Table 1.

Current protocols employed to decellularize plant material.

Decellularization Treatment	Compounds	Time	Decellularized Plants	Advantages	Limitations
Chemical	SDS (0.1 to 10%, depending on plant material); Triton X-100; bleach (10%) Hexane pre- treatment can be performed when wax cuticle present	12 h to 3 weeks, depending on the plant material	Amazon sword [55], Anthurium [35], Anthurium (queen) [35], Apple interior [24,32,33,53,56], Asparagus [56], Bamboo [31,35], Basil [55], Broccoli stem [52,56], Cabbage [57], Calathea zebrina [35], Carrot [52,56], Celery [53,56,58], Cucumber [56], Ficus hispida [59], Garcinia [59], Green onion [56,60], Impatiens capensis [35], Jujube [52], Leek [61], Lucky bamboo [55], Orchid pseudobulb [35], Pachira aquatica [59], Parsley stem [34,58], Peanut hairy root [34], Persimmon [52], Potato [56], Solenostemon “wasabi” [35], Spinach [34,54,55,58,60,62,63], Sweet yellow bell pepper [52], Sweet wormwood [34], summer lilac [35], tomato [55], Ubuçu Palm fibers [64], Vanilla [35]	Gold standard, well characterized; demonstrated ability to decellularize a multitude of plant materials with different structural and chemical compositions	Use of harsh chemicals; potential toxic residue thus, requires intense washing steps; time consuming; chemicals are environmentally toxic [46]
Detergent-Free [59]	Heated bleach and NaHCO3 solutions or bleach with surfactant	Minutes to hours, depending on the plant material	Bamboo stem, Ficus hispida, Garcinia, Pachira aquatica	Oxidation may enhance cellulose breakdown	Strong chemicals; able to degrade scaffold when heated [59]
Freeze/Enzymatic [65]	Lyophilization, DNAse I	24 h	Transgenic plant cultured cell lines: Hairy root, Tobacco bright yellow (BY-2), Monocot rice cells (Oryza sativa L.)	Retains native proteins	Additional clearing with surfactant might be needed to remove debris [1]
Supercritical Fluid (scCO2) [58]	scCO2 (2500 psi at 33 °C); PAA as cosolvent (2%); bleach if scaffold clearing required; Hexane pre- treatment can be performed when wax cuticle present	3 h (+6 h if clearing required)	Celery, Parsley stem, Spinach leaf, Sweet mint leaf	Fast; use of soft approach with minimal amount of chemicals; sterilization step included	Needs to be characterized on a larger diversity of plants; specialized equipment required [58]