Tunable ciprofloxacin delivery through personalized electrospun patches for tympanic membrane perforations.

Approximately 740 million symptomatic patients are affected by otitis media every year. Being an inflammatory disease affecting the middle ear, it is one of the primary causes of tympanic membrane (TM) perforations, often resulting in impaired hearing abilities. Antibiotic therapy using broad-spectrum fluoroquinolones, such as ciprofloxacin (CIP), is frequently employed and considered the optimal route to treat otitis media. However, patients often get exposed to high dosages to compensate for the low drug concentration reaching the affected site. Therefore, this study aims to integrate tissue engineering with drug delivery strategies to create biomimetic scaffolds promoting TM regeneration while facilitating a localized release of CIP. Distinct electrospinning (ES) modalities were designed in this regard either by blending CIP into the polymer ES solution or by incorporating nanoparticles-based co-ES/electrospraying. The combination of these modalities was investigated as well. A broad range of release kinetic profiles was achieved from the fabricated scaffolds, thereby offering a wide spectrum of antibiotic concentrations that could serve patients with diverse therapeutic needs. Furthermore, the incorporation of CIP into the TM patches demonstrated a favorable influence on their resultant mechanical properties. Biological studies performed with human mesenchymal stromal cells confirmed the absence of any cytotoxic or anti-proliferative effects from the released antibiotic. Finally, antibacterial assays validated the efficacy of CIP-loaded scaffolds in suppressing bacterial infections, highlighting their promising relevance for TM applications.

Joint knee loads during squat with constant or variable resistance in males. A clinical trial.

BACKGROUND: In the squat movement, the use of constant resistance (CR) generates greater compression and shear forces close to 90° of knee flexion, increasing joint overload. However, when used variable resistance (VR) there is no consensus about the effect of knee joint load. The aim of this study was to compare knee torques using constant or variable resistance during the squat exercise. METHODS: Twenty-one healthy male subjects (mean age, 24 [SD, 3] years; height, 1.76 [SD, 0.04] m), who practice squats during strength training routine. Were simultaneously record data from the platform force and tridimensional kinematic to obtain torques around knee. 15 repetitions were performed up to maximum knee flexion with the use of variable (RV) or constant (CR) resistance in a single session. RESULTS: Significant differences regarding the angles only in the sagittal plane at the end of the ascending phase of the squat, with less knee extension in the VR condition. In the sagittal and frontal planes, lower values of extensor and abductor torque were found in the VR condition at the angles of greater knee flexion. CONCLUSION: The use of variable resistance compared to constant resistance seems to be an alternative to be considered when the objective is to minimize the internal loads on the knee joint in exercises such as the squat in amplitudes of greater flexion. This study indicated that VR helps subjects who are learning the squat movement, enabling the application of this knowledge in physical therapy or physical training clinics.

3D Printed Tubulointerstitium Chip as an In Vitro Testing Platform.

Chronic kidney disease (CKD) ranks as the twelfth leading cause of death worldwide with limited treatment options. The development of in vitro models replicating defined segments of the kidney functional units, the nephrons, in a physiologically relevant and reproducible manner can facilitate drug testing. The aim of this study was to produce an in vitro organ-on-a-chip platform with extrusion-based three-dimensional (3D) printing. The manufacturing of the tubular platform was produced by printing sacrificial fibers with varying diameters, providing a suitable structure for cell adhesion and proliferation. The chip platform was seeded with primary murine tubular epithelial cells and human umbilical vein endothelial cells. The effect of channel geometry, its reproducibility, coatings for cell adhesion, and specific cell markers were investigated. The developed chip presents single and dual channels, mimicking segments of a renal tubule and the capillary network, together with an extracellular matrix gel analogue placed in the middle of the two channels, envisioning the renal tubulointerstitium in vitro. The 3D printed platform enables perfusable circular cross-section channels with fully automated, rapid, and reproducible manufacturing processes at low costs. This kidney tubulointerstitium on-a-chip provides the first step toward the production of more complex in vitro models for drug testing.

Matrix metalloproteinase degradable, in situ photocrosslinked nanocomposite bioinks for bioprinting applications.

