Micro-computed Tomography Reconstruction Research Articles

Cortico-cancellous osseointegration into additively manufactured titanium implants using a load-bearing femoral ovine model.

Introduction: Titanium-based implants can be used to fill voids in bone reconstruction surgery. Through additive manufacturing (AM), it is possible to produce titanium implants with osteoconductive properties such as high porosity and low stiffness. AM facilitates a level of design flexibility and personalization that is not feasible with traditional techniques. Methods: In this study, osseointegration into titanium alloy (Ti-6Al-4V) lattices was investigated for 12weeks post-implantation using a novel bicortical load-bearing ovine model. The objective was to assess the safety and efficacy of AM-fabricated implants using two lattice structures of contrasting stiffness spanning the full width of the femoral condyle. Results: This was achieved by evaluating implant osseointegration and bone-implant contact properties by histomorphometry, scoring local implant tissue responses via histopathology, and micro-computed tomography reconstruction. Discussion: We found that Ti-6Al-4V implants facilitated widespread and extensive osseointegration, with bone maturation ongoing at the conclusion of the trial period. Following the implantation period, no adverse clinical indications that could be directly ascribed to the presence of the implanted device were identified, as determined by macroscopic and microscopic observation.

Identification of Root Canal Morphology in Fused-rooted Mandibular Second Molars From X-ray Images Based on Deep Learning

Introduction: Understanding the intricate anatomical morphology of fused-rooted mandibular second molars (MSMs) is essential for root canal treatment. The present study utilized a deep learning approach to identify the three-dimensional root canal morphology of MSMs from two-dimensional X-ray images. MethodsA total of 271 fused-rooted MSMs were included in the study. Micro-computed tomography reconstruction images and two-dimensional X-ray projection images were obtained. The ground truth of three-dimensional root canal morphology was determined through micro-computed tomography images, which were classified into merging, symmetrical, and asymmetrical types. To amplify the X-ray image dataset, traditional augmentation techniques from the python package Augmentor and a multiangle projection method were employed. Identification of root canal morphology was conducted using the pretrained VGG19, ResNet18, ResNet50, and EfficientNet-b5 on X-ray images. The classification results from convolutional neural networks (CNNs) were then compared with those performed by endodontic residents. ResultsThe multiangle projection augmentation method outperformed the traditional approach in all CNNs except for EfficientNet-b5. ResNet18 combined with the multiangle projection method outperformed all other combinations, with an overall accuracy of 79.25%. In specific classifications, accuracies of 81.13%, 86.79%, and 90.57% were achieved for merging, symmetrical, and asymmetrical types, respectively. Notably, CNNs surpassed endodontic residents in classification performance; the average accuracy for endodontic residents was only 60.38% (P < .05). ConclusionsCNNs were more effective than endodontic residents in identifying the three-dimensional root canal morphology of MSMs. The result indicates that CNNs possess the capacity to employ two-dimensional images effectively in aiding three-dimensional diagnoses.

