title: Glucan HBP-A increase type II collagen expression of chondrocytes in vitro and tissue engineered cartilage in vivo.
abstract: OBJECTIVE
  Although chondroprotective activities have been documented for polysaccharides, the potential target of different polysaccharide may differ. The study was aimed to explore the effect of glucan HBP-A in chondrocyte monolayer culture and chondrocytes-alginate hydrogel constructs in vivo, especially on the expression of type II collagen.
  
  
  METHODS
  Chondrocytes isolated from rabbit articular cartilage were cultured and verified by immunocytochemical staining of type II collagen. Chondrocyte viability was assessed after being treated with HBP-A in different concentrations. Morphological status of chondrocytes-alginate hydrogel constructs in vitro was observed by scanning electron microscope (SEM). The constructs were treated with HBP-A and then injected to nude mice subcutaneously. Six weeks after transplantation, the specimens were observed through transmission electron microscopy (TEM). The mRNA expressions of disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTs-5), aggrecan and type II collagen in both monolayer culture and constructs were determined by real time polymerase chain reaction (PCR). The expression of type II collagen and matrix metalloproteinases-3 (MMP-3) in chondrocyte monolayer culture was also tested through Western blot and enzyme linked immunosorbent assay (ELISA), respectively.
  
  
  RESULTS
  MMP-3 secretion and ADAMTs-5 mRNA expression in vitro were inhibited by HBP-A at 0.3 mg/mL concentration. In morphological study, there were significant appearance of collagen in those constructs treated by HBP-A. Accordingly, in both chondrocyte monolayer culture and chondrocytes-alginate hydrogel constructs, the expression of type II collagen was increased significantly in HBP-A group when compared with control group (P<0.001).
  
  
  CONCLUSIONS
  The study documented that the potential pharmacological target of glucan HBP-A in chondrocytes monolayer culture and tissue engineered cartilage in vivo may be concerned with the inhibition of catabolic enzymes MMP-3, ADAMTs-5, and increasing of type II collagen expression.
authors.affiliation: Research Institute of Orthopaedics, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China, ningtcm@126.com.
authors.forename: Yue-long
authors.initials: YL
authors.lastname: Cao
authors.forename: Ting
authors.initials: T
authors.lastname: Liu
authors.forename: Jian
authors.initials: J
authors.lastname: Pang
authors.forename: Ning-yang
authors.initials: NY
authors.lastname: Gao
authors.forename: Hong-sheng
authors.initials: HS
authors.lastname: Zhan
authors.forename: Yin-yu
authors.initials: YY
authors.lastname: Shi
authors.forename: Xiang
authors.initials: X
authors.lastname: Wang
authors.forename: Shun-chun
authors.initials: SC
authors.lastname: Wang
mesh_terms: D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism; D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000464:Alginates / Q000494:pharmacology; D000818:Animals; D002358:Cartilage, Articular / Q000187:drug effects / Q000502:physiology*; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005936:Glucans / Q000494:pharmacology*; D020723:Glucuronic Acid / Q000494:pharmacology; D006603:Hexuronic Acids / Q000494:pharmacology; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000494:pharmacology; D007150:Immunohistochemistry; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D008819:Mice, Nude; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D053674:Aggrecans; D000464:Alginates; D024043:Collagen Type II; D005936:Glucans; D006603:Hexuronic Acids; D012333:RNA, Messenger; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D020723:Glucuronic Acid; D051722:ADAM Proteins; D019278:Matrix Metalloproteinase 3