Bioprinting of miRNA-Induced Spheroids for Vascularized, Heterocellular Bone Regeneration

  • Chem Eng J. 2026 Sep 15:544:178521. doi: 10.1016/j.cej.2026.178521.
Nazmiye Celik  1  2  3 ,  Amar Yeware  4 ,  Vaibhav Pal  2  5 ,  Miji Yeo  1  2 ,  Myoung Hwan Kim  2  4 ,  Logan Haugh  4 ,  Ibrahim T Ozbolat  1  2  4  6  7 ,  Daniel J Hayes  2  4  6
Affiliations
  • 1. Department of Engineering Science and Mechanics, Pennsylvania State University, 212 Earth-Engineering Sciences Bldg., University Park, PA 16802, USA.
  • 2. The Huck Institutes of the Life Sciences, Penn State University, University Park, PA 16802, USA.
  • 3. The Translational Tissue Engineering Center, Wilmer Eye Institute and Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD, 21231 USA.
  • 4. Department of Biomedical Engineering, Pennsylvania State University, Chemical and Biomedical Engineering Bldg., University Park, PA 16802, USA.
  • 5. Department of Chemistry, Penn State University, University Park, PA 16802, USA.
  • 6. Materials Research Institute, Penn State University, University Park, PA 16802, USA.
  • 7. Department of Neurosurgery, Penn State College of Medicine, Hershey, PA 17033, USA.
PMID: 42559531 DOI: 10.1016/j.cej.2026.178521
Abstract

Successful bone regeneration requires coupled osteogenic and vascular development; however, achieving simultaneous multicellular differentiation within engineered tissues remains challenging. Here, we developed a MicroRNA (miR)-guided spheroid platform to induce dual osteogenic and endothelial differentiation of human adipose-derived stem cells (hASCs) for vascularized bone regeneration. hASCs were transfected with miR-148b or miR-210 to promote osteogenic and vascular-associated phenotypes, respectively, and assembled into spheroids that were bioprinted within an nHA-containing GelMA microgel environment using aspiration-assisted bioprinting (AAB). The integrated platform combined miR-guided osteogenic and endothelial differentiation, spatially organized AAB-based spheroid assembly, and an nHA-containing GelMA microgel environment to support vascularized bone tissue regeneration. The engineered constructs maintained high cell viability (> 90%) and supported active cell spreading and migration within the microgel matrix, together with increased osteogenic and endothelial gene expression. To further verify their in vivo regenerative potential, the constructs were implanted into mouse critical-size calvarial defects, where those containing miR-transfected hASCs improved bone regeneration, achieving ~91% of defect closure, and promoted the formation of vessel-like CD31-positive structures compared to controls. Together, these findings demonstrate that combining miR-mediated dual-lineage differentiation with spatially organized spheroid assembly and a supportive microgel environment provides a promising strategy for vascularized bone tissue engineering.

Keywords
bioprinting; miRNA; osteogenesis; spheroids; vascularization.
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