Innovative encapsulation for enhanced drug delivery using stingless bee propolis and honey Tetragonisca angustula (Hymenoptera: Meliponini), and their antimicrobial activity

Authors

DOI:

https://doi.org/10.70099/BJ/2025.02.03.10

Keywords:

Tetragonisca angustula, stingless bee, polymeric nanoparticles, controlled released, antimicrobial activity

Abstract

Propolis, known for its immunological and therapeutic properties due to flavonoids and phenolic acids, has been used in alternative medicine for many years. In Ecuador, the commercialization of stingless bee honey has been increasing recently, although the physical-chemical and biological characteristics of honey from each species are unknown due to their different behaviors and diets. The objective of this research was to synthesize chitosan nanoparticles by encapsulating commercial honey (NPMC), honey (NPMA), and propolis (NPP) from Tetragonisca angustula by ionic gelation and glutaraldehyde cross-linking. The polymeric nanoparticles were structurally characterized (TGA and FT-IR), and their antimicrobial activity against E. coli and S. aureus, cytotoxicity, and controlled release were evaluated in two phases at different pH levels, one gastric (pH 1.8) and intestinal (pH 7.4). FT-IR analysis of the polymeric nanoparticles revealed the presence of hydroxyl and alkyl groups. NPMA and NPP showed characteristic vibrations of flavonoids and C=O, regardless of the molecular weight of chitosan. In the TGA, the NPMC had decomposition efficiencies of 97.22% (LMW) and 92.10% (HMW). Propolis nanoparticles achieved encapsulation efficiencies of 94.31% and 89.21%. NPPA (HMW-chitosan-based nanoparticles encapsulating propolis from stingless bees) was the most effective against S. aureus. NPPB (LMW-chitosan-based nanoparticles encapsulating propolis from stingless bees) at 10 μg/mL showed higher cytotoxicity than NPMAA (HMW-chitosan-based nanoparticles encapsulating honey from stingless bees). In summary, propolis nanoparticles maintained efficiencies above 76%, while NPMC and NPMA exhibited loading capacities greater than 27%. In contrast, formulations with high molecular weight chitosan showed slightly lower values, although these differences were not statistically significant.

References

1. Villalobos J HM, Córdova A L V, Perú L, Espinosa E EA, Guanoluisa R CE, Villacís YJ. Impact of bees (Apis mellifera L.) as Pollinating Agents in the Performance of the Zucchini Crop (Cucurbita pepo L.) in the Canton Riobamba, Province of Chimborazo. Vol. 15. 2019.

2. Michener C. The Bees of the World. 2nd ed. 2007.

3. González J, Quezada J. Biodiversidad y Desarrollo Humano en Yucatán I Uso de la Flora y Fauna Silvestre [Internet]. 2016 [cited 2024 May 1]. Available from: https://www.cicy.mx/documentos/cicy/sitios/biodiversidad/pdfs/cap7/19%20produccion%20tradicional%20de%20miel.pdf

4. Paulo S. Variabilidade genética de Tetragonisca angustula (Hymenoptera, Apidae, Meliponini) de meliponários. Genetic variability of Tetragonisca angustula (Hymenoptera, Apidae, Meliponini) from meliponaries. 2013.

5. Rojas DG. Diversidad morfológica y genética de Tetragonisca angustula (Hymenoptera: Apidae) en Cundinamarca, Colombia. 2022.

6. Bravo E. La biodiversidad en el Ecuador. Abya-Yala. Quito; 2014.

7. Nates-Parra G. Neotropical Braconidae Wasps-233 Campos Biota Colombiana. Vol. 2, Biota Colombiana. 2001.

8. Kerr JT. Butterfly Species Richness Patterns in Canada: Energy, Heterogeneity, and the Potential Consequences of Climate Change [Internet]. Vol. 5, Ecology. 2001. Available from: http://www.npwrc.usgs.gov/resource/distr/lepid/bflyusa/bflyusa.htm

9. Manrique AJ, Santana WC. Flavonoides, actividades antibacteriana y antioxidante de propóleos de abejas sin aguijón, Melipona quadrifasciata, Melipona compressipes, Tetragonisca angustula y Nannotrigona sp. de Brasil y Venezuela. Vol. 26, Zootecnia Trop. 2008.

