In vitro study of the biological activity of a recombinant chondroitin sulfate (CS) produced by fermentation in Escherichia coli

Abstract

Chondroitin Sulfate (CS) is a natural glycosaminoglycan found in cartilage and other connective tissues, and its therapeutic potential has been explored in various diseases. The current literature suggests CS to have significant impact an anti-inflammatory drug, as well as a preventative drug for several cancers, such as colorectal cancer. It achieves this by downregulating inflammatory responses and decreasing cytokine secretion. CS is currently extracted by animal-based production which has many drawbacks due to the high demand and low yield and the used extraction techniques. One of the most suggested alternatives to animal-based production of CS is its recombinant biosynthesis through microbial engineering. Currently there are no records of a recombinant CS (rCS) and its biological effect on Colorectal cancer cells. The primary objective of the study was to evaluate the dose and time-dependent effects of recombinant and animal-derived Chondroitin Sulfate on HCT116 cell viability in an in vitro analysis. The results showed no significant impact of rCS or animal-derived CS on cell viability 24 hours after treatment; however, at 48 hours, rCS demonstrated a reduction cell proliferation, indicating a possible dose- or time-dependent effect, though not statistically significant. The effect of rCS treatment on the gene expression of inflammation markers, IL-6, IL-8, IL-10, and MMP9 were also evaluated. IL-6, IL-8, and IL-10 were selected due to CS’s well-established role in modulating inflammation by altering the gene expression of these genes. MMP9 was selected based on its’ role in tumor progression and the tumor microenvironment. While a reduction in IL-6 and IL-8 expression was observed, no significant changes in expression were revealed for any studied gene. This suggests that neither rCS nor animal- derived CS had a pronounced impact on inflammation pathways in HCT116 cells.