title: Study of tissue engineered bone nodules by Fourier transform infrared spectroscopy.
abstract: The key criteria for assessing the success of bone tissue engineering are the quality and quantity of the produced minerals within the cultured constructs. The accumulation of calcium ions and inorganic phosphates in culture medium serves as nucleating agents for the formation of hydroxyapatite, which is the main inorganic component of bone. Bone nodule formation is one of the hallmarks of mineralization in such cell cultures. In this study, we developed a new two-step procedure to accelerate bone formation in which mouse bone cell aggregates were produced first on various chemically treated non-adhesive substrates. After this step, the bone cells' growth and mineralization were followed in conventional culture plates. The number and size of cell aggregates were studied with light microscopy. The minerals' formation in the form of nodules produced by the cell aggregates and the bone crystal quality were studied with Fourier Transform Infrared (FTIR) spectroscopy. The FTIR spectra of the ash specimens (mineral phase only) from thermal gravimetric analysis (TGA) provided valuable information of the quality of the minerals. The υ(4) PO(4) region (550-650 cm(-1)), which reveals apatitic and non-apatitic HPO(4) or PO(4) environments, and phosphate region (910-1180 cm(-1)) were examined for the minerals produced in the form of nodules. The peak position and intensity of the spectra demonstrate that the quality of the bone produced by cell aggregates, especially from the bigger ones, which were formed on Plunoric treated substrates, exhibit a composition more similar to that of native bone. This work establishes a new protocol for high quality bone formation and characterization, with the potential to be applied to bone tissue engineering.
authors.affiliation: Institute for Science and Technology in Medicine, School of Medicine, Keele University, Stoke-on-Trent, UK.
authors.forename: Halil Murat
authors.initials: HM
authors.lastname: Aydin
authors.forename: Bin
authors.initials: B
authors.lastname: Hu
authors.forename: Josep Sulé
authors.initials: JS
authors.lastname: Suso
authors.forename: Alicia
authors.initials: A
authors.lastname: El Haj
authors.forename: Ying
authors.initials: Y
authors.lastname: Yang
mesh_terms: D000818:Animals; D001842:Bone and Bones / Q000166:cytology* / Q000502:physiology*; D002113:Calcification, Physiologic; D002449:Cell Aggregation; D018929:Cell Culture Techniques; D002460:Cell Line; D051379:Mice; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D023822:Tissue Engineering / Q000379:methods*