Kiwi-derived N-doped green carbon dots: Antioxidant and anti-inflammatory potential for hyperglycemia control

Document Type : Original Article

Authors

Department of Pharmaceutics, Faculty of Pharmacy, Alexandria University

Abstract

Diabetes mellitus is a major global health problem characterized by chronic hyperglycemia, which can lead to serious complications if left untreated. In the recent years, the development of novel diagnostic and therapeutic tools for diabetes has become increasingly important. Among emerging approaches, the green synthesis of nanomaterials has drawn significant interest due to its eco-friendly nature. This study investigates the use of kiwifruit as a sustainable carbon precursor with triethanolamine as a nitrogen dopant for the phyto-mediated production of carbon dots (K-NCDs) utilizing a simple solvothermal technique. The produced K-NCDs were thoroughly characterized using UV–Visible spectroscopy, fluorescence spectrophotometry, FT-IR, X-ray diffraction (XRD), transmission electron microscopy, and others. The K-NCDs had an average size of 9.2±3 nm, a ζ-potential of -26.1±1.85 mV, and a quasi-spherical morphology. They exhibited an intense green emission under the excitation of 275 nm, with FT-IR analysis proving nitrogen doping and XRD verifying their amorphous structure. Biological activity evaluation demonstrated potent antioxidant and hypoglycemic effects.
At a concentration of 5 mg/mL, K-NCDs achieved 99.6 % 2,2-diphenyl-1-picrylhydrazyl inhibition and a dose-dependent catalase-like activity. They also inhibited α-amylase, a key target for managing type 2 diabetes, by 79.7±2.66%. In vitro insulin-sensitizing assays on insulin-resistant HepG2 and PANC-1 cells revealed enhanced glucose at concentrations of 25-100 μg/mL. Moreover, oral hypoglycemic efficacy was confirmed in a streptozocin-induced diabetic rat model following a single 400 mg/kg oral dose. The study findings support the use of K-NCDs as a promising, eco-friendly therapeutic option for hyperglycemia.

Highlights

•    Sustainable and eco-friendly N-doped carbon dots were prepared from kiwifruit
•    K-NCDs showed a high relative fluorescence quantum yield of 37.9 %
•    Dose-dependent antioxidant, α-amylase inhibition, and catalase-like effects detected 
•    K-NCDs showed insulin resistance reversal in vitro in HepG2/IRM and PANC-1 cells
•    Oral hypoglycemic efficacy following K-NCDs oral administration in diabetic rat model

