title: Raman Spectroscopic Analyses of Jaw Periosteal Cell Mineralization.
abstract: To achieve safer patient treatments, serum-free cell culture conditions have to be established for cell therapies. In previous studies, we demonstrated that serum-free culture favored the proliferation of MSCA-1+osteoprogenitors derived from the jaw periosteum. In this study, the in vitro formation of bone-specific matrix by MSCA-1+jaw periosteal cells (JPCs, 3 donors) was assessed and compared under serum-free and serum-containing media conditions using the marker-free Raman spectroscopy. Based on a standard fluorescence assay, JPCs from one patient were not able to mineralize under serum-containing culture conditions, whereas the other cells showed similar mineralization levels under both conditions. Raman spectra from mineralizing MSCA-1+JPCs revealed higher levels of hydroxyapatite formation and higher mineral to matrix ratios under serum-free culture conditions. Higher carbonate to phosphate ratios and higher crystallinity in JPCs cultured under serum-containing conditions indicated immature bone formation. Due to reduced collagen production under serum-free conditions, we obtained significant differences in collagen maturity and proline to hydroxyproline ratios compared to serum-free conditions. We conclude that Raman spectroscopy is a useful tool for the assessment and noninvasive monitoring of in vitro mineralization of osteoprogenitor cells. Further studies should extend this knowledge and improve JPC mineralization by optimizing culture conditions.
authors.affiliation: Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), Department of Cell and Tissue Engineering, Nobelstr. 12, 70569 Stuttgart, Germany; Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University, Tübingen, Silcherstr. 7/1, 72076 Tübingen, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Brauchle
authors.affiliation: Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University, Tübingen, Silcherstr. 7/1, 72076 Tübingen, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Carvajal Berrio
authors.affiliation: Department of Oral and Maxillofacial Surgery, University Hospital of Tübingen, 72076 Tübingen, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Rieger
authors.affiliation: Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), Department of Cell and Tissue Engineering, Nobelstr. 12, 70569 Stuttgart, Germany; Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University, Tübingen, Silcherstr. 7/1, 72076 Tübingen, Germany; Department of Medicine/Cardiology, Cardiovascular Research Laboratories, David Geffen School of Medicine at UCLA, 675 Charles E. Young Drive South, MRL 3645, Los Angeles, CA, USA.
authors.forename: Katja
authors.initials: K
authors.lastname: Schenke-Layland
authors.affiliation: Department of Oral and Maxillofacial Surgery, University Hospital of Tübingen, 72076 Tübingen, Germany.
authors.forename: Siegmar
authors.initials: S
authors.lastname: Reinert
authors.affiliation: Department of Oral and Maxillofacial Surgery, University Hospital of Tübingen, 72076 Tübingen, Germany.
authors.forename: Dorothea
authors.identifier: 0000-0001-7467-6802
authors.initials: D
authors.lastname: Alexander