Alginate-modified graphene oxide anchored with lactoperoxidase as a novel bioactive nanocombination for colorectal cancer therapy.

It is imperative to explore new biocompatible drugs with low toxicity for use in medicinal fields such
as fighting tumors. Bovine lactoperoxidase (BLPO) stems from the most important enzymes in the
bovine whey that provide a proper pattern for nano-formulation with nanomaterials. LPO is a suitable
protein to be coated or adsorbed to alginate modified graphene oxide (GO-SA), which forms the
modified GO-SA-LPO hybrid structure. This novel combination provides LPO stability with strong
anticancer effects and boosts immunity response. The characterization results obtained from different
techniques confirmed a successful LPO adsorption on the GO-SA composite surface. Moreover, nanoformulation
of LPO with GO-SA composite exhibited a reduction in its size and overall charge. In
addition, the experimental results showed greater LPO activity stability in the modified GO-SA-LPO
nanocombination than free LPO after storage for 10 weeks at 4 °C. The in vitro study, a crucial step
in the validation of our approach, demonstrated that the modified GO-SA-LPO nanocombination
showed a potent anticancer selectivity toward colon cancer cell lines more than GO-SA composite
or free form of LPO, which enhanced in a dose-dependent manner with high safety manner against
normal cells. The apoptotic effect of this novel nanocombination was confirmed by the greatest
variations in the expression of both well-known apoptosis genes (p53 and Bcl-2), severe changes in the
cellular morphology, DNA fragmentation, and nuclear staining with fluorescence yellow and orange
of the target cancer cells. Also, this superior efficacy of the modified GO-SA-LPO nanocombination
was induced by suppressing some pro-inflammatory cytokines, including tumor necrosis factor-alpha
(TNF-α), interleukin (IL-6), and necrosis factor-kappa B (NF-ĸB). Our observations presented that the
modified nanocombination of LPO may offer a novel remedy for treating colon tumors via induced
apoptosis pathway, inflammation reduction, and immune response improvement.