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Sajnani G.,Neural Repair and Biomaterials Laboratory | Requena J.R.,University of Santiago de Compostela
Prion | Year: 2012

It has been described that the breakdown of β-sheets in PrP Sc by denaturation results in loss of infectivity and PK-sensitivity, suggesting a relationship between the structure and PK-resistance. It is also known that an important fraction of total PrPSc is PK-sensitive and can be isolated by the method we already described. Consequently, we decided to employ the PK-sensitive fraction of PrPSc as a potential and useful tool for structural studies. Thus, two essential questions were addressed in our recent article. First, the difference in the infectivity between the sensitive and resistant fractions and second, whether sensitive and resistant PrP Sc shared the same conformation or were only different size multimers with the same basic conformation. Here we discuss our latest data in light of recent infectivity studies and their possible implications on the conformation of the prion. © 2012 Landes Bioscience.


Sajnani G.,University of Santiago de Compostela | Sajnani G.,Neural Repair and Biomaterials Laboratory | Silva C.J.,Albany Research Center | Ramos A.,University of Santiago de Compostela | And 11 more authors.
PLoS Pathogens | Year: 2012

One of the main characteristics of the transmissible isoform of the prion protein (PrPSc) is its partial resistance to proteinase K (PK) digestion. Diagnosis of prion disease typically relies upon immunodetection of PK-digested PrPSc following Western blot or ELISA. More recently, researchers determined that there is a sizeable fraction of PrPSc that is sensitive to PK hydrolysis (sPrPSc). Our group has previously reported a method to isolate this fraction by centrifugation and showed that it has protein misfolding cyclic amplification (PMCA) converting activity. We compared the infectivity of the sPrPSc versus the PK-resistant (rPrPSc) fractions of PrPSc and analyzed the biochemical characteristics of these fractions under conditions of limited proteolysis. Our results show that sPrPSc and rPrPSc fractions have comparable degrees of infectivity and that although they contain different sized multimers, these multimers share similar structural properties. Furthermore, the PK-sensitive fractions of two hamster strains, 263K and Drowsy (Dy), showed strain-dependent differences in the ratios of the sPrPSc to the rPrPSc forms of PrPSc. Although the sPrPSc and rPrPSc fractions have different resistance to PK-digestion, and have previously been shown to sediment differently, and have a different distribution of multimers, they share a common structure and phenotype.


Vara H.,Neural Repair and Biomaterials Laboratory | Collazos-Castro J.E.,Neural Repair and Biomaterials Laboratory
ACS Applied Materials and Interfaces | Year: 2015

Carbon microfibers (MFs) coated with conducting polymers may provide a solution for long-term recording of activity from individual or small groups of neurons. Attaching cell adhesion molecules to the electro-sensitive surface might further improve electrode-neuron contact, thus enhancing signal stability and fidelity. We fabricated biofunctionalized microelectrodes consisting of 7-∼m diameter carbon MFs coated with poly(3,4-ethylenedioxythiophene) doped with poly[(4-styrenesulfonic acid)-co-(maleic acid)] (PEDOT:PSS-co-MA), and linked N-Cadherin to the polymer surface. These electrodes were tested for recording artificially generated electric potentials, as well as multiunit activity (MUA), sharp wave-ripple complexes (SWRs), and field excitatory postsynaptic potentials (fEPSPs) in rat hippocampal slices. The effects of electrode length and functionalization were compared. PEDOT:PSS-co-MA coating improved electric current detection and reduced the electrical noise but had no significant effect on the amplitude of recorded biopotentials. Surface biofunctionalization lowered the electric current flow, and further reduced the electrical noise. Additionally, it increased the amplitude of the recorded MUA, finally doubling the signal-to-noise ratio achieved with bare carbon MFs. Biofunctionalization benefits were apparent only for potentials from cells putatively adjacent to the microelectrode. Analysis of fEPSPs excluded adverse effects of functionalized electrodes in basal synaptic transmission. These results demonstrate the possibility of enhancing the amplitude and signal-to-noise ratio of neural recordings by coating the microelectrodes with conducting polymers modified with neural cell adhesion molecules, and support the use of biofunctionalized MFs in advanced neuroprosthetic devices. © 2015 American Chemical Society.


