In Vivo Behavior of Hydroxyapatite Evaluated by Attenuated Total Reflection Infrared Microscopy (ATR-FTIR)

Abstract:

The study aimed the evaluation of the effect of osseous repair in the synthetic hydroxyapatite cylinder (HA) implanted in rabbit’s tibia. Fifteen adult animals, weighing around 2.5 and 3.0 Kg, sourced by the Fluminense Federal University Animal Lab were anesthetized with spinal anesthesia, and two perforations were created in right tibia and filled with cylinders (2 x 6 mm) of HA. The animals were killed after 28 days for evaluating the HA cylinder through attenuated total reflection infrared microscopy (ATR-FTIR). The two fragments of tibia with the cylinder were collected and fixed in alcohol 70° prior to dehydration in successive alcohol solutions and then impregnated and embedded in methyl-methacrylate. Not stained neither pasted to lamina, sections of 200 µm thickness were analyzed in a Shimadzu IRPrestige-21 Automatic Infrared Microscope (AIM-8800) with ATR Objective Mirror (ATR-8800M). Prior to implantation, HA cylinder was characterized by X-Ray diffraction (XRD), Attenuated Total Reflection- Infra Red microscopy (ATR-FTIR) and X-Ray Fluorescence (XRF). After the implantation, the spectrum of the ATR-FTIR at the interface to neoformed bone was acquired. The hydroxyapatite synthesized was crystalline and stoichiometric with Ca/P ratio of 1.66, becoming carbonated after 28 days of implantation. We conclude that attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) is a useful tool for biomaterial evaluation and able to detect the modification of the chemical HA cylinder pattern occurred probably by ions migration in the biological environment.

• Periodical: Key Engineering Materials (Volumes 396-398)
• Pages: 61-64
• DOI: https://doi.org/10.4028/www.scientific.net/KEM.396-398.61
• Online since: October 2008
• Authors: Monica Calasans-Maia, Silvia R. A. Santos, Antonella M. Rossi, José Mauro Granjeiro
• Keywords: ATR, FT-IR, Hydroxyapatite (HAP), Rabbit Tibia

References

[1] E.A. Munhoz, J.O. Ferreira, R.Y. Yaedu, J.M. Granjeiro. Radiographic assessment of impacted mandibular third molar sockets filled with composite xenogenic bone graft. Dentomaxillofac Radiol. Vol. 35(2006), p.371.

DOI: 10.1259/dmfr/64880289

[2] R.C. Oliveira, M.C. Sicca, T.L. Silva, T.M. Cestari, D.T. Oliveira, M.A.R. Buzalaf, R. Taga, E.M. Taga, J.M. Granjeiro. Effect of deproteinization temperature on the preparation of microgranular bovine cortical bone. Microscopic and biochemical analysis in rat subcutaneous tissue. J Appl Oral Sci (formerly Rev Fac Odontol Bauru) Vol. 7 (1999).

DOI: 10.1590/s1678-77572005000400013

[3] J.T. Sanada, J.G.R. Rodrigues, G.C. Canova, T.M. Cestari, E.M. Taga, R. Taga, M.A.R. Buzalaf, J.M. Granjeiro. Histologic, radiographic and imunoglobuline profile analysis after implantation blocks of demineralized bovine cancellous bone graft in muscle of rats. J Appl Oral Sci Vol. 11 (2003).

DOI: 10.1590/s1678-77572003000300010

[4] T. Accorsi-Mendonça, M.B. Conz, T.C. Barros. et al. Physicochemical characterization of two deproteinized bovine xenografts. Braz Oral Res, Vol. 22 (2008), pp.5-10.

DOI: 10.1590/s1806-83242008000100002

[5] A. Uschida, N. Araki, Y. Shinto, H. Yoshikawa, E. Kurisaki, K. Ono. The use of calcium hydroxyapatite ceramic in bone human surgery. J Bone Joint Surg Br. Vol. 72(1990), p.298302.

DOI: 10.1302/0301-620x.72b2.2155908

[6] A. S. B. Moreira, M. T. Pastoreli, L. H. F. Damasceno, H. L. A. Estudo experimental da influência das dimensões dos grânulos de hidroxiapatita na integração óssea. Acta Ortop Bras. Vol. 11 (2003), pp.240-250.

DOI: 10.1590/s1413-78522003000400007

[7] D. McNaughton. Synchrotron Infrared spectroscopy in Biology and Biochemistry. Australian Biochemist. Vol. 36 (2005), pp.55-58.

[8] L. Brancaleon, M. P. Bamberg, T. Sakamaki, and N. Kollias. Attenuated Total RefectionFourier Transform Infrared Spectroscopy as a Possible Method to Investigate Biophysical Parameters of Stratum Corneum In Vivo. The Journal of Investigative Dermatology. Vol. 116 (2001).

DOI: 10.1046/j.1523-1747.2001.01262.x

[9] J.A. Mendez, A. González-Couchón, M. Salvado, F. Collia, J.A. de Pedro, B. Levenfeld, M. Fernández, B. Vasquez and J. San Román. In vitro and in vivo Behavior of Bioactive Glass Composite Bearing a NSAID. Key Engineering Materials. Vol. 254-256 (2004).

DOI: 10.4028/www.scientific.net/kem.254-256.177

[10] F. Picard, T. Buffeteau, B. Desbat, MAuger, M. Pezolet. Biophysical Journal. Vol. 76 (1999), pp.539-551. Acknowledgements Special thanks to Fluminense Federal University, CNPq, FINEP, FAPERJ and Capes by the financial support.