title: [The application of comparative proteomic analysis to screen proteins associated with mechanical properties of engineered cartilage: a preliminary study].
abstract: OBJECTIVE
  To study proteins correlated with the mechanical properties of engineered cartilage by screening significantly changed proteins during cartilage formation by comparative proteomic analysis.
  
  
  METHODS
  Human chondrocyte, cultured and expanded, were seeded onto a polyglycolic acid/polylactic acid (PGA/PLA) scaffolds. After 4 weeks of culture in vitro, the constructs were divided into three groups. There were 6 specimens in each group. For the regular in vitro culture group (A), the constructs were kept in culture at the original condition for an additional 6 weeks. For in vivo groups, the constructs were implanted subcutaneously into nude mice for either 6 weeks (B) or 12 weeks (C). All specimens were harvested for gross observation, average wet weight and volume measurement, histology, immunohistochemistry and biomechanics to evaluate the results. Meanwhile, comparative proteomic analysis was performed for each group, and those proteins involved in extracellular matrix with at least 2 folds up-regulation were chosen for further exploration. The correlations between Young's modulus and the relative content of the selected proteins were analyzed by Pearson correlation coefficient.
  
  
  RESULTS
  All these samples in the three groups eventually formed hyaline-like cartilage structure. Specimens in C and B groups were similar with adult articular cartilage in appearance, and had multiple mature lacuna in histology. However, those specimens in A group had loose texture with irregular hypertrophy lacuna. Specimens implanted for 12 weeks in vivo had better wet weight (372.5 +/- 35.4) mg and Young's modulus (8.68 +/- 2.65) MPa than those cultured in vivo for 6 weeks (346 +/- 34.5) mg, (3.25 +/- 1.24) MPa (P < 0.01). In group A, they were (184.4 +/- 12.28) mg and (0.7 +/- 0.23) MPa. This study had detected 44 proteins in ECM by comparative proteomic analysis, then chosing the greatest ratio of 6 up-regulation proteins compared between C and A groups. The correlation results indicated the content of Decorin, Chondroadherin and Fibromodulin were linear correlation with the mechanical properties of engineered cartilage (P < 0.05).
  
  
  CONCLUSIONS
  Comparative proteomic analysis could provide large scale information of associated proteins, making it profit for advanced research on the relationship between extracellular matrix and mechanical properties of engineered cartilage by combination with tissue reconstruction techniques.
authors.affiliation: Department of Plastic and Reconstructive Laboratory of Tissue Engineering, Shanghai 200011, China.
authors.forename: Jin-Ping
authors.initials: JP
authors.lastname: Ding
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: Liu
authors.forename: Dan
authors.initials: D
authors.lastname: Yan
authors.forename: Ming-Liang
authors.initials: ML
authors.lastname: Ye
authors.forename: Han-Fa
authors.initials: HF
authors.lastname: Zou
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: Wen-Jie
authors.initials: WJ
authors.lastname: Zhang
authors.forename: Yi-Lin
authors.initials: YL
authors.lastname: Cao
authors.forename: Guang-Dong
authors.initials: GD
authors.lastname: Zhou
mesh_terms: D000818:Animals; D002356:Cartilage / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism*; D005333:Fetus / Q000166:cytology; D006801:Humans; D008819:Mice, Nude; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
chemical_list: D020543:Proteome