306 Intraoral Spectroscopy for the Identification and Study of Molar Hypomineralization

OBJECTIVES/GOALS: Molar hypomineralization (MH) is a highly prevalent dental disease that leads to rapid enamel decay even with preventative measures. While the harm is apparent, the etiology of the condition is not. Further, MH is difficult to study due to limited intact extracted teeth. Therefore, we need a method to study MH non-destructively and intraorally. METHODS/STUDY POPULATION: Recent work has shown excess proteins not commonly found in enamel are present in teeth with MH. This is theorized to be due to disruption and infiltration in the cells that form enamel leading to leftover protein and under mineralized enamel. We hypothesizethese proteins have specific spectroscopic signatures that can be detected using light. We further hypothesize that modern fiber optic probes can provide a method for non-destructive, pain-free, and rapid intraoral examination of MH. Initially, Micro Raman spectroscopy was used to detect specific vibrational bands associated with organics in teeth with MH followed by the collection of spectra of teeth with MH and heathy enamel using a spectrophotometer. These spectra were examined for any obvious differences. RESULTS/ANTICIPATED RESULTS: Currently 12 teeth were collected, and micro-computed tomography reconstructions confirmed location in 3D of MH lesions. Micro Raman of a MH-affected tooth revealed clear organic associated Amide I and III vibrational bands when compared to a synthetic hydroxyapatite (mineral in enamel) powder standard. We determined the wavelength of light that can be used to detect spectral differences between healthy enamel and teeth with MH. The next steps include optimization of the protocol of the intraoral spectrometer with the determined wavelength for implementation in clinic to allow for collection of spectra without the need for tooth extraction. DISCUSSION/SIGNIFICANCE: We hope that this work will lead to advancements in our understanding of the mechanism of MH as well as act as a proof of concept for a MH diagnostic tool. In the long term, the goal is to ideally lead to improvements in dental health care, decrease dental costs, and improve overall quality of life for the children with this condition.

Multiphase micro-computed tomography reconstructions provide dynamic respiratory function in a mouse lung fibrosis model

Micro-computed tomography derived functional biomarkers used in lung disease research can significantly complement end-stage histomorphometric measures while also allowing for longitudinal studies. However, no approach for visualizing lung dynamics across a full respiratory cycle has yet been described. Using bleomycin-induced lung fibrosis and the antifibrotic drug nintedanib as a test model, we implemented a four-dimensional (4D) micro-CT imaging approach consisting of 30 reconstructed volumes per respiratory cycle, coupled with deep-learning-assisted segmentation of lung volumes. 4D micro-CT provided an accurate description of inhalatory and exhalatory lung dynamics under resting conditions and revealed an inflammation-related obstructive pattern at day 7, followed by a restrictive pattern associated with fibrosis development at day 21. A milder restriction and fibrotic pathology resulted from nintedanib treatment. The similarity of 4D micro-CT data with those produced by diagnostic measurements, also points to its great potential as an exploratory tool for the discovery of clinically relevant therapeutic compounds.

Numerical simulations to determine the stimulation of the crista ampullaris during the Head Impulse Test

The Head Impulse Test, the most widely accept test to assess the vestibular function, comprises rotations of the head based on idealized orientations of the semicircular canals, instead of their individual arrangement specific for each patient. In this study, we show how computational modelling can help personalize the diagnosis of vestibular diseases. Based on a micro-computed tomography reconstruction of the human membranous labyrinth and their simulation using Computational Fluid Dynamics and Fluid-Solid Interaction techniques, we evaluated the stimulus experienced by the six cristae ampullaris under different rotational conditions mimicking the Head Impulse Test. The results show that the maximum stimulation of the crista ampullaris occurs for directions of rotation that are more aligned with the orientation of the cupulae (average deviation from alignment of 4.7°, 9.8°, and 19.4° for the horizontal, posterior, and superior maxima, respectively) than with the planes of the semicircular canals (average deviation from alignment of 32.4°, 70.5°, and 67.8° for the horizontal, posterior, and superior maxima, respectively). A plausible explanation is that when rotations are applied with respect to the center of the head, the inertial forces acting directly over the cupula become dominant over the endolymphatic fluid forces generated in the semicircular canals. Our results indicate that it is necessary to consider cupulae orientation to ensure optimal conditions for testing the vestibular function.

Modeling anabolic and antiresorptive therapies for fracture healing in a mouse model of osteogenesis imperfecta.

