In JoVE (2)
Other Publications (8)
- Journal of Biomedical Materials Research. Part A
- Pharmaceutical Research
- Biotechnology and Bioengineering
- Tissue Engineering. Part A
- Acta Biomaterialia
- Matrix Biology : Journal of the International Society for Matrix Biology
- The Journal of Neuroscience : the Official Journal of the Society for Neuroscience
- Journal of Neurochemistry
Articles by Idalis Villanueva in JoVE
Use of Galvanic Skin Responses, Salivary Biomarkers, and Self-reports to Assess Undergraduate Student Performance During a Laboratory Exam Activity Idalis Villanueva1, Maria Valladares1, Wade Goodridge1 1Department of Engineering Education, Utah State University Research on undergraduate students' academic achievement emotions have primarily relied on self-reports in laboratory settings. Studies rarely include bio-physiological measures to these self-reports. This protocol will describe a methodology that integrates self-reports with and bio-physiological measures to assess student response and performance during a laboratory examination activity.
Utilizing Electroencephalography Measurements for Comparison of Task-Specific Neural Efficiencies: Spatial Intelligence Tasks Benjamin J. Call1, Wade Goodridge1, Idalis Villanueva1, Nicholas Wan2, Kerry Jordan2 1Department of Engineering Education, Utah State University, 2Department of Psychology, Utah State University This manuscript describes an approach to measure neural activity of humans while solving spatially focused engineering problems. The electroencephalogram methodology helps interpret beta brain wave measurements in terms of neural efficiency, with the aim of ultimately enabling comparisons of task performance both between problem types and between participants.
Other articles by Idalis Villanueva on PubMed
Mechanical Stimulation of TMJ Condylar Chondrocytes Encapsulated in PEG Hydrogels Journal of Biomedical Materials Research. Part A. Nov, 2007 | Pubmed ID: 17437304 Temporomandibular joint (TMJ) disorders are most commonly associated with TMJ disc dislocation and osteoarthritis, which can cause erosion of the articular cartilage on the head of the mandibular condyle. There has been little attention focused on treating the damaged condylar cartilage. Therefore, the overall goal of this research is to create a tissue engineering therapy for resurfacing the damaged cartilage of the condylar process with healthy living tissue. Initially, bovine condylar cartilage explants were studied to understand the tissue structure, composition, and gene expression of the native tissue. The cell response of isolated condylar chondrocytes encapsulated in photopolymerized poly(ethylene glycol) hydrogels as a tissue engineering scaffold was examined in the presence and absence of dynamic loading for up to three days of culture. Condylar chondrocyte viability was maintained within the PEG hydrogel constructs over the culture period and loading conditions. Cell response was examined through real-time RTPCR for collagen types I and II and aggrecan, nitric oxide production, cell proliferation, proteoglycan (PG) synthesis, and spatial distribution of extracellular matrix through histology. This study demonstrates that PEG hydrogel constructs are suitable for condylar chondrocyte encapsulation in the absence of loading. However, dynamic compressive strains resulted in inhibition of gene expression, cell proliferation, and PG synthesis.
Designing 3D Photopolymer Hydrogels to Regulate Biomechanical Cues and Tissue Growth for Cartilage Tissue Engineering Pharmaceutical Research. Oct, 2008 | Pubmed ID: 18509600 Synthetic hydrogels fabricated from photopolymerization are attractive for tissue engineering for their controlled macroscopic properties, the ability to incorporate biological functionalities, and cell encapsulation. The goal of the present study was to exploit the attractive features of synthetic hydrogels to elucidate the role of gel structure and chemistry in regulating biomechanical cues.
Cross-linking Density Alters Early Metabolic Activities in Chondrocytes Encapsulated in Poly(ethylene Glycol) Hydrogels and Cultured in the Rotating Wall Vessel Biotechnology and Bioengineering. Mar, 2009 | Pubmed ID: 18949761 In designing a tissue engineering strategy for cartilage repair, selection of both the bioreactor, and scaffold is important to the development of a mechanically functional tissue. The hydrodynamic environment associated with many bioreactors enhances nutrient transport, but also introduces fluid shear stress, which may influence cellular response. This study examined the combined effects of hydrogel cross-linking and the hydrodynamic environment on early chondrocyte response. Specifically, chondrocytes were encapsulated in poly(ethylene glycol) (PEG) hydrogels having two different cross-linked structures, corresponding to a low and high cross-linking density. Both cross-linked gels yielded high water contents (92% and 79%, respectively) and mesh sizes of 150 and 60 A respectively. Cell-laden PEG hydrogels were cultured in rotating wall vessels (RWV) or under static cultures for up to 5 days. Rotating cultures yielded low fluid shear stresses (< or = 0.11 Pa) at the hydrogel periphery indicating a laminar hydrodynamic environment. Chondrocyte response was measured through total DNA content, total nitric oxide (NO) production, and matrix deposition for glycosaminoglycans (GAG). In static cultures, gel cross-linking had no effect on DNA content, NO production, or GAG production; although GAG production increased with culture time for both cross-linked gels. In rotating cultures, DNA content increased, NO production decreased, and overall GAG production decreased when compared to static controls for the low cross-linked gels. For the high cross-linked gels, the hydrodynamic environment had no effect on DNA content, but exhibited similar results to the low cross-linked gel for NO production, and matrix production. Our findings demonstrated that at early culture times, when there is limited matrix production, the hydrodynamic environment dramatically influences cell response in a manner dependent on the gel cross-linking, which may impact long-term tissue development.