The development of suitable bioinks with high printability, mechanical strength, biodegradability, and biocompatibility is a key challenge for the clinical translation of 3D constructs produced with bioprinting technologies. In this work, we developed a new type of nanocomposite bioinks containing thiolated mesoporous silica nanoparticles (MSN) that act as active fillers within norbornene-functionalized hydrogels. The MSNs could rapidly covalently crosslink the hydrogels upon exposure to UV light. The mechanical properties of the gels could be modulated from 9.3 to 19.7 kPa with increasing concentrations of MSN. The ability of the MSN to covalently crosslink polymeric networks was, however, significantly influenced by polymer architecture and the number of functional groups. Modification of the outer surface of MSNs with matrix metalloproteinase (MMP) sensitive peptides (MSN-MMPs) resulted in proteinase K and MMP-9 enzyme responsive biodegradable bioinks. Additional cysteine modified RGD peptide incorporation enhanced cell-matrix interactions and reduced the gelation time for bioprinting. The nanocomposite bioinks could be printed by using extrusion-based bioprinting. Our nanocomposite bioinks preserved their shape during in vitro studies and encapsulated MG63 cells preserved their viability and proliferated within the bioinks. As such, our nanocomposite bioinks are promising bioinks for creating bioprinted constructs with tunable mechanical and degradation properties.

Hypotrochoidal scaffolds for cartilage regeneration.

The main function of articular cartilage is to provide a low friction surface and protect the underlying subchondral bone. The extracellular matrix composition of articular cartilage mainly consists of glycosaminoglycans and collagen type II. Specifically, collagen type II fibers have an arch-like organization that can be mimicked with segments of a hypotrochoidal curve. In this study, a script was developed that allowed the fabrication of scaffolds with a hypotrochoidal design. This design was investigated and compared to a regular 0-90 woodpile design. The mechanical analyses revealed that the hypotrochoidal design had a lower component Young's modulus while the toughness and strain at yield were higher compared to the woodpile design. Fatigue tests showed that the hypotrochoidal design lost more energy per cycle due to the damping effect of the unique microarchitecture. In addition, data from cell culture under dynamic stimulation demonstrated that the collagen type II deposition was improved and collagen type X reduced in the hypotrochoidal design. Finally, Alcian blue staining revealed that the areas where the stress was higher during the stimulation produced more glycosaminoglycans. Our results highlight a new and simple scaffold design based on hypotrochoidal curves that could be used for cartilage tissue engineering.

Comparison of Zinc Oxide Nanoparticle Integration into Non-Woven Fabrics Using Different Functionalisation Methods for Prospective Application as Active Facemasks.

The development of advanced facemasks stands out as a paramount priority in enhancing healthcare preparedness. In this work, different polypropylene non-woven fabrics (NWF) were characterised regarding their structural, physicochemical and comfort-related properties. The selected NWF for the intermediate layer was functionalised with zinc oxide nanoparticles (ZnO NPs) 0.3 and 1.2wt% using three different methods: electrospinning, dip-pad-dry and exhaustion. After the confirmation of ZnO NP content and distribution within the textile fibres by morphological and chemical analysis, the samples were evaluated regarding their antimicrobial properties. The functionalised fabrics obtained via dip-pad-dry unveiled the most promising data, with 0.017 ± 0.013wt% ZnO NPs being mostly located at the fibre's surface and capable of total eradication of Staphylococcus aureus and Escherichia coli colonies within the tested 24 h (ISO 22196 standard), as well as significantly contributing (**** p < 0.0001) to the growth inhibition of the bacteriophage MS2, a surrogate of the SARS-CoV-2 virus (ISO 18184 standard). A three-layered structure was assembled and thermoformed to obtain facemasks combining the previously chosen NWF, and its resulting antimicrobial capacity, filtration efficiency and breathability (NP EN ISO 149) were assessed. The developed three-layered and multiscaled fibrous structures with antimicrobial capacities hold immense potential as active individual protection facemasks.

Flexible, Suturable, and Leak-free Scaffolds for Vascular Tissue Engineering Using Melt Spinning.

