title: In situ atomic force microscopy of partially demineralized human dentin collagen fibrils.
abstract: Dentin collagen fibrils were studied in situ by atomic force microscopy (AFM). New data on size distribution and the axial repeat distance of hydrated and dehydrated collagen type I fibrils are presented. Polished dentin disks from third molars were partially demineralized with citric acid, leaving proteins and the collagen matrix. At this stage collagen fibrils were not resolved by AFM, but after exposure to NaOCl(aq) for 100-240 s, and presumably due to the removal of noncollagenous proteins, individual collagen fibrils and the fibril network of dentin connected to the mineralized substrate were revealed. High-aspect-ratio silicon tips in tapping mode were used to image the soft fibril network. Hydrated fibrils showed three distinct groups of diameters: 100, 91, and 83 nm and a narrow distribution of the axial repeat distance at 67 nm. Dehydration resulted in a broad distribution of the fibril diameters between 75 and 105 nm and a division of the axial repeat distance into three groups at 67, 62, and 57 nm. Subfibrillar features (4 nm) were observed on hydrated and dehydrated fibrils. The gap depth between the thick and thin repeating segments of the fibrils varied from 3 to 7 nm. Phase mode revealed mineral particles on the transition from the gap to the overlap zone of the fibrils. This method appears to be a powerful tool for the analysis of fibrillar collagen structures in calcified tissues and may aid in understanding the differences in collagen affected by chemical treatments or by diseases.
authors.affiliation: Department of Preventive and Restorative Dental Sciences, University of California, 707 Parnassus Avenue D-2260, San Francisco, CA 94143, USA. shabeli@itsa.ucsf.edu
authors.forename: Stefan
authors.initials: S
authors.lastname: Habelitz
authors.forename: Mehdi
authors.initials: M
authors.lastname: Balooch
authors.forename: Sally J
authors.initials: SJ
authors.lastname: Marshall
authors.forename: Guive
authors.initials: G
authors.lastname: Balooch
authors.forename: Grayson W
authors.initials: GW
authors.lastname: Marshall
mesh_terms: D000328:Adult; D003094:Collagen / Q000737:chemistry*; D003804:Dentin / Q000648:ultrastructure*; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods*; D008963:Molar / Q000648:ultrastructure
chemical_list: D003094:Collagen