A comparative in vitro study of an innovative hybrid scaffold for bone regeneration: Alginate-based 3D-printed composites incorporating 58S Bioactive glass, Hydroxyapatite, and Hardystonite, modified with Gelatin nanofibers

Dec 19, 2025International journal of biological macromolecules

Lab comparison of a new 3D-printed alginate scaffold with bioactive glass, bone minerals, and gelatin nanofibers for bone healing

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Abstract

The GE-Alg/HAD scaffolds achieved a compressive strength of 13.4 MPa ± 4.2.

Hybrid scaffolds were created using a blend of ceramic phases and alginate.
Chemical interactions between polymer and ceramic components were confirmed through infrared spectroscopy.
GE-Alg/BG scaffolds demonstrated a biodegradation rate of 80%, while GE-Alg/HA had a swelling ratio of 220%.
All bioceramic-containing scaffolds showed improved bioactivity, cell adhesion, and proliferation compared to alginate alone.
The osteogenic potential of the scaffolds was validated using specific staining and gene expression analyses.

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The fabrication of scaffolds with appropriate properties is a critical aspect of bone tissue engineering (BTE). Hybrid scaffolds have recently emerged as a promising strategy for achieving optimal structural, mechanical, and biological performance. In this study, we prepared and characterized hybrid scaffolds using 3D printing and electrospinning techniques to compare three widely used ceramic phases, 58S Bioactive glass (BG), Hydroxyapatite (HA), and Hardystonite (HAD), allowing informed selection of bioceramics based on targeted scaffold properties. Scaffold fabrication involved blending each ceramic phase with Alginate (Alg) at a 50:50 weight ratio to create printable bioinks, which were subsequently used to produce 3D scaffolds via direct ink writing (DIW). Gelatin (GE) nanofibers were then electrospun onto the printed structures. All scaffolds were characterized for their chemical, mechanical, rheological, morphological, and biological properties. Fourier transform infrared spectroscopy (FTIR) analysis of the hybrid scaffolds confirmed chemical interactions between the polymer and ceramic components. The GE-Alg/HAD scaffolds exhibited the highest layer height (426.5 μm ± 30.6) and compressive strength (13.4 MPa ± 4.2), while GE-Alg/BG and GE-Alg/HA showed the highest biodegradation (80 %) and swelling ratio (220 %), respectively. All bioceramic-containing scaffolds demonstrated enhanced bioactivity, cell adhesion, and proliferation compared to Alg alone. The osteogenic potential of the scaffolds was confirmed in vitro using Alizarin Red staining and real-time PCR. Overall, our results show that scaffold properties were strongly influenced by the composition, particle size, and surface charge of the incorporated bio-ceramics. These findings indicate that the prepared hybrid scaffolds exhibit strong potential for applications in BTE.

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A comparative in vitro study of an innovative hybrid scaffold for bone regeneration: Alginate-based 3D-printed composites incorporating 58S Bioactive glass, Hydroxyapatite, and Hardystonite, modified with Gelatin nanofibers

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