gms | German Medical Science

Deutscher Kongress für Orthopädie und Unfallchirurgie
74. Jahrestagung der Deutschen Gesellschaft für Unfallchirurgie
96. Tagung der Deutschen Gesellschaft für Orthopädie und Orthopädische Chirurgie
51. Tagung des Berufsverbandes der Fachärzte für Orthopädie und Unfallchirurgie

26. - 29.10.2010, Berlin

Stem cell loaded functionalized fibrin-based gels for tissue regeneration

Meeting Abstract

  • C. Vater - University Hospital Carl Gustav Carus, Department of Orthopaedics, Dresden, Germany
  • A. Jacobi - University Hospital Carl Gustav Carus, Department of Orthopaedics, Dresden, Germany
  • P. Kasten - University Hospital Carl Gustav Carus, Department of Orthopaedics, Dresden, Germany
  • M. Schieker - Ludwig-Maximilians-University, Experimental Surgery and Regenerative Medicine, Munich, Germany
  • K.-P. Günther - University Hospital Carl Gustav Carus, Department of Orthopaedics, Dresden, Germany
  • M. Stiehler - University Hospital Carl Gustav Carus, Department of Orthopaedics, Dresden, Germany

Deutscher Kongress für Orthopädie und Unfallchirurgie. 74. Jahrestagung der Deutschen Gesellschaft für Unfallchirurgie, 96. Tagung der Deutschen Gesellschaft für Orthopädie und Orthopädische Chirurgie, 51. Tagung des Berufsverbandes der Fachärzte für Orthopädie. Berlin, 26.-29.10.2010. Düsseldorf: German Medical Science GMS Publishing House; 2010. DocIN17-824

DOI: 10.3205/10dkou102, URN: urn:nbn:de:0183-10dkou1025

Published: October 21, 2010

© 2010 Vater et al.
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Outline

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Objective: Instant cell-based therapy denotes a promising, clinically relevant alternative to classical two-step tissue engineering strategies including cell expansion prior to implantation [Kasten et al. 2008]. Multipotent mesenchymal stromal cells (MSCs) can be easily isolated and culture expanded from bone marrow aspirates and provide an excellent source of progenitor cells [Stiehler et al. 2006]. In this context, practicable approaches require immediate progenitor cell immobilization in a three-dimensional, biocompatible matrix supporting cellular proliferation and differentiation. Fibrin is a physiologically occuring protein that polymerizes during haemostasis. The aim of this study was to establish a protocol for three-dimensional fibrin-based immobilization and osteogenic stimulation of MSCs. Cell-loaded gels were evaluated for cell viability and osteogenic differentiation. Furthermore the release of bone morphogenetic protein-4 (BMP-4) from fibrin gels was investigated.

Methods: For preparation of gels Tissucol™ fibrin sealant kit (FC) or platelet-rich plasma (PRP) in combination with or without tricalciumphosphat (TCP) granules were used at different fibrin concentrations. BMP-4 release from the gels was measured using BMP-4 ELISA kit. Human immortalized single-cell derived MSCs [Böcker et al. 2008] were added to the fibrin or PRP solution and cultured for 7 days with and without osteogenic supplements. Cell viability was assessed by use of live/dead staining. Lactate dehydrogenase (LDH) activity was determined to investigate cellular proliferation. For evaluation of osteogenic differentiation cell-specific alkaline phosphatase (ALP) activity was quantified.

Results and conclusions: Gel viscosity was optimized by modification of fibrin concentrations. The amount of fibrinogen concentration used was positively correlated to the rate of rhBMP-4 release from the gels. MSCs showed sufficient viability in all types of fibrinogen and PRP-based gels, respectively and no differences concerning cell viability could be observed between FC- and PRP-gels. Cells were able to proliferate and differentiate towards osteogenic lineage as detected by cell-specific ALP-activity. Thereby highest cell numbers and cell-specific ALP activity could be observed for scaffolds made of TCP granules and PRP (p<0.05).

We conclude that fibrin gels can serve as a naturally occuring matrix for the immobilization of stem cells enabling cell proliferation and differentiation at the defect site. We will further pursue this approach for therapy of musculoskeletal defects.