Self-crosslinked fibrous collagen/chitosan blends: Processing, properties evaluation and monitoring of degradation by bi-fluorescence imaging.

Porous collagen/chitosan scaffolds with different Collagen:Chitosan (Coll:Ch) ratios were prepared by freeze-drying followed by self-crosslinking via dehydrothermal treatment (DHT) and characterized as biomaterials for tissue engineering. Cy7 and Cy5.5 fluorochromes were covalently grafted to collagen and chitosan, respectively. Thus, it was possible, using optical fluorescence imaging of the two fluorochromes, to simultaneously track their in vivo biodegradation, in a blend scaffold form. The fluorescence signal evolution, due to the bioresorption, corroborated with histological analysis. In vitro cytocompatibility of Coll:Ch blend scaffolds were evaluated with standardized tests. In addition, the scaffolds showed a highly interconnected porous structure. Extent of crosslinking was analyzed by convergent analysis using thermogravimetry, Fourier Transform Infrared Spectroscopy and PBS uptake. The variations observed with these techniques indicate strong interactions between collagen and chitosan (covalent and hydrogen bonds) promoted by the DHT. The mechanical properties were characterized to elucidate the impact of the different processing steps in the sample preparation (DHT, neutralization and sterilization by ß-irradiation) and showed a robust processing scheme with low impact of Coll:Ch composition ratio.

3D MRI heart segmentation of mouse embryos.

MRI has become an effective tool for anatomical mice studies. Currently, embryologists study the development of mouse embryos in order to understand the mechanisms of human development. The aim of the research presented in this paper, is to develop a semi-automatic image segmentation framework based 3D deformable models to identify cardiac malformations which are a major cause of death in children. The segmentation systems have been used to segment 3D mouse embryos heart structures. Results on the ventricles and on the heart muscle are presented and compared with manually segmented models.

Measurement of knee cartilage thickness using MRI: a reproducibility study in a meniscectomized guinea pig model of osteoarthritis.

The in vivo precision (reproducibility) of quantitative MRI is of particular importance in osteoarthritis (OA) progression of small magnitude and response to therapy. In this study, three-dimensional high-resolution MRI performed at 7 T was used to assess the short-term reproducibility of measurements of mean tibial cartilage thickness in a meniscectomized guinea pig model of OA. MR image acquisition was repeated five times in nine controls (SHAM) and 10 osteoarthritic animals 3 months after meniscectomy (MNX), in vivo. The animals were then killed for histomorphometric assessment and correlation with the MRI-based measurements. Medial tibial cartilage thickness was measured on MR images using semi-automatic dedicated 3D software developed in-house. The reproducibility of measurements of cartilage thickness was assessed by five repeated MRI examinations with a short recovery delay between examinations (48 h). The computed coefficients of variation were 8.9% for the SHAM group and 8.2% for the MNX group. The coefficients of variation were compatible with expected thickness variations between normal and pathological animals. A positive agreement and significant partial correlation (Spearman r' = 0.74; P < 0.01) between the MRI and histomorphometric data was established. Three-dimensional high-resolution MRI is a promising non-invasive research tool for in vivo follow-up. This modality could be used for staging and monitoring therapy response in small-animal models of OA.

Non-invasive in vivo quantification of the medial tibial cartilage thickness progression in an osteoarthritis rabbit model with quantitative 3D high resolution micro-MRI.

OBJECTIVE: To develop a quantitative non-invasive in vivo three-dimensional (3D) high resolution (HR) micro-magnetic resonance imaging (microMRI) protocol to measure the medial tibial cartilage thickness (MT.ThC) in the normal rabbit and in the anterior cruciate ligament transection (ACLT) rabbit model of osteoarthritis and quantify the progression of MT.ThC. METHODS: The left knee of 10 control and 40 operated rabbits was imaged in vivo with a 7T microMRI system at 3 and 5 months after ACLT. A 3D fast low angle short (FLASH) fat-suppressed MRI protocol was implemented leading to 44x176 microm(3) spatial resolution and to 44 microm(3) isotropic voxel after cubic interpolation. Semi-automatic MT.ThC measurements were made in 3D, in four different locations, in vivo and longitudinally in both groups. At 5 months, gross macroscopy, visual analogical evaluation of the cartilage and histology were compared to the MR-based MT.ThC. RESULTS: At 3 and 5 months, the MT.ThC measured in the minimum interbone distance area was the thinnest MR-based MT.ThC. It was significantly lower in the operated group and among the four evaluated MT.ThC, it was the most discriminative between the normal and the operated groups (P<0.05). The MT.ThC measured in the minimum interbone distance area was also the most sensitive to change in the operated group (66.4% MT.ThC loss, P=0.003) while no significant changes were observed in the control group. CONCLUSION: Quantitative 3D HR microMRI allowed for non-invasive longitudinal MT.ThC measurements in four different locations in both the normal and the operated rabbits. We concluded the MT.ThC measured in the minimum interbone distance area reflected the severity of the disease and was the most effective to measure the progression of the medial tibial cartilage destruction.