Osteogenesis imperfecta (OI) is a genetic bone fragility disorder that features frequent fractures. Bone healing outcomes are contingent on a proper balance between bone formation and resorption, and drugs such as bone morphogenetic proteins (BMPs) and bisphosphonates (BPs) have shown to have utility in modulating fracture repair. While BPs are used for OI to increase BMD and reduce pain and fracture rates, there is little evidence for using BMPs as local agents for fracture healing (alone or with BPs). In this study, we examined wild-type and OI mice (Col1a2+/G610C ) in a murine tibial open fracture model with (i) surgery only/no treatment, (ii) local BMP-2 (10 µg), or (iii) local BMP-2 and postoperative zoledronic acid (ZA; 0.1 mg/kg total dose). Microcomputed tomography reconstructions of healing fractures indicated BMP-2 was less effective in an OI setting, however,BMP-2 +ZA led to considerable increases in bone volume (+193% WT, p < 0.001; +154% OI, p < 0.001) and polar moment of inertia (+125% WT, p < 0.01; +248% OI, p < 0.05). Tissue histology revealed a thinning of the neocortex of the callus in BMP-2 treated OI bone, but considerable retention of woven bone in the healing callus with BMP + ZA specimens. These data suggest a cautious approach may be warranted with the sole application of BMP-2 in an OI surgical setting as a bone graft substitute. However, this may be overcome by off-label BPadministration.

Ancient mitochondrial genomes recovered from small vertebrate bones through minimally destructive DNA extraction: Phylogeography of the New Zealand gecko genus Hoplodactylus.

Methodological and technological improvements are continually revolutionizing the field of ancient DNA. Most ancient DNA extraction methods require the partial (or complete) destruction of finite museum specimens, which disproportionately impacts small or fragmentary subfossil remains, and future analyses. We present a minimally destructive ancient DNA extraction method optimized for small vertebrate remains. We applied this method to detect lost mainland genetic diversity in the large New Zealand diplodactylid gecko genus Hoplodactylus, which is presently restricted to predator-free island and mainland sanctuaries. We present the first mitochondrial genomes for New Zealand diplodactylid geckos, recovered from 19 modern, six historical/archival (1898-2011) and 16 Holocene Hoplodactylus duvaucelii sensu latu specimens, and one modern Woodworthia sp. specimen. No obvious damage was observed in post-extraction micro-computed tomography reconstructions. All "large gecko" specimens examined from extinct populations were found to be conspecific with extant Hoplodactylus species, suggesting their large relative size evolved only once in the New Zealand diplodactylid radiation. Phylogenetic analyses of Hoplodactylus samples recovered two genetically (and morphologically) distinct North and South Island clades, probably corresponding to distinct species. Finer phylogeographical structuring within Hoplodactylus spp. highlighted the impacts of Late Cenozoic biogeographical barriers, including the opening and closure of Pliocene marine straits, fluctuations in the size and suitability of glacial refugia, and eustatic sea-level change. Recent mainland extinction obscured these signals from the modern tissue-derived data. These results highlight the utility of minimally destructive DNA extraction in genomic analyses of less well studied small vertebrate taxa, and the conservation of natural history collections.

Craniomaxillofacial morphology in a murine model of ephrinB1 conditional deletion in osteoprogenitor cells

ObjectiveEFNB1 mutation causes craniofrontonasal dysplasia (CFND), a congenital syndrome associated with craniomaxillofacial anomalies characterised by coronal craniosynostosis, orbital hypertelorism, and midface dysplasia. The aim of this murine study was to investigate the effect of the EfnB1 conditional gene deletion in osteoprogenitor cells on the craniomaxillofacial skeletal morphology. DesignThe skulls of male and female mice, in which EfnB1 was deleted by Cre (a site-specific DNA recombinase) under the control of the Osterix (Osx) promoter (EfnB1OB-/-), were compared to those without EfnB1 deletion (Osx:Cre control) at two ages (4 and 8 weeks; n = 6 per group). The three-dimensional micro-computed tomography reconstructions were prepared to calculate 17 linear measurements in the cranial vault (brain box), midface and mandible. Coronal and sagittal sutures from the 8-week-old mice were also subjected to histological examination. ResultsEfnB1OB-/- mice displayed significantly larger cranial height, larger interorbital and nasal widths, smaller maxillary width than controls by 8 weeks (p < 0.05), but mandibular size was not significantly different (p > 0.05). Binomial testing showed significantly smaller EfnB1OB-/- skulls at 4 weeks but larger at 8 weeks (p < 0.05). Histological examination revealed increased bony fusion and fibrous connective tissue deposition at the coronal suture of EfnB1OB-/- mice compared with controls. ConclusionsCraniofacial phenotype of the murine model of EfnB1 deletion in osteoprogenitor cells partially represents the human CFND phenotype, with implications for better understanding mechanisms involved in skeletal morphogenesis and malocclusion.

