for 48h. The leaves incubated in deionized water were acted as controls. Finally, the leaves were bleached in 4% NaOCl solution for 12 h. Histology, DAPI staining, and SEM study of scaffold showed complete decellularization with significantly reduced DNA quantity. Water vapor transmission rate and swelling percentage increased significantly and the scaffolds were hemocompatible. Contact angle revealed a significant increase in hydrophilicity and at maximum load of 2.83 N, percent elongation at break was 0.094 mm/mm. The FT-IR bands were not disturbed, and average roughness (Ra) and root mean square roughness (RMS) (Rq) of NCCS was 193.27 nm and 153.48 nm, respectively. Specific surface area, pore volume, and pore radius of NCCS were increased. The Madin-Darby canine kidney (MDCK) cells were adhered on NCCS and cell viability on decellularized leaf was 88.18% ± 16.12%. Subcutaneously implanted NCCS revealed infiltrated host cells. These characteristics establish NCCS as a sustainable, ethical and cost-effective substitute to mammalian tissue-derived scaffolds for biomedical applications.