Medium Osmolarity and Pericellular Matrix Development Improves Chondrocyte Survival when Photoencapsulated in Poly(ethylene Glycol) Hydrogels at Low Densities Tissue Engineering. Part A. Oct, 2009 | Pubmed ID: 19331581 The ability to encapsulate cells over a range of cell densities is important toward mimicking cell densities of native tissues and rationally designing strategies where cell source and/or cell numbers are clinically limited. Our preliminary findings demonstrate that survival of freshly isolated adult bovine chondrocytes dramatically decreases when photoencapsulated in poly(ethylene glycol) hydrogels at low densities (4 million cells/mL). During enzymatic digestion of cartilage, chondrocytes undergo a harsh change in their microenvironment. We hypothesize that the absence of exogenous antioxidants, the hyposmotic environment, and the loss of a protective pericellular matrix (PCM) increase chondrocytes' susceptibility to free radical damage during photoencapsulation. Incorporation of antioxidants and serum into the encapsulation medium improved cell survival twofold compared to phosphate-buffered saline. Increasing medium osmolarity from 330 to 400 mOsm (physiological) improved cell survival by 40% and resulted in approximately 2-fold increase in adenosine triphosphate (ATP) production 24 h postencapsulation. However, cell survival was only temporary. Allowing cells to reproduce some PCM before photoencapsulation in 400 mOsm medium resulted in superior cell survival during and postencapsulation for up to 15 days. In summary, the combination of antioxidants, physiological osmolarity, and the development of some PCM result in an improved robustness against free radical damage during photoencapsulation.
Cell-matrix Interactions and Dynamic Mechanical Loading Influence Chondrocyte Gene Expression and Bioactivity in PEG-RGD Hydrogels Acta Biomaterialia. Oct, 2009 | Pubmed ID: 19508905 The pericellular matrix (PCM) surrounding chondrocytes is thought to play an important role in transmitting biochemical and biomechanical signals to the cells, which regulates many cellular functions including tissue homeostasis. To better understand chondrocytes interactions with their PCM, three-dimensional poly(ethylene glycol) (PEG) hydrogels containing Arg-Gly-Asp (RGD), the cell-adhesion sequence found in fibronectin and which is present in the PCM of cartilage, were employed. RGD was incorporated into PEG hydrogels via tethers at 0.1, 0.4 and 0.8 mM concentrations. Bovine chondrocytes were encapsulated in the hydrogels and subjected to dynamic compressive strains (0.3 Hz, 18% amplitude strain) for 48h, and their response assessed by cell morphology, ECM gene expression, cell proliferation and matrix synthesis. Incorporation of RGD did not influence cell morphology under free swelling conditions. However, the level of cell deformation upon an applied strain was greater in the presence of RGD. In the absence of dynamic loading, RGD appears to have a negative effect on chondrocyte phenotype, as seen by a 4.7-fold decrease in collagen II/collagen I expressions in 0.8mM RGD constructs. However, RGD had little effect on early responses of chondrocytes (i.e. cell proliferation and matrix synthesis/deposition). When isolating RGD as a biomechanical cue, cellular response was very different. Chondrocyte phenotype (collagen II/collagen I ratio) and proteoglycan synthesis were enhanced with higher concentrations of RGD. Overall, our findings demonstrate that RGD ligands enhance cartilage-specific gene expression and matrix synthesis, but only when mechanically stimulated, suggesting that cell-matrix interactions mediate chondrocyte response to mechanical stimulation.
