AccScience Publishing / IJB / Online First / DOI: 10.36922/IJB026350379
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RESEARCH ARTICLE

Boronate ester-crosslinked GelMA composite bioink for 3D bioprinting of cell-laden corneal stroma and in vivo evaluation in rabbits

Yi-Jen Hsueh1,2† ,  Chung-An Chen3,4,5† ,  Chin-Yu Yang2 ,  Chang-Hong Lin1 ,  Chun-Yuan Cheng5 ,  Chih-Hsin Lin6,7,8 ,  Shih-Jung Liu5,9 ,  Tsung-Ting Tsai3,4,5* ,  Hung-Chi Chen1,2,3*
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1 Department of Ophthalmology, Chang Gung Memorial Hospital, Linkou Branch, Taoyuan , Taiwan
2 Center for Tissue Engineering, Chang Gung Memorial Hospital, Linkou Branch, Taoyuan , Taiwan
3 Department of Medicine, College of Medicine, Chang Gung University, Taoyuan , Taiwan
4 Department of Orthopaedic Surgery, Chang Gung Memorial Hospital, Linkou Branch, Taoyuan , Taiwan
5 Spine Section and Bone and Joint Research Center, Chang Gung Memorial Hospital, Taoyuan , Taiwan
6 Institute of Molecular Medicine and Bioengineering, National Yang Ming Chiao Tung University, Hsinchu , Taiwan
7 Department of Biological Science and Technology, College of Engineering Bioscience, National Yang Ming Chiao Tung University, Hsinchu , Taiwan
8 Department of Biological Science and Technology, College of Engineering Bioscience, National Yang Ming Chiao Tung University, Hsinchu , Taiwan
9 Department of Mechanical Engineering, College of Engineering, Chang Gung University, Taoyuan , Taiwan
†These authors contributed equally to this work.
IJB, 026350379 https://doi.org/10.36922/IJB026350379
Received: 3 February 2026 | Revised: 12 September 2026 | Accepted: 15 September 2026 | Published online: 15 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

Corneal transplantation restores vision by replacing damaged corneal tissue with a donor graft, but the limited supply and variable quality of donor corneas remain major challenges. We therefore developed an engineered corneal substitute using gelatin methacryloyl (GelMA), a modified gelatin. Phenylboronic acid-functionalized gelatin (PBA@G) and poly(vinyl alcohol) (PVA) were incorporated to introduce dynamic boronate ester crosslinking and modify the rheological behavior of the hydrogel. The formulation optimized for bioprinting contained 10% GelMA, 10% PBA@G, and 1% PVA (10G10P1PVA). Scanning electron microscopy revealed an interconnected porous microstructure. The rheological behavior, optical transmittance, swelling, degradation, and printability of 10G10P1PVA were characterized. Human corneal stromal cells (CSCs) remained viable through day 7, and the CCK-8 assay showed significantly higher metabolic activity in 10G10P1PVA than in 10G. Phalloidin staining showed cell spreading and longitudinal F-actin organization. RT-qPCR showed no marked ACTA2 induction, with expression remaining below that of the myofibroblast-like reference group. The corneal stromal cell marker ALDH3A1 was expressed at higher levels in cells cultured in 10G10P1PVA than in either the 10G or 10G10P group. These findings support the short-term cytocompatibility of the newly developed bioink and selected stromal-associated features. An exploratory rabbit ALK evaluation was performed to document short-term ocular response, epithelial recovery, and stromal organization following implantation over 7 days. Together, the material and biological results support further evaluation of 10G10P1PVA as a GelMA-based composite bioink for corneal stromal bioprinting.

Keywords
Artificial corneal stroma
Bioink
Bioprinting
Biomaterials
Photocrosslinking
3D cell culture
Funding
This study was partially supported by Chang Gung Memorial Hospital (CMRPG3Q0481) for Chen H.C. and the Taiwan National Science and Technology Council Grants (NSTC 114-2314-B-182-002 and 114-2811-B-182-014) for Tsai TT. The funding organization played no role in the design or conduct of this research.
Conflict of interest
Chung-An Chen is an employee of AngenMed Therapeutics, Inc., but was not involved in any activities that could constitute a conflict of interest in relation to this study. The other authors declare that there is no conflict of interest.
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International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing
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