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Open Access Research article

Collagen type I and decorin expression in tenocytes depend on the cell isolation method

Markus U Wagenhäuser1, Matthias F Pietschmann1*, Birte Sievers1, Denitsa Docheva2, Matthias Schieker2, Volkmar Jansson1 and Peter E Müller1

Author Affiliations

1 Department of Orthopaedic Surgery, Ludwig-Maximilians-University Munich - Campus Grosshadern, Marchioninistr 15, 81377, Munich, Germany

2 Experimental Surgery and Regenerative Medicine, Department of Surgery, Ludwig-Maximilians-University – Campus Innenstadt, Nußbaumstr 20, 80336, Munich, Germany

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BMC Musculoskeletal Disorders 2012, 13:140  doi:10.1186/1471-2474-13-140

Published: 8 August 2012

Abstract

Backround

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.

Conclusion

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.

Keywords:
Tenocytes; Tissue engineering; Isolation method; Gene expression