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<p>Collagen is the most abundant structural protein in the extracellular matrix, and its biocompatibility can be enhanced by chemical modification into a negatively charged matrix. This study investigates the effect of alkaline hydrolysis time (24, 48, 72, and 96`&#xa0;h) on the rheological behavior and processability of anionic concentrated collagen solutions extracted from bovine tendon. By systematically modulating hydrolysis time, we demonstrate how increasing negative charge density drives a rearrangement of the collagen network, resulting in more elastic and shear-thinning behavior, as evidenced by higher storage moduli, increased zero-shear viscosity, and enhanced creep-recovery response. Among the tested formulations, the 72-h hydrolyzed collagen exhibited the most balanced rheological profile and enabled extrusion-based 3D printing with high fidelity, achieving geometry preservation of 104 ± 4% (star angle), 98 ± 2% (scaffold area), and effective macroporosity of 84%. Longer hydrolysis led to decreased scaffold pore size and increased absorption of phosphate-buffered saline. All materials were non-cytotoxic and supported cell adhesion (58.74—74.89%). Overall, this work highlights how rheological characterization can guide the design and processability of collagen-based bioinks, linking structural and mechanical properties to 3D printing performance and biological outcomes.</p> Graphical Abstract <p></p>

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Anionic collagen processed by alkaline hydrolysis: rheological, biological, and 3D printability assessment

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年4月13日

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  • Lívia C. Massimino,
  • Mirella R. V. Bertolo-Cagnoto,
  • Larissa Tessaro,
  • Virginia da C. A. Martins,
  • Bianca Chieregato Maniglia,
  • Ana Maria de G. Plepis,
  • Monica B. Mathor

摘要

Collagen is the most abundant structural protein in the extracellular matrix, and its biocompatibility can be enhanced by chemical modification into a negatively charged matrix. This study investigates the effect of alkaline hydrolysis time (24, 48, 72, and 96` h) on the rheological behavior and processability of anionic concentrated collagen solutions extracted from bovine tendon. By systematically modulating hydrolysis time, we demonstrate how increasing negative charge density drives a rearrangement of the collagen network, resulting in more elastic and shear-thinning behavior, as evidenced by higher storage moduli, increased zero-shear viscosity, and enhanced creep-recovery response. Among the tested formulations, the 72-h hydrolyzed collagen exhibited the most balanced rheological profile and enabled extrusion-based 3D printing with high fidelity, achieving geometry preservation of 104 ± 4% (star angle), 98 ± 2% (scaffold area), and effective macroporosity of 84%. Longer hydrolysis led to decreased scaffold pore size and increased absorption of phosphate-buffered saline. All materials were non-cytotoxic and supported cell adhesion (58.74—74.89%). Overall, this work highlights how rheological characterization can guide the design and processability of collagen-based bioinks, linking structural and mechanical properties to 3D printing performance and biological outcomes.

Graphical Abstract