Micro CT visualization of silver nanoparticles in the middle and inner ear of rat and transportation pathway after transtympanic injection.

BACKGROUND: Silver nanoparticles (Ag NPs) displayed strong activities in anti-bacterial, anti-viral, and anti-fungal studies and were reportedly efficient in treating otitis media. Information on distribution of AgNPs in different compartments of the ear is lacking. OBJECTIVE: To detect distribution of Ag NPs in the middle and inner ear and transportation pathways after transtympanic injection. METHODS: Contrast effect of Ag NPs in the micro CT imaging was assessed in a phantom. AgNPs at various concentrations (1.85 mM, 37.1 mM, and 370.7 mM) were administered to rat middle ear using transtympanic injection and cadaver heads were imaged using micro CT at several time points. RESULTS: The lowest concentration of Ag NPs that could be visualized using micro CT was 37.1 mM. No difference was observed between the solvents, deionized H2O and saline. Ag NPs at 37.1 mM were visible in the middle ear on 7 d post-administration. Ag NPs at 370.7 mM generated signals in the middle ear, ossicular chain, round window membrane, oval window, scala tympani, and Eustachian tube for both 4 h and 24 h time points. A gradient distribution of Ag NPs from the middle ear to the inner ear was detected. The pathways for Ag NPs to be transported from the middle ear into the inner ear are round and oval windows. CONCLUSION: This study provided the imaging evidence that Ag NPs are able to access the inner ear in a dose-dependent manner after intratympanic administration, which is relevant to design the delivery concentration in the future clinic application in order to avoid adverse inner ear effect.

New insights to the role of aryl hydrocarbon receptor in bone phenotype and in dioxin-induced modulation of bone microarchitecture and material properties.

Bone is a target for high affinity aryl hydrocarbon receptor (AHR) ligands, such as dioxins. Although bone morphology, mineral density and strength are sensitive endpoints of dioxin toxicity, less is known about effects on bone microarchitecture and material properties. This study characterizes TCDD-induced modulations of bone tissue, and the role of AHR in dioxin-induced bone toxicity and for normal bone phenotype. Six AHR-knockout (Ahr(-/-)) and wild-type (Ahr(+/+)) mice of both genders were exposed to TCDD weekly for 10 weeks, at a total dose of 200µg/kgbw. Bones were examined with micro-computed tomography, nanoindentation and biomechanical testing. Serum levels of bone remodeling markers were analyzed, and the expression of genes related to osteogenic differentiation was profiled using PCR array. In Ahr(+/+) mice, TCDD-exposure resulted in harder bone matrix, thinner and more porous cortical bone, and a more compact trabecular bone compartment. Bone remodeling markers and altered expression of a number of osteogenesis related genes indicated imbalanced bone remodeling. Untreated Ahr(-/-) mice displayed a slightly modified bone phenotype as compared with untreated Ahr(+/+) mice, while TCDD exposure caused only a few changes in bones of Ahr(-/-) mice. Part of the effects of both TCDD-exposure and AHR-deficiency were gender dependent. In conclusion, exposure of adult mice to TCDD resulted in harder bone matrix, thinner cortical bone, mechanically weaker bones and most notably, increased trabecular bone volume fraction in Ahr(+/+) mice. AHR is involved in bone development of a normal bone phenotype, and is crucial for manifestation of TCDD-induced bone alterations.

Patient setup accuracy in DIBH radiotherapy of breast cancer with lymph node inclusion using surface tracking and image guidance.

Studying setup accuracy in breast cancer patients with axillary lymph node inclusion in deep inspiration breath-hold (DIBH) after patient setup with surface-guided radiotherapy (SGRT) and image-guided radiotherapy (IGRT). Breast cancer patients (N = 51) were treated (50 Gy in 25 fractions) with axillary lymph nodes within the planning target volume (PTV). Patient setup was initiated with tattoos and lasers, and further adjusted with SGRT. The DIBH guidance was based on SGRT. Orthogonal and/or tangential imaging was analyzed for residual position errors of bony landmarks, the breath-hold level (BHL), the skin outline, and the heart; and setup margins were calculated for the PTV. The calculated PTV margins were 4.3 to 6.3 and 2.8 to 4.6 mm before and after orthogonal imaging, respectively. The residual errors of the heart were 3.6 ± 2.2 mm and 2.5 ± 2.4 mm before and 3.0 ± 2.5 and 2.9 ± 2.3 mm after orthogonal imaging in the combined anterior-posterior/lateral and the cranio-caudal directions, respectively, in tangential images. The humeral head did not benefit from daily IGRT, but SGRT guided it to the correct location. We presented a slightly complicated but highly accurate workflow for DIBH treatments. The residual position errors after both SGRT and IGRT were excellent compared to previous literature. With well-planned SGRT, IGRT brings only slight improvements to systematic accuracy. However, with the calculated PTV margins and the number of outliers, imaging cannot be omitted despite SGRT, unless the PTV margins are re-evaluated.