Coronary artery disease affects millions worldwide. Bypass surgery remains the gold standard; however, autologous tissue is not always available. Hence, the need for an off-the-shelf graft to treat these patients remains extremely high. Using melt spinning, we describe here the fabrication of tubular scaffolds composed of microfibers aligned in the circumferential orientation mimicking the organized extracellular matrix in the tunica media of arteries. By variation of the translational extruder speed, the angle between fibers ranged from 0 to ∼30°. Scaffolds with the highest angle showed the best performance in a three-point bending test. These constructs could be bent up to 160% strain without kinking or breakage. Furthermore, when liquid was passed through the scaffolds, no leakage was observed. Suturing of native arteries was successful. Mesenchymal stromal cells were seeded on the scaffolds and differentiated into vascular smooth muscle-like cells (vSMCs) by reduction of serum and addition of transforming growth factor beta 1 and ascorbic acid. The scaffolds with a higher angle between fibers showed increased expression of vSMC markers alpha smooth muscle actin, calponin, and smooth muscle protein 22-alpha, whereas a decrease in collagen 1 expression was observed, indicating a positive contractile phenotype. Endothelial cells were seeded on the repopulated scaffolds and formed a tightly packed monolayer on the luminal side. Our study shows a one-step fabrication for ECM-mimicking scaffolds with good handleability, leak-free property, and suturability; the excellent biocompatibility allowed the growth of a bilayered construct. Future work will explore the possibility of using these scaffolds as vascular conduits in in vivo settings.

Cellular uptake of modified mRNA occurs via caveolae-mediated endocytosis, yielding high protein expression in slow-dividing cells.

Nucleic acids have clear clinical potential for gene therapy. Plasmid DNA (pDNA) was the first nucleic acid to be pursued as a therapeutic molecule. Recently, mRNA came into play as it offers improved safety and affordability. In this study, we investigated the uptake mechanisms and efficiencies of genetic material by cells. We focused on three main variables (1) the nucleic acid (pDNA, or chemically modified mRNA), (2) the delivery vector (Lipofectamine 3000 or 3DFect), and (3) human primary cells (mesenchymal stem cells, dermal fibroblasts, and osteoblasts). In addition, transfections were studied in a 3D environment using electrospun scaffolds. Cellular internalization and intracellular trafficking were assessed by using enhancers or inhibitors of endocytosis and endosomal escape. The polymeric vector TransIT-X2 was included for comparison purposes. While lipoplexes utilized several entry routes, uptake via caveolae served as the main route for gene delivery. pDNA yielded higher expression levels in fast-dividing fibroblasts, whereas, in slow-dividing osteoblasts, cmRNA was responsible for high protein production. In the case of mesenchymal stem cells, which presented an intermediate doubling time, the combination vector/nucleic acid seemed more relevant than the nucleic acid per se. In all cases, protein expression was higher when the cells were seeded on 3D scaffolds.

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 biomimetic arch-like 3D bioprinted construct for cartilage regeneration using gelatin methacryloyl and silk fibroin-gelatin bioinks.

In recent years, engineering biomimetic cellular microenvironments have been a top priority for regenerative medicine. Collagen II, which is arranged in arches, forms the predominant fiber network in articular cartilage. Due to the shortage of suitable microfabrication techniques capable of producing 3D fibrous structures,in vitroreplication of the arch-like cartilaginous tissue constitutes one of the major challenges. Hence, in the present study, we report a 3D bioprinting approach for fabricating arch-like constructs using two types of bioinks, gelatin methacryloyl (GelMa) and silk fibroin-gelatin (SF-G). The bioprinted SF-G constructs displayed increased proliferation of the encapsulated human bone marrow-derived mesenchymal stem cells compared to the GelMA constructs. Biochemical assays, gene, and protein expression exhibited the superior role of SF-G in forming the fibrous collagen network and chondrogenesis. Protein-protein interaction study using Metascape evaluated the function of the proteins involved. Further GeneMANIA and STRING analysis using Col 2A1, SOX 9, ACAN, and the genes upregulated on day 21 in RT-PCR, i.e.ß-catenin, TGFßR1, Col 1A1 in SF-G and PRG4, Col 10A1, MMP 13 in GelMA validated ourin vitroresults. These findings emphasized the role of SF-G in regulating the Wnt/ß-catenin and TGF-ßsignaling pathways. Hence, the 3D bioprinted arch-like constructs possess a substantial potential for cartilage regeneration.

