gms | German Medical Science

Deutscher Kongress für Orthopädie und Unfallchirurgie, 75. Jahrestagung der Deutschen Gesellschaft für Unfallchirurgie, 97. Tagung der Deutschen Gesellschaft für Orthopädie und Orthopädische Chirurgie, 52. Tagung des Berufsverbandes der Fachärzte für Orthopädie und Unfallchirurgie

25. - 28.10.2011, Berlin

Novel polyurethane scaffolds for meniscus tissue engineering: Cytotoxicity and cell culture

Meeting Abstract

  • L. Chaoxu - Orthopedic Trauma Dept., Hannover Medical School, Hannover, Germany
  • R. Meister - Orthopedic Trauma Dept., Hannover Medical School, Hannover, Germany
  • C. Haasper - Orthopedic Trauma Dept., Hannover Medical School, Hannover, Germany
  • C. Krettek - Orthopedic Trauma Dept., Hannover Medical School, Hannover, Germany
  • M. Jagodzinski - Orthopedic Trauma Dept., Hannover Medical School, Hannover, Germany

Deutscher Kongress für Orthopädie und Unfallchirurgie. 75. Jahrestagung der Deutschen Gesellschaft für Unfallchirurgie, 97. Tagung der Deutschen Gesellschaft für Orthopädie und Orthopädische Chirurgie, 52. Tagung des Berufsverbandes der Fachärzte für Orthopädie. Berlin, 25.-28.10.2011. Düsseldorf: German Medical Science GMS Publishing House; 2011. DocPO14-970

DOI: 10.3205/11dkou607, URN: urn:nbn:de:0183-11dkou6079

Veröffentlicht: 18. Oktober 2011

© 2011 Chaoxu et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielfältigt, verbreitet und öffentlich zugänglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Gliederung

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Questionnaire: One of the most important requirements for scaffolds used to reconstruct meniscus is biocompatibility. A novel meniscus implant (ActifitTM) made from polyurethane is manufactured to replace the meniscus. Bone marrow stromal cells (BMSC) are investigated widely in the field of tissue engineering. The objective of this study was to investigate the biocompatibility of polyurethane meniscus implant on BMSC.

Methods: 20-80 ml bone marrow aspirates from the iliac crest were collected from 7. Cultivation of human BMSC were performed according to a modified protocol as previously described [1]. Scaffold extraction from 3 day to 21 days was collected according to a standardized protocol [2]. After culturing 1×105 cells with extraction for 48 h, cell proliferation was investigated using the MTS assay, and Live/Dead assay was utilized to assess cell viability. 6×106 cells of third passage resuspended in 1ml culture medium were seeded into one ActifitTM scaffold using a 27G syringe and cultured for 4 weeks. The histological assay was performed every 1 week. The data of all groups were compared using one-way ANOVA at each time point, a significance level of 0.05 was used (SPSS 15.0).

Results and Conclusions: MTS results showed there was no cell growth inhibition detected after cultured in extract. (Figure 1 [Fig. 1]). BMSC were able to metabolize the MTS into a brown formazan product after a 24 h incubation period with the collected extracts. This result was demonstrated in the Calcein AM/EthD-1 staining as well (Fig.2a,b). Few cells were stained by EthD-1 in negative control and extracted culture groups. In the direct contact study, light microscopy pictures showed cells were distributed in the central part of the scaffolds after seeding. The cell proliferation was not inhabited after adhesion on the scaffolds.

As a degradable material, polyurethane has been used in clinic for several years. However commonly used polyurethanes based on aromatic diisocyanates are lack biocompatibility, because degradation products from the aromatic segment are toxic. In our study, the meniscus scaffolds were made from polyurethanes that is based on 1,4-butane diisocyanate (BDI) instead of aromatic diisocyanates. As the degradation products of BDI are non-toxic, we believed our meniscus scaffold is biocompatible, which has no negative effect on cell viability and proliferation. And this hypothesis was proved through MTS assay and Calcein A/EthD-1 staining. The result reveled the extract obtained from scaffolds did not cause cell death among BMSCs, which was further confirmed by MTS assay. After seeded into the scaffolds, BMSC proliferated well in the pores of the scaffold and formed tissue engineered constructs with the scaffolds finally. Above all, the polymer scaffold did not affect hBMSC proliferation and viability, so the scaffold can be considered noncytotoxic. In future, studies in vitro and in vivo need to clarify the effect of BMSC loaded scaffolds on meniscus regeneration.