Electron Microprobe Analysis in Guided Tissue Regeneration: A Case Report

ABSTRACT

Objectives: Several procedures have been advocated as regenerative procedures in periodontology, but one of the most widely used techniques up to now is guided tissue regeneration (GTR). Likewise, different assessment methods based on clinical, radiographic or histological measurements have been proposed for the evaluation of these regenerative procedures. However, none of the methods used for human material incorporates quantitative X-ray microanalysis to assess the degree of mineralization of the regenerated periodontal hard tissues. The objective of this report was to evaluate, using quantitative X-ray microprobe analysis, the newly-formed hard tissue in a periodontal infrabony defect.

Methods: Electron microprobe analysis was used to study the nature of the newly-formed hard tissue 3 years after treatment with guided tissue regeneration in a patient with localized aggressive periodontitis.

Results: Our quantitative analyses, using the peak-to-background method, showed calcium/phosphorus mass ratio of 1.50±0.38 in the newly-formed hard tissue around the affected tooth root.

Conclusion: Quantitative X-ray microprobe analysis is a useful tool that may provide an accurate assessment of the degree of mineralization in an extremely small tissue sample. (Eur J Dent 2007;1:40- 44)

Key words

Guided tissue regeneration - Bone regeneration - Periodontal regeneration - Quantitative X-ray microprobe analysis - Mineralization

