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RESEARCH ARTICLE

Sequential multi-material 3D bioprinting of dual-stiffness peptide scaffolds for compartmentalized, multicellular bone marrow niche modeling

Dana M. Alhattab1,2† ,  Salwa Alshehri3† ,  Manar M. Farooqui1 ,  Alexander U. Valle-Pérez1 ,  Manola Moretti1,4 ,  Panayiotis Bilalis1 ,  Antonio Cárdenas-Calvario1 ,  Hanan Jafar5 ,  Yiting Lei6,7 ,  Christian Baumgartner8 ,  Robert Hoehndorf2 ,  Charlotte A. E. Hauser1,8
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1 Laboratory for Nanomedicine, Bioengineering Program, Division of Biological & Environmental Science & Engineering (BESE), King Abdullah University of Science and Technology (KAUST), Thuwal , Saudi Arabia
2 Computer, Electrical and Mathematical Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), Thuwal , Saudi Arabia
3 Biological Sciences Department, College of Science, University of Jeddah, Jeddah , Saudi Arabia
4 KAUST Catalysis Platform, Physical Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), Thuwal , Saudi Arabia
5 Cell Therapy Center, The University of Jordan, Amman , Jordan
6 Department of Orthopedic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing , China
7 Department of Biomedical Engineering, The Chinese University of Hong Kong, Hong Kong SAR , China
8 Institute of Healthcare Engineering, Graz University of Technology , Austria
†These authors contributed equally to this work.
IJB, 026350380 https://doi.org/10.36922/IJB026350380
Received: 31 December 2025 | Revised: 28 August 2026 | Accepted: 17 September 2026 | Published online: 28 September 2026
© 2026 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

Three-dimensional (3D) bioprinting holds great potential for tissue engineering, yet reproducing the bone marrow (BM) microenvironment remains challenging: it requires biomaterials that support the diverse BM cell types while reproducing the mechanical heterogeneity of its two major niche compartments. Here we developed an in vitro BM model representing both the endosteal and central marrow niches, using ultrashort self-assembling peptide hydrogels as bioinks and scaffolding material. Rheological characterization showed that Ac-IIZK-NH₂ (IIZK) hydrogels were stiffer than Ac-IIFK-NH₂ (IIFK); IIZK was therefore used for the endosteal compartment and IIFK for the central marrow compartment. Both peptides showed excellent printability and shape fidelity using a robotic arm-based bioprinter with a customized coaxial nozzle. The compartments were fabricated sequentially: the stiffer IIZK compartment was printed first as a self-supporting structure, and the separately cellularized IIFK compartment was then printed into the cavity it defines. This order lets each compartment's composition be set by the tissue it represents rather than by the demands of extrusion, while in-line coaxial gelation preserves the interface. Cell viability exceeded 90% for mesenchymal stromal cells (MSCs), BM endothelial cells (BMECs) and the leukemia lines KG1a and HL-60. BMECs spontaneously formed capillary-like networks, and MSCs upregulated RUNX2, BGLAP and SP7 even without induction medium, depositing a calcium-rich mineralized matrix upon osteogenic differentiation. Tri-culture experiments demonstrated multicellular interaction and spatial organization. Looking ahead, the CD34-positive KG1a line used here as a hematopoietic surrogate can be replaced by primary HSCs without altering the scaffold design or the printing workflow, opening the way to studies of niche-driven hematopoiesis, leukemic drug resistance and patient-specific BM modeling.

Keywords
Ultrashort self-assembling peptides
3D bioprinting
Peptide bioink
Osteogenic differentiation
Bone marrow niche-like model
Funding
This work was financially supported by King Abdullah University of Science and Technology under the Smart-Health Center of Excellence (KCSH) project number 4938. Graz University of Technology (TU Graz) provided additional funding for this research project.
Conflict of interest
Both Charlotte A. E. Hauser and Yiting Lei serve as Editorial Board Members of this journal, but were not in any way involved in the editorial and peer-review process conducted for this paper, directly or indirectly. The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper.
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International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing
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