Dynamic Loading Stimulates Chondrocyte Biosynthesis when Encapsulated in Charged Hydrogels Prepared from Poly(ethylene Glycol) and Chondroitin Sulfate Matrix Biology : Journal of the International Society for Matrix Biology. Jan, 2010 | Pubmed ID: 19720146 This study aimed to elucidate the role of charge in mediating chondrocyte response to loading by employing synthetic 3D hydrogels. Specifically, neutral poly(ethylene glycol) (PEG) hydrogels were employed where negatively charged chondroitin sulfate (ChS), one of the main extracellular matrix components of cartilage, was systematically incorporated into the PEG network at 0%, 20% or 40% to control the fixed charge density. PEG hydrogels were employed as a control environment for extracellular events which occur as a result of loading, but which are not associated with a charged matrix (e.g., cell deformation and fluid flow). Freshly isolated bovine articular chondrocytes were embedded in the hydrogels and subject to dynamic mechanical stimulation (0.3Hz, 15% amplitude strains, 6h) and assayed for nitric oxide production, cell proliferation, proteoglycan synthesis, and collagen deposition. In the absence of loading, incorporation of charge inhibited cell proliferation by approximately 75%, proteoglycan synthesis by approximately 22-50% depending on ChS content, but had no affect on collagen deposition. Dynamic loading had no effect on cellular responses in PEG hydrogels. However, dynamically loading 20% ChS gels inhibited nitrite production by 50%, cell proliferation by 40%, but stimulated proteoglycan and collagen deposition by 162% and 565%, respectively. Dynamic loading of 40% ChS hydrogels stimulated nitrite production by 62% and proteoglycan synthesis by 123%, but inhibited cell proliferation by 54% and collagen deposition by 52%. Upon removing the load and culturing under free-swelling conditions for 36h, the enhanced matrix synthesis observed in the 20% ChS gels was not maintained suggesting that loading is necessary to stimulate matrix production. In conclusion, extracellular events associated with a charged matrix have a dramatic affect on how chondrocytes respond to mechanical stimulation within these artificial 3D matrices suggesting that streaming potentials and/or dynamic changes in osmolarity may be important regulators of chondrocytes while cell deformation and fluid flow appear to have less of an effect.
Comparative Impact of Voltage-gated Calcium Channels and NMDA Receptors on Mitochondria-mediated Neuronal Injury The Journal of Neuroscience : the Official Journal of the Society for Neuroscience. May, 2012 | Pubmed ID: 22573686 Glutamate excitotoxicity, a major component of many neurodegenerative disorders, is characterized by excessive calcium influx selectively through NMDARs. However, there is a substantial uncertainty concerning why other known routes of significant calcium entry, in particular, VGCCs, are not similarly toxic. Here, we report that in the majority of neurons in rat hippocampal and cortical cultures, maximal L-type VGCC activation induces much lower calcium loading than toxic NMDAR activation. Consequently, few depolarization-activated neurons exhibit calcium deregulation and cell death. Activation of alternative routes of calcium entry induced neuronal death in proportion to the degree of calcium loading. In a small subset of neurons, depolarization evoked stronger calcium elevations, approaching those induced by toxic NMDA. These neurons were characterized by elevated expression of VGCCs and enhanced voltage-gated calcium currents, mitochondrial dysfunction and cell death. Preventing VGCC-dependent mitochondrial calcium loading resulted in stronger cytoplasmic calcium elevations, whereas inhibiting mitochondrial calcium clearance accelerated mitochondrial depolarization. Both observations further implicate mitochondrial dysfunction in VGCC-mediated cell death. Results indicate that neuronal vulnerability tracks the extent of calcium loading but does not appear to depend explicitly on the route of calcium entry.
The Interactive Roles of Zinc and Calcium in Mitochondrial Dysfunction and Neurodegeneration Journal of Neurochemistry. Oct, 2013 | Pubmed ID: 24127746 Zinc has been implicated in neurodegeneration following ischemia. In analogy with calcium, zinc has been proposed to induce toxicity via mitochondrial dysfunction, but the relative role of each cation in mitochondrial damage remains unclear. Here, we report that under conditions mimicking ischemia in hippocampal neurons - normal (2 mM) calcium plus elevated (> 100 μM) exogenous zinc - mitochondrial dysfunction evoked by glutamate, kainate or direct depolarization is, despite significant zinc uptake, primarily governed by calcium. Thus, robust mitochondrial ion accumulation, swelling, depolarization, and reactive oxygen species generation were only observed after toxic stimulation in calcium-containing media. This contrasts with the lack of any mitochondrial response in zinc-containing but calcium-free medium, even though zinc uptake and toxicity were strong under these conditions. Indeed, abnormally high, ionophore-induced zinc uptake was necessary to elicit any mitochondrial depolarization. In calcium- and zinc-containing media, depolarization-induced zinc uptake facilitated cell death and enhanced accumulation of mitochondrial calcium, which localized to characteristic matrix precipitates. Some of these contained detectable amounts of zinc. Together these data indicate that zinc uptake is generally insufficient to trigger mitochondrial dysfunction, so that mechanism(s) of zinc toxicity must be different from that of calcium. Zinc and calcium are both implicated in ischemic injury. While calcium toxicity is mediated by mitochondrial dysfunction, mechanisms of zinc toxicity are unclear. Here we show that under conditions mimicking ischemia in hippocampal neurons, glutamate-induced mitochondrial dysfunction - swelling (red mitochondrion), depolarization and increased ROS generation - is triggered only by calcium, while zinc mainly facilitates calcium-dependent mitochondrial damage.