10. Vit P, González I, Sorroza L, Pedro RMS. Caracterización físicoquímica de miel de angelita Tetragonisca angustula (Latreille, 1811) producida en Esmeraldas, Ecuador. Revista Ciencia UNEMI [Internet]. 2016 [cited 2024 May 1];9(20):77–84. Available from: https://www.redalyc.org/articulo.oa?id=582663826011

11. Galindo-Pérez M, Morales-Ibarría M, Reyes-Duarte D, Hernández-Arana A, Campos-Terán J, Arroyo-Maya I. NANOPARTÍCULAS DE C-FICOCIANINA: AUTOENSAMBLADO, ACTIVIDAD ANTIOXIDANTE Y ESTUDIOS DE ESTABILIDAD. Vol. 2023, Año.

12. Stark WJ. Nanoparticles in biological systems. Vol. 50, Angewandte Chemie - International Edition. 2011. p. 1242–58.

13. Mohammed MA, Syeda JTM, Wasan KM, Wasan EK. An overview of chitosan nanoparticles and its application in non-parenteral drug delivery. Vol. 9, Pharmaceutics. MDPI AG; 2017.

14. Jafernik K, Ładniak A, Blicharska E, Czarnek K, Ekiert H, Wiącek AE, et al. Chitosan-Based Nanoparticles as Effective Drug Delivery Systems—A review. Vol. 28, Molecules. MDPI; 2023.

15. Rashki S, Asgarpour K, Tarrahimofrad H, Hashemipour M, Ebrahimi MS, Fathizadeh H, et al. Chitosan-based nanoparticles against bacterial infections. Vol. 251, Carbohydrate Polymers. Elsevier Ltd; 2021.

16. Liu SH, Chiu CY, Shi CM, Chiang MT. Functional comparison of high and low molecular weight chitosan on lipid metabolism and signals in high-fat diet-fed rats. Mar Drugs. 2018 Jul 30;16(8).

17. Wang W, Meng Q, Li Q, Liu J, Zhou M, Jin Z, et al. Chitosan derivatives and their application in biomedicine. Vol. 21, International Journal of Molecular Sciences. MDPI AG; 2020.

18. Kono H, Kato T. Elucidation of substituent distribution states for carboxymethyl chitosan by detailed NMR analysis. Carbohydrate Polymer Technologies and Applications. 2021 Dec 25;2.

19. Mansour A, Romani M, Acharya AB, Rahman B, Verron E, Badran Z. Drug Delivery Systems in Regenerative Medicine: An Updated Review. Vol. 15, Pharmaceutics. MDPI; 2023.

20. Garg U, Chauhan S, Nagaich U, Jain N. Current advances in chitosan nanoparticles based drug delivery and targeting. Vol. 9, Advanced Pharmaceutical Bulletin. Tabriz University of Medical Sciences; 2019. p. 195–204.

21. Temmerman R, Garmyn A, Antonissen G, Vanantwerpen G, Vanrobaeys M, Haesebrouck F, et al. Evaluation of fluoroquinolone resistance in clinical avian pathogenic escherichia coli isolates from flanders (Belgium). Antibiotics. 2020 Nov 1;9(11):1–14.

22. Kong YT, Abdullah V, Mokhtar SU, Kutty RV. Preliminary studies on extraction of propolis using vitamin E d-α-Tocopheryl polyethylene glycol succinate (vitamin E TPGS) and compare their antimicrobial activities. Malays J Microbiol. 2020;16(5):346–52.

23. Hudzicki Jan. Kirby-Bauer Disk Diffusion Susceptibility Test Protocol [Internet]. 2009. Available from: www.atcc.org

24. Sigma-Aldrich. Cell Proliferation Kit I (MTT) [Internet]. 2020 [cited 2023 Oct 8]. Available from: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/ documents/191/466/11465007001.pdf

25. Cavalu S, Prokisch J, Laslo V, Vicas S. Preparation, structural characterisation and release study of novel hybrid microspheres entrapping nanoselenium, produced by green synthesis. IET Nanobiotechnol. 2017 Jun 1;11(4):426–32.

26. Jayakumar R, Reis RL, Mano JF. Synthesis and characterization of pH-sensitive thiol-containing chitosan beads for controlled drug delivery applications. Drug Deliv. 2007 Jan 1;14(1):9–17.

27. Ceballos S, Lozano C, Aspiroz C, Ruiz-Ripa L, Eguizábal P, Campaña-Burguet A, et al. Beyond CC398: Characterisation of Other Tetracycline and Methicillin-Resistant Staphylococcus aureus Genetic Lineages Circulating in Spanish Hospitals. Pathogens. 2022 Mar 1;11(3).

28. Macario Melitón Fierro Martínez. Ecología de Tetragonisca angustula (Apidae, Meliponini): comunicación de recursos y áreas de congregación de zánganos. 2011.

29. Kuen CY, Masarudin MJ. Chitosan Nanoparticle-Based System: A New Insight into the Promising Controlled Release System for Lung Cancer Treatment. Vol. 27, Molecules. MDPI; 2022.