Reproducibility and agreement of micro-CT and histomorphometry in human trabecular bone with different metabolic status.

The use of micro-computed tomography (micro-CT) to study bone microstructure is continuously increasing. Thus, it is important to ensure that micro-CT can differentiate healthy and pathological bone. This study aimed to determine whether the reproducibility of bone histomorphometry and micro-CT, and agreement between the techniques, vary in bone samples with different metabolic status. Iliac crest biopsies (n = 36) were obtained from healthy subjects (n = 10) and from patients with osteoporosis (OP) (n = 15) or renal osteodystrophy (ROD) (n = 11). Micro-CT and histomorphometry analyses were repeated twice. Results were analyzed in separate groups and after pooling the data. Bone histomorphometry detected generally known differences between the diseases, whereas micro-CT did not detect differences between normal and ROD samples as effectively. Repeated measurements for BV/TV, Tb.Th, Tb.N, and Tb.Sp exhibited linear correlation coefficients (ρ) of 0.87-0.92 [coefficients of variations (CV), 8.3-27.2%] for histomorphometry and of 0.66-0.94 (CV, 4.4-23.4%) for micro-CT. There were no significant differences in reproducibility among samples from different study groups. Correlations between BV/TV (micro-CT) and mineralized bone volume (Md.V/TV, histomorphometry) were weaker than between BV/TV (micro-CT) and BV/TV (histomorphometry). When comparing the techniques, BV/TV, Tb.Th, and Tb.N displayed moderate correlations (ρ = 0.39-0.62, P < 0.05), and the agreement for BV/TV was highest in OP samples. The agreement between the techniques using clinical bone samples was moderate. Especially, micro-CT was less effective than bone histomorphometry in differentiating ROD from normal samples. The reproducibility was not affected by the health status of bone. Histomorphometry is still needed in clinical practice to study the remodeling balance in bone, and the methods are complementary.

A tube-source X-ray microtomography approach for quantitative 3D microscopy of optically challenging cell-cultured samples.

Development and study of cell-cultured constructs, such as tissue-engineering scaffolds or organ-on-a-chip platforms require a comprehensive, representative view on the cells inside the used materials. However, common characteristics of biomedical materials, for example, in porous, fibrous, rough-surfaced, and composite materials, can severely disturb low-energy imaging. In order to image and quantify cell structures in optically challenging samples, we combined labeling, 3D X-ray imaging, and in silico processing into a methodological pipeline. Cell-structure images were acquired by a tube-source X-ray microtomography device and compared to optical references for assessing the visual and quantitative accuracy. The spatial coverage of the X-ray imaging was demonstrated by investigating stem-cell nuclei inside clinically relevant-sized tissue-engineering scaffolds (5x13 mm) that were difficult to examine with the optical methods. Our results highlight the potential of the readily available X-ray microtomography devices that can be used to thoroughly study relative large cell-cultured samples with microscopic 3D accuracy.

Gas-foamed poly(lactide-co-glycolide) and poly(lactide-co-glycolide) with bioactive glass fibres demonstrate insufficient bone repair in lapine osteochondral defects.

Deep osteochondral defects may leave voids in the subchondral bone, increasing the risk of joint structure collapse. To ensure a stable foundation for the cartilage repair, bone grafts can be used for filling these defects. Poly(lactide-co-glycolide) (PLGA) is a biodegradable material that improves bone healing and supports bone matrix deposition. We compared the reparative capacity of two investigative macroporous PLGA-based biomaterials with two commercially available bone graft substitutes in the bony part of an intra-articular bone defect created in the lapine femur. New Zealand white rabbits (n = 40) were randomized into five groups. The defects, 4 mm in diameter and 8 mm deep, were filled with neat PLGA; a composite material combining PLGA and bioactive glass fibres (PLGA-BGf); commercial beta-tricalcium phosphate (ß-TCP) granules; or commercial bioactive glass (BG) granules. The fifth group was left untreated for spontaneous repair. After three months, the repair tissue was evaluated with X-ray microtomography and histology. Relative values comparing the operated knee with its contralateral control were calculated. The relative bone volume fraction (∆BV/TV) was largest in the ß-TCP group (p ≤ 0.012), which also showed the most abundant osteoid. BG resulted in improved bone formation, whereas defects in the PLGA-BGf group were filled with fibrous tissue. Repair with PLGA did not differ from spontaneous repair. The PLGA, PLGA-BGf, and spontaneous groups showed thicker and sparser trabeculae than the commercial controls. We conclude that bone repair with ß-TCP and BG granules was satisfactory, whereas the investigational PLGA-based materials were only as good as or worse than spontaneous repair.

