Tissue Eng Regen Med.
2018 Jun;15(3):311-319. 10.1007/s13770-018-0117-y.
Nondestructive Assessment of Glycosaminoglycans in Engineered Cartilages Using Hexabrix-Enhanced Micro-Computed Tomography
- Affiliations
-
- 1Department of Orthopedic Surgery, School of Medicine, Ajou University, 206 World Cup-ro, Yeongtong-gu, Suwon 16499, Korea. dr.bhmin@gmail.com
- 2Department of Biomedical Sciences, College of Medicine, Inha University, 100 Inha-ro, Nam-gu, Incheon 22212, Korea.
- 3Cell Therapy Center, Ajou University Medical Center, 206 World Cup-ro, Yeongtong-gu, Suwon 16499, Korea.
- 4Department of Molecular Science and Technology, Ajou University, 206 World Cup-ro, Yeongtong-gu, Suwon 16499, Korea.
- 5Present Address: Department of Orthopedic Surgery, Yanbian University Hospital, Yanji, China.
- KMID: 2411879
- DOI: http://doi.org/10.1007/s13770-018-0117-y
Abstract
- It is very useful to evaluate the content and 3D distribution of extracellular matrix non-destructively in tissue engineering. This study evaluated the feasibility of using micro-computed tomography (µCT) with Hexabrix to measure quantitatively sulfated glycosaminoglycans (GAGs) of engineered cartilage. Rabbit chondrocytes at passage 2 were used to produce artificial cartilages in polyglycolic acid scaffolds in vitro. Engineered cartilages were incubated with Hexabrix 320 for 20 min and analyzed via µCT scanning. The number of voxels in the 2D and 3D scanning images were counted to estimate the amount of sulfated GAGs. The optimal threshold value for quantification was determined by regression analysis. The 2D µCT images of an engineered cartilage showed positive correlation with the histological image of Safranin-O staining. Quantitative data obtained with the 3D µCT images of 14 engineered cartilages showed strong correlation with sulfated GAGs contents obtained by biochemical analysis (R² = 0.883, p < 0.001). Repeated exposure of engineered cartilages to Hexabrix 320 and µCT scanning did not significantly affect cell viability, total DNA content, or the total content of sulfated GAGs. We conclude that µCT imaging using Hexabrix 320 provides high spatial resolution and sensitivity to assess the content and 3D distribution of sulfated GAGs in engineered cartilages. It is expected to be a valuable tool to evaluate the quality of engineered cartilage for commercial development in the future.
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