Bioprinting of Stem Cell Spheroids Followed by Post-Printing Chondrogenic Differentiation for Cartilage Tissue Engineering.

Cartilage tissue presents low self-repair capability and lesions often undergo irreversible progression. Structures obtained by tissue engineering, such as those based in extrusion bioprinting of constructs loaded with stem cell spheroids may offer valuable alternatives for research and therapeutic purposes. Human mesenchymal stromal cell (hMSC) spheroids can be chondrogenically differentiated faster and more efficiently than single cells. This approach allows obtaining larger tissues in a rapid, controlled and reproducible way. However, it is challenging to control tissue architecture, construct stability, and cell viability during maturation. Herein, this work reports a reproducible bioprinting process followed by a successful post-bioprinting chondrogenic differentiation procedure using large quantities of hMSC spheroids encapsulated in a xanthan gum-alginate hydrogel. Multi-layered constructs are bioprinted, ionically crosslinked, and post chondrogenically differentiated for 28 days. The expression of glycosaminoglycan, collagen II and IV are observed. After 56 days in culture, the bioprinted constructs are still stable and show satisfactory cell metabolic activity with profuse extracellular matrix production. These results show a promising procedure to obtain 3D models for cartilage research and ultimately, an in vitro proof-of-concept of their potential use as stable chondral tissue implants.

Development of a device useful to reproducibly produce large quantities of viable and uniform stem cell spheroids with controlled diameters.

Three-dimensional cellular aggregates can mimic the natural microenvironment of tissues and organs and obtaining them through controlled and reproducible processes is mandatory for scaling up and implementing drug cytotoxicity and efficacy tests, as well as tissue engineering protocols. The purpose of this work was to develop and evaluate the performance of a device with two different geometries fabricated by additive manufacturing. The methodology was based on casting a microwell array insert using a non-adhesive hydrogel to obtain highly regular microcavities to standardize spheroid formation and morphology. Spheroids of dental pulp stem cells, bone marrow stromal cells and embryonic stem cells showing high cell viability and average diameters of around 253, 220, and 500 µm, respectively, were produced using the device with the geometry considered most adequate. The cell aggregates showed sphericity indexes above 0.9 and regular surfaces (solidity index higher than 0.96). Around 1000 spheroids could be produced in a standard six-well plate. Overall, these results show that this method facilitates obtaining a large number of uniform, viable spheroids with pre-specified average diameters and through a low-cost and reproducible process for a myriad of applications.

Development and characterization of the InVesalius Navigator software for navigated transcranial magnetic stimulation.

BACKGROUND: Neuronavigation provides visual guidance of an instrument during procedures of neurological interventions, and has been shown to be a valuable tool for accurately positioning transcranial magnetic stimulation (TMS) coils relative to an individual's anatomy. Despite the importance of neuronavigation, its high cost, low portability, and low availability of magnetic resonance imaging facilities limit its insertion in research and clinical environments. NEW METHOD: We have developed and validated the InVesalius Navigator as the first free, open-source software for image-guided navigated TMS, compatible with multiple tracking devices. A point-based, co-registration algorithm and a guiding interface were designed for tracking any instrument (e.g. TMS coils) relative to an individual's anatomy. RESULTS: Localization, precision errors, and repeatability were measured for two tracking devices during navigation in a phantom and in a simulated TMS study. Errors were measured in two commercial navigated TMS systems for comparison. Localization error was about 1.5 mm, and repeatability was about 1 mm for translation and 1° for rotation angles, both within limits established in the literature. COMPARISON WITH EXISTING METHODS: Existing TMS neuronavigation software programs are not compatible with multiple tracking devices, and do not provide an easy to implement platform for custom tools. Moreover, commercial alternatives are expensive with limited portability. CONCLUSIONS: InVesalius Navigator might contribute to improving spatial accuracy and the reliability of techniques for brain interventions by means of an intuitive graphical interface. Furthermore, the software can be easily integrated into existing neuroimaging tools, and customized for novel applications such as multi-locus and/or controllable-pulse TMS.

Surgical planning for resection of an ameloblastoma and reconstruction of the mandible using a selective laser sintering 3D biomodel.

Ameloblastoma is a benign locally aggressive infiltrative odontogenic lesion. It is characterized by slow growth and painless swelling. The treatment for ameloblastoma varies from curettage to en bloc resection, and the reported recurrence rates after treatment are high; the safety margin of resection is important to avoid recurrence. Advances in technology brought about great benefits in dentistry; a new generation of computed tomography scanners and 3-dimensional images enhance the surgical planning and management of maxillofacial tumors. The development of new prototyping systems provides accurate 3D biomodels on which surgery can be simulated, especially in cases of ameloblastoma, in which the safety margin is important for treatment success. A case of mandibular follicular ameloblastoma is reported where a 3D biomodel was used before and during surgery.