title: Scaffold preferences of mesenchymal stromal cells and adipose-derived stem cells from green fluorescent protein transgenic mice influence the tissue engineering of bone.
abstract: We have analysed the growth and differentiation of mesenchymal stromal cells (MSC) from bone marrow, and of adipose derived stem cells (ASC) from murine abdominal fat tissue, of green fluorescent protein (GFP) transgenic animals grown directly on two types of hydroxyapatite ceramic bone substitutes. BONITmatrix® and NanoBone® have specific mechanical and physiochemical properties such as porosity and an inner surface that influence cellular growth. Both MSC and ASC were separately seeded on 200mg of each biomaterial and cultured for 3 weeks under osteogenic differentiation conditions. The degree of mineralisation was assessed by alizarin red dye and the specific alkaline phosphatase activity of the differentiated cells. The morphology of the cells was examined by scanning electron microscopy and confocal microscopy. The osteoblastic phenotype of the cells was confirmed by analysing the expression of bone-specific genes (Runx2, osteocalcin, osteopontin, and osteonectin) by semiquantitative reverse transcriptase polymerase chain reaction (PCR). Comparison of BONITmatrix® and NanoBone® showed cell type-specific preferences in terms of osteogenic differentiation. MSC-derived osteoblast-like cells spread optimally on the surface of NanoBone® but not BONITmatrix® granules. In contrast BONITmatrix® granules conditioned the growth of osteoblast-like cells derived from ASC. The osteoblastic phenotype of the cultured cells on all matrices was confirmed by specific gene expression. Our results show that the in vitro growth and osteogenic differentiation of murine MSC or ASC of GFP transgenic mice are distinctly influenced by the ceramic substratum. While NanoBone® granules support the proliferation and differentiation of murine MSC isolated from bone marrow, the growth of murine ASC is supported by BONITmatrix® granules. NanoBone® is therefore recommended for use as scaffold in tissue engineering that requires MSC, whereas ASC can be combined with BONITmatrix® for in vitro bone engineering.
authors.affiliation: Clinic of Oral and Maxillofacial Surgery, University of Technology Dresden, University Hospital Carl Gustav Carus, Fetscherstr. 74, D-01307 Dresden, Germany; Department of Bioinformatics, University of Applied Sciences Mittweida, Technikumplatz 17, D-09648 Mittweida, Germany; Centre for Translational Bone, Joint and Soft Tissue Research, Fetscherstr. 74, D-01307 Dresden, Germany. Electronic address: gretel.wittenburg@uniklinikum-dresden.de.
authors.forename: Gretel
authors.initials: G
authors.lastname: Wittenburg
authors.affiliation: Clinic of Oral and Maxillofacial Surgery, University of Technology Dresden, University Hospital Carl Gustav Carus, Fetscherstr. 74, D-01307 Dresden, Germany.
authors.forename: Viktoria
authors.initials: V
authors.lastname: Flade
authors.affiliation: Institute of Physiological Chemistry, University of Technology Dresden, Medical Faculty Carl Gustav Carus, Fetscherstr. 74, D-01307 Dresden, Germany.
authors.forename: Annette I
authors.initials: AI
authors.lastname: Garbe
authors.affiliation: Clinic of Oral and Maxillofacial Surgery, University of Technology Dresden, University Hospital Carl Gustav Carus, Fetscherstr. 74, D-01307 Dresden, Germany; Centre for Translational Bone, Joint and Soft Tissue Research, Fetscherstr. 74, D-01307 Dresden, Germany.
authors.forename: Günter
authors.initials: G
authors.lastname: Lauer
authors.affiliation: Department of Bioinformatics, University of Applied Sciences Mittweida, Technikumplatz 17, D-09648 Mittweida, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Labudde
mesh_terms: D050153:Abdominal Fat / Q000166:cytology*; D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D018786:Bone Substitutes / Q000737:chemistry; D002130:Calcium Phosphates / Q000737:chemistry; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D048430:Cell Shape; D050659:Core Binding Factor Alpha 1 Subunit / Q000032:analysis; D004338:Drug Combinations; D017886:Durapatite / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D010006:Osteoblasts / Q000502:physiology; D015675:Osteocalcin / Q000032:analysis; D010012:Osteogenesis / Q000502:physiology; D015676:Osteonectin / Q000032:analysis; D010641:Phenotype; D016062:Porosity; D012822:Silicon Dioxide / Q000737:chemistry; D013234:Stem Cells / Q000502:physiology*; D013499:Surface Properties; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: C519914:BONITmatrix; D018786:Bone Substitutes; D002130:Calcium Phosphates; D050659:Core Binding Factor Alpha 1 Subunit; D004338:Drug Combinations; C534915:NanoBone; D015676:Osteonectin; C493716:Runx2 protein, mouse; D015675:Osteocalcin; D049452:Green Fluorescent Proteins; D012822:Silicon Dioxide; D017886:Durapatite; D000469:Alkaline Phosphatase
keywords: Adipose-derived stem cells; BONITmatrix(®); Bone tissue engineering; Green fluorescent protein; Hydroxyapatite; Mesenchymal stromal cells; NanoBone(®); Osteoblast-like cells