Graphene oxide incorporated sodium alginate-gelatin composite hydrogel loaded with Moringa oleifera leaf extract for bifunctional tissue regeneration and anticancer applications

This study presents the development of sodium alginate–gelatin (SG) composite hydrogel scaffolds incorporated with graphene oxide (GO) and loaded with Moringa oleifera leaf extract (MLE) for tissue regeneration and anticancer applications. The MLE was loaded in varying concentration 2.5 g/100 mL, 5 g/100 mL and 10 g/100 mL to SGG hydrogels (SGG@I, SGG@II and SGG@III) respectively. The prepared GO, SGG, SGG@I, SGG@II and SGG@III were characterized by X-ray diffraction (XRD), UV visible spectroscopy and Fourier Transform Infrared (FTIR) spectroscopy to evaluate the physiochemical properties. The swelling and degradation behavior of developed hydrogels were analyzed which stimulated under acidic pH 5.0 conditions. Also, The cumulative MLE release % from SGG@I, SGG@II and SGG@III hydrogels was observed to be enhanced under acidic pH 5.0 PBS medium. Mechanical testing exhibited the stability of hydrogel membranes by incorporating the GO and MLE extract by using the Universal Testing Machine (UTM). The MLE extract and MLE extract loaded hydrogel showed the scavenging of free radicals in dose dependent manner by following the DPPH assay. The hydrogels SGG, SGG@I, SGG@II and SGG@III were evaluated for their biocompatibility and tissue regeneration capability against HUVEC cell lines by following the MTT assay. All the hydrogel membranes showed significant biocompatibility for 5 days with the highest concentration of MLE demonstrated 71.00 ± 3.2 % cell viability. Furthermore, SGG and loaded MLE hydrogels SGG@I, SGG@II and SGG@III were analyzed against MCF-7 and BT-549 breast cancer cell lines by MTT assay. The increasing concentration of MLE stimulated the anticancer potential of hydrogels as SGG@III showed the cell viability of 39.00 ± 3.15 % and 54.55 ± 2.95 % by day 5th against MCF-7 and BT-549 cell lines respectively. The intracellular reactive oxygen species (ROS) generation potential of the hydrogel was evaluated using the DCFH-DA assay, revealing a time-dependent increase in green fluorescence intensity in SGG@III-treated cells over 72 h. The sustained elevation in ROS levels suggested the developed MLE loaded hydrogels ability to induce oxidative stress in cancer cells which support its potential as an effective therapeutic agent for post-surgical breast cancer treatment with good biocompatibility.