Biofabrication of engineered dento-alveolar tissue.

Oral health is essential for a good overall health. Dento-alveolar conditions have a high prevalence, ranging from tooth decay periodontitis to alveolar bone resorption. However, oral tissues exhibit a limited regenerative capacity, and full recovery is challenging. Therefore, regenerative therapies for dento-alveolar tissue (e.g., alveolar bone, periodontal membrane, dentin-pulp complex) have gained much attention, and novel approaches have been proposed in recent decades. This review focuses on the cells, biomaterials and the biofabrication methods used to develop therapies for tooth root bioengineering. Examples of the techniques covered are the multitude of additive manufacturing techniques and bioprinting approaches used to create scaffolds or tissue constructs. Furthermore, biomaterials and stem cells utilized during biofabrication will also be described for different target tissues. As these new therapies gradually become a reality in the lab, the translation to the clinic is still minute, with a further need to overcome multiple challenges and broaden the clinical application of these alternatives.

Chitin nanofibrils modulate mechanical response in tympanic membrane replacements.

The tympanic membrane (TM), is a thin tissue lying at the intersection of the outer and the middle ear. TM perforations caused by traumas and infections often result in a conductive hearing loss. Tissue engineering has emerged as a promising approach for reconstructing the damaged TM by replicating the native material characteristics. In this regard, chitin nanofibrils (CN), a polysaccharide-derived nanomaterial, is known to exhibit excellent biocompatibility, immunomodulation and antimicrobial activity, thereby imparting essential qualities for an optimal TM regeneration. This work investigates the application of CN as a nanofiller for poly(ethylene oxide terephthalate)/poly(butylene terephthalate) (PEOT/PBT) copolymer to manufacture clinically suitable TM scaffolds using electrospinning and fused deposition modelling. The inclusion of CN within the PEOT/PBT matrix showed a three-fold reduction in the corresponding electrospun fiber diameters and demonstrated a significant improvement in the mechanical properties required for TM repair. Furthermore, in vitro biodegradation assay highlighted a favorable influence of CN in accelerating the scaffold degradation over a period of one year. Finally, the oto- and cytocompatibility response of the nanocomposite substrates corroborated their biological relevance for the reconstruction of perforated eardrums.

Modeling a Dynamic Printability Window on Polysaccharide Blend Inks for Extrusion Bioprinting.

Extrusion-based bioprinting is one of the most widespread technologies due to its affordability, wide range of processable materials, and ease of use. However, the formulation of new inks for this technique is based on time-consuming trial-and-error processes to establish the optimal ink composition and printing parameters. Here, a dynamic printability window was modeled for the assessment of the printability of polysaccharide blend inks of alginate and hyaluronic acid with the intent to build a versatile predictive tool to speed up the testing procedures. The model considers both the rheological properties of the blends (viscosity, shear thinning behavior, and viscoelasticity) and their printability (in terms of extrudability and the ability of forming a well-defined filament and detailed geometries). By imposing some conditions on the model equations, it was possible to define empirical bands in which the printability is ensured. The predictive capability of the built model was successfully verified on an untested blend of alginate and hyaluronic acid chosen to simultaneously optimize the printability index and minimize the size of the deposited filament.

Manufacturing of scaffolds with interconnected internal open porosity and surface roughness.

