A versatile micromodel technology to explore biofilm development in porous media flows.

Bacterial biofilms that grow in porous media are critical to ecosystem processes and applications ranging from soil bioremediation to bioreactors for treating wastewater or producing value-added products. However, understanding and engineering the complex phenomena that drive the development of biofilms in such systems remains a challenge. Here we present a novel micromodel technology to explore bacterial biofilm development in porous media flows. The technology consists of a set of modules that can be combined as required for any given experiment and conveniently tuned for specific requirements. The core module is a 3D-printed micromodel where biofilm is grown into a perfusable porous substrate. High-precision additive manufacturing, in particular stereolithography, is used to fabricate porous scaffolds with precisely controlled architectures integrating flow channels with diameters down to several hundreds of micrometers. The system is instrumented with: ultraviolet-C light-emitting diodes; on-line measurements of oxygen consumption and pressure drop across the porous medium; camera and spectrophotometric cells for the detection of biofilm detachment events at the outlet. We demonstrate how this technology can be used to study the development of Pseudomonas aeruginosa biofilm for several days within a network of flow channels. We find complex dynamics whereby oxygen consumption reaches a steady-state but not the pressure drop, which instead features a permanent regime with large fluctuations. We further use X-ray computed microtomography to image the spatial distribution of biofilms and computational fluid dynamics to link biofilm development with local flow properties. By combining the advantages of additive manufacturing for the creation of reproducible 3D porous microarchitectures with the flow control and instrumentation accuracy of microfluidics, our system provides a platform to study the dynamics of biofilm development in 3D porous media and to rapidly test new concepts in process engineering.

Diffusion properties of asymptomatic lumbar intervertebral discs in a pediatric cohort: a preliminary study of apparent diffusion coefficient.

PURPOSE: To explore the apparent diffusion coefficients of intervertebral discs in an asymptomatic pediatric cohort. METHODS: We conducted a prospective MRI study of the lumbar spine from below the thoracolumbar junction to the lumbosacral junction on 12 subjects (mean age 13 y.o.) with no spinal pathology or spinal posture disorder. MRI was carried out using a 1.5 T machine with acquisitions realized both in sagittal and coronal planes. First, disc hydration was determined, and then, diffusion-weighted images were obtained using an SE single-shot echo-planar sequence. Apparent diffusion coefficients (ADC) of anterior annulus fibrosus (AAF), nucleus pulposus (NP) and posterior annulus fibrosus (PAF) were measured in the sagittal plane. RESULTS: Averaged hydration of 0.27 SD 0.03 confirmed the asymptomatic nature of discs. Average scaled values of ADC were 0.46 SD 0.01, 0.22 SD 0.09 and 0.18 SD 0.03 for NP, AAF and PAF, respectively. ADC of NP were almost constant along the spine; PAF values show a slight increase in the thorax-sacrum direction, while AAF values showed a pronounced decrease. Locally, ADC of AAF was higher compared to ADC PAF values below the thoracolumbar junction and it reversed for subjacent discs. CONCLUSIONS: In our knowledge, our study provided the first diffusive properties of asymptomatic intervertebral discs in an adolescent cohort. ADC of NP was slightly higher than adults'. ADC evolutions of AAF were correlated with lordosis concavity which pointed out the role of compressive strain on fluid transport properties. This study could furnish information about segment homeostasis for exploration of pediatric spinal pathologies.

Does a combined screw and dowel construct improve tibial fixation during anterior cruciate ligament reconstruction?

PURPOSE: The aims of the present study were to compare the biomechanical properties of tibial fixation in hamstring-graft ACL reconstruction using interference screw and a novel combination interference screw and dowel construct. MATERIAL AND METHODS: We compared the fixation of 30 (2- and 4-stranded gracilis and semitendinosis tendons) in 15 fresh-frozen porcine tibiae with a biocomposite resorbable interference screw (Group 1) and a screw and dowel construct (Group 2). Each graft was subjected to load-to-failure testing (50 mm/min) to determine maximum load, displacement at failure and pullout strength. RESULTS: There were no significant differences between the biomechanical properties of the constructs. Multivariate analysis demonstrated that combination constructs (ß = 140.20, p = 0.043), screw diameter (ß = 185, p = 0.006) and 4-strand grafts (ß = 51, p = 0.050) were associated with a significant increase in load at failure. Larger screw diameter was associated with increased construct stiffness (ß = 20.15, p = 0.020). CONCLUSION: The screw and dowel construct led to significantly increased fixation properties compared to interference screws alone in a porcine model. Increased screw diameter and utilization of 4-strand ACL grafts also led to improvement in load-to-failure of the construct. However, this is an in vitro study and additional investigations are needed to determine whether the results are reproducible in vivo. LEVEL OF EVIDENCE: Level V; Biomechanical study.

