Pinghui Zhou1,2, Bangguo Wei1,2, Jingjing Guan1, Yu Chen2,3,4, Yansong Zhu3, Yuchen Ye1, Yue Meng1, Jianzhong Guan5,6, Yingji Mao7,8,9. 1. Department of Orthopedics, First Affiliated Hospital, Bengbu Medical College, Bengbu, 233004, China. 2. Anhui Province Key Laboratory of Tissue Transplantation, Bengbu Medical College, Bengbu, 233030, China. 3. School of Life Sciences, Bengbu Medical College, Bengbu, 233030, China. 4. Department of Plastic Surgery, the First Affiliated Hospital of Bengbu Medical College, Bengbu, 233004, China. 5. Department of Orthopedics, First Affiliated Hospital, Bengbu Medical College, Bengbu, 233004, China. jzguan2002@163.com. 6. Anhui Province Key Laboratory of Tissue Transplantation, Bengbu Medical College, Bengbu, 233030, China. jzguan2002@163.com. 7. Department of Orthopedics, First Affiliated Hospital, Bengbu Medical College, Bengbu, 233004, China. myj123@bbmc.edu.cn. 8. Anhui Province Key Laboratory of Tissue Transplantation, Bengbu Medical College, Bengbu, 233030, China. myj123@bbmc.edu.cn. 9. School of Life Sciences, Bengbu Medical College, Bengbu, 233030, China. myj123@bbmc.edu.cn.
Abstract
BACKGROUND: Degeneration of the annulus fibrosus (AF), an important structure of the intervertebral disc, is one of the main causes of degenerative disc disease. Fabrication of scaffolds replicating the stratified microstructure of the AF is critical for the successful regeneration of AF. METHODS: In this study, we cultured rabbit AF-derived stem cells (AFSCs) using fabricated electrospun fibrous poly-L-lactic acid scaffolds with different diameters. We applied cyclic tensile strain (CTS) on the scaffolds to regulate the differentiation of AFSCs into specific cell types that resided at the inner, middle, and outer zones of the AF. RESULTS: We found that the morphologies of AFSCs on the smaller-fiber-diameter scaffolds were nearly round, whereas spindle-like cells morphologies were observed on large-diameter scaffolds. CTS enhanced these phenomena and made the cells slender. The expression levels of collagen-I in cells increased as a function of the fiber diameter, whereas collagen-II and aggrecan exhibited opposite trends. Moreover, the application of CTS upregulated the gene expressions of collagen-I, collagen-II, and aggrecan. CONCLUSION: Overlaying the scaffolds with different CTS-stimulated cells could eventually lead to engineered AF tissues with hierarchical structures that approximated the native AF tissue. Thus, the proposed methodologies could be potentially applied for AF regeneration.
BACKGROUND:Degeneration of the annulus fibrosus (AF), an important structure of the intervertebral disc, is one of the main causes of degenerative disc disease. Fabrication of scaffolds replicating the stratified microstructure of the AF is critical for the successful regeneration of AF. METHODS: In this study, we cultured rabbit AF-derived stem cells (AFSCs) using fabricated electrospun fibrous poly-L-lactic acid scaffolds with different diameters. We applied cyclic tensile strain (CTS) on the scaffolds to regulate the differentiation of AFSCs into specific cell types that resided at the inner, middle, and outer zones of the AF. RESULTS: We found that the morphologies of AFSCs on the smaller-fiber-diameter scaffolds were nearly round, whereas spindle-like cells morphologies were observed on large-diameter scaffolds. CTS enhanced these phenomena and made the cells slender. The expression levels of collagen-I in cells increased as a function of the fiber diameter, whereas collagen-II and aggrecan exhibited opposite trends. Moreover, the application of CTS upregulated the gene expressions of collagen-I, collagen-II, and aggrecan. CONCLUSION: Overlaying the scaffolds with different CTS-stimulated cells could eventually lead to engineered AF tissues with hierarchical structures that approximated the native AF tissue. Thus, the proposed methodologies could be potentially applied for AF regeneration.
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