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Bone repair/regeneration is usually investigated through X-ray computed microtomography (μCT) supported by histology of extracted samples, to analyse biomaterial structure and new bone formation processes. Magnetic resonance imaging (μMRI) shows a richer tissue contrast than μCT, despite at lower resolution, and could be combined with μCT in the perspective of conducting non-destructive 3D investigations of bone. A pipeline designed to combine μMRI and μCT images of bone samples is here described and applied on samples of extracted human jawbone core following bone graft. We optimized the coregistration procedure between μCT and μMRI images to avoid bias due to the different resolutions and contrasts. Furthermore, we used an Adaptive Multivariate Clustering, grouping homologous voxels in the coregistered images, to visualize different tissue types within a fused 3D metastructure. The tissue grouping matched the 2D histology applied only on 1 slice, thus extending the histology labelling in 3D. Specifically, in all samples, we could separate and map 2 types of regenerated bone, calcified tissue, soft tissues, and/or fat and marrow space. Remarkably, μMRI and μCT alone were not able to separate the 2 types of regenerated bone. Finally, we computed volumes of each tissue in the 3D metastructures, which might be exploited by quantitative simulation. The 3D metastructure obtained through our pipeline represents a first step to bridge the gap between the quality of information obtained from 2D optical microscopy and the 3D mapping of the bone tissue heterogeneity and could allow researchers and clinicians to non-destructively characterize and follow-up bone regeneration., (Copyright © 2017 John Wiley & Sons, Ltd.)

Aims: It has been accepted that bone damage and subsequent resorption occurs in cases of adverse interfacial bone strain. Fractured implants represents an important opportunity for evaluating the bone response to overstrain. The objective of the study was to analyze the bone morphology and morphometry around retrieved fractured implants in humans. Five fractured implants retrieved after 22.4 (± 2.8) years were used in the present study. The investigation was conducted in a transmitted brightfield and circularly polarized light Microscope. The parameters considered were the total bone area, total bone remodeling area, bone-remodeling rate (BRR), and bone transverse collagen fibers orientation (CFO)., Findings: For grouped samples, the BRR was 51.9% (± 10) while the transverse CFO was 13.0% (± 9.7). The interthread BRR was 58.1% (± 4.0) for level 1 and 40.4% (± 12.8) for level 2 with a significant decrease in BRR (p = 0.019) as a function of depth along the implant length., Conclusions: Both bone level and bone quality attains stability to a certain level with respect to implant neck fracture following physiologic interaction between mechanical and biological inputs. The BRR significantly decrease as function of threads depth., (© 2014 Wiley Periodicals, Inc.)

Direct laser fabrication (DLF) allows solids with complex geometry to be produced by sintering metal powder particles in a focused laser beam. In this study, 10 Ti6Al4V alloy model dental root implants were obtained by DLF, and surface characterization was carried out using stereo scanning electron microscopy to produce 3D reconstructions. The surfaces were extremely irregular, with approximately 100 microm deep, narrow intercommunicating crevices, shallow depressions and deep, rounded pits of widely variable shape and size, showing ample scope for interlocking with the host bone. Roughness parameters were as follows: R(t), 360.8 microm; R(z), 358.4 microm; R(a), 67.4 microm; and R(q), 78.0 microm. Disc specimens produced by DLF with an identically prepared surface were used for biocompatibility studies with rat calvarial osteoblasts: After 9 days, cells had attached and spread on the DLF surface, spanning across the crevices, and voids. Cell density was similar to that on a commercial rough microtextured surface but lower than on commercial smooth machined and smooth-textured grit-blasted, acid-etched surfaces. Human fibrin clot extension on the DLF surface was slightly improved by inorganic acid etching to increase the microroughness. With further refinements, DLF could be an economical means of manufacturing implants from titanium alloys., ((c) 2008 Wiley Periodicals, Inc.)

