PMID- 26455178
OWN - NLM
STAT- MEDLINE
DCOM- 20160302
LR  - 20181202
IS  - 1002-1892 (Print)
IS  - 1002-1892 (Linking)
VI  - 29
IP  - 1
DP  - 2015 Jan
TI  - [CONDITIONS OF SYNOVIAL MESENCHYMAL STEM CELLS DIFFERENTIATING INTO 
      FIBROCARTILAGE CELLS].
PG  - 81-91
AB  - OBJECTIVE: To explore the conditions of synovial derived mesenchymal stem cells 
      (SMSCs) differentiating into the fibrocartilage cells by using the orthogonal 
      experiment. METHODS: The synovium was harvested from 5 adult New Zealand white 
      rabbits, and SMSCs were separated by adherence method. The flow cytometry and 
      multi-directional differentiation method were used to identify the SMSCs. The 
      conditions were found from the preliminary experiment and literature review. The 
      missing test was carried out to screen the conditions and then 12 conditions were 
      used for the orthogonal experiment, including transforming growth factor beta1 
      (TGF-beta1), bone morphogenic protein 2 (BMP-2), dexamethasone (DEX), proline, 
      ascorbic acid (ASA), pyruvic acid, insulin + transferrin + selenious acid 
      pre-mixed solution (ITS), bovin serum albumin (BSA), basic fibroblast growth 
      factor (bFGF), intermittent hydraulic pressure (IHP), bone morphogenic protein 7 
      (BMP-7), and insulin-like growth factor (IGF). The L60 (212) orthogonal 
      experiment was designed using the SPSS 18.0 with 2 level conditions and the cells 
      were induced to differentiate on the small intestinal submucosa (SIS)-3D 
      scaffold. The CD151+/CD44+ cells were detected with the flow cytometry and then 
      the differentiation rate was recorded. The immumohistochemical staining, cellular 
      morphology, toluidine blue staining, and semi-quantitative RT-PCR examination for 
      the gene expressions of sex determining region Y (SRY)-box 9 gene (Sox9), 
      aggrecan gene (AGN), collagen type I gene (Col I), collagen type II gene (Col 
      II), collagen type IX gene (Col IX) were used for result confirmation. The 
      differentiation rate was calculated as the product of CD151/CD44+ cells and cells 
      with Col I high expression. The grow curve was detected with the DNA abundance 
      using the PicoGreen Assay. The visual observation and the variances analysis 
      among the variable were used to evaluate the result of the orthogonal experiment, 
      1 level interaction was considered. The q-test and the least significant 
      difference (LDS) were used for the variance analysis with a type III calibration 
      model. The test criteria (a) was 0.05. RESULTS: The cells were certified as 
      SMSCs, the double-time of the cells was 28 hours. During the differentiation into 
      the fibrocartilage, the volume of the SIS-3D scaffold enlarged double every 5 
      days. The scaffolds were positively stained by toluidine blue at 14 days. The 
      visual observation showed that high levels of TGF-beta1 and BMP-7 were optimum for 
      the differentiation, and BMP-7 showed the interaction with BMP-2. The conditions 
      of DEX, ASA, ITS, transferrin, bFGF showed decreasing promotional function by 
      degrees, and the model showed the perfect relevance. P value was 0.000 according 
      to the variance analysis. The intercept analysis showed different independent 
      variables brought about variant contribution; the TGF-beta1, ASA, bFGF, IGF, and 
      BMP-7 were more remarkable, which were similar to the visual observation. 
      CONCLUSION: In the process of the SMSCs differentiation into the fibrocartilage, 
      the concentrations of TGF-beta1, ASA, bFGF, and IGF reasonably can improve the 
      conversion rate of the fibrocartilage cells. The accurate conditions of the 
      reaulatory factor should be explored further.
FAU - Fu, Peiliang
AU  - Fu P
FAU - Cong, Ruijun
AU  - Cong R
FAU - Chen, Song
AU  - Chen S
FAU - Zhang, Lei
AU  - Zhang L
FAU - Ding, Zheru
AU  - Ding Z
FAU - Zhou, Qi
AU  - Zhou Q
FAU - Li, Lintao
AU  - Li L
FAU - Xu, Zhenyu
AU  - Xu Z
FAU - Wu, Yuli
AU  - Wu Y
FAU - Wu, Haishan
AU  - Wu H
LA  - chi
PT  - Journal Article
PL  - China
TA  - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi
JT  - Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = 
      Chinese journal of reparative and reconstructive surgery
JID - 9425194
RN  - 0 (Aggrecans)
RN  - 0 (Bone Morphogenetic Protein 2)
RN  - 0 (Bone Morphogenetic Proteins)
RN  - 0 (Collagen Type I)
RN  - 0 (Collagen Type II)
RN  - 0 (Transforming Growth Factor beta1)
RN  - 0 (Transforming Growth Factor beta3)
RN  - 103107-01-3 (Fibroblast Growth Factor 2)
SB  - IM
MH  - Aggrecans
MH  - Animals
MH  - Bone Morphogenetic Protein 2/administration & dosage/*pharmacology
MH  - Bone Morphogenetic Proteins
MH  - Cell Differentiation
MH  - Collagen Type I
MH  - Collagen Type II/metabolism
MH  - Fibroblast Growth Factor 2
MH  - *Fibrocartilage
MH  - Mesenchymal Stem Cells/*cytology/drug effects/metabolism
MH  - Rabbits
MH  - Synovial Membrane/*cytology
MH  - Tissue Engineering
MH  - Transforming Growth Factor beta1
MH  - Transforming Growth Factor beta3/administration & dosage/*pharmacology
EDAT- 2015/10/13 06:00
MHDA- 2016/03/05 06:00
CRDT- 2015/10/13 06:00
PHST- 2015/10/13 06:00 [entrez]
PHST- 2015/10/13 06:00 [pubmed]
PHST- 2016/03/05 06:00 [medline]
PST - ppublish
SO  - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2015 Jan;29(1):81-91.