AFRL Additive Manufacturing Modeling Series: Challenge 4, In Situ Mechanical Test of an IN625 Sample with Concurrent High-Energy Diffraction Microscopy Characterization

We describe 3D characterization of an additively manufactured Inconel 625 nickel-base superalloy specimen conducted during a uniaxial tension test using a suite of nondestructive x-ray techniques. High-energy diffraction microscopy in both near- and far-field modalities are employed in situ to track evolution of the material orientation and stress–strain fields at six points during the mechanical test, and these data streams are registered with micro-computed tomography reconstructions which probe the material density. This data volume was matched to a multi-modal serial sectioning characterization of the specimen taken after loading, described in this article’s companion. Twenty-eight grains which were monitored throughout the experiment were selected to form the basis for AFRL AM Modeling Series Challenge 4, Microscale Structure-to-Properties.

Effect of Trabecular Microstructure of Spinous Process on Spinal Fusion and Clinical Outcomes After Posterior Lumbar Interbody Fusion: Bone Surface/Total Volume as Independent Favorable Indicator for Fusion Success

We assessed the trabecular microarchitecture of the spinous process as an autograft and investigated its correlations with fusion success and clinical outcomes for patients undergoing posterior lumbar interbody fusion. Micro-computed tomography reconstruction techniques were used to scan cancellous bone specimens from spinous processes. We then measured the microarchitectural parameters for 105 subjects. The patient cohort included 44 older men and 61 postmenopausal women with a minimum of 2-year follow-up data available. The complete fusion rate was 87.6% (92 of 105) at the last follow-up. When stratified by fusion status, the union group had significantly greater bone surface/total volume (BS/TV) and trabecular number but significantly lower trabecular separation than the nonunion group. No statistically significant differences were observed between the 2 groups in the clinical variables, except for the bone mineral density at the femoral neck (P= 0.028). On binomial logistic regression analysis, BS/TV was identified as an independent predictor for fusion success (odds ratio, 8.532; P= 0.032). The receiver operating characteristic curve showed that BS/TV had excellent performance in predicting successful fusion (area under the curve, 0.807). Using a cutoff value for BS/TV of 3.145, a greater BS/TV was significantly associated with visual analog scale scores for lower back pain 6 months postoperatively and lower Oswestry disability index scores at 12 and 24 months postoperatively but not with visual analog scale scores for leg pain. Our data suggest that microstructural deterioration of the spinal process as an autograft has detrimental effects on spinal fusion and clinical outcomes for patients undergoing instrumented posterior lumbar interbody fusion. Specifically, the microstructural parameter BS/TV has good potential for assessing lumbar bone quality and predicting fusion success.

Integrated additive design and manufacturing approach for the bioengineering of bone scaffolds for favorable mechanical and biological properties

