Study of tissue engineered bone nodules by Fourier transform infrared spectroscopy

摘要

The key criteria for assessing the success of bone tissue engineering are the quality and quantity of thenproduced minerals within the cultured constructs. The accumulation of calcium ions and inorganicnphosphates in culture medium serves as nucleating agents for the formation of hydroxyapatite, which isnthe main inorganic component of bone. Bone nodule formation is one of the hallmarks ofnmineralization in such cell cultures. In this study, we developed a new two-step procedure to acceleratenbone formation in which mouse bone cell aggregates were produced first on various chemically treatednnon-adhesive substrates. After this step, the bone cells’ growth and mineralization were followed innconventional culture plates. The number and size of cell aggregates were studied with light microscopy.nThe minerals’ formation in the form of nodules produced by the cell aggregates and the bone crystalnquality were studied with Fourier Transform Infrared (FTIR) spectroscopy. The FTIR spectra of thenash specimens (mineral phase only) from thermal gravimetric analysis (TGA) provided valuableninformation of the quality of the minerals. The y4 PO4 region (550–650 cmu00021), which reveals apatitic andnnon-apatitic HPO4 or PO4 environments, and phosphate region (910–1180 cmu00021) were examined for thenminerals produced in the form of nodules. The peak position and intensity of the spectra demonstratenthat the quality of the bone produced by cell aggregates, especially from the bigger ones, which werenformed on Plunoric treated substrates, exhibit a composition more similar to that of native bone. Thisnwork establishes a new protocol for high quality bone formation and characterization, with thenpotential to be applied to bone tissue engineering.