ANGIOGENIC POTENTIAL OF THE HYDROXYAPATITE-BASED COMPOSITES LOADED WITH PLANT EXTRACT AND NANOPARTICLES

Viktoriia  Holubnycha; Oleksandr  Govorun; Veronika  Pron; Marharyta  Holubnycha; Ganna  Yanovska

doi:10.21272/eumj.2025;13(4);1052-1058

Viktoriia Holubnycha Public Health Department, Sumy State University, Sumy, Ukraine https://orcid.org/0000-0002-1241-2550
Oleksandr Govorun Department of Biology and Biology Teaching Methodology, Sumy State Pedagogical University named after A.S. Makarenko, Sumy, Ukraine https://orcid.org/0000-0002-6626-1241
Veronika Pron Academic and Research Medical Institute, Sumy State University, Sumy, Ukraine
Marharyta Holubnycha Lviv Polytechnic National University, Lviv, Ukraine https://orcid.org/0009-0008-7909-8755
Ganna Yanovska Institute of Agrophysics, Polish Academy of Sciences, Lublin, Poland https://orcid.org/0000-0001-8040-7457

DOI: https://doi.org/10.21272/eumj.2025;13(4);1052-1058

Keywords: bone regeneration, neovascularization, pomegranate, silver, health

Abstract

Introduction. The growing ageing population and increasing incidence of bone-related disorders have intensified the demand for advanced regenerative materials. Hydroxyapatite-based composites enriched with metal nanoparticles and plant extracts show great promise due to their biocompatibility, antimicrobial activity, and ability to promote tissue regeneration. This study aimed to evaluate how such functionalized composites influence vessel formation using the chick chorioallantoic membrane (CAM) model.

Methods. Hydroxyapatite-based composites were prepared by incorporating silver nanoparticles (10 µg/g) and pomegranate peel extract (30 mg/g) into hydroxyapatite, followed by stirring and drying. Supernatants of the obtained composites were applied onto the chick chorioallantoic membrane (CAM) of fertilized eggs on embryonic day 10 to assess angiogenic activity. After 24–48 h incubation, CAM images were taken and analyzed morphometrically using ImageJ to quantify vessel density and bifurcation points. Statistical significance was evaluated with GraphPad Prism (p < 0.05). The study was approved by the Local Ethical Committee (Decision No. 3/10, 16 October 2025).

Results. Treatment with AgNPs loaded hydroxyapatite caused vessel dilation and tortuosity, whereas composites containing pomegranate extract—alone or combined with AgNPs—showed minimal vascular alterations. By day 12, a significant rise (p < 0.05) in vessel number was detected only in the group treated with composite containing pomegranate, highlighting its angiogenic activity, while the proportion of bifurcated vessels remained unchanged across all groups.

Discussion. This study evaluated hydroxyapatite-based composites functionalized with silver nanoparticles (AgNPs) and pomegranate peel extract. Overall, the study demonstrates that the angiogenic response of hydroxyapatite-based composites depends on their functional components: silver nanoparticles suppress vessel formation, whereas pomegranate peel extract enhances it, highlighting the importance of composition in designing biomaterials for regenerative applications.

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References

Lee SS, Du X, Kim I, Ferguson SJ. Scaffolds for bone-tissue engineering. Matter. 2022;5(9):2722-2759. https://doi.org/10.1016/j.matt.2022.06.003

Noori A, Hoseinpour M, Kolivand S, Lotfbakhshaiesh N, Ebrahimi‐Barough S, Ai J, Azami M. Exploring the various effects of Cu doping in hydroxyapatite nanoparticle. Sci Rep. 2024;14:3421. https://doi.org/10.1038/s41598-024-53704-x

Ielo I, Calabrese G, De Luca G, Conoci S. Recent advances in hydroxyapatite-based biocomposites for bone tissue regeneration in orthopedics. Int J Mol Sci. 2022;23(17):9721. https://doi.org/10.3390/ijms23179721

Spilmont M, Léotoing L, Davicco MJ, et al. Pomegranate peel extract prevents bone loss in a preclinical model of osteoporosis and stimulates osteoblastic differentiation in vitro. Nutrients. 2015;7(11):9265-9284. https://doi.org/10.3390/nu7115465

Munir MU, Salman S, Ihsan A, Elsaman T. Synthesis, characterization, functionalization and bio-applications of hydroxyapatite nanomaterials: an overview. Int J Nanomedicine. 2022;17:1903-1925. https://doi.org/10.2147/IJN.S360670

Díaz-Puertas R, Álvarez-Martínez FJ, Falco A, Barrajón-Catalán E, Mallavia R. Phytochemical-based nanomaterials against antibiotic-resistant bacteria: an updated review. Polymers (Basel). 2023;15(6):1392. https://doi.org/10.3390/polym15061392

Siafaka PI, Miliotou AN, Okur ME, Karaotmarlı Güven G, Karantas ID, Üstündağ Okur N. Nanoformulations loaded with phytochemicals for combating wound infections and promoting wound healing: current applications and innovations. Appl Sci (Basel). 2025;15(10):5413. https://doi.org/10.3390/app15105413

Díaz L, Zambrano E, Flores ME, et al. Ethical considerations in animal research: the principle of 3Rs. Rev Invest Clin. 2021;73(4):20000380. https://doi.org/10.24875/ric.20000380

Holubnycha V, Pshenychnyi R, Bolshanina S, Derevianko T, Deineka V, Skwarek Y, Husak Y, Kowalska K, Yanovska A. ZnO NPs-Hydroxyapatite Composite Loaded with Phytoextract for Bone Defect Healing. 2025 IEEE 15th International Conference Nanomaterials: Applications & Properties (NAP) 7-12 Sept. Bratislava, Slovakia. 2025;NRA07:1-6. https://doi.org/10.1109/NAP68437.2025.11216223.

Owji N, Kohli N, Frost OG, Sawadkar P, Snow M, Knowles JC, García-Gareta E. Ex ovo chorioallantoic membrane assay as a model of bone formation by biomaterials. ACS Macro Lett. 2024;13(10):1362-1368. https://doi.org/10.1021/acsmacrolett.4c00343

Wang Y, Wang X, Zhou D, Xia X, Zhou H, Wang Y, Ke H. Synthesis and biological evaluation of silver–copper nanoparticles with antibacterial activity. ACS Omega. 2023;8(23):20323-20331. https://doi.org/10.1021/acsomega.2c08180

Bruna T, Maldonado-Bravo F, Jara P, Caro N. Silver nanoparticles and their antibacterial applications. Int J Mol Sci. 2021;22(13):7202. https://doi.org/10.3390/ijms22137202

Baharara J, Namvar F, Mousavi Nejad M, Ramezani Farzin T, Mohamad R. Anti-angiogenesis effect of biogenic silver nanoparticles synthesized using Salvia officinalis on chick chorioallantoic membrane (CAM). Molecules. 2014;19(9):13498-13508. https://doi.org/10.3390/molecules190913498

Tornese R, Montefusco A, Placì R, et al. Antiangiogenic potential of pomegranate extracts. Plants (Basel). 2024;13(23):3350. https://doi.org/10.3390/plants13233350