title: Collagen type I and decorin expression in tenocytes depend on the cell isolation method.
abstract: BACKGROUND
  The treatment of rotator cuff tears is still challenging. Tendon tissue engineering (TTE) might be an alternative in future. Tenocytes seem to be the most suitable cell type as they are easy to obtain and no differentiation in vitro is necessary. The aim of this study was to examine, if the long head of the biceps tendon (LHB) can deliver viable tenocytes for TTE. In this context, different isolation methods, such as enzymatic digestion (ED) and cell migration (CM), are investigated on differences in gene expression and cell morphology.
  
  
  METHODS
  Samples of the LHB were obtained from patients, who underwent surgery for primary shoulder arthroplasty. Using ED as isolation method, 0.2% collagenase I solution was used. Using CM as isolation method, small pieces of minced tendon were put into petri-dishes. After cell cultivation, RT-PCR was performed for collagen type I, collagen type III, decorin, tenascin-C, fibronectin, Scleraxis, tenomodulin, osteopontin and agreccan.
  
  
  RESULTS
  The total number of isolated cells, in relation to 1 g of native tissue, was 14 times higher using ED. The time interval for cell isolation was about 17 hours using ED and approximately 50 days using CM. Cell morphology in vitro was similar for both isolation techniques. Higher expression of collagen type I could be observed in tenocyte-like cell cultures (TLCC) using ED as isolation method (p < 0.05), however decorin expression was higher in TLCC using CM as isolation method (p < 0.05). Dedifferentiation potential seemed to be similar for both isolation techniques.
  
  
  CONCLUSIONS
  In summary tenocyte-like cells can be obtained with both isolation methods (ED and CM) from the LHB. As no obvious disadvantage could be seen using ED, this method is more suitable for clinical use, as time for cell isolation is shorter and a remarkably higher number of cells can be obtained. However, both isolation methods can further be improved.
authors.affiliation: Department of Orthopaedic Surgery, Ludwig-Maximilians-University Munich-Campus Grosshadern, Marchioninistr 15, 81377 Munich, Germany.
authors.forename: Markus U
authors.initials: MU
authors.lastname: Wagenhäuser
authors.forename: Matthias F
authors.initials: MF
authors.lastname: Pietschmann
authors.forename: Birte
authors.initials: B
authors.lastname: Sievers
authors.forename: Denitsa
authors.initials: D
authors.lastname: Docheva
authors.forename: Matthias
authors.initials: M
authors.lastname: Schieker
authors.forename: Volkmar
authors.initials: V
authors.lastname: Jansson
authors.forename: Peter E
authors.initials: PE
authors.lastname: Müller
mesh_terms: D000368:Aged; D002465:Cell Movement / Q000235:genetics; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D024042:Collagen Type I / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D058575:Decorin / Q000096:biosynthesis* / Q000235:genetics; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008297:Male; D008875:Middle Aged; D013710:Tendons / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism*
chemical_list: D024042:Collagen Type I; C546878:DCN protein, human; D058575:Decorin