Keywords

Main Subjects


(1)    International Diabetes Federation. International Diabetes Federation Diabetes Atlas. 2023. Available from: https://diabetesatlas.org/.
(2)    Heller SR, Geybels MS, Iqbal A, Liu L, Wagner L, Chow E. A higher non-severe hypoglycaemia rate is associated with an increased risk of subsequent severe hypoglycaemia and major adverse cardiovascular events in individuals with type 2 diabetes in the LEADER study. Diabetologia. 2022;65:55-64.
(3)    Bhatia S. Nanoparticles types, classification, characterization, fabrication methods and drug delivery applications. Natural polymer drug delivery systems: Nanoparticles, plants, and algae. 2016:33-93.
(4)    Cui L, Ren X, Sun M, Liu H, Xia L. Carbon dots: Synthesis, properties and applications. Nanomaterials. 2021;11(12):3419.
(5)    Sahana S, Gautam A, Singh R, Chandel S. A recent update on development, synthesis methods, properties and application of natural products derived carbon dots. Natural Products and Bioprospecting. 2023;13(1):51.
(6)    Yao L, Zhao M-M, Luo Q-W, Zhang Y-C, Liu T-T, Yang Z, et al. Carbon quantum dots-based nanozyme from coffee induces cancer cell ferroptosis to activate antitumor immunity. ACS nano. 2022;16(6):9228-39.
(7)    Qureshi WA, Vivekanandan B, Jayaprasath JA, Ali D, Alarifi S, Deshmukh K. Antimicrobial Activity and Characterization of Pomegranate Peel‐Based Carbon Dots. Journal of Nanomaterials. 2021;2021(1):9096838.
(8)    Prakash A, Muthu M, Raja G, Gopal J. Preparation and Characterization of Carbon Nanodots from turmeric soot for anti-coliform and anti-oral bacterial applications and as anti-staphylococcal coatings. Nanotheranostics. 2025;9(1):31.
(9)    Humaera NA, Fahri AN, Armynah B, Tahir D. Natural source of carbon dots from part of a plant and its applications: a review. Luminescence. 2021;36(6):1354-64.
(10)    Zeng M, Wang Y, Liu M, Wei Y, Wen J, Zhang Y, et al. Potential efficacy of herbal medicine-derived Carbon Dots in the treatment of diseases: from mechanism to clinic. International Journal of Nanomedicine. 2023:6503-25.
(11)    Guo S, Zhang R, Liu Y, Zhang Q, Liu X, Wu X, et al. Synthesis, applications in therapeutics, and bioimaging of traditional Chinese medicine-derived carbon dots. Carbon Letters. 2024;34(2):545-64.
(12)    Lu F, Zhang Y, Cheng J, Zhang M, Luo J, Qu H, et al. Maltase and sucrase inhibitory activities and hypoglycemic effects of carbon dots derived from charred Fructus crataegi. Materials Research Express. 2019;6(12):125005.
(13)    Sun Z, Lu F, Cheng J, Zhang M, Zhu Y, Zhang Y, et al. Hypoglycemic bioactivity of novel eco-friendly carbon dots derived from traditional Chinese medicine. Journal of Biomedical Nanotechnology. 2018;14(12):2146-55.
(14)    El Azab EF, Alakilli SYM, Saleh AM, Alhassan HH, Alanazi HH, Ghanem HB, et al. Actinidia deliciosa Extract as a Promising Supplemental Agent for Hepatic and Renal Complication-Associated Type 2 Diabetes (In Vivo and In Silico-Based Studies). Int J Mol Sci. 2023;24(18).
(15)    Satpal D, Kaur J, Bhadariya V, Sharma K. Actinidia deliciosa (Kiwi fruit): A comprehensive review on the nutritional composition, health benefits, traditional utilization, and commercialization. Journal of Food processing and Preservation. 2021;45(6):e15588.
(16)    Mechchate H, Es-Safi I, Haddad H, Bekkari H, Grafov A, Bousta D. Combination of Catechin, Epicatechin, and Rutin: Optimization of a novel complete antidiabetic formulation using a mixture design approach. The Journal of Nutritional Biochemistry. 2021;88:108520.
(17)    Arul V, Sethuraman MG. Facile green synthesis of fluorescent N-doped carbon dots from Actinidia deliciosa and their catalytic activity and cytotoxicity applications. Optical Materials. 2018;78:181-90.
(18)    Atchudan R, Kishore SC, Gangadaran P, Edison TNJI, Perumal S, Rajendran RL, et al. Tunable fluorescent carbon dots from biowaste as fluorescence ink and imaging human normal and cancer cells. Environmental Research. 2022;204:112365.
(19)    Wang K, Li M, Han Q, Fu R, Ni Y. Inhibition of α-amylase activity by insoluble and soluble dietary fibers from kiwifruit (Actinidia deliciosa). Food Bioscience. 2021;42:101057.
(20)    Aboul-Enein A, Salama DZ, Gaafar A, Aly H, Ahmed H. Comparative hypoglycemic effect of acetone extract of banana (Musa paradisiaca), kiwi (Actinidia deliciosa planch) and olive (Olea europaea L.), byproducts. Bioscience Research. 2019;16:854-69.
(21)    Rehman G, Hamayun M, Iqbal A, Ul Islam S, Arshad S, Zaman K, et al. In Vitro Antidiabetic Effects and Antioxidant Potential of Cassia nemophila Pods. Biomed Res Int. 2018;2018:1824790.
