PMID- 23981625
OWN - NLM
STAT- MEDLINE
DCOM- 20140624
LR  - 20181202
IS  - 2542-5641 (Electronic)
IS  - 0366-6999 (Linking)
VI  - 126
IP  - 16
DP  - 2013 Aug
TI  - In vitro cartilage production using an extracellular matrix-derived scaffold and 
      bone marrow-derived mesenchymal stem cells.
PG  - 3130-7
AB  - BACKGROUND: Cartilage repair is a challenging research area because of the 
      limited healing capacity of adult articular cartilage. We had previously 
      developed a natural, human cartilage extracellular matrix (ECM)-derived scaffold 
      for in vivo cartilage tissue engineering in nude mice. However, before these 
      scaffolds can be used in clinical applications in vivo, the in vitro effects 
      should be further explored. METHODS: We produced cartilage in vitro using a 
      natural cartilage ECM-derived scaffold. The scaffolds were fabricated by 
      combining a decellularization procedure with a freeze-drying technique and were 
      characterized by scanning electron microscopy (SEM), micro-computed tomography 
      (micro-CT), histological staining, cytotoxicity assay, biochemical and 
      biomechanical analysis. After being chondrogenically induced, the induction 
      results of BMSCs were analyzed by histology and Immunohisto-chemistry. The 
      attachment and viability assessment of the cells on scaffolds were analyzed using 
      SEM and LIVE/DEAD staining. Cell-scaffold constructs cultured in vitro for 1 week 
      and 3 weeks were analyzed using histological and immunohistochemical methods. 
      RESULTS: SEM and micro-CT revealed a 3-D interconnected porous structure. The 
      majority of the cartilage ECM was found in the scaffold following the removal of 
      cellular debris, and stained positive for safranin O and collagen II. Viability 
      staining indicated no cytotoxic effects of the scaffold. Biochemical analysis 
      showed that collagen content was (708.2-44.7) microg/mg, with GAG (254.7 +/- 25.9) 
      microg/mg. Mechanical testing showed the compression moduli (E) were (1.226 +/- 0.288) 
      and (0.052 +/- 0.007) MPa in dry and wet conditions, respectively. Isolated canine 
      bone marrow-derived stem cells (BMSCs) were induced down a chondrogenic pathway, 
      labeled with PKH26, and seeded onto the scaffold. Immunofluorescent staining of 
      the cell-scaffold constructs indicated that chondrocyte-like cells were derived 
      from seeded BMSCs and excreted ECM. The cell-scaffold constructs contained pink, 
      smooth and translucent cartilage-like tissue after 3 weeks of culture. We 
      observed evenly distributed cartilage ECM proteoglycans and collagen type II 
      around seeded BMSCs on the surface and inside the pores throughout the scaffold. 
      CONCLUSION: This study suggests that a cartilage ECM scaffold holds much promise 
      for in vitro cartilage tissue engineering.
FAU - Zhao, Yan-hong
AU  - Zhao YH
AD  - Chinese People's Liberation Army General Hospital, Beijing, China.
FAU - Yang, Qiang
AU  - Yang Q
FAU - Xia, Qun
AU  - Xia Q
FAU - Peng, Jiang
AU  - Peng J
FAU - Lu, Shi-bi
AU  - Lu SB
FAU - Guo, Quan-yi
AU  - Guo QY
FAU - Ma, Xin-long
AU  - Ma XL
FAU - Xu, Bao-shan
AU  - Xu BS
FAU - Hu, Yong-cheng
AU  - Hu YC
FAU - Zhao, Bin
AU  - Zhao B
FAU - Zhang, Li
AU  - Zhang L
FAU - Wang, Ai-yuan
AU  - Wang AY
FAU - Xu, Weng-jing
AU  - Xu WJ
FAU - Miao, Jun
AU  - Miao J
FAU - Liu, Yue
AU  - Liu Y
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China
TA  - Chin Med J (Engl)
JT  - Chinese medical journal
JID - 7513795
SB  - IM
MH  - Animals
MH  - Biomechanical Phenomena
MH  - Cartilage/*cytology
MH  - Cell Survival
MH  - Cells, Cultured
MH  - Dogs
MH  - Extracellular Matrix/*physiology
MH  - Humans
MH  - Immunohistochemistry
MH  - Male
MH  - Mesenchymal Stem Cells/*cytology
MH  - Tissue Engineering/*methods
MH  - *Tissue Scaffolds
EDAT- 2013/08/29 06:00
MHDA- 2014/06/25 06:00
CRDT- 2013/08/29 06:00
PHST- 2013/08/29 06:00 [entrez]
PHST- 2013/08/29 06:00 [pubmed]
PHST- 2014/06/25 06:00 [medline]
PST - ppublish
SO  - Chin Med J (Engl). 2013 Aug;126(16):3130-7.