Aptamer-engineered extracellular vesicles enable targeted celecoxib delivery for in vitro breast and colon cancer therapy

Cancer remains a leading cause of death worldwide, largely due to limitations in current therapies that often suffer from poor selectivity, systemic toxicity, and drug resistance. Innovative drug delivery platforms that can selectively target tumor cells while sparing healthy tissue are urgently needed to improve treatment outcomes. In this study, we address these challenges by developing a novel targeted delivery system using extracellular vesicles (EVs) loaded with celecoxib (Cel), a potent anti-inflammatory drug with anticancer properties, and functionalized with DNA aptamer AS1411 (Apt) to enhance tumor targeting. Celecoxib was efficiently encapsulated into EVs derived wharton’s jelly (WJ) mesenchymal stem cells (MSCs) via sonication. Subsequently, the EV surfaces were chemically conjugated with amine-modified Apt using EDC/NHS chemistry, creating a targeted nanosystem (Cel-apt-EVs) designed for enhanced recognition and uptake by cancer cells. Comprehensive in vitro analyses demonstrated that Cel-apt-EVs significantly improved cellular binding affinity and cytotoxicity in MCF-7 and HCT116 cancer cell lines compared to free celecoxib and non-functionalized EV formulations, while exhibiting minimal toxicity towards normal WI38 fibroblasts. Cell cycle analysis revealed that Cel-apt-EVs induced significant G0/G1 phase arrest in cancer cells, indicating inhibition of cell cycle progression. Furthermore, Annexin V/propidium iodide (PI)-based flow cytometry confirmed enhanced apoptotic cell death, with a marked increase in early and late apoptotic populations following Cel-apt-EVs treatment. Cel-apt-EVs effectively inhibited cancer cell migration in wound healing assays and markedly suppressed angiogenesis in the chick chorioallantoic membrane (CAM) model. These results highlight that aptamer-functionalized EVs provide a promising and biocompatible platform for precision delivery of celecoxib, improving anticancer efficacy while reducing off-target effects. This strategy offers new avenues for the development of safer and more effective targeted therapies in oncology.