Quantification of Periapical Bone Destruction in Mice by Micro-computed Tomography

Abstract

Bacterial infections of the dental pulp result in tissue destruction and periapical bone resorption. The availability of genetically engineered mouse strains is a major advantage in the use of this model system for studies of periapical pathogenesis. The main limitation of the mouse model is its small size, and the necessity for laborious histologic analyses to quantify periapical bone destruction. In the present study, we evaluated the use of a new technology, high-resolution micro-computed tomography (micro-CT), for the rapid and non-invasive quantification of periapical bone destruction. Periapical lesions were induced in the lower first molars of mice by exposing the pulp to the oral environment. Mandibles were harvested on day 21 after pulp exposure, and were subjected to micro-CT analysis, with 17-μm-thick radiographic sections. Samples were then decalcified, embedded, and sectioned for histology. The cross-sectional area of periapical lesions was determined by image analysis of corresponding micro-CT and histologic sections. The results showed a highly significant correlation between micro-CT and histology (p < 0.0001), with mean differences of 4.1% (range, 0.9 to 7.2%) between the two methods. The mean error associated with image analysis was 4.9% for images obtained by both micro-CT and histology. The variability of replicate (n = 5) independent micro-CT determinations was 3.4%, less than that associated with the image analysis error. These results demonstrate that micro-CT imaging is a rapid, reproducible, and non-invasive method, that gives results that are closely comparable with those obtained by histology. Micro-CT appears to have utility for the accurate quantification of changes in bone architecture in small biological specimens.

Keywords

micro-computed tomography bone resorption periapical lesion.

Get full access to this article

View all access options for this article.

References

Bender IB , Seltzer S. (1961). Roentgenographic and direct observation of experimental lesions in bone. J Am Dent Assoc 62:708-716.

Bergenholtz G. (1990). Pathogenic mechanisms in pulpal disease. J Endodont 16:98-101.

Bodner L. , Bar-Ziv J. (1996). Radiographic features of central giant cell granuloma of the jaws in children. Pediatr Radiol 26:148-151.

Bodner L. , Sarnat H. , Bar-Ziv J. , Kaffe I. (1996). Computed tomography in pediatric oral and maxillofacial surgery. J Dent Child 63:32-38.

Bonse U. , Busch F. , Günnewig 0 , Beckmann F. , Pahl R. , Delling G. , et al. (1994). 3D computed X ray tomography of human cancellous bone at 8 μm spatial and 10-4 energy resolution. Bone Miner 25:25-38.

Cardoso AS , Mitchell DF (1971). Progression of pulpitis to necrosis and periapical disease in deciduous and permanent teeth of monkeys. J Dent Res 50:934-938.

Engelke K. , Graeff W. , Meiss L. , Hahn M. , Delling G. (1993). High spatial resolution imaging of bone mineral using computed microtomography. Comparison with microradiography and undecalcified histologic sections. Invest Radiol 28:341-349.

Erausquin J. , Muruzabal M. (1968). Tissue reaction to root canal cements in the rat molar. Oral Surg 26:360-373.

Feldkamp LA , Goldstein SA , Parfitt AM , Jesion G. , Kleerekoper M. (1989). The direct examination of three-dimensional bone architecture in vitro by computed tomography. J Bone Miner Res 4:3-11.

10.

Kinney JH , Lane NE , Haupt DL (1995). In vivo, three-dimensional microscopy of trabecular bone. J Bone Miner Res 10:264-270.

11.

Krennmair G. , Lenglinger F. (1995). Imaging of mandibular cysts with a dental computed tomography software program. Int J Oral Maxillofac Surg 24:48-52.

12.

Kullendorff B. , Grondahl K. , Rohlin M. , Henrikson CO (1988). Subtraction radiography for the detection of periapical bone lesions. Endodont Dent Trauma 4:253-259.

13.

Li XJ , Muller R. , Stevens ML , Golden J. , Seeherman H. , Bouxstein ML (1998). Evaluation of trabecular bone morphometry: comparison of micro-computed tomography and histomorphometry in the rat proximal tibia (abstract). Bone 23:SA 465.

14.

Minami M. , Kaneda T. , Yamamoto H. , Ozawa K. , Itai Y. , Ozawa M. , et al. (1992). Ameloblastoma in the maxillomandibular region-MR imaging. Radiology 184:389-393.

15.

Minami M. , Kaneda T. , Ozawa K. , Yamamoto H. , Itai Y. , Ozawa M. , et al. (1996). Cystic lesions of the maxillomandibular region: MR imaging distinction of odontogenic keratocysts and ameloblastomas from other cysts. Am J Roentgenol 166:943-949.

16.

Muller R. , Ruegsegger P. (1997). Micro-tomographic imaging for the nondestructive evaluation of trabecular bone architecture. In: Bone research in biomechanics. Lowet G , Ruegsegger P , Weinans H , Meunier A , editors. Amsterdam: IOS Press, pp. 61-79.

17.

Muller R. , Hildebrand T. , Hauselmann HJ , Ruegsegger P. (1996). In vivo reproducibility of three-dimensional structural properties of noninvasive bone biopsies using 3D-pQCT. J Bone Miner Res 11:1745-1750.

18.

Muller R. , Van Campenhout H. , Van Damme B. , Van Der Perre G. , Dequeker J. , Hildebrand T. , et al. (1998). Morphometric analysis of human bone biopsies: a quantitative structural comparison of histologic sections and micro-computed tomography. Bone 23:59-66.

19.

rstavik D. (1991). Radiographic evaluation of apical periodontitis and endodontic treatment results: a computer approach. Int Dent J 41:89-98.

20.

rstavik D. , Farrants G. , Wahl T. , Kerekes K. (1990). Image analysis of endodontic radiographs: digital subtraction and quantitative densitometry. Endodont Dent Trauma 6:6-11.

21.

Ramadan A. , Mitchell D. (1962). A Roentgenographic study of experimental bone destruction. Oral Surg Oral Med Oral Pathol 15:934-943.

22.

Ruegsegger P. , Koller B. , Muller R. (1996). A microtomographic system for the nondestructive evaluation of bone architecture. Calcif Tissue Int 58:24-29.

23.

Serota K. (1981). Detection of periapical lesions in rats using radiopharmaceuticals (MS thesis). Boston, MA: Harvard School of Dental Medicine.

24.

Sorenson JA , Cameron JR (1967). A reliable in vivo measurement of bone mineral content. J Bone Jt Surg 49:481-497.

25.

Stashenko P. , Yu SM , Wang CY ( 1992). Kinetics of immune and cytokine resposes to endodontic infections. J Endodont 18:422-428.

26.

Teles R. , Wang CY , Stashenko P. (1997). Increased susceptibility of RAG-2 SCID mice to dissemination of endodontic infections. Infect Immun 65:3781-3787.

27.

Tyndall DA , Kapa SF , Bagnell CP (1990). Digital subtraction radiography for detecting cortical and cancellous bone changes in the periapical region. J Endodont 16:173-178.

28.

Wang CY , Stashenko P. (1991). Kinetics of bone-resorbing activity in induced periapical lesions. J Dent Res 70 :1362-1366.

29.

Wenzel A. , Grondahl HG (1995). Direct digital radiography in the dental office . Int Dent J 119:27-34.

30.

White SC , Gratt BM (1979). Clinical trials of intraoral dental xeroradiography . J Am Dent Assoc 99:810-816.

31.

Yu SM , Stashenko P. (1987). Identification of inflammatory cells in developing rat periapical lesions. J Endodont 13:535-540.