title: Induction of mesenchymal stem cell chondrogenic differentiation and functional cartilage microtissue formation for in vivo cartilage regeneration by cartilage extracellular matrix-derived particles.
abstract: We propose a method of preparing a novel cell carrier derived from natural cartilage extracellular matrix (ECM), designated cartilage ECM-derived particles (CEDPs). Through a series of processes involving pulverization, sieving, and decellularization, fresh cartilage was made into CEDPs with a median diameter of 263 ± 48 μm. Under microgravity culture conditions in a rotary cell culture system (RCCS), bone marrow stromal cells (BMSCs) can proliferate rapidly on the surface of CEDPs with high viability. Histological evaluation and gene expression analysis indicated that BMSCs were differentiated into mature chondrocytes after 21 days of culture without the use of exogenous growth factors. Functional cartilage microtissue aggregates of BMSC-laden CEDPs formed as time in culture increased. Further, the microtissue aggregates were directly implanted into trochlear cartilage defects in a rat model (CEDP+MSC group). Gait analysis and histological results indicated that the CEDP+MSC group obtained better and more rapid joint function recovery and superior cartilage repair compared to the control groups, in which defects were treated with CEDPs alone or only fibrin glue, at both 6 and 12 weeks after surgery. In conclusion, the innovative cell carrier derived from cartilage ECM could promote chondrogenic differentiation of BMSCs, and the direct use of functional cartilage microtissue facilitated cartilage regeneration. This strategy for cell culture, stem cell differentiation and one-step surgery using cartilage microtissue for cartilage repair provides novel prospects for cartilage tissue engineering and may have further broad clinical applications.
  
  
  CONCLUSIONS
  We proposed a method to prepare a novel cell carrier derived from natural cartilage ECM, termed cartilage ECM-derived particles (CEDPs), which can support proliferation of MSCs and facilitate their chondrogenic differentiation. Further, the direct use of functional cartilage microtissue of MSC-laden CEDP aggregates for cartilage repair in vivo induced hyaline-like articular cartilage repair. This strategy for cell culture, stem cell differentiation and the one-step surgery for cartilage repair provide novel prospects for cartilage tissue engineering and may have further broad clinical applications.
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China; School of Medicine, Nankai University, 94th Weijin Road, Nankai District, Tianjin 300071, PR China.
authors.forename: Heyong
authors.initials: H
authors.lastname: Yin
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Sun
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China; School of Medicine, Nankai University, 94th Weijin Road, Nankai District, Tianjin 300071, PR China.
authors.forename: Xun
authors.initials: X
authors.lastname: Sun
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China.
authors.forename: Yichi
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China; Hebei Medical University, 361th Zhongshan East Road, Shijiazhuang 050017, PR China.
authors.forename: Pan
authors.initials: P
authors.lastname: Li
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China.
authors.forename: Haoye
authors.initials: H
authors.lastname: Meng
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Yu
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China.
authors.forename: Bo
authors.initials: B
authors.lastname: Xiao
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China.
authors.forename: Tian
authors.initials: T
authors.lastname: Fan
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China.
authors.forename: Yiguo
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Xu
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China.
authors.forename: Aiyuan
authors.initials: A
authors.lastname: Wang
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China.
authors.forename: Quanyi
authors.initials: Q
authors.lastname: Guo
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China. Electronic address: pengjiang301@126.com.
authors.forename: Jiang
authors.initials: J
authors.lastname: Peng
authors.affiliation: Institute of Orthopedics, Chinese PLA General Hospital, 28th Fuxing Road, Beijing 100850, PR China.
authors.forename: Shibi
authors.initials: S
authors.lastname: Lu
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena; D002358:Cartilage, Articular / Q000000981:diagnostic imaging / Q000187:drug effects / Q000473:pathology / Q000502:physiology*; D002448:Cell Adhesion; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D058948:Cell Tracking; D020219:Chondrogenesis; D004247:DNA / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D005684:Gait; D006041:Goats; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010316:Particle Size; D017207:Rats, Sprague-Dawley; D012038:Regeneration; D023822:Tissue Engineering / Q000639:trends*; D055114:X-Ray Microtomography
chemical_list: D049452:Green Fluorescent Proteins; D004247:DNA
keywords: Cartilage extracellular matrix; Cartilage regeneration; Mesenchymal stem cell; Microtissue; Tissue engineering