Characterization of Macrophages and Osteoclasts in the Osteosarcoma Tumor Microenvironment at Diagnosis: New Perspective for Osteosarcoma Treatment?

Biological and histopathological techniques identified osteoclasts and macrophages as targets of zoledronic acid (ZA), a therapeutic agent that was detrimental for patients in the French OS2006 trial. Conventional and multiplex immunohistochemistry of microenvironmental and OS cells were performed on biopsies of 124 OS2006 patients and 17 surgical ("OSNew") biopsies respectively. CSF-1R (common osteoclast/macrophage progenitor) and TRAP (osteoclast activity) levels in serum of 108 patients were correlated to response to chemotherapy and to prognosis. TRAP levels at surgery and at the end of the protocol were significantly lower in ZA+ than ZA- patients (padj = 0.0011; 0.0132). For ZA+-patients, an increase in the CSF-1R level between diagnosis and surgery and a high TRAP level in the serum at biopsy were associated with a better response to chemotherapy (p = 0.0091; p = 0.0251). At diagnosis, high CD163+ was associated with good prognosis, while low TRAP activity was associated with better overall survival in ZA- patients only. Multiplex immunohistochemistry demonstrated remarkable bipotent CD68+/CD163+ macrophages, homogeneously distributed throughout OS regions, aside osteoclasts (CD68+/CD163-) mostly residing in osteolytic territories and osteoid-matrix-associated CD68-/CD163+ macrophages. We demonstrate that ZA not only acts on harmful osteoclasts but also on protective macrophages, and hypothesize that the bipotent CD68+/CD163+ macrophages might present novel therapeutic targets.

An energy approach describes spine equilibrium in adolescent idiopathic scoliosis.

The adolescent idiopathic scoliosis (AIS) is a 3D deformity of the spine whose origin is unknown and clinical evolution unpredictable. In this work, a mixed theoretical and numerical approach based on energetic considerations is proposed to study the global spine deformations. The introduced mechanical model aims at overcoming the limitations of computational cost and high variability in physical parameters. The model is constituted of rigid vertebral bodies associated with 3D effective stiffness tensors. The spine equilibrium is found using minimization methods of the mechanical total energy which circumvents forces and loading calculation. The values of the model parameters exhibited in the stiffness tensor are retrieved using a combination of clinical images post-processing and inverse algorithms implementation. Energy distribution patterns can then be evaluated at the global spine scale to investigate given time patient-specific features. To verify the reliability of the numerical methods, a simplified model of spine was implemented. The methodology was then applied to a clinical case of AIS (13-year-old girl, Lenke 1A). Comparisons of the numerical spine geometry with clinical data equilibria showed numerical calculations were performed with great accuracy. The patient follow-up allowed us to highlight the energetic role of the apical and junctional zones of the deformed spine, the repercussion of sagittal bending in sacro-illiac junctions and the significant role of torsion with scoliosis aggravation. Tangible comparisons of output measures with clinical pathology knowledge provided a reliable basis for further use of those numerical developments in AIS classification, scoliosis evolution prediction and potentially surgical planning.

Water-in-PDMS Emulsion Templating of Highly Interconnected Porous Architectures for 3D Cell Culture.

The development of advanced techniques of fabrication of three-dimensional (3D) microenvironments for the study of cell growth and proliferation has become one of the major motivations of material scientists and bioengineers in the past decade. Here, we present a novel residueless 3D structuration technique of poly(dimethylsiloxane) (PDMS) by water-in-PDMS emulsion casting and subsequent curing process in temperature-/pressure-controlled environment. Scanning electron microscopy and X-ray microcomputed tomography allowed us to investigate the impact of those parameters on the microarchitecture of the porous structure. We demonstrated that the optimized emulsion casting process gives rise to large-scale and highly interconnected network with pore size ranging from 500 µm to 1.5 mm that turned out to be nicely adapted to 3D cell culture. Experimental cell culture validations were performed using SaOS-2 (osteosarcoma) cell lines. Epifluorescence and deep penetration imaging techniques as two-photon confocal microscopy unveiled information about cell morphology and confirmed a homogeneous cell proliferation and spatial distribution in the 3D porous structure within an available volume larger than 1 cm3. These results open alternative scenarios for the fabrication and integration of porous scaffolds for the development of 3D cell culture platforms.