Investigation of the effect mechanisms of Moringa oleifera loaded three-dimensional printed patches on In Vitro colon cancer cells

Colorectal cancer (CRC) is a form of cancer that initiates in the rectum. It stands as one of the most prevalent malignancies within the digestive system, characterized by high morbidity and mortality rates, and continues to pose a significant global health challenge. In the present day, the primary approach to treating CRC is through surgical intervention; however, this method frequently yields undesirable outcomes. To address these challenges, systemically administered chemotherapy has emerged as a treatment option. While chemotherapy is recognized as a pivotal strategy for inhibiting tumor growth or metastasis, it often brings about severe side effects, including local inflammation, cardiac toxicity, and anaphylactic reactions. The objective of this study is to examine the effects of in vitro local chemotherapy and herbal-derived local treatment on colon cancer cells by loading moringa oleifera (MO) extract and oxaliplatin (OX) drug into separate 3D printed patches. The goal is to minimize systemic toxicity, ensure the necessary and sufficient dosage of the drug, reduce serious side effects through effective transmission, and prevent the risk of possible recurrence of cancer cells in vitro after treatment by ensuring sustainable drug release. As part of the study, moringa oleifera (MO) extract and oxaliplatin (OX) drug were loaded into polycaprolactone and gelatin polymers at varying doses (50-100-250-500 µg/ml). Characterization analyses were conducted on the resulting 3D printed patches. Additionally, assessments were made on cell viability, lactate dehydrogenase, RNA isolation, cDNA, and apoptosis in the HCT 116 colon cancer cell line. The characterization results of the 3D printed patches, as outcomes of the study, demonstrated the successful production of MO and OX loaded patches. Furthermore, in the controlled release analysis of these patches, it was observed that the drug release in a pH 4 acidic environment was significantly higher compared to the pH 7.4 environment. The significance of this lies in the acidic environment present in cancerous tissue. Our in vitro studies on colon cancer cells found that 3D printed patches loaded separately with MO and OX exhibited antiproliferative effects on the HCT 116 cell line. Notably, in the cell cytotoxicity analysis, LDH activity increased more, especially in the group where the MO loaded 3D printed patch was applied compared to OX. Additionally, a decrease in the expression levels of apoptotic genes BCL2, BAX, CASP8, CASP9, BCL-xL, and p53 was detected. In conclusion, applying active ingredient loaded 3D printed patches locally in cancerous tissue for CRC treatment holds promise. This approach not only reduces systemic toxicity but also allows for the targeted release of the active ingredient in the desired tissue, thereby minimizing serious side effects. Moreover, our findings suggest that the MO loaded 3D printed patch exhibits greater efficacy on colon cancer cells in vitro than the OX loaded 3D printed patch.