Bio-inspired plant leaf skeleton based three dimensional scaffold for three dimensional cell culture

Abstract Large quantities of natural fibers are available in the plant biomass that can be utilized for various purposes including three dimensional cell culture and tissue engineering due to their biocompatibility, ecofriendly, easy availability and cost effective. Especially, leaf skeletons (venation architecture) have a complex hierarchical architecture with novel properties. In this present invention describes about developing three dimensional scaffold from plant leaf skeletons for three dimensional cell culture. Plant leaf skeleton is prepared by simple and rapid method which using sodium hydroxide pretreatment under pressurized condition at 120°C for 1 h. The prepared plant skeleton has microporous surface topography. Also, the plant leaf skeleton is mainly composed by hemicellulose, cellulose and lignin. The microscopic analysis clearly indicates that human mesenchymal stem cells (hMSCs) attached and proliferated on plant leaf skeleton. Interestingly, cell density of hMSCs is increased on plant leaf skeleton by incubation time-dependent manner. Our study confirmed that sodium hydroxide via surface modified Ficus religiosa leaf skeleton enhances the attachment and proliferation of the human mesenchymal stem cells because of their biocompatibility and porous nature. The chemical nature and venation architecture of leaf skeleton has facilitated nutrient and oxygen absorption to promote cell-cell interaction, long term cell culture, and possible scope for induction of cell differentiation. Thus, leaf venation architecture can be applied as three dimensional (3D) cell-culture platform for multi-layer cell culture, cell-based assay model, high-throughput drug screening, cell-replacement therapy, wound healing and substitute for skin. Moreover, this scaffold could also be well-suited to co-culture screening strategies and stem cell differentiation for tissue engineering. Our present invention highlighted agro-wastes as precursor for novel scaffold materials to construct the 3D cell culture platform.