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<p>In this study, we introduce a one-step semi-solid extrusion 3D printing strategy to fabricate gelatin-polyvinylpyrrolidone (GAG-PVP) scaffolds loaded with a low dose (0.5 wt%) of alendronate (ALN) and crosslinked in situ with 1 wt% genipin. The genipin-crosslinked ALN scaffold (GAG-PVP-GEN-ALN) demonstrated enhanced functional performance compared to both non-crosslinked GAG-PVP and GAG-PVP-ALN controls. Its peak swelling reached 462% at 5&#xa0;h, surpassing the 362% of the unmodified scaffold and preventing the rapid dissolution observed for GAG-PVP-ALN, before gradually deswelling for 96&#xa0;h. Water contact angle measurements confirmed that genipin fully restored surface hydrophobicity (101.9°), counteracting the pronounced wettability induced by ALN (47.8°) and exceeding the 78.2° of the GAG-PVP matrix, which is consistent with swelling ratio. Differential scanning calorimetry (DSC) indicated enhanced thermal stability of the crosslinked gelatin, with shifts in both glass transition and denaturation temperatures reflecting greater molecular rigidity despite the presence of glycerol as a plasticizer. Mechanical testing showed that while alendronate alone reduced mechanical performance, the combined inclusion of alendronate and genipin significantly enhanced scaffold properties compared to gelatin-polyvinyl pyrrolidone blend: tensile strength increased from 19.7&#xa0;MPa to 39.8&#xa0;MPa, elastic modulus rose from 805&#xa0;MPa to 1174&#xa0;MPa, and microhardness improved from 9.24&#xa0;MPa to 22.3&#xa0;MPa, values nearing those of native cancellous bone. The sustained ALN release profile extended from an abrupt 3&#xa0;h burst in GAG-PVP-ALN to a controlled 48&#xa0;h delivery in GAG-PVP-GEN-ALN, following first-order kinetics. Both direct and indirect cytotoxicity assays confirmed high cell viability (&gt; 85%) without morphological abnormalities. These results highlight that embedding low-dose ALN within a genipin-crosslinked gelatin-PVP network results in a mechanically robust, biocompatible scaffold with tunable swelling and prolonged drug release, offering a versatile platform for localized bone tissue engineering.</p>

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Synergistic Effects of Genipin and Alendronate in 3D-Bioprinted Gelatin-Polyvinylpyrrolidone Scaffolds

  • 期刊论文

  • 发表日期：

    2025年11月10日

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  • Marija Jovanović,
  • Ivana Stajčić,
  • Miloš Petrović,
  • Dušica Stojanović,
  • Nataša Radulović,
  • Svetlana Ibrić,
  • Petar Uskoković

摘要

In this study, we introduce a one-step semi-solid extrusion 3D printing strategy to fabricate gelatin-polyvinylpyrrolidone (GAG-PVP) scaffolds loaded with a low dose (0.5 wt%) of alendronate (ALN) and crosslinked in situ with 1 wt% genipin. The genipin-crosslinked ALN scaffold (GAG-PVP-GEN-ALN) demonstrated enhanced functional performance compared to both non-crosslinked GAG-PVP and GAG-PVP-ALN controls. Its peak swelling reached 462% at 5 h, surpassing the 362% of the unmodified scaffold and preventing the rapid dissolution observed for GAG-PVP-ALN, before gradually deswelling for 96 h. Water contact angle measurements confirmed that genipin fully restored surface hydrophobicity (101.9°), counteracting the pronounced wettability induced by ALN (47.8°) and exceeding the 78.2° of the GAG-PVP matrix, which is consistent with swelling ratio. Differential scanning calorimetry (DSC) indicated enhanced thermal stability of the crosslinked gelatin, with shifts in both glass transition and denaturation temperatures reflecting greater molecular rigidity despite the presence of glycerol as a plasticizer. Mechanical testing showed that while alendronate alone reduced mechanical performance, the combined inclusion of alendronate and genipin significantly enhanced scaffold properties compared to gelatin-polyvinyl pyrrolidone blend: tensile strength increased from 19.7 MPa to 39.8 MPa, elastic modulus rose from 805 MPa to 1174 MPa, and microhardness improved from 9.24 MPa to 22.3 MPa, values nearing those of native cancellous bone. The sustained ALN release profile extended from an abrupt 3 h burst in GAG-PVP-ALN to a controlled 48 h delivery in GAG-PVP-GEN-ALN, following first-order kinetics. Both direct and indirect cytotoxicity assays confirmed high cell viability (> 85%) without morphological abnormalities. These results highlight that embedding low-dose ALN within a genipin-crosslinked gelatin-PVP network results in a mechanically robust, biocompatible scaffold with tunable swelling and prolonged drug release, offering a versatile platform for localized bone tissue engineering.