• References

• 1 American Academy of Periodontology. Glossary of Periodontal Terms. 4th Edition. Chicago: Illinois; The American Academy of Periodontology 2001: 44
• 2 Isidor F, Karring T, Nyman S, Lindhe J. New attachmentreattachment following reconstructive periodontal surgery. J Clin Periodontol 1985; 12: 728-735
• 3 Lynch SE. Methods for evaluation of regenerative procedures.. J Periodontol 1992; 63: 1085-1092
• 4 Isaka J, Ohazama A, Kobayashi M. et al Participation of periodontal ligament cells with regeneration of alveolar bone. J Periodontol 2001; 72: 314-323
• 5 Marks SCJr, Miller SC. Local delivery of prostaglandin E1 induces periodontal regeneration in adult dogs. J Periodontal Res 1994; 29: 103-108
• 6 Ripamonti U, Reddi AH. Tissue engineering, morphogenesis, and regeneration of the periodontal tissues by bone morphogenetic proteins.. Crit Rev Oral Biol Med 1997; 8: 154-163
• 7 Murakami S, Takayama S, Ikezawa K. et al Regeneration of periodontal tissues by basic fibroblast growth factor.. J Periodontal Res 1999; 34: 424-430
• 8 Karring T. Regenerative periodontal therapy. J Int Acad Periodontol 2000; 2: 101-109
• 9 Cochran DL, King GN, Schoolfield J, Velasquez-Plata D, Melloning JT, Jones A. The effect of enamel matrix proteins on periodontal regeneration as determined by histological analyses.. J Periodontol 2003; 74: 1043-1055
• 10 Pontoriero R, Lindhe J, Nyman S, Karring T, Rosenberg E, Sanavi R. Guided tissue regeneration in degree II furcation involved mandibular molars. J Clin Periodontol 1988; 15: 247-254
• 11 Aukhil I, Pettersson E, Suggs C. Guided tissue regeneration. An experimental procedure in beagle dogs.. J Periodontol 1986; 57: 727-734
• 12 Gottlow J, Nyman S, Lindhe J, Karring T, Wennstrom J. New attachment formation in the human periodontium by guided tissue regeneration. Case reports. J Clin Periodontol 1986; 6: 604-616
• 13 Mardas N, Kostopoulos L, Stavropoulos A, Karring T. Osteogenesis by guided tissue regeneration and demineralized bone matrix. J Clin Periodontol 2003; 30: 176-183
• 14 Silvestri M, Sartori S, Rasperini G, Ricci G, Rota C, Cattaneo V. Comparison of infrabony defects treated with enamel matrix derivative versus guided tissue regeneration with a nonresorbable membrane. J Clin Periodontol 2003; 30: 386-393
• 15 Christgau M, Schmalz G, Reich E, Wenzel A. Clinical and radiographical split-mouth-study on resorbable versus non-resorbable GTR-membranes. J Clin Periodontol 1995; 22: 306-315
• 16 Lekovic V, Klokkevold PR, Kenney EB, Dimitrijelic B, Nedic M, Weinlaender M. Histologic evaluation of guided tissue regeneration using 4 barrier membranes: a comparative furcation study in dogs.. Periodontol 1998; 69: 54-61
• 17 Unsal B, Ozcan G, Tuter G, Kurtis B, Yalim M. Evaluation of initial attachment of human gingival fibroblast cells to biodegradable membranes in vitro by light and scanning electron microscopy.. J Oral Sci 1999; 41: 57-60
• 18 Schantz JT, Hutmacher DW, Ng KW, Khor HL, Lim MT, Teoh SH. Evaluation of a tissue-engineered membranecell construct for guided bone regeneration. Int J Oral Maxillofac Implants 2002; 17: 161-174
• 19 Campos A, González-Jaranay M, Moreu G, Sánchez-Quevedo MC. Electron microprobe analysis in periodontal guided tissue regeneration.. Cell Biol Int 1993; 17: 695-696
• 20 Boyde A, Maconnachie E, Reid SA, Delling G, Mundy GR. Scanning electron microscopy in bone pathology: review of methods, potential and applications. Scann Electron Microsc 1986; 4: 1537-1564
• 21 Sánchez-Quevedo MC, Crespo PV, García JM, Campos A. Xray microanlytical histochemistry of human circumpulpar and mantle dentine. Bone Miner 1989; 6: 323-329
• 22 Zambrano M, Nikitakis NG, Sanchez-Quevedo MC, Sauk JJ, Sedano H, Rivera H. Oral and dental manifestations of vitamin D-dependent rickets type I: Report of a pediatric case. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2003; 95: 705-709
• 23 Armitage GC. Development of a classification system for periodontal diseases and conditions. Ann Periodontol 1999; 4: 1-6
• 24 Warley A. Standards for the application of X-ray microanalysis to biological specimens. J Microscopy 1990; 157: 135-147
• 25 Warley A. X-ray microanalysis for biologists. London: Portland Press; 1977: 0
• 26 O’Leary TJ, Drake RB, Naylor JE. The plaque control record. J Periodontol 1972; 43: 38
• 27 Clark DC, Chin Quee T, Bergeron MJ, Chan EC, Lautar- Lemay C, de Gruchy K. Reliability of attachment level measurements using the cementoenamel junction and a plastic stent. J Periodontol 1987; 58: 115-118
• 28 Bowers GM, Chadroff B, Carnevale R. et al Histologic evaluation of new attachment apparatus formation in humans.. J Periodontol 1989; 60: 664-674
• 29 Statham PJ, Pawley JB. A new method for particle X-ray microanalysis on peak to background measurements. Scann Electron Microsc 1978; 1: 469-478
• 30 Small JA, Heinrich KFJ, Newbury DE, Myklebust RL. Progress in the development of the peak-to-background method for the quantitative analysis of single particles with electron probe. Scann Electron Microsc 1979; 11: 807-816
• 31 Campos A, Rodríguez IA, Sánchez-Quevedo MC, García JM, Nieto-Albano OH, Gómez de Ferraris ME. Mineralization of human premolar occlusal fissures. A quantitative histochemical microanalysis.. Histol Histopathol 2000; 15: 499-502..
• 32 López-Escámez JA, Crespo PV, Cañizares F, Campos A. Standards for quantification of element in the otolithic membrane by electron probe x-ray microanalysis: calibration curves and electron beam sensitivity.. J Micros 1993; 171: 215-222
• 33 López-Escámez JA, Cañizares FJ, Crespo PV, Baeyens JM, Campos A. Electron probe microanalysis of gentamicininduced changes on ionic composition of the vestibular gelatinous membrane. Hear Res 1994; 76: 60-66.
• 34 López-Escámez JA, Campos A. Standards for X-ray microanalysis of calcified structures.. Scann Electron Microsc 1994; 8: 171-185..
• 35 Sánchez-Quevedo MC, Ceballos G, García JM, Rodríguez IA, Gómez de Ferraris ME, Campos AE. Scanning electron microscopy and calcification in amelogenesis imperfecta in anterior and posterior human teeth.. Histol Histopathol 2001; 16: 827-832
• 36 Sánchez-Quevedo MC, Ceballos G, García JM, Luna JD, Rodriguez IA, Campos AE. Dentine structure and mineralization in hypocalcified amelogenesis imperfecta: a quantitative X-ray histochemical study.. Oral Dis 2004; 10: 94-98
• 37 Hall TA, Anderson HC, Appleton TC. The use of thin specimens for X-ray microanalysis in biology.. J Microsc 1973; 99: 177-182.
• 38 Armstrong JT. Quantitative elemental analysis of individual microparticles with electron beam instruments. In: Heinrich KFJ, Newbury DE, eds. Electron probe quantitation. New York. Plenum Press 1991; 261: 315
• 39 Glimcher MJ. The nature of the mineral component of bone and the mechanism of calcification. In: Coe FL, Favus MJ, eds. Disorders of bone and mineral metabolism. New York. Raven Press Ltd 1992; 265: 286