In vitro gastrointestinal digestion and cytotoxic effect of ovalbumin-conjugated linoleic acid nanocomplexes

Abstract

The aim of this work was to examine the behavior of conjugated linoleic acid (CLA) delivery systems based on ovalbumin (OVA) and their derived nanoparticles (OVAn1 and OVAn2), under static in vitro gastrointestinal digestion model. In addition, potential cytotoxic effect of these inclusion complexes on a human colon cancer cell line (HT-29) was evaluated. OVA was resistant to gastric and intestinal digestion, while OVA nanoparticles were very susceptible to digestive enzymes hydrolysis. Particle size distribution (PDS) and sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) for OVA evidenced the presence of a protein fragment of similar size after simulated digestive process. Conversely, for nanoparticles, partial and total hydrolysis in gastric and intestinal phases, respectively, was evidenced. After in vitro gastrointestinal digestion, released CLA (RCLA) was assayed. In case of OVA, as CLA carrier, RCLA was 37%, while for OVA nanoparticles, lower RCLA values (~10–20%) were obtained. From cytotoxic assays, it was observed that CLA molecule was responsible for cell death, whereas OVA or their derived nanoparticles were not cytotoxic on HT-29 cells. On the other hand, flow cytometry analysis revealed that main death mechanism for CLA, and their inclusion complexes was apoptosis. OVA-CLA and OVAn1-CLA inclusion complexes displayed the highest potential cytotoxic activity and apoptotic index. Information derived from this work could be relevant for the design of CLA delivery systems as promising nanosupplements for production of new functional and excipient foods for both prevention and control of colon cancer.