Osteogenic differentiation of intact human amniotic membrane

Andrea Lindenmair, Susanne Wolbank, Guido Stadler, Alexandra Meinl, Anja Peterbauer-Scherb, Johann Eibl, Helene Polin, Christian Gabriel, Martijn van Griensven, Heinz Redl

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

Tissue engineering strategies usually require cell isolation and combination with a suitable biomaterial. Human amniotic membrane (AM) represents a natural two-layered sheet comprising cells with proven stem cell characteristics. In our approach, we evaluated the differentiation potential of AM in toto with its sessile stem cells as alternative to conventional approaches requiring cell isolation and combination with biomaterials. For this, AM-biopsies were differentiated in vitro using two osteogenic media compared with control medium (CM) for 28 days. Mineralization and osteocalcin expression was demonstrated by (immuno)histochemistry. Alkaline phosphatase (AP) activity, calcium contents and mRNA expression of RUNX2, AP, osteopontin, osteocalcin, BMP-2 (bone morphogenetic protein), and BMP-4 were quantified and AM viability was evaluated. Under osteogenic conditions, AM-biopsies mineralized successfully and by day 28 the majority of cells expressed osteocalcin. This was confirmed by a significant rise in calcium contents (up to 27.4 ± 6.8 mg/dl d28), increased AP activity, and induction of RUNX2, AP, BMP-2 and BMP-4 mRNA expression. Relatively high levels of viability were retained, especially in osteogenic media (up to 78.3 ± 19.0% d14; 62.9 ± 22.3% d28) compared to CM (42.2 ± 15.2% d14; 35.1 ± 8.6% d28). By this strategy, stem cells within human AM can successfully be driven along the osteogenic pathways while residing within their natural environment.

Original languageEnglish
Pages (from-to)8659-8665
Number of pages7
JournalBiomaterials
Volume31
Issue number33
DOIs
StatePublished - Nov 2010
Externally publishedYes

Keywords

  • Biomineralization
  • Bone tissue engineering
  • Human amniotic membrane
  • Osteogenesis
  • Sessile stem cells

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