Manufacturing of three-dimensional scaffolds with multiple levels of porosity are an advantage in tissue regeneration approaches to influence cell behavior. Three-dimensional scaffolds with surface roughness and intra-filament open porosity were successfully fabricated by additive manufacturing combined with chemical foaming and porogen leaching without the need of toxic solvents. The decomposition of sodium citrate, a chemical blowing agent, generated pores within the scaffold filaments, which were interconnected and opened to the external environment by leaching of a water-soluble sacrificial phase, as confirmed by micro-CT and buoyancy measurements. The additional porosity did not result in lower elastic modulus, but in higher strain at maximum load, i.e. scaffold ductility. Human mesenchymal stromal cells cultured for 24 h adhered in greater numbers on these scaffolds when compared to plain additive-manufactured ones, irrespectively of the scaffold pre-treatment method. Additionally, they showed a more spread and random morphology, which is known to influence cell fate. Cells cultured for a longer period exhibited enhanced metabolic activity while secreting higher osteogenic markers after 7 days in culture. STATEMENT OF SIGNIFICANCE: Inspired by the function of hierarchical cellular structures in natural materials, this work elucidates the development of scaffolds with multiscale porosity by combining in-situ foaming and additive manufacturing, and successive porogen leaching. The resulting scaffolds displayed enhanced mechanical toughness and multiscale pore network interconnectivity, combined with early differentiation of adult mesenchymal stromal cells into the osteogenic lineage.

Assessment of motor function in individuals with hemiplegia post-stroke / Evaluación de la función motriz en hemipléjicos posaccidente cerebrovascular / Avaliação da função motora em hemiplégicos pós-acidente vascular encefálico

ABSTRACT This study aims to compare the performance of the sit-to-stand test and walking speed in individuals with chronic hemiplegia post-stroke and a control group (CG). Moreover, we will investigate whether lower limb resistance, measured based on the sit-to-stand test, is related to walking speed in individuals with chronic hemiplegia and a CG. Finally, we will verify if there are intra-group differences for the tests by dividing the hemiplegia group (HG) according to motor and sensorimotor function assessment classification. A cross-sectional design was used among a group with chronic hemiplegia (n=28) and a healthy CG (n=22). The HG was classified by the Fugl-Meyer scale, and both groups were evaluated using the 1-minute sit-to-stand test. The walking speed was calculated using a 3D kinematics system. Lower limb resistance among HG differed significantly from the CG, as well as walking speed. We found a strong correlation between the tests (ρ=0.773; p<0.001). No differences were found for the sit-to-stand tests and walking speed when dividing the HG into individuals with greater or lesser motor and sensory impairment, using the Fugl-Meyer scale. Therefore, individuals with hemiplegia, regardless of having a more pronounced classification of motor and sensory impairment on the Fugl-Meyer scale, showed lower limb resistance and lower walking speed compared with individuals without hemiplegia post-stroke.

RESUMEN El objetivo de este estudio es comparar el desempeño del test de levantarse y sentarse y la velocidad de marcha en individuos con hemiplejía crónica debido a accidente cerebrovascular (ACV) y un grupo control (GC). Además, se investigará si existe asociación entre la resistencia de los miembros inferiores, medida desde el test de levantarse y sentarse, y la velocidad de marcha en individuos con hemiplejía crónica y un GC. Por último, se verificará si existen diferencias intragrupales en las pruebas al dividir el grupo hemiplejía (GH) según la clasificación de evaluación de deterioro motor y sensorial. El método utilizado fue el transversal en un grupo con hemiplejía crónica (n=28) y un GC sin ninguna patología (n=22). El GH se clasificó mediante la escala de Fugl-Meyer, y ambos grupos se evaluaron mediante el test de levantarse y sentarse de un minuto. La velocidad de marcha se calculó mediante el sistema cinemático tridimensional. Entre los resultados obtenidos, se observó que la resistencia de los miembros inferiores entre GH difería significativamente del GC, así como la velocidad de marcha. Se demostró una fuerte correlación entre las pruebas (ρ=0,773; p<0,001). No se encontraron diferencias en las pruebas de levantarse y sentarse y la velocidad de la marcha al dividir el GH en individuos con mayor o menor deterioro motor y sensorial, utilizando la escala de Fugl-Meyer. Por lo tanto, las personas con hemiplejía, independientemente de tener un mayor deterioro motor y sensorial según la escala de Fugl-Meyer, tuvieron una menor resistencia de las extremidades inferiores y una menor velocidad de marcha en comparación con las personas sin hemiplejía pos-ACV.