(22)    Wickramaratne MN, Punchihewa JC, Wickramaratne DB. In-vitro alpha amylase inhibitory activity of the leaf extracts of Adenanthera pavonina. BMC Complement Altern Med. 2016;16(1):466.
(23)    Soliman MO, El-Kamel AH, Shehat MG, Bakr BA, El-Moslemany RM. Lactoferrin decorated bilosomes for the oral delivery of quercetin in type 2 diabetes: In vitro and in vivo appraisal. Int J Pharm. 2023;647:123551.
(24)    Gao F, Jian L, Zafar MI, Du W, Cai Q, Shafqat RA, et al. 4-Hydroxyisoleucine improves insulin resistance in HepG2 cells by decreasing TNF-α and regulating the expression of insulin signal transduction proteins. Mol Med Rep. 2015;12(5):6555-60.
(25)    Dwivedi S, Gottipati A, Ganugula R, Arora M, Friend R, Osburne R, et al. Oral Nanocurcumin Alone or in Combination with Insulin Alleviates STZ-Induced Diabetic Neuropathy in Rats. Mol Pharm. 2022;19(12):4612-24.
(26)    Richardson DP, Ansell J, Drummond LN. The nutritional and health attributes of kiwifruit: a review. Eur J Nutr. 2018;57(8):2659-76.
(27)    Yu M, Man Y, Lei R, Lu X, Wang Y. Metabolomics Study of Flavonoids and Anthocyanin-Related Gene Analysis in Kiwifruit (Actinidia chinensis) and Kiwiberry (Actinidia arguta). Plant Molecular Biology Reporter. 2020;38(3):353-69.
(28)    Atchudan R, Edison TNJI, Lee YR. Nitrogen-doped carbon dots originating from unripe peach for fluorescent bioimaging and electrocatalytic oxygen reduction reaction. Journal of Colloid and Interface Science. 2016;482:8-18.
(29)    Prathap N, Balla P, Shivakumar MS, Periyasami G, Karuppiah P, Ramasamy K, et al. Prosopis juliflora hydrothermal synthesis of high fluorescent carbon dots and its antibacterial and bioimaging applications. Scientific Reports. 2023;13(1):9676.
(30)    Kundu A, Maity B, Basu S. Orange Pomace-Derived Fluorescent Carbon Quantum Dots: Detection of Dual Analytes in the Nanomolar Range. ACS Omega. 2023;8(24):22178-89.
(31)    Georgakilas V, Perman JA, Tucek J, Zboril R. Broad Family of Carbon Nanoallotropes: Classification, Chemistry, and Applications of Fullerenes, Carbon Dots, Nanotubes, Graphene, Nanodiamonds, and Combined Superstructures. Chemical Reviews. 2015;115(11):4744-822.
(32)    Afshary H, Amiri M, Bezaatpour A, Wark M. Electrochemiluminescence sensor based on N-doped carbon quantum dots for determination of ceftazidime in real samples. J Electrochem Soc. 2022;169(2):026523.
(33)    John BK, John N, Korah BK, Thara C, Abraham T, Mathew B. Nitrogen-doped carbon quantum dots as a highly selective fluorescent and electrochemical sensor for tetracycline. Journal of Photochemistry and Photobiology A: Chemistry. 2022;432:114060.
(34)    Manioudakis J, Victoria F, Thompson CA, Brown L, Movsum M, Lucifero R, et al. Effects of nitrogen-doping on the photophysical properties of carbon dots. Journal of Materials Chemistry C. 2019;7(4):853-62.
(35)    Bhatti JS, Sehrawat A, Mishra J, Sidhu IS, Navik U, Khullar N, et al. Oxidative stress in the pathophysiology of type 2 diabetes and related complications: Current therapeutics strategies and future perspectives. Free Radical Biology and Medicine. 2022;184:114-34.
(36)    Dong C, Wang S, Ma M, Wei P, Chen Y, Wu A, et al. Inhibition of oxidative stress in vivo through enzyme-like activity of carbon dots. Applied Materials Today. 2021;25:101178.
(37)    Wang Y, Li L, Liu H, Zhao T, Meng CW, Liu Z, et al. Bioactive compounds and in vitro antioxidant activities of peel, flesh and seed powder of kiwi fruit. International Journal of Food Science & Technology. 2018;53:2239–45.
(38)    Baliyan S, Mukherjee R, Priyadarshini A, Vibhuti A, Gupta A, Pandey RP, et al. Determination of Antioxidants by DPPH Radical Scavenging Activity and Quantitative Phytochemical Analysis of Ficus religiosa. Molecules. 2022;27(4):1326.
(39)    Murru C, Badía-Laíño R, Díaz-García ME. Synthesis and Characterization of Green Carbon Dots for Scavenging Radical Oxygen Species in Aqueous and Oil Samples. Antioxidants. 2020;9(11):1147.
(40)    Yang D, Li L, Cao L, Chang Z, Mei Q, Yan R, et al. Green Synthesis of Lutein-Based Carbon Dots Applied for Free-Radical Scavenging within Cells. Materials. 2020;13(18):4146.
(41)    Innocenzi P, Stagi L. Carbon dots as oxidant-antioxidant nanomaterials, understanding the structure-properties relationship. A critical review. Nano Today. 2023;50:101837.
(42)    Leonida MD, Kumar I, Elshaer MR, Mahmoud Z, Lozanovska B, Bijja UK, et al. Ecofriendly approaches to efficiently enhance catalase performance. International Journal of Biological Macromolecules. 2024;280:135597.
