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Fibroblast Matrix Enhanced Three-Dimensional-Bioprinted Hydrogel for Osteochondral Regeneration
Devy F Garna1,2, Aryan S Shet1, Levi Randall Morgan1
1Centre for Oral, Clinical and Translational Sciences, Faculty of Dentistry, Oral and Craniofacial Sciences, King's College London, London, United Kingdom.
Tissue Engineering. Part A
|December 30, 2025
Summary
This study optimized a decellularization protocol using 10% nonidet P-40 (NP-40) for 3 hours to create cell-derived decellularized extracellular matrix (dECM) for tissue engineering. The dECM enhanced hydrogel properties, improving cell viability and differentiation for bone and cartilage applications.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Cell Biology
Background:
- Decellularized extracellular matrix (dECM) is crucial for tissue engineering, providing structural and biochemical cues while minimizing immune responses.
- Donor shortages necessitate alternative sources of dECM, particularly for bone and cartilage regeneration.
- Effective removal of deoxyribonucleic acid (DNA) is vital to prevent immunogenicity in cell-derived dECM.
Purpose of the Study:
- To develop a standardized and reproducible protocol for producing cell-derived dECM from human dermal fibroblasts.
- To optimize the decellularization process for complete DNA removal while preserving ECM components.
- To evaluate the impact of dECM incorporation into 3D-bioprinted hydrogels on cell viability, differentiation, and mechanical properties.
Main Methods:
- Human dermal fibroblasts were decellularized using varying concentrations (1%, 10%) and durations (1h, 3h) of nonidet P-40 (NP-40) lysis buffer.
- Decellularization efficacy was quantified using dsDNA Qubit assay, gel electrophoresis, and protein assays.
- Hydrogels (alginate-gelatin) incorporating dECM were fabricated via extrusion bioprinting, and their structural, mechanical, and biological properties were assessed.
Main Results:
- A 3-hour incubation with 10% NP-40 effectively removed DNA (<50 ng dsDNA) and RNA, yielding the highest protein content.
- Incorporation of dECM into alginate-gelatin hydrogels significantly enhanced cell viability and glycosaminoglycan (GAG) synthesis.
- dECM-containing hydrogels exhibited a 33% increase in Young's modulus and confirmed dECM integration via Raman spectroscopy.
Conclusions:
- An optimized decellularization protocol (10% NP-40, 3h) provides a robust source of cell-derived dECM below immunogenic thresholds.
- Fibroblast-derived dECM significantly improves the performance of alginate-gelatin hydrogels for bone and cartilage tissue engineering.
- This standardized dECM production method offers a promising alternative to tissue-derived matrices for bioink development.