RESUMO O objetivo deste estudo é comparar os desempenhos no teste de sentar e levantar e a velocidade de caminhada de indivíduos com hemiplegia crônica decorrente de acidente vascular encefálico (AVE) e um grupo-controle (GC). Além disso, será investigado se existe associação entre a resistência de membros inferiores, mensurada a partir do teste de sentar e levantar, e a velocidade de caminhada em indivíduos com hemiplegia crônica e um GC. Por fim, será verificado se existem diferenças intragrupo para os testes ao dividir o grupo hemiplegia (GH) de acordo com a classificação de avaliação do comprometimento motor e sensorial. O método utilizado foi o delineamento transversal entre um grupo com hemiplegia crônica (n=28) e um GC sem nenhuma patologia (n=22). O GH foi classificado a partir da escala de Fugl-Meyer, e ambos os grupos foram avaliados por meio do teste de sentar e levantar de um minuto. A velocidade de caminhada foi calculada a partir de um sistema de cinemetria tridimensional. Entre os resultados obtidos, foi percebido que a resistência de membros inferiores do GH diferiu significativamente do GC, assim como a velocidade de caminhada. Foi demonstrada uma correlação forte entre os testes (ρ=0,773; p<0,001). Não foram encontradas diferenças nos testes de sentar e levantar e velocidade de caminhada ao dividir o GH em indivíduos com maior ou menor comprometimento motor e sensorial, com a escala de Fugl-Meyer. Portanto, indivíduos com hemiplegia, independentemente de ter uma classificação de comprometimento motor e sensorial mais acentuada na escala de Fugl-Meyer, apresentaram menor resistência de membros inferiores e menor velocidade de caminhada comparados com indivíduos sem hemiplegia pós-AVE.

Effect of high content nanohydroxyapatite composite scaffolds prepared via melt extrusion additive manufacturing on the osteogenic differentiation of human mesenchymal stromal cells.

The field of bone tissue engineering seeks to mimic the bone extracellular matrix composition, balancing the organic and inorganic components. In this regard, additive manufacturing (AM) of high content calcium phosphate (CaP)-polymer composites holds great promise towards the design of bioactive scaffolds. Yet, the biological performance of such scaffolds is still poorly characterized. In this study, melt extrusion AM (ME-AM) was used to fabricate poly(ethylene oxide terephthalate)/poly(butylene terephthalate) (PEOT/PBT)-nanohydroxyapatite (nHA) scaffolds with up to 45 wt% nHA, which presented significantly enhanced compressive mechanical properties, to evaluate their in vitro osteogenic potential as a function of nHA content. While osteogenic gene upregulation and matrix mineralization were observed on all scaffold types when cultured in osteogenic media, human mesenchymal stromal cells did not present an explicitly clear osteogenic phenotype, within the evaluated timeframe, in basic media cultures (i.e. without osteogenic factors). Yet, due to the adsorption of calcium and inorganic phosphate ions from cell culture media and simulated body fluid, the formation of a CaP layer was observed on PEOT/PBT-nHA 45 wt% scaffolds, which is hypothesized to account for their bone forming ability in the long term in vitro, and osteoconductivity in vivo.

The degradation of gelatin/alginate/fibrin hydrogels is cell type dependent and can be modulated by targeting fibrinolysis.

In tissue engineering, cell origin is important to ensure outcome quality. However, the impact of the cell type chosen for seeding in a biocompatible matrix has been less investigated. Here, we investigated the capacity of primary and immortalized fibroblasts of distinct origins to degrade a gelatin/alginate/fibrin (GAF)-based biomaterial. We further established that fibrin was targeted by degradative fibroblasts through the secretion of fibrinolytic matrix-metalloproteinases (MMPs) and urokinase, two types of serine protease. Finally, we demonstrated that besides aprotinin, specific targeting of fibrinolytic MMPs and urokinase led to cell-laden GAF stability for at least forty-eight hours. These results support the use of specific strategies to tune fibrin-based biomaterials degradation over time. It emphasizes the need to choose the right cell type and further bring targeted solutions to avoid the degradation of fibrin-containing hydrogels or bioinks.

