# Protocol for fabricating immune-regulatory bone organoids via plant-based bioinks

Article · Wang T, Gao G, Zhang X, Huo K, Liu C, Geng Z, Bai L, Zhang H, Liu H, Xu K, Lai Y, Wang J, Jing Y, He C, Wang G, Su J, Ren X

- Archive id: doi:10.1016/j.isci.2026.117866
- Published: 2026-10-12
- Venue: iScience
- DOI: https://doi.org/10.1016/j.isci.2026.117866
- PMID: 42904200
- PMCID: PMC13682476
- Citations: 0
- Source URL: https://doi.org/10.1016/j.isci.2026.117866
- PDF: https://europepmc.org/articles/PMC13682476/pdf
- Provenance: europepmc

## Abstract

Bone organoids offer three-dimensional systems for studying skeletal development, disease, and regeneration. However, many existing bone organoid systems do not adequately incorporate immune-regulatory cues, despite their central role in bone repair. Their translational potential is further limited by the use of animal-derived matrices such as Matrigel, which are xenogeneic, poorly defined, and subject to batch-to-batch variation. To address these limitations, we developed a Matrigel-free and exogenous cytokine-free bioink composed of Aloe vera gel (AV), gelatin methacryloyl (GelMA), and Laponite. This formulation supports the fabrication of immune-regulatory bone organoids by providing both a printable extracellular matrix-like scaffold and intrinsic immune-regulatory signals. In vitro , the bioink promotes sustained macrophage reprogramming and helps establish an immune microenvironment that supports bone organoid maturation. Here, we describe a step-by-step protocol for preparing the plant-containing bioink, fabricating immune-regulatory bone organoids, and evaluating their performance in cranial bone defect models. This defined and reproducible platform provides a practical alternative to conventional Matrigel-based systems and may be adapted for the engineering of other immune-responsive tissues.

_Abstract reproduced under CC-BY (article license as recorded by Europe PMC; abstract reproduced unmodified apart from markup removal)._

Topics: [Organoids & Organ-on-Chip](https://space-kitty.com/topics/organoids)

---
Space Kitty Archive · https://space-kitty.com · API: https://space-kitty.com/api/v1 · MCP: https://space-kitty.com/mcp · Premium: $0.20/request via x402 (Base Sepolia (testnet)). Metadata: arXiv (CC0), OpenAlex (CC0), Europe PMC (CC BY/CC0 works), ClinicalTrials.gov.