30. Magnavacca A, Sangiovanni E, Racagni G, Dell’Agli M. The antiviral and immunomodulatory activities of propolis: An update and future perspectives for respiratory diseases. Vol. 42, Medicinal Research Reviews. John Wiley and Sons Inc; 2022. p. 897–945.

31. Cavalu S, Bisboaca S, Mates IM, Pasca PM, Laslo V, Costea T, et al. Novel Formulation Based on Chitosan-Arabic Gum Nanoparticles Entrapping Propolis Extract Production, physico-chemical and structural characterization [Internet]. Available from: http://www.revistadechimie.ro

32. Matos BN, Lima AL, Cardoso CO, Cunha-Filho M, Gratieri T, Gelfuso GM. Follicle-Targeted Delivery of Betamethasone and Minoxidil Co-Entrapped in Polymeric and Lipid Nanoparticles for Topical Alopecia Areata Treatment. Pharmaceuticals. 2023 Sep 1;16(9).

33. Vică ML, Glevitzky M, Heghedűş-Mîndru RC, Dumitrel GA, Heghedűş-Mîndru G, Popa M, et al. Phyto-Inhibitory and Antimicrobial Activity of Brown Propolis from Romania. Antibiotics. 2023 Jun 1;12(6).

34. Torres AR, Sandjo LP, Friedemann MT, Tomazzoli MM, Maraschin M, Mello CF, et al. Chemical characterization, antioxidant and antimicrobial activity of propolis obtained from melipona quadrifasciata quadrifasciata and tetragonisca angustula stingless bees. Brazilian Journal of Medical and Biological Research. 2018;51(6).

35. Przybyłek I, Karpiński TM. Antibacterial properties of propolis. Vol. 24, Molecules. MDPI AG; 2019.

36. Yilmaz Atay H. Antibacterial activity of chitosan-based systems. In: Functional Chitosan: Drug Delivery and Biomedical Applications. Springer Singapore; 2020. p. 457–89.

37. Kravanja G, Primožič M, Knez Ž, Leitgeb M. Chitosan-based (Nano)materials for Novel Biomedical Applications. Vol. 24, Molecules. MDPI AG; 2019.

38. Divya K, Vijayan S, George TK, Jisha MS. Antimicrobial properties of chitosan nanoparticles: Mode of action and factors affecting activity. Fibers and Polymers. 2017 Feb 1;18(2):221–30.

39. Aibani N, Rai R, Patel P, Cuddihy G, Wasan EK. Chitosan nanoparticles at the biological interface: implications for drug delivery. Vol. 13, Pharmaceutics. MDPI; 2021.

40. Ortiz M, Rodríguez H, Lucci E, Coro J, Pernía B, Montero-Calderon A, et al. Serological Cross-Reaction between Six Thiadiazine by Indirect ELISA Test and Their Antimicrobial Activity. Methods Protoc. 2023 Apr 1;6(2).

41. Sarangapani S, Patil A, Ngeow YK, Elsa Mohan R, Asundi A, Lang MJ. Chitosan nanoparticles' functionality as redox active drugs through cytotoxicity, radical scavenging and cellular behaviour. Integrative Biology (United Kingdom). 2018 May 1;10(5):313–24.

42. Sonin D, Pochkaeva E, Zhuravskii S, Postnov V, Korolev D, Vasina L, et al. Biological safety and biodistribution of chitosan nanoparticles. Nanomaterials. 2020 Apr 1;10(4).

43. Cardoso MJ, Costa RR, Mano JF. Marine origin polysaccharides in drug delivery systems. Vol. 14, Marine Drugs. MDPI AG; 2016.

44. Sukri N, Putri TTM, Mahani, Nurhadi B. Characteristics of propolis encapsulated with gelatin and sodium alginate by complex coacervation method. Int J Food Prop [Internet]. 2023 Sep 22 [cited 2025 Aug 24];26(1):696–707. Available from: https://www.tandfonline.com/doi/pdf/10.1080/10942912.2023.2179635

45. Amrane S, Chaalal M, Bouriche S, Ydjedd S, Rezgui F, Amessis-Ouchemoukh N, et al. Optimisation of microencapsulation efficiency of propolis phenolic compounds by double emulsion method using response surface methodology. Acta Aliment [Internet]. 2024 Mar 11 [cited 2025 Aug 24];53(1):34–47. Available from: https://doi.org/10.1556/066.2023.00244

46. Saroglu O, Karadag A. Propolis-loaded liposomes: characterization and evaluation of the in vitro bioaccessibility of phenolic compounds. ADMET DMPK [Internet]. 2024 [cited 2025 Aug 24];12(1):209–24. Available from: https://doi.org/10.5599/admet.2204

