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Fabrication of Decellularized Engineered Extracellular Matrix through Bioreactor-Based Environment for Bone Tissue
Hanieh Nokhbatolfoghahaei1,2, Zahrasadat Paknejad3,4, Mahboubeh Bohlouli2
1Dental Research Center, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran 1983969413, Iran.
ACS Omega
|December 21, 2020
Summary
Developing extracellular matrix (ECM)-contained grafts for bone tissue engineering is crucial. This study optimized decellularization protocols for engineered scaffolds, finding Triton X-100 and bioreactor use enhance ECM retention and osteoinductivity.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Regenerative Medicine
Background:
- Native bone grafts have limitations, driving interest in in vitro extracellular matrix (ECM) bioscaffolds for bone tissue engineering.
- Engineered scaffolds offer a promising alternative for creating ECM-contained bone grafts.
Purpose of the Study:
- To develop an ECM-contained construct by decellularizing an engineered gelatin-coated β-tricalcium phosphate (gTCP) scaffold.
- To optimize decellularization protocols for cell-loaded gTCP scaffolds using stem cells.
- To evaluate the efficacy of bioreactor-based culture in enhancing ECM deposition and osteoinductivity.
Main Methods:
- Seeding gTCP scaffolds with buccal fat pad-derived stem cells.
- Comparing four decellularization protocols (sodium dodecyl sulfate, trypsin, Triton X-100, combined) for cell removal and ECM preservation.
- Evaluating decellularization efficacy under static and dynamic (rotating/perfusion bioreactor) conditions over 24 days.
- Assessing cytotoxicity and osteoinductive capability of reseeded scaffolds.
Main Results:
- All protocols effectively removed cells, but Triton X-100 yielded significantly more remnant ECM.
- Bioreactor-incubated scaffolds showed higher levels of ECM proteins, including collagen and glycosaminoglycans.
- Reseeded scaffolds cultured in bioreactors demonstrated enhanced osteoinductivity compared to static cultures.
Conclusions:
- Triton X-100 is a suitable decellularization agent for preserving ECM components.
- Bioreactor cultivation significantly enhances ECM production and osteoinductivity in engineered bone scaffolds.
- Combining optimized decellularization with bioreactor technology is a promising strategy for developing ECM-contained grafts for bone tissue engineering.
