Characterization and in ovo pre-angiogenic evaluation of royal jelly enriched jellyfish collagen sponges

Abstract Marine-derived collagen has gained significant attention in biomedical applications due to its biocompatibility and low immunogenicity. In this study, bioactive porous sponges were fabricated by incorporating royal jelly (RJ) into jellyfish ( Rhizostoma pulmo ) collagen matrices. The sponges were prepared via lyophilization and chemically crosslinked using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS). Three different RJ concentrations (0.5, 1.0, and 1.5 mg ml ⁻¹ ) were evaluated and compared against an RJ-free crosslinked control. The physicochemical and morphological properties of the sponges were characterized using Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy, swelling analysis, and mechanical testing. Mechanical evaluations showed that while numerical variations in the initial compressive moduli were observed, the final load-bearing capacity remained consistent across all groups. Additionally, a reduction in swelling capacity was recorded at the highest RJ concentration (1.5 mg ml ⁻¹ ). The pre-angiogenic potential of the sponges was assessed using the in ovo chorioallantoic membrane assay. Macroscopic and quantitative analyses revealed that the 1.0 mg ml ⁻¹ formulation supported vessel formation and tissue integration more effectively than the control and other experimental groups. However, increasing the concentration to 1.5 mg ml ⁻¹ did not result in further enhancement, indicating a saturation effect. These findings suggest that RJ-enriched jellyfish collagen sponges hold potential as functional biomaterials for biomedical applications.