Development of an In Vitro Biomimetic Peripheral Neurovascular Platform.

Nerves and blood vessels are present in most organs and are indispensable for their function and homeostasis. Within these organs, neurovascular (NV) tissue forms congruent patterns and establishes vital interactions. Several human pathologies, including diabetes type II, produce NV disruptions with serious consequences that are complicated to study using animal models. Complex in vitro organ platforms, with neural and vascular supply, allow the investigation of such interactions, whether in a normal or pathological context, in an affordable, simple, and direct manner. To date, a few in vitro models contain NV tissue, and most strategies report models with nonbiomimetic representations of the native environment. To this end, we have established here an NV platform that contains mature vasculature and neural tissue, composed of human microvascular endothelial cells (HMVECs), induced pluripotent stem cell (iPSCs)-derived sensory neurons, and primary rat Schwann cells (SCs) within a fibrin-embedded polymeric scaffold. First, we show that SCs can induce the formation of and stabilize vascular networks to the same degree as the traditional and more thoroughly studied human dermal fibroblasts (HDFs). We also show that through SC prepatterning, we are able to control vessel orientation. Using our NV platform, we demonstrate the concomitant formation of three-dimensional neural and vascular tissue, and the influence of different medium formulations and cell types on the NV tissue outcome. Finally, we propose a protocol to form mature NV tissue, via the integration of independent neural and vascular constituents. The platform described here provides a versatile and advanced model for in vitro research of the NV axis.

Fabrication of a mimetic vascular graft using melt spinning with tailorable fiber parameters.

Smooth muscle cells play a pivotal role in maintaining blood pressure and remodeling of the extracellular matrix. These cells have a characteristic spindle shape and are aligned in the radial direction to aid in the constriction of any artery. Tissue engineered grafts have the potential to recreate this alignment and offer a viable alternative to non-resorbable or autologous grafts. Specifically, with melt spinning small diameter fibers can be created that can align circumferentially on the scaffolds. In this study, a set of simplified equations were formulated to predict the final fiber parameters. Smooth muscle cell alignment was monitored on the fabricated scaffolds. Finally, a co-culture of smooth muscle cells in direct contact with endothelial cells was performed to assess the influence of the smooth muscle cell alignment on the morphology of the endothelial cells. The results show that the equations were able to accurately predict the fiber diameter, distance and angle. Primary vascular smooth muscle cells aligned according to the fiber direction mimicking the native orientation. The co-culture with endothelial cells showed that the aligned smooth muscle cells did not have an influence on the morphology of the endothelial cells. In conclusion, we formulated a series of equations that can predict the fiber parameters during melt spinning. Furthermore, the method described here can create a vascular graft with smooth muscle cells aligned circumferentially that morphologically mimics the native orientation.

Center of pressure position in the touches of the star excursion balance test in healthy individuals.

BACKGROUND: The Star Excursion Balance Test (SEBT) is a clinical test that aims to assess postural control. Its interpretation is related to the understanding of the motor specificities required. Adjustments must be made to the center of pressure (COP) to maintain balance during testing movements. Comprehend the specifics of these adjustments for each direction can allow the development of more suitable exercises for balance training. The aim was to compare the positions of the COP on the different directions of the SEBT and correlate the reachs obtained in the SEBT with the distances from the COP to the borders of the base of support (BOS). METHODS: Sixteen healthy subjects participated in the study. Measurements were made by performing the SEBT over the force platform. The Kruskal-Wallis test followed by Bonferroni's post hoc test was used to compare directions. The Pearson correlation test was used to check the correlation of parametric variables and Spearman correlation test for the nonparametric ones. RESULTS: The position of the COP at the touch differs from the anterior direction to the other directions of the SEBT and the performance in this direction is correlated with the proximity of the COP to the anterior limit of the support base. The performances in the other directions did not correlate with the COP position. CONCLUSION: The requirements of the compensation mechanisms for postural control are different between the directions of the SEBT.