title: Suppression of osteoblast-related genes during osteogenic differentiation of adipose tissue derived stromal cells.
abstract: Recent studies indicated a lower osteogenic differentiation potential of adipose tissue-derived stromal cells (ASCs) compared to bone marrow derived mesenchymal stromal cells. The aim of this study was to evaluate the effects of potent combinations of highly osteogenic bone morphogenetic proteins (BMPs) in order to enhance the osteogenic differentiation potential of ASCs. Human ASCs were cultured for 10 days in the presence of osteogenic medium consisting of dexamethasone, ß-glycerophosphate and ascorbat-2-phosphate (OM) supplemented with BMP-2, BMP-6, BMP-9+IGF-2 and BMP-2,-6,-9 (day 1+2: 50 ng/ml, days 3-6: 100 ng/ml, days 7-10: 200 ng/ml). The formation of the osteoblast phenotype was evaluated by quantification of osteoblast-related marker genes using real-time polymerase chain reaction (RT-PCR). Matrix mineralization was assessed by Alizarin Red S staining. Statistical analysis was carried out using the one-way analysis of variance (ANOVA) followed by the Scheffe's post hoc procedure. Osteogenic medium (OM) significantly increased the expression of alkaline phosphatase (ALP) and osteocalcin (p < 0.05) and led to a stable matrix mineralization. Under the influence of BMP-9+IGF-2 and BMP-2,-6,-9 the ALP expression further increased compared to ASCs cultured with OM only (p < 0.01). However, multiple osteogenic markers showed no change or decreased under the influence of OM and BMP combinations (p < 0.05). The current results indicate a restricted osteogenic differentiation potential of ASCs and suggest careful reconsideration of their use in bone tissue engineering applications.
authors.affiliation: Department of Oral & Maxillofacial Surgery (Head of Scientific Laboratories: Prof. Yahya Açil, Head: Prof. Dr. Jörg Wiltfang), Christian-Albrechts University, Kiel, Germany.
authors.forename: Yahya
authors.initials: Y
authors.lastname: Açil
authors.affiliation: Department of Oral & Maxillofacial Surgery (Head of Scientific Laboratories: Prof. Yahya Açil, Head: Prof. Dr. Jörg Wiltfang), Christian-Albrechts University, Kiel, Germany.
authors.forename: Amir Alexander
authors.initials: AA
authors.lastname: Ghoniem
authors.affiliation: Center for Oral and Dental Health, Kars, Turkey. Electronic address: aydingulses@gmail.com.
authors.forename: Aydin
authors.initials: A
authors.lastname: Gülses
authors.affiliation: Department of Plastic Surgery, University of Lübeck, Lübeck, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Kisch
authors.affiliation: Department of Plastic Surgery, University of Lübeck, Lübeck, Germany.
authors.forename: Felix
authors.initials: F
authors.lastname: Stang
authors.affiliation: Department of Oral & Maxillofacial Surgery (Head of Scientific Laboratories: Prof. Yahya Açil, Head: Prof. Dr. Jörg Wiltfang), Christian-Albrechts University, Kiel, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Wiltfang
authors.affiliation: Department of Plastic Surgery, University of Lübeck, Lübeck, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Gierloff
mesh_terms: D000273:Adipose Tissue / Q000166:cytology* / Q000502:physiology; D000469:Alkaline Phosphatase / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology; D055418:Bone Morphogenetic Protein 6 / Q000494:pharmacology; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002454:Cell Differentiation / Q000502:physiology*; D005786:Gene Expression Regulation / Q000187:drug effects; D055427:Growth Differentiation Factor 2; D055412:Growth Differentiation Factors / Q000494:pharmacology; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000494:pharmacology; D010006:Osteoblasts / Q000378:metabolism / Q000502:physiology*; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction; D017154:Stromal Cells / Q000502:physiology*
chemical_list: C525718:BMP2 protein, human; C525816:BMP6 protein, human; D055396:Bone Morphogenetic Protein 2; D055418:Bone Morphogenetic Protein 6; D019485:Bone Morphogenetic Proteins; C525887:GDF2 protein, human; D055427:Growth Differentiation Factor 2; D055412:Growth Differentiation Factors; D015675:Osteocalcin; D007335:Insulin-Like Growth Factor II; D000469:Alkaline Phosphatase
keywords: ASC; Adipose derived stromal cells; BMP; Bone tissue engineering; Growth factors; Osteogenic differentiation