title: Role of Cortico-Cancellous Heterologous Bone in Human Periodontal Ligament Stem Cell Xeno-Free Culture Studied by Synchrotron Radiation Phase-Contrast Microtomography.
abstract: This study was designed to quantitatively demonstrate via three-dimensional (3D) images, through the Synchrotron Radiation Phase-Contrast Microtomography (SR-PhC-MicroCT), the osteoinductive properties of a cortico-cancellous scaffold (Osteobiol Dual Block-DB) cultured with human Periodontal Ligament Stem Cells (hPDLSCs) in xeno-free media. In vitro cultures of hPDLSCs, obtained from alveolar crest and horizontal fibers of the periodontal ligament, were seeded onto DB scaffolds and cultured in xeno-free media for three weeks. 3D images were obtained by SR-PhC-microCT after one and three weeks from culture beginning. MicroCT data were successively processed with a phase-retrieval algorithm based on the Transport of Intensity Equation (TIE). The chosen experimental method, previously demonstratively applied for the 3D characterization of the same constructs in not xeno-free media, quantitatively monitored also in this case the early stages of bone formation in basal and differentiating conditions. Interestingly, it quantitatively showed in the xeno-free environment a significant acceleration of the mineralization process, regardless of the culture (basal/differentiating) medium. This work showed in 3D that the DB guides the osteogenic differentiation of hPDLSCs in xeno-free cultures, in agreement with 2D observations and functional studies previously performed by some of the authors. Indeed, here we fully proved in 3D that expanded hPDLSCs, using xeno-free media formulation, not only provide the basis for Good Manufacturing Practice (preserving the stem cells' morphological features and their ability to differentiate into mesenchymal lineage) but have to be considered, combined to DB scaffolds, as interesting candidates for potential clinical use in new custom made tissue-engineered constructs.
authors.affiliation: Department of Clinical Sciences-Unit of Biochemistry, Biology and Physics, Polytechnic University of Marche, Via Brecce Bianche 1, 60131 Ancona, Italy. s.mazzoni@alisf1.univpm.it.
authors.forename: Serena
authors.initials: S
authors.lastname: Mazzoni
authors.affiliation: Sincrotrone Trieste S.C.p.A., Strada Statale 14 km 163.5 in AREA Science Park, 34149 Trieste, Italy. patezian@gmail.com.
authors.forename: Sara
authors.initials: S
authors.lastname: Mohammadi
authors.affiliation: Sincrotrone Trieste S.C.p.A., Strada Statale 14 km 163.5 in AREA Science Park, 34149 Trieste, Italy. giuliana.tromba@elettra.trieste.it.
authors.forename: Giuliana
authors.initials: G
authors.lastname: Tromba
authors.affiliation: Department of Medical, Oral and Biotechnological Sciences, University of Chieti-Pescara, 66100 Chieti, Italy. francesca.diomede@unich.it.
authors.forename: Francesca
authors.initials: F
authors.lastname: Diomede
authors.affiliation: Department of Medical, Oral and Biotechnological Sciences, University of Chieti-Pescara, 66100 Chieti, Italy. apiattelli@unich.it.
authors.forename: Adriano
authors.initials: A
authors.lastname: Piattelli
authors.affiliation: Department of Medical, Oral and Biotechnological Sciences, University of Chieti-Pescara, 66100 Chieti, Italy. trubiani@unich.it.
authors.forename: Oriana
authors.initials: O
authors.lastname: Trubiani
authors.affiliation: Department of Clinical Sciences-Unit of Biochemistry, Biology and Physics, Polytechnic University of Marche, Via Brecce Bianche 1, 60131 Ancona, Italy. a.giuliani@univpm.it.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Giuliani
mesh_terms: D000328:Adult; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D006801:Humans; D008858:Microscopy, Phase-Contrast / Q000379:methods; D010012:Osteogenesis; D010513:Periodontal Ligament / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*; D017356:Synchrotrons; D023822:Tissue Engineering; D054457:Tissue Scaffolds
keywords: biomaterial; osteogenesis; periodontal ligament stem cells; phase-contrast microtomography; xeno-free medium