47. Ramli NA, Ali N, Hamzah S, Yatim NI. Physicochemical characteristics of liposome encapsulation of stingless bees' propolis. Heliyon [Internet]. 2021 Apr 1 [cited 2025 Aug 24];7(4). Available from: https://doi.org/10.1016/j.heliyon.2021.e06649

48. Jansen-Alves C, Victoria FN, Borges CD, Zambiazi RC. Encapsulation of propolis extract in ovalbumin protein particles: characterization and in vitro digestion. Nat Prod Res [Internet]. 2024 [cited 2025 Aug 24];38(10):1766–70. Available from: https://doi.org/10.1080/14786419.2023.2214945

49. Jansen-Alves C, Fernandes KF, Crizel-Cardozo MM, Krumreich FD, Borges CD, Zambiazi RC. Microencapsulation of Propolis in Protein Matrix Using Spray Drying for Application in Food Systems. Food Bioproc Tech [Internet]. 2018 Jul 1 [cited 2025 Aug 24];11(7):1422–36. Available from: https://doi.org/10.1007/s11947-018-2115-4

50. Zhang H, Fu Y, Xu Y, Niu F, Li Z, Ba C, et al. One-step assembly of zein/caseinate/alginate nanoparticles for encapsulation and improved bioaccessibility of propolis. Food Funct [Internet]. 2019 Feb 20 [cited 2025 Aug 24];10(2):635–45. Available from: https://pubs.rsc.org/en/content/articlehtml/2019/fo/c8fo01614c

51. Nori MP, Favaro-Trindade CS, Matias de Alencar S, Thomazini M, de Camargo Balieiro JC, Contreras Castillo CJ. Microencapsulation of propolis extract by complex coacervation. Lwt - Food Science and Technology [Internet]. 2011 [cited 2025 Aug 24];44(2):429–35. Available from: https://doi.org/10.1016/J.LWT.2010.09.010

52. Cea-Pavez I, Manteca-Bautista D, Morillo-Gomar A, Quirantes-Piné R, Quiles JL. Influence of the Encapsulating Agent on the Bioaccessibility of Phenolic Compounds from Microencapsulated Propolis Extract during In Vitro Gastrointestinal Digestion. Foods [Internet]. 2024 Feb 1 [cited 2025 Aug 24];13(3). Available from: https://doi.org/10.3390/foods13030425

53. Salehi A, Rezaei A, Damavandi MS, Kharazmi MS, Jafari SM. Almond gum-sodium caseinate complexes for loading propolis extract: Characterization, antibacterial activity, release, and in-vitro cytotoxicity. Food Chem [Internet]. 2023 Mar 30 [cited 2025 Aug 24];405:134801. Available from: https://www.sciencedirect.com/science/article/abs/pii/S0308814622027637?via%3Dihub

54. Shakoury N, Aliyari MA, Salami M, Emam-Djomeh Z, Vardhanabhuti B, Moosavi-Movahedi AA. Encapsulation of propolis extract in whey protein nanoparticles. LWT [Internet]. 2022 Mar 15 [cited 2025 Aug 24];158:113138. Available from: https://www.sciencedirect.com/science/article/pii/S0023643822000731?via%3Dihub

55. Spanidi E, Karapetsas A, Voulgaridou GP, Letsiou S, Aligiannis N, Tsochantaridis I, et al. A New Controlled Release System for Propolis Polyphenols and Its Biochemical Activity for Skin Applications. Plants [Internet]. 2021 Feb 1 [cited 2025 Aug 24];10(2):1–27. Available from: https://doi.org/10.3390/plants10020420

Downloads

Published

2025-09-15

How to Cite

Otuna-Hernández, D., Hernández Hernández, A. D., Espinoza-Castro, L., Spencer, L. M., & Rodríguez, H. (2025). Innovative encapsulation for enhanced drug delivery using stingless bee propolis and honey Tetragonisca angustula (Hymenoptera: Meliponini), and their antimicrobial activity. BioNatura Journal: Ibero-American Journal of Biotechnology and Life Sciences, 2(3), 19. https://doi.org/10.70099/BJ/2025.02.03.10

Issue

Vol. 2 No. 3 (2025)

Section

Research Articles

Categories

License

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Authors who publish with BioNatura Journal agree to the following terms: Authors retain copyright and grant the BioNatura Institutional Publishing Consortium (BIPC) right of first publication with the work simultaneously licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0). This allows others to share the work with an acknowledgment of the work's authorship and initial publication in this journal.This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.