KMID : 1377020220190051099
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Tissue Engineering and Regenerative Medicine 2022 Volume.19 No. 5 p.1099 ~ p.1111
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In Situ-Forming Collagen/poly-¥ã-glutamic Acid Hydrogel System with Mesenchymal Stem Cells and Bone Morphogenetic Protein-2 for Bone Tissue Regeneration in a Mouse Calvarial Bone Defect Model
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Cho Sun-Hee
Shin Keun-Koo Kim Sun-Young Cho Mi-Young Oh Doo-Byoung Lim Yong-Taik
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Abstract
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Background: Bone marrow-derived mesenchymal stem cells (BMSCs) and bone morphogenetic protein-2 (BMP-2) have been studied for bone repair because they have regenerative potential to differentiate into osteoblasts. The development of injectable and in situ three-dimensional (3D) scaffolds to proliferate and differentiate BMSCs and deliver BMP-2 is a crucial technology in BMSC-based tissue engineering.
Methods: The proliferation of mouse BMSCs (mBMSCs) in collagen/poly-¥ã-glutamic acid (Col/¥ã-PGA) hydrogel was evaluated using LIVE/DEAD and acridine orange and propidium iodide assays. In vitro osteogenic differentiation and the gene expression level of Col/¥ã-PGA(mBMSC/BMP-2) were assessed by alizarin red S staining and quantitative reverse-transcription polymerase chain reaction. The bone regeneration effect of Col/¥ã-PGA(mBMSC/BMP-2) was evaluated in a mouse calvarial bone defect model. The cranial bones of the mice were monitored by micro-computed tomography and histological analysis.
Results: The developed Col/¥ã-PGA hydrogel showed low viscosity below ambient temperature, while it provided a high elastic modulus and viscous modulus at body temperature. After gelation, the Col/¥ã-PGA hydrogel showed a 3D and interconnected porous structure, which helped the effective proliferation of BMSCs with BMP-2. The Col/¥ã-PGA (mBMSC/BMP-2) expressed more osteogenic genes and showed effective orthotopic bone formation in a mouse model with a critical-sized bone defect in only 3?4 weeks.
Conclusion: The Col/¥ã-PGA(mBMSC/BMP-2) hydrogel was suggested to be a promising platform by combining collagen as a major component of the extracellular matrix and ¥ã-PGA as a viscosity reducer for easy handling at room temperature in BMSC-based bone tissue engineering scaffolds.
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KEYWORD
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Hydrogel scaffold, Bone morphogenetic protein-2, Mesenchymal stem cell, Calvarial defect, Bone regeneration
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