The Venue, Aartselaar: See 37 unbiased reviews of The Venue, rated 3 of 5 on Tripadvisor and ranked #18 of Boomsesteenweg 92 Hoek Pierstraat, Aartselaar Belgium. Website Date of visit: September Great food, good drinks (prices are okay) and they have an amazing terrace when the sun is shining.
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However, the majority of the experimental reports lack details of the protocol applied for micro-CT image analysis.
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In particular, protocols aimed at quantifying the rodent jawbone have been poorly described and lack information, thereby impeding or even excluding the generation of the replicate data. Image acquisition and processing, especially segmentation and filtration due to the different software programmes, were often not clearly explained. In some studies, the ROIs used were not scaled to the dimensions of the bone size, causing inaccurate measurements of the trabecular bone and resulting in over-sampling between the compared studies.
This lack of consistency makes it very difficult to interpret the reported outcomes and to compare the findings across the different studies. Thus, there is a need to use standardized terminologies and report the parameters evaluated, particularly with respect to micro-CT analysis of rodent specimens. Moreover, the results can be biased if the procedures are not standardized, as the system is extremely user-sensitive.
Thus, the present paper proposes a methodology for standardized, accurate, and reproducible quantitative analysis of the rat jawbone microarchitecture using micro-CT imaging. It primarily focuses on the standardization of the procedures for evaluating trabecular bone in the rat jaw using both manual and semi-automatic delineation as well as the appropriate positioning of the volumes of interest to be analysed.
The main purpose was to define a robust protocol for the rodent jawbone based on the guidelines established by Bouxsein et al. Some important considerations to keep in mind while quantifying rodent jawbones are summarized as follows. The choice of voxel size is critical in order to make reliable measurements. Ideally, the smallest voxel size highest scan resolution is used for scanning; however, this results in longer acquisition times, as they need to collect more projections and create large data sets.
Thus, an appropriate trade-off between voxel size and scanning time should be taken into account. In such scenarios, the ratio of the voxels to object size is important, as the higher the ratio, the more accurate the morphologic measurements. This ensures that high-resolution imaging can be achieved through higher photon counts, thereby reducing time costs, which should not exceed the ratio of The ex vivo micro-CT applied in the present study has a voxel size of 7.
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In the current protocol, the ring reduction was set at 15 to remove artefacts caused by the scanning. Gaussian filtering using appropriate thresholding should be adopted to remove image noise. It is of utmost importance to compare the images of the original with the segmented ones to ensure that the extracted bone is representative of the original structure, as any error will systematically impact the results.
The protocol applied in the present case study made use of global thresholding, which employs a fixed range of greyscales lower and upper scales set at 70 and for both foreground white and the pixels outside of the range area, which are set as the background black. There are also other thresholding options available, such as adaptive, automatic, and multilevel thresholding; however, in the current protocol, since the region of interest comprises trabecular bone, the global thresholding option is able to achieve the desired image filtration.
Finally, v selecting the appropriate VOI at a particular site is essential. The contouring method used for the delineation of the bone can either be automatic as proposed in upgrades of the Skyscan software , a self-defined fixed ellipsoid or rectangular area that captures the bone of interest or an ROI with an irregular contour drawn manually on a slice-by-slice method.
For example, in the current methodology, the suture-forming elements between the maxilla and sphenoid as well as the mandibular canal the regular circular opening in the ROI of most jawed species is omitted during the analysis of the trabecular bone. This is because the sutures of the maxilla are not connected by bone and the mandibular canal contains a neuro-vascular bundle enclosed around a compact bone shell, which is devoid of trabecular bone.
Such considerations are very valuable to keep in mind during analysis, as it demonstrates the importance of the VOI selection and the risk of sampling across several adjacent bones. Thus, depending on the anatomic skeletal site of interest, the appropriate type of contouring method can be selected. Since some of the trabecular parameters are influenced by changes in VOI size and location, caution must be taken when interpreting the significant differences between the different trabecular parameters.
Thus, it is recommended that researchers try to understand the existing biases in their studies by testing the effect of different VOI sizes and location to accurately interpret the functional differences of trabecular parameters. The choice of VOI size and location should be guided by an understanding of the function of the bone for example, loading and the anatomical region in different species.
Trabecular analyses of irregularly shaped bones are often constricted by anatomy, and in these cases, it is often better to determine biomechanically rather than anatomically homologous VOIs. A limitation encountered while formulating the protocol involved the extraction of the cortical bone in the rodent jaw. The cortical bone was difficult to visualize due to its similarity in grey values with the dentin and enamel in the jawbone.
One way to measure the cortical bone was to include the whole mandible or maxilla, subsequently exclude the teeth, and then deduct the trabecular region that was measured earlier in the molar region. However, this proved to be difficult to implement due to the extraction of the teeth and roots in the molar region and the presence of the sinus and nasal cavities in the maxilla. The excision of irregularly shaped bones of the teeth and their roots from the image of interest proved to be arduous. It made it very challenging to acquire a standard and reproducibly shaped VOI for all specimens to be analysed and to obtain a consistent shape between the different animals.
These difficulties in measurements may explain the lack of studies that consistently quantify cortical bone changes in the rodent jaw.
Nevertheless, micro-CT and nano-CT cannot be used in a clinical setting to measure the structural parameters of the bone, and this is where cone beam computed tomography CBCT can help. Another preferred choice for image quantification is in vivo micro-CT. It can monitor bone changes that occur in real time over a period of several months. It also has the possibility to employ radiopaque contrast agents, which can enhance image visibility in different conditions, such as micro damage, 38 vascular morphology, 39 , 40 , 41 , 42 and cartilage degradation. Moreover, the key outcome of this study is the establishment of a method that is able to define the VOI in the rodent jawbone in a standardized and reproducible manner for the evaluation of trabecular bone morphology.
Ultimately, the protocol developed in this work provides a robust and reproducible methodology for micro-CT analysis of relevant rat jawbone ROIs and is intended to ensure the global, accurate, and consistent reporting of micro-CT-derived jawbone morphometry. By including various ROIs most susceptible to bone changes as a result of experimental interventions within the jawbone, one can have a wide range of ROIs to choose from, depending on the experimental study.