Collazos-Castro J.E.,Neural Repair and Biomaterials Laboratory | Garcia-Rama C.,Neural Repair and Biomaterials Laboratory | Alves-Sampaio A.,Neural Repair and Biomaterials Laboratory
Acta Biomaterialia | Year: 2016

Electroactive systems that promote directional axonal growth and migration of glial progenitor cells (GPC) are needed for the treatment of neurological injuries. We report the functionalization of electroconducting microfibers with multiple biomolecules that synergistically stimulate the proliferation and migration of GPC, which in turn induce axonal elongation from embryonic cerebral cortex neurons. PEDOT doped with poly[(4-styrenesulfonic acid)-co-(maleic acid)] was synthesized on carbon microfibers and used for covalent attachment of molecules to the electroactive surface. The molecular complexes that promoted GPC proliferation and migration, followed by axonal extension, were composed of polylysine, heparin, basic fibroblast growth factor (bFGF), and matricellular proteins; the combination of bFGF with vitronectin or fibronectin being indispensable for sustained glial and axonal growth. The rate of glial-induced axonal elongation was about threefold that of axons growing directly on microfibers functionalized with polylysine alone. Electrical stimuli applied through the microfibers released bFGF and fibronectin from the polymer surface, consequently reducing GPC proliferation and promoting their differentiation into astrocytes, without causing cell detachment or toxicity. These results suggest that functionalized electroactive microfibers may provide a multifunctional tool for controlling neuron-glia interactions and enhancing neural repair. Statement of Significance: We report a multiple surface functionalization strategy for electroconducting microfibers (MFs), in order to promote proliferation and guided migration of glial precursor cells (GPC) and consequently create a permissive substrate for elongation of central nervous system (CNS) axons. GPC divided and migrated extensively on the functionalized MFs, leading to fast elongation of embryonic cerebral cortex axons. The application of electric pulses thorough the MFs controlled glial cell division and differentiation. The functionalized MFs provide an advanced tool for neural tissue engineering and for controlling neuron-glial interactions. CNS axonal growth associated to migratory glial precursors, together with the possibility of directing glial differentiation by electrical stimuli applied through the MFs, open a new research avenue to explore for CNS repair. © 2016 Acta Materialia Inc.


Alves-Sampaio A.,Neural Repair and Biomaterials Laboratory | Garcia-Rama C.,Neural Repair and Biomaterials Laboratory | Collazos-Castro J.E.,Neural Repair and Biomaterials Laboratory
Biomaterials | Year: 2016

Poly(3, 4-ethylenedioxythiophene)-coated carbon microfibers (PEDOT-MFs) hold promise for developing advanced neuroprostheses and neural repair devices. We investigated the chronic cellular responses to PEDOT-MFs implanted into the uninjured and the transected rat spinal cord, and compared the effects of polymer surface biofunctionalization with covalently attached polylysine (PLL) or a multimolecular complex of PLL, heparin, basic fibroblast growth factor (bFGF), and fibronectin. An alginate gel was used to facilitate microfiber implantation and reduce connective tissue scarring after spinal cord injury (SCI). PLL/heparin/bFGF/fibronectin-functionalized PEDOT-MFs showed excellent integration within the uninjured and injured spinal cord, frequently establishing contact with neuronal somas, axons, dendrites and glial cells, accompanied by very little or absent scarring response. On the contrary, non-functionalized and PLL-functionalized microfibers provoked inflammation and fibrosis with loss of neural elements in the surrounding tissue. Within the lesion, the PEDOT-MFs by themselves facilitated longitudinal alignment of migratory cells and growing axons, and their modification with PLL/heparin/bFGF/fibronectin promoted tissue healing, enhancing blood vessel formation and axonