Abstract
This study developed a 3D-printed, triphasic (subchondral bone, calcified, and articular cartilage) scaffold using biological macromolecule-based bioinks to support the chondrogenic differentiation of bone marrow-derived mesenchymal stem cells (BM-MSCs). A subchondral bone layer was formed by blending various concentrations of graphene oxide (GO) (1% and 2% w/w) into an alginate (Alg) and gelatin (Gel) bioink, two natural biopolymers known for their biocompatibility and biodegradability. Following mechanical and biocompatibility assessments, the 1% GO concentration was selected and applied consistently through the subchondral and calcified cartilage layers. In contrast, the gradient of lyophilized platelet-rich plasma (PRP) powder was adjusted to 1%, 2%, and 3% (w/v) to more accurately replicate the characteristics of calcified and articular cartilage. Triphasic scaffolds with different PRP gradients were evaluated for water absorption, biodegradability, rheological behavior, stem cell viability, and chondroinductive activity. The results indicated that 3D-printed triphasic scaffolds containing 1% or 2% PRP exhibited favorable biomechanical properties, with no significant differences between the two concentrations. However, scaffolds with 2% PRP facilitated the attachment, proliferation, and survival of BM-MSCs, as indicated by an increase in the expression of cartilage-related genes and enhanced production of glycosaminoglycan (GAG), as confirmed through real-time PCR and Alcian Blue staining, respectively.
Data availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
The authors would like to express their gratitude to the Cellular and Molecular Research Center, Iran University of Medical Sciences, for supporting this research under Grant Number 24717.
Funding
This research did not receive any external financial funding. The authors acknowledge that a grant number (24717) was assigned by the Cellular and Molecular Research Center, Iran University of Medical Sciences, but no funds were disbursed. The study was conducted using shared resources and voluntary contributions.
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Faezeh Ghobadi: Conceptualization, Methodology, Formal Analysis, Investigation, Visualization, and Writing Original Draft. Maryam Mohammadi: Resources. Rooja Kalantarzadeh: Formal Analysis. Arezoo Ashrafnia Menarbazari: Formal Analysis. Jila Majidi: Reviewing and Editing. Ehsan Lotfi: Formal Analysis. Shokoufeh Borhan: Resources. Yousef Fatahi: Visualization (Graphical Abstract). Narendra Pal Singh Chauhan: Reviewing. Ghazaleh Salehi: Supervision, Methodology, Writing Original Draft, and Resources. Sara Simorgh: Supervision, Project Administration, Reviewing and Editing, and Resources.
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Ghobadi, F., Mohammadi, M., Kalantarzadeh, R. et al. Optimized gradient of lyophilized platelet-rich plasma in biomimetic 3D-printed triphasic scaffold based on alginate and gelatin for osteochondral tissue engineering. Sci Rep (2026). https://doi.org/10.1038/s41598-026-37615-7
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DOI: https://doi.org/10.1038/s41598-026-37615-7