X-ray microtomography of collagen and polylactide scaffolds in liquids.

Methods to image and assess the microstructure of polymer based biomaterials in liquid phase, for example cell culture medium, are well warranted. X-ray microtomography could provide a mean to visualize and analyze such structures. However, the density of such polymers is close to that of water and hence the X-ray contrast is poor. Moreover, if the biomaterials contain cells and are dried, the cell morphology may be distorted. In this paper we test phosphotungstic acid (PTA) staining to improve the contrast. We imaged collagen and PLA samples with µCT in air, water and alcohol. The methods were compared visually and with contrast to noise ratio calculated from the images. Our results demonstrate that with alcohol the PLA can be imaged also in liquid phase. PTA staining seems to be a good method to increase the contrast for collagen in µCT imaging.

X-ray microtomographic confirmation of the reliability of CBCT in identifying the scalar location of cochlear implant electrode after round window insertion.

Cone-beam computed tomography (CBCT) plays a key role in cochlear implantation in both planning implantation before surgery and quality control during surgery due to the high spatial resolution and convenience of application in the operation theater. We recently designed a novel, highresolution cone-beam acquisition system that has been tested in temporal bones with cochlear implantation to identify the scalar localization of the electrode arrays. The current study aimed to verify the reliability of the experimental CBCT set-up using high-resolution in vitro X-ray microtomography (µCT) imaging as a reference. Nine human temporal bones were studied by inserting a straight electrode of a cochlear implant using the round window approach followed by sequential imaging using experimental CBCT and µCT with and without 1% iodine as the contrast agent. In the CBCT images, the electrodes were located in the scala tympani and near the lateral wall in all temporal bones. In the µCT images, the cochlear fine structures, including Reissner's membrane, stria vascularis, spiral ligament, basilar membrane, spiral limbus, osseous spiral lamina, and Rosenthal's canal that hosts the spiral ganglion cells, were clearly delineated; the electrode array avoided the lateral wall of the scala tympani in the hook region and then ran along the lateral wall of the scala tympani without any exception, a feature that was also detected in a temporal bone with ruptures in the basilar and Reissner's membranes. In conclusion, the current in vitro µCT imaging system produced high-quality images that could demonstrate the fine cochlear structures faithfully and verify the reliability of a novel experimental CBCT set-up aimed for clinical application in identifying the scalar localization of the electrode array. The straight electrode is safe for cochlear structures with low risk of translocation and is suitable for atraumatic implantation, although a large gap between the contacts and the modiolus exists.

Delayed Computed Tomography Arthrography of Human Knee Cartilage In Vivo.

OBJECTIVE: We investigated the feasibility of delayed computed tomography (CT) arthrography for evaluation of human knee cartilage in vivo. Especially, the diffusion of contrast agent out of the joint space and the optimal time points for imaging were determined. DESIGN: Two patients were imaged using delayed CT arthrography and delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) techniques. RESULTS: Two hours after injection, the concentration of contrast agent in the joint space was still high enough (20% to 24.5% of the initial concentration at 0 minutes) to allow delayed CT arthrography. The half-life of the contrast agent in the joint space varied from 30 to 60 minutes. The contrast agent concentration in patellar and femoral cartilage reached the maximum after 30 and 60 minutes, respectively. According to dGEMRIC, there were no differences between patients. However, in delayed CT arthrography, the penetration of the contrast agent was higher in the osteoarthritic knee cartilage. CONCLUSIONS: Contrast agent remained in the joint space long enough to enable delayed CT arthrography of cartilage. After 30 minutes, the normalized contrast agent concentration was higher in the cartilage of the osteoarthritic knee in comparison with the healthy knee. To conclude, delayed CT arthrography exhibited potential for use in the clinical evaluation of cartilage integrity.