Additive manufacturing (AM) presents the possibility of personalized bone scaffolds with unprecedented structural and functional designs. In contrast to earlier conventional design concepts, e.g. raster-angle, a workflow was established to produce scaffolds with triply periodic minimal surface (TPMS) architecture. A core challenge is the realization of such structures using melt-extrusion based 3D printing. This study presents methods for generation of scaffold design files, finite element (FE) analysis of scaffold Young’s moduli, AM of scaffolds with polycaprolactone (PCL), and a customized in vitro assay to evaluate cell migration. The reliability of FE analysis when using computer-aided designed models as input may be impeded by anomalies introduced during 3D printing. Using micro-computed tomography reconstructions of printed scaffolds as an input for numerical simulation in comparison to experimentally obtained scaffold Young’s moduli showed a moderate trend (R2 = 0.62). Interestingly, in a preliminary cell migration assay, adipose-derived mesenchymal stromal cells (AdMSC) migrated furthest on PCL scaffolds with Diamond, followed by Gyroid and Schwarz P architectures. A similar trend, but with an accelerated AdMSC migration rate, was observed for PCL scaffolds surface coated with calcium-phosphate-based apatite. We elaborate on the importance of start-to-finish integration of all steps of AM, i.e. design, engineering and manufacturing. Using such a workflow, specific biological and mechanical functionality, e.g. improved regeneration via enhanced cell migration and higher structural integrity, may be realized for scaffolds intended as temporary guiding structures for endogenous tissue regeneration.

Effect of micro-computed tomography reconstruction protocols on bone fractal dimension analysis.

To evaluate the influence of the level of three micro-CT reconstruction tools: beam-hardening correction (BHC), smoothing filter (SF), and ring artefact correction (RAC) on the fractal dimension (FD) analysis of trabecular bone. Five Wistar rats' maxillae were individually scanned in a SkyScan 1174 micro-CT device, under the following settings: 50 kV, 800 µA, 10.2 µm voxel size, 0.5 mm Al filter, rotation step 0.5°, two frames average, 180° rotation and scan time of 35 min. The raw images were reconstructed under the standard protocol (SP) recommended by the manufacturer, a protocol without any artefact correction tools (P0) and 35 additional protocols with different combinations of SF, RAC and BHC levels. The same volume of interest was established in all reconstructions for each maxilla and the FD was calculated using the Kolmogorov (box counting) method. One-way ANOVA with Dunnet's post-hoc test was used to compare the FD of each reconstruction protocol (P0-P35) with the SP (α = 5%). Multiple linear regression verified the dependency of reconstruction tools in FD. Overall, FD values are not dependent on RAC (p = 0.965), but increased significantly when the level of BHC and SF increased (p < 0.001). FD values from protocols with BHC at 45% combined with SF of 2, and BHC at 30% combined with SF of 4 or 6 had no statistical difference compared to SP. BHC and SF tools affect the FD values of micro-CT images of the trabecular bone. Therefore, these reconstruction parameters should be standardized when the FD is analyzed.

Impact of micro-computed tomography reconstruction protocols on bone microarchitecture analysis

The aim of this study was to assess the influence of reconstruction protocols of micro-computed tomography (micro-CT) images on analyses of bone microarchitecture. Micro-CT images of the maxillae of 5 Wistar rats were reconstructed with different protocols by varying the levels of the following tools: smoothing filter (SF; 2-6); ring artifact correction (RAC; 5-15); and beam hardening correction (BHC; 15%-60%). A control protocol (P0; without any correction tool) and a standard protocol (SP; according to the manufacturer's recommendation: SF = 2; RAC = 5; BHC = 45%) were also obtained. For each protocol, 8 bone microarchitecture parameters were calculated (bone volume/total volume [BV/TV], bone surface/total volume [BS/TV], trabecular number [Tb.N], trabecular thickness [Tb.Th], trabecular separation [Tb.Sp], degree of anisotropy [DA], connectivity density [Conn.D], and total porosity [Po-tot]) by using CTAn software. Test protocols were compared with the SP by using 1-way analysis of variance and the Dunnet post hoc test (α = 0.05). An inverse relationship was observed between BHC tool levels and all microarchitecture parameters except BV/TV and Tb.Th. The combination of BHC and SF significantly influenced all microarchitecture parameters except for DA, for which all protocols were similar to the SP (P > .05). Calculation of bone microarchitecture parameters is influenced by the applied levels of artifact correction tools, mainly BHC and SF. It is necessary to standardize such tool levels for correct data interpretation.

