Chitosan-PEI passivated carbon dots for plasmid DNA and miRNA-153 delivery in cancer cells
Saloni Thakur,
Reena V. Saini,
Neelam Thakur,
Rohit Sharma,
Joydeep Das,
Petr Slama,
Hardeep Singh Tuli,
Shafiul Haque,
Hatoon A. Niyazi,
Mohammed Moulay,
Steve Harakeh,
Adesh K. Saini
Affiliations
Saloni Thakur
Faculty of Applied Sciences and Biotechnology, Shoolini University, Solan, 173229, India
Reena V. Saini
Department of Biotechnology, MMEC, Maharishi Markandeshwar (Deemed to Be University), Mullana, 133207, India
Neelam Thakur
School of Advance Chemical Sciences, Shoolini University, Solan, 173229, India
Rohit Sharma
Faculty of Applied Sciences and Biotechnology, Shoolini University, Solan, 173229, India; Corresponding author.
Joydeep Das
Department of Chemistry, Physical Sciences, Mizoram University, Aizawl, 796004, India; Corresponding author.
Petr Slama
Laboratory of Animal Immunology and Biotechnology, Department of Animal Morphology, Physiology and Genetics, Faculty of AgriSciences, Mendel University in Brno, 61300, Brno, Czech Republic
Hardeep Singh Tuli
Department of Biotechnology, MMEC, Maharishi Markandeshwar (Deemed to Be University), Mullana, 133207, India
Shafiul Haque
Research and Scientific Studies Unit, College of Nursing and Allied Health Sciences, Jazan University, Jazan, 45142, Saudi Arabia; Gilbert and Rose-Marie Chagoury School of Medicine, Lebanese American University, Beirut, Lebanon; Centre of Medical and Bio-Allied Health Sciences Research, Ajman University, Ajman, United Arab Emirates
Hatoon A. Niyazi
Department of Clinical Microbiology and Immunology, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia
Mohammed Moulay
Embryonic Stem Cells Research Unit, King Fahd Medical Research Center. King Abdul Aziz University, Jeddah, Saudi Arabia; Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia
Steve Harakeh
King Fahd Medical Research Center, and Yousef Abdullatif Jameel Chair of Prophetic Medicine Application, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia
Adesh K. Saini
Department of Biotechnology, MMEC, Maharishi Markandeshwar (Deemed to Be University), Mullana, 133207, India; Corresponding author.
These days carbon dots have been developed for multiple biomedical applications. In the current study, the transfection potential of synthesized carbon dots from single biopolymers such as chitosan, PEI-2kDa, and PEI-25kDa (CS-CDs, PEI2-CDs, and PEI25-CDs) and by combining two biopolymers (CP2-CDs and CP25-CDs) through a bottom-up approach have been investigated. The characterization studies revealed successful synthesis of fluorescent, positively charged carbon dots <20 nm in size. Synthesized carbon dots formed a stable complex with plasmid DNA (EGFP-N1) and miRNA-153 that protected DNA/miRNA from serum-induced degradation. In-vitro cytotoxicity analysis revealed minimal cytotoxicity in cancer cell lines (A549 and MDA-MB-231). In-vitro transfection of EGFP-N1 plasmid DNA with PEI2-CDs, PEI25-CDs and CP25-CDs demonstrated that these CDs could strongly transfect A549 and MDA-MB-231 cells. The highest EGFP-N1 plasmid transfection efficiency was observed with PEI2-CDs at a weight ratio of 32:1. PEI25-CDs polyplex showed maximum transfection at a weight ratio of 8:1 in A549 at a weight ratio of 16:1 in MDA-MB-231 cells. CP25-CDs exhibited the highest transfection at a weight ratio of 16:1 in both cell lines. The in-vitro transfection of target miRNA, i.e., miR-153 in A549 and MDA-MB-231 cells with PEI2-CDs, PEI25-CDs, and CP25-CDs suggested successful transfer of miR-153 into cells which induced significant cell death in both cell lines. Importantly, CS-CDs and CP2-CDs could be tolerated by cells up to 200 μg/mL concentration, while PEI2-CDs, PEI25-CDs, and CP25-CDs showed non-cytotoxic behavior at low concentrations (25 μg/mL). Together, these results suggest that a combination of carbon dots synthesized from chitosan and PEI (CP25-CDs) could be a novel vector for transfection nucleic acids that can be utilized in cancer therapy.