(43)    Chavez J, Khan A, Watson KR, Khan S, Si Y, Deng AY, et al. Carbon Nanodots Inhibit Tumor Necrosis Factor-α-Induced Endothelial Inflammation through Scavenging Hydrogen Peroxide and Upregulating Antioxidant Gene Expression in EA.hy926 Endothelial Cells. Antioxidants. 2024;13(2):224.
(44)    Muhammad P, Hanif S, Li J, Guller A, Rehman FU, Ismail M, et al. Carbon dots supported single Fe atom nanozyme for drug-resistant glioblastoma therapy by activating autophagy-lysosome pathway. Nano Today. 2022;45:101530.
(45)    Rehman G, Hamayun M, Iqbal A, Islam S, Arshad S, Zaman K, et al. In Vitro Antidiabetic Effects and Antioxidant Potential of Cassia nemophila Pods. BioMed Research International. 2018;2018:6.
(46)    Harish M, Ahmed F, Urooj A. In vitro hypoglycemic effects of Butea monosperma Lam. leaves and bark. J Food Sci Technol. 2014;51(2):308-14.
(47)    Parvathy C, P.K P. Evaluation of Anti-diabetic Potential of Anti-microbial Carbon Quantum Dots from Vitis Vinifera Seeds. Nano Biomedicine and Engineering. 2023;15.
(48)    Kaur N, Kumar V, Nayak SK, Wadhwa P, Kaur P, Sahu SK. Alpha-amylase as molecular target for treatment of diabetes mellitus: A comprehensive review. Chemical Biology & Drug Design. 2021;98(4):539-60.
(49)    Gong L, Feng D, Wang T, Ren Y, Liu Y, Wang J. Inhibitors of α-amylase and α-glucosidase: Potential linkage for whole cereal foods on prevention of hyperglycemia. Food Science & Nutrition. 2020;8(12):6320-37.
(50)    Dhiman VK, Chauhan V, Kanwar SS, Singh D, Pandey H. Purification and characterization of actinidin from Actinidia deliciosa and its utilization in inactivation of α-amylase. Bulletin of the National Research Centre. 2021;45(1):213.
(51)    Kota S, Dumpala P, Sajja R, Anantha R. Heteroatom-doped carbon dots from medicinal plants as novel biomaterials for as-use biomedical applications in comparison with synthetic drug, zaltoprofen. Scientific Reports. 2024;14(1):13160.
(52)    Prasada P, Nirmalab PV. Green Carbon Dots Derived From Acetosa Sagittata Aerial Parts And In Vitro Antidiabetic, Antioxidant And Cytotoxic Properties. Revista Argentina de Clínica Psicológica     2023;XXXII:63-71 
(53)    Park Y, Kim Y, Chang H, Won S, Kim H, Kwon W. Biocompatible nitrogen-doped carbon dots: synthesis, characterization, and application. Journal of Materials Chemistry B. 2020;8(39):8935-51.
(54)    Magalhaes CM, Ribeiro E, Fernandes S, Esteves da Silva J, Vale N, Pinto da Silva L. Safety Evaluation of Carbon Dots in UM-UC-5 and A549 Cells for Biomedical Applications. Cancers (Basel). 2024;16(19).
(55)    Patel DK, Won S-Y, Jung E, Deb Dutta S, Patil TV, Lim K-T, et al. Assessment of the biocompatibility and bioimaging potential of fluorescent carbon dots derived from waste biomass. Materials Letters. 2024;361:136152.
(56)    Salvatore T, Marfella R, Rizzo MR, Sasso FC. Pancreatic cancer and diabetes: A two-way relationship in the perspective of diabetologist. Int J Surg. 2015;21 Suppl 1:S72-7.
(57)    Kumar M, Chinnathambi S, Bakhori N, Abu N, Etezadi F, Thangavel V, et al. Biomass-derived carbon dots as fluorescent quantum probes to visualize and modulate inflammation. Sci Rep. 2024;14(1):12665.
(58)    Sharma A, Choi HK, Lee HJ. Carbon Dots for the Treatment of Inflammatory Diseases: An Appraisal of In Vitro and In Vivo Studies. Oxid Med Cell Longev. 2023;2023:3076119.
(59)    Dong C, Ma X, Huang Y, Zhang Y, Gao X. Carbon dots nanozyme for anti-inflammatory therapy via scavenging intracellular reactive oxygen species. Front Bioeng Biotechnol. 2022;10:943399.
(60)    Kong B, Yang T, Cheng F, Qian Y, Li C, Zhan L, et al. Carbon dots as nanocatalytic medicine for anti-inflammation therapy. J Colloid Interface Sci. 2022;611:545-53.
(61)    Shao T, Yuan P, Zhu L, Xu H, Li X, He S, et al. Carbon Nanoparticles Inhibit ?-Glucosidase Activity and Induce a Hypoglycemic Effect in Diabetic Mice. Molecules. 2019;24(18):3257.
(62)    Camlik G, Ozakca I, Bilakaya B, Ozcelikay AT, Velaro AJ, Wasnik S, et al. Development of composite carbon quantum dots-insulin formulation for oral administration. Journal of Drug Delivery Science and Technology. 2022;76:103833.
(63)    Zhao J, Zhang Y, Zhao Y, Wu T, Chen Y, Zhang Y, et al. Protective Effects of Zingiberis Carbonisata-Based Carbon Dots on Diabetic Liver Injury in Mice. Journal of Biomedical Nanotechnology. 2022;18(8):1975-85.
(64)    Rochette L, Zeller M, Cottin Y, Vergely C. Diabetes, oxidative stress and therapeutic strategies. Biochim Biophys Acta, Gen Subj. 2014;1840(9):2709-29.