Micro-computed tomography reconstructions of tibiae of stem cell transplanted osteogenesis imperfecta mice

Micro-computed tomography (micro-CT) is commonly used to assess bone quality and to evaluate the outcome of experimental therapies in animal models of bone diseases. Generating large datasets is however challenging and data are rarely made publicly available through shared repositories. Here we describe a dataset of micro-CT reconstructed scans of the proximal part of 21 tibiae from wild-type mice, osteogenesis imperfecta mice (homozygous oim/oim) and oim/oim mice transplanted with human amniotic fluid stem cells. The dataset contains, for each sample, 991 8-bit Bitmap reconstructed images and a 3D reconstruction of the bone in the PLY format, available at the online repository Figshare. In line with the increasing effort to make scientific datasets open-access, our data can be downloaded and used by other researchers to compare their observations with ours and to directly test scientific questions on osteogenesis imperfecta bones without the need to generate complete datasets.

Extensive phenotyping of the orofacial and dental complex in Crouzon syndrome

ObjectivesFibroblast growth factor receptor 2 (FGFR2) C342Y/+ mutation is a known cause of Crouzon syndrome that is characterised by craniosynostosis and midfacial hypoplasia. Our aim was to conduct extensive phenotyping of the maxillary, mandibular and dental morphology associated with this mutation. Materials and methodsMorphometric data were obtained from 40 mice, representing two genotypes (Crouzon and wild-type) and two sexes (males and females) (n=10 in each group). Dental analysis further categorised the first molars into the two jaws (maxillary and mandibular) (n=20 in each group). Maxillary, mandibular and dental morphology was compared by analysing 23 linear landmark-based dimensions in three-dimensional micro-computed tomography reconstructions. ResultsCompared with wild-type, Crouzon (FGFR2C342Y/+) maxillae were significantly shorter in maximum height, anterior and posterior lengths and middle width, but larger in posterior width (p<0.05 for height; p<0.001 for other comparisons). In the Crouzon mandible, the ascending and descending heights, effective and mandibular lengths, and intercoronoid and intercondylar widths were significantly shorter, whereas intergonial width was larger (p<0.01 for intercondylar width; p<0.001 for other comparisons). Crouzon teeth were significantly smaller mesiodistally, but larger in crown height (p<0.001 for each comparison). All Crouzon mice presented with bifid mandibular condyles and a quarter presented with expansive bone lesions in the mandibular incisor alveolus. ConclusionsOur findings of hypoplasia in all three planes in Crouzon maxillae and mandibles, together with the presence of bifid mandibular condyles and expansive bone lesions, may be relevant to maxillofacial surgery and orthodontics. Beyond skeletal effects, the FGFR2C342Y/+ mutation is now implicated in affecting tooth development. This study’s skeletal phenomics data also provides baseline data against which the effect of various treatments can now be assessed.

Xenograft Bone Inclusion Improves Incorporation of Hydroxyapatite Cement Into Cancellous Defects

Hydroxyapatite cement (HAC) is biocompatible and osteoconductive, but its slow resorption limits new bone formation. The purpose of this investigation was to determine the effects of adding partially demineralized xenograft cortical bone in an established animal model. Eight 6-month-old female New Zealand white rabbits were used. Drill-hole defects (8.0 mm long, 5.0 mm diameter) were prepared and filled with either HAC alone or HAC mixed with processed xenograft bone particles from young pigs (XBC) at a volumetric ratio of approximately 25%. The particles were approximately 5 mm long and 1 mm diameter and were extensively washed, demineralized in dilute hydrochloric acid, and rewashed. Microcomputed tomography scanning and histology were performed after 10 weeks. New bone and inflammatory/immune response were graded on a 0 to 3 scale and calcein labeling was quantified as percent area new bone. Statistical analyses were by Student's t tests. XBC showed significantly more new bone formation than HAC throughout the defect (P < 0.05). XBC also showed significantly more inflammatory/immune response than HAC (P < 0.05). The three-dimensional microcomputed tomography reconstructions showed that the HAC was basically inert, whereas the XBC took on an appearance suggestive of more extensive incorporation. Adding xenograft to HAC creates a bioactive composite that is more rapidly incorporated, resorbed, and replaced by new bone. The presence of processed xenograft particles creates a vigorous inflammatory response, but there may be some benefit to the resorption rate of the HAC as a result of the infiltration of cells. Future research should focus on the longer-term incorporation and remodeling of XBC.

