Intact human amniotic membrane differentiated towards the chondrogenic lineage
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ORIGINAL PAPER
Intact human amniotic membrane differentiated towards the chondrogenic lineage Andrea Lindenmair • Sylvia Nu¨rnberger • Guido Stadler • Alexandra Meinl Christa Hackl • Johann Eibl • Christian Gabriel • Simone Hennerbichler • Heinz Redl • Susanne Wolbank
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Received: 31 July 2013 / Accepted: 29 April 2014 / Published online: 15 May 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract Human amniotic membrane (hAM) represents a tissue that is well established as biomaterial in the clinics with potential for new applications in regenerative medicine. For tissue engineering (TE) strategies, cells are usually combined with inductive factors and a carrier substrate. We have previously recognized that hAM represents a natural, preformed sheet including highly potent stem cells. In the present approach for cartilage regeneration we have induced chondrogenesis in hAM in vitro. For this, hAM biopsies were cultured for up to 56 days under chondrogenic conditions. The induced hAM was characterized for remaining viability, glycosaminoglycan (GAG) accumulation using histochemical analysis, and a quantitative assay. Collagen I, II and X was immunohistochemically determined and cartilage-specific mRNA expression of (sex determining region Y-) box 9, cartilage oligomeric matrix protein
(COMP), aggrecan (AGC1), versican (CSPG2), COL 1A1, COL9A2, melanoma inhibitory activity (MIA), and cartilage-linking protein 1 (CRTL1) analyzed by quantitative real-time polymerase chain reaction. Human AM was successfully induced to accumulate GAG, as demonstrated by Alcianblue staining and a significant (p \ 0.001) increase of GAG/viability under chondrogenic conditions peaking in a 29.9 ± 0.9-fold induction on day 56. Further, upon chondrogenic induction collagen II positive areas were identified within histological sections and cartilage-specific markers including COMP, AGC1, CSPG2, COL1A1, COL9A2, MIA, and CRTL1 were found upregulated at mRNA level. This is the first study, demonstrating that upon in vitro induction viable human amnion expresses cartilage-specific markers and accumulates GAGs within the biomatrix. This is a promising first step towards a potential use of living hAM for cartilage TE.
A. Lindenmair S. Nu¨rnberger (&) G. Stadler A. Meinl H. Redl S. Wolbank Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center, Donaueschingenstraße 13, 1200 Vienna, Austria e-mail: [email protected]
S. Nu¨rnberger A. Meinl Bernhard Gottlieb University School of Dentistry, Sensengasse 2a, 1090 Vienna, Austria
A. Lindenmair C. Hackl C. Gabriel S. Hennerbichler Red Cross Blood Transfusion Service of Upper Austria, Krankenhausstraße 7, 4017 Linz, Austria
S. Nu¨rnberger Department of Traumatology, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria J. Eibl Bio-Products and Bio-Engineering AG, Schottenring 10, 1010 Vienna, Austria
A. Lindenmair S. Nu¨rnberger G. Stadler A. Meinl C. Hackl C. Gabriel S. Hennerbichler
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