This study is the first to investigate the collagen fiber orientation in human bone around titanium dental implants after loading. Birefringence under circularly polarized light (CPL) and scanning electron microscopy (SEM) investigation were used to assess both collagen orientation and density. Twenty osseointegrated dental implants were retrieved from the mandible bone of 10 patients and were used in the present study. The implants were retrieved after 6 months of function. The birefringence measurements were performed on digitized images using both the peri-implant bone (PB) and the alveolar bone (AB) retrieved at the moment of implant placing. All images detected at 100x were measured using a software image analysis. The results showed that 32.96% (3.208 +/- 0.435 mm(2)) of the PB area was composed of transverse collagen fibers while only 19.70% (1.957 +/- 0.253 mm(2)) was composed of longitudinally collagen fibers. In the AB 26.99% (2.620 +/- 0.520 mm(2)) of the examined area was composed of transverse collagen fibers while 22.25% (2.160 +/- 0.320 mm(2)) was composed of longitudinally collagen fibers. The CPL measurements of the birefringence indicated that the difference in area between the two collagen fibers orientations was statistically significant (p = 0.03) for both AB and PB. In PB the transverse collagen fibers were significantly increased (p < 0.01) than in AB; also the longitudinal collagen fibers differ significantly in the two groups (p = 0.02). The transverse collagen fibers in PB are more present under the lower flank of the threads where the load acts with a compression vectors, while outside the tip of the implant threads the collagen fibers run more longitudinally due to the loads that act with a tensile vectors. SEM observations showed, in the first collagen layers facing the implant surface, a random direction of the collagen fibers, while in the areas away from the implant surface the collagen fibers were well oriented and run parallel. In conclusion, in these loaded dental implants, the bone adjacent to the first two implant threads shows a significant increase of the amount of transverse collagen fibers that were mainly associated with the lower flank of the threads where compressive load acts. The longitudinal collagen fibers also differs significantly from those found in the AB., (Copyright 2005 Wiley Periodicals, Inc.)

The aim of the present study was a quantitative evaluation of the in vitro fibrin clot extension on different implant surfaces. Forty-five disk-shaped commercially pure Grade 2 titanium samples with three different surface topographies (machined, DPS, and Plus) were used in the present study. For the quantitative evaluation of the fibrin clot, 30 specimens were used (10 per group); human whole blood was employed. Venous blood was drawn from three healthy adult volunteers, and 0.2 mL were immediately dropped onto the surface of each specimen. Contact time was 5 min at room temperature; then the samples were rinsed with saline solution and fixed in a buffered solution of glutaraldehyde and paraformaldehyde. Samples were washed again with buffer and dehydrated in an ascending alcohol series. Specimens belonging to all groups were observed under SEM at a magnification of 1000x. From each sample, 50 random micrographs were collected in .tif format with an N x M 1024 x 768 grid of pixels. Quantitative analysis of fibrin clot extension showed the following results: in machined samples fibrin clot extension was 345987.2 +/- 63747.7 pixels(2) (mean +/- SD), in DPS samples fibrin clot extension was 375930.9 +/- 54726.86 pixels(2) (mean +/- SD), and in Plus samples, fibrin clot extension was 612333.6 +/- 46268.42 pixels(2) (mean +/- SD). With ANOVA it was possible to find that there were significant differences among the groups. The Tukey test revealed that the extension of the fibrin clot of Plus samples was statistically higher compared to both machined and DPS samples. The results of this in vitro study indicate that there is a correlation between implant surface morphology and fibrin clot extension. Improvement in surface microtexture complexity seems to determine the formation of a more extensive and three dimensionally complex fibrin scaffold. Further investigations are necessary to explain in more detail the mechanisms that regulate the fibrin clot formation on different implant surfaces., (Copyright 2005 Wiley Periodicals, Inc.)

Biological interactions occurring at the bone-biomaterial interface are critical for long-term clinical success. Bio-Oss is a deproteinized, sterilized bovine bone that has been extensively used in bone regeneration procedures. The aim of the present study was a comparative light, scanning, and electron microscopy evaluation of the interface between Bio-Oss and bone in specimens retrieved after sinus augmentation procedures. Under light microscopy, most of the particles were surrounded by newly formed bone, while in a few cases, at the interface of some particles it was possible to observe marrow spaces and biological fluids. Under scanning electron microscopy, in most cases, the particle perimeter appeared lined by bone that was tightly adherent to the biomaterial surface. Transmission electron microscopy showed that the bone tissue around the biomaterial showed all the phases of the bone healing process. In some areas, randomly organized collagen fibers were present, while in other areas, newly formed compact bone was present. In the first bone lamella collagen fibers contacting the Bio-Oss surface were oriented at 243.73 +/- 7.12 degrees (mean +/- SD), while in the rest of the lamella they were oriented at 288.05 +/- 4.86 degrees (mean +/- SD) with a statistically significant difference of 44.32 degrees (p < 0.001). In the same areas the intensity of gray value was 172.56 +/- 18.15 (mean +/- SD) near the biomaterial surface and 158.71 +/- 21.95 (mean +/- SD) in the other part of the lamella with an unstatistically significant difference of 13.79 (p = 0.071). At the bone-biomaterial interface there was also an electron-dense layer similar to cement lines. This layer had a variable morphology being, in some areas, a thin line, and in other areas, a thick irregular band. The analyses showed that Bio-Oss particles do not interfere with the normal osseous healing process after sinus lift procedures and promote new bone formation. In conclusion, this study serves as a better understanding of the morphologic characteristics of Bio-Oss and its interaction with the surrounding tissues., (Copyright 2005 Wiley Periodicals, Inc.)