In-vivo high-resolution X-ray microtomography for liver and spleen tumor assessment in mice.

The present study sought to validate the use of glycery1-2-oley-1,3-bis-[7-(3-amino-2,4,6-triiodophenyl)- heptanoate] (DHOG) contrast agent for mouse spleen tumor and liver metastasis imaging by high-resolution X-ray microtomography. Three groups of female nude mice were compared: controls (n = 5), and mice injected with 2.5 x 10(6) STC1 tumor cells in the spleen, imaged at 15 days (group G15, n = 5) and at 30 days (group G30, n = 5, of which one died before imaging). Micro-CT scans (X-ray voltage, 50 kVp; anode current, 200 microA; exposure time, 632 ms; 180 rotational steps resulting in 35 microm isotropic spatial resolution) were acquired at 0, 0.75, 2 and 4 h after i.v. injection of DHOG. CT number (Hounsfield units: HU) and contrast-to-noise ratios (CNR) were determined in three organs. Statistical analysis was performed by Mann-Whitney U-test. Contrast enhancement in normal spleen and liver increased, respectively to 1020 +/- 159 and 351 +/- 27 HU over baseline at 4 h, and 482 +/- 3 and 203 +/- 14 HU on day 6 after a single contrast injection. Automated three-dimensional reconstruction and modeling of the spleen provided accurate and quantifiable images. Spleen tumor and liver metastases did not take up DHOG, making them detectable in contrast to the increased signal in normal tissue. The smallest liver metastasis detected measured 0.3 mm in diameter. High-resolution X-ray micro-CT in living mice using DHOG contrast agent allowed visualization and volume quantification of normal spleen and of spleen tumor and its liver metastases.

Knee cartilage thickness measurements using MRI: a 4(1/2)-month longitudinal study in the meniscectomized guinea pig model of OA.

OBJECTIVE: The aim of this study was to follow, over a 4(1/2)-month period, the medial tibia cartilage thickness on a meniscectomy (MNX) guinea pig osteoarthritis (OA) model and to compare with control animals, using three-dimensional high-resolution magnetic resonance imaging (3D HR-MRI). METHODS: MRI experimentations were performed in vivo at 7 T on guinea pig knee joints. 3D HR-MR images were acquired in 60 controls (SHAM) and 45 osteoarthritic animals (MNX) at four time-points (15, 45, 90 and 135 days) after surgery. Medial tibial cartilage thickness was measured from MRI images using in-house dedicated 3D software followed by a statistical analysis. At each time-point 15 SHAM and 15 MNX animals were sacrificed for histomorphometric assessments. RESULTS: No significant difference of mean cartilage thickness between the groups was found at early stage (D45) using MRI; however, significant differences were found between the groups at D90 (P<0.001) and D135 (P<0.001). Histomorphometry data confirmed the pathological status of the animals and was well correlated with MRI at D15 (r=0.79, P<0.01), D45 (r=0.67, P<0.01), and D135 (r=0.39, P<0.05) for SHAM, and at D45 (r=0.63, P<0.01), and D135 (r=0.81, P<0.01) for MNX. CONCLUSION: Medial tibial cartilage measurement based on HR-MR images enables the monitoring of longitudinal cartilage thickness changes. This technique showed significant differences between SHAM and MNX as from D90 after surgery. It could be used as a noninvasive and reproducible tool to monitor therapeutic response in this OA model.