Attenuation in trabecular bone: A comparison between numerical simulation and experimental results in human femur

Numerical simulations (finite-difference time domain) are compared to experimental results of ultrasound wave propagation through human trabecular bones. Three-dimensional high-resolution microcomputed tomography reconstructions served as input geometry for the simulation. The numerical simulation took into account scattering, but not absorption. Simulated and experimental values of the attenuation coefficients (alpha, dB/cm) and the normalized broadband ultrasound attenuation (nBUA, dB/cm/MHz) were measured and compared on a set of 28 samples. While experimental and simulated nBUA values were highly correlated (R(2)=0.83), and showed a similar dependence with bone volume fraction, the simulation correctly predicted experimental nBUA values only for low bone volume fraction (BV/TV). Attenuation coefficients were underestimated by the simulation. The absolute difference between experimental and simulated alpha values increased with both BV/TV and frequency. As a function of frequency, the relative difference between experimental and simulated alpha values decreased from 60% around 400 kHz to 30% around 1.2 MHz. Under the assumption that the observed discrepancy expresses the effect of the absorption, our results suggests that nBUA and its dependence on BV/TV can be mostly explained by scattering, and that the relative contribution of scattering to alpha increases with frequency, becoming predominant (>50 %) over absorption for frequencies above 600 kHz.

Numerical and experimental structural analysis of trabecular architectures

This paper presents the application of the Cell Method (CM) to the static analysis of 3D structures obtained from micro-computed tomography reconstructions of trabecular bone. The CM is a recently introduced numerical method, based on a direct discrete formulation of equilibrium equations, which is particularly promising for the analysis of complex structures. In fact, due to the direct discrete approach employed, no restriction is imposed by differentiability conditions and the characteristic length of the elementary cell of the discretization can be of the same order of magnitude as the heterogeneities of the structure. The same 3D microstructures used for the numerical simulations were reproduced by means of a rapid prototyping process, by selective laser sintering of polyamide powder. The compression elastic modulus of the replicas was experimentally determined and used as parameter for comparison with the simulations results. The experimental values are in good agreement with the numerical ones, thus validating the methodology employed.

NEW TRENDS IN TISSUE ENGINEERED CARTILAGE: MICROFLUID DYNAMICS IN 3-D ENGINEERED CELL SYSTEMS

Bioreactors allowing culture medium direct-perfusion overcome diffusion limitations associated with static culturing and provide flow-mediated mechanical stimuli. The hydrodynamic stress imposed on chondrocytes will depend not only on the culture medium flow rate, but also on the scaffold three-dimensional (3D) micro-architecture. We performed computational fluid-dynamic (CFD) simulations of the flow of culture medium through a 3D porous scaffold, with the aim of predicting the shear stress acting on the cells as a function of parameters that can be set in a tissue-engineering experiment, such as the medium flow rate and the diameter of the perfused scaffold section. We developed two CFD models: the first model (Model 1) was built from micro-computed tomography reconstruction of the actual scaffold geometry, while the second model (Model 2) was based on a simplification of the actual scaffold microstructure. The two models showed comparable results in terms of the distribution of the shear stresses acting on the inner surfaces of the scaffold walls. Models 1 and 2 gave a median shear stress of 3 mPa at a flow rate of 0.5 cm3 min-1 through a 15 mm diameter scaffold. Our results provide a basis for the completion of more exhaustive quantitative studies to further assess the relationship between perfusion at known micro-fluid dynamic conditions and tissue growth in vitro.