Triple‑Phytochemical Nanoparticle for CRC: Formulation, In Vitro Release, Safety, and Gene‑Expression Signals

Investigators report developing a triple-payload nanoparticle carrying resveratrol, acetyl-11-keto-β-boswellic acid (AKBA), and quercetin—referred to as the 3X nanoparticle formulation—for colorectal cancer–relevant in vitro evaluation.

The abstract describes physicochemical characterization and encapsulation efficiency, in vitro pharmaceutical characteristics (including release/permeation), cytotoxicity testing in Caco-2 cells, and gene-expression analyses; it also reports minimal hemolysis as an in vitro safety-related result. Findings are organized across physicochemical characterization, payload incorporation, in vitro release/permeation behavior, hemocompatibility testing, cytotoxicity in Caco-2 cells, and gene-expression readouts after exposure. Across these sections, the authors frame the work as an early feasibility effort to assemble and screen a combination phytochemical nanoformulation under laboratory conditions.

Physical characterization is presented as baseline stability-related metrics for the 3X nanosuspension. The authors report a mean particle diameter of 198.5 nm, a polydispersity index (PDI) of 0.492, and a zeta potential of −32.7 mV. In the article’s narrative, these measurements describe the size distribution and surface charge profile of the formulation as prepared and tested, without extending claims beyond the reported measurements. Together, these values define the physical profile carried forward into subsequent in vitro assays.

Payload incorporation is reported as compound-specific encapsulation performance within the single formulation. The authors describe quantifying encapsulation efficiencies for each phytochemical in 3X and report values of 90% for AKBA, 80% for resveratrol, and 75% for quercetin. These loading results are presented as the study’s accounting of how much of each agent was retained within the nanoparticle system at the time of measurement. The investigators position these reported efficiencies as the basis for subsequent release/transport experiments and cell-based testing.

Release and transport behavior are summarized in the context of how the multi-agent system behaves under the study’s in vitro conditions. In describing in vitro permeation studies, the authors report that the formulation demonstrated a “controlled release” mechanism for the encapsulated phytochemicals.

Hemocompatibility and viability-related screening are reported as separate assay readouts. For hemolysis testing, the authors report minimal hemolysis of approximately 3% in vitro. In parallel, cytotoxicity testing in the Caco-2 colorectal cancer cell line is reported with an IC50 of 365 µg in the study’s cytotoxicity assay (reported in the abstract without additional unit context). Together, these values are presented as in vitro safety/compatibility and cell-viability readouts for the 3X formulation.

Gene-expression results are described as directionally altered transcripts in Caco-2 cells following exposure to 3X. The investigators report downregulation of TGFβ and COX-2, reduced IL-1β expression, and upregulation of TNFα compared with control cells; the abstract also reports increased nitric oxide (NO) alongside these gene-expression changes. In their conclusions, the authors characterize the overall findings as preliminary and state that further in vivo studies are warranted to validate these findings and clarify underlying molecular mechanisms.

Questions that follow from the reported dataset include how the in vitro release/transport behavior might map onto in vivo exposure and whether the observed transcript signals would persist in more complex tumor-relevant models. The study’s closing message remains centered on what was measured in vitro and what the authors propose to evaluate next.

Key Takeaways:

• The authors report a baseline physical characterization profile for 3X using mean diameter, PDI, and zeta potential as the primary presented metrics.
• Investigators describe controlled-release behavior in permeation-related in vitro testing as part of the formulation’s reported pharmaceutical characteristics.
• The paper reports low hemolysis and a Caco-2 cytotoxicity IC50 alongside directionally changed gene-expression signals (including TGFβ, COX-2, TNFα, NO, and IL-1β) after 3X exposure.