Sustainable processing from native vegetation such as agave, nopal and aloe, is mainly centered on the components of lignin, cellulose, and hemicellulose, defined generally as lignocellulosic. Cytotoxicity of lignocellulosic biopolymer membranes (LBMs) of mucilages of Opuntia ficus-indica, Aloe barbadensis Miller, fructans, and residual Agave tequilana Weber fibers at different concentrations (30, 50 and 70%), was evaluated in contact with human fibroblast cells (ATCC® PCS-201-010), cultured in vitro. Long and short LMBs were formed from the fibrous material. The interaction showed mild cytotoxicity according to ISO 10993-5: 2009. Cytotoxic activity demonstrated in the presence of fibroblasts implied a maximum cytotoxicity of close to 46% and a minimum of 7% in LBMs. UV light absorption results of the large fiber LMBs showed the highest prevention of passage of light, having a protective effect mechanism of delaying the response of hypersensitivity as a result of exposure to UV radiation. The morphology and structure were characterized using SEM, FT-IR, and image texture analysis. According to the LBMs reactivity measured, the feasible uses of these membranes can be suggested as materials for biomedical applications over a short period of time, with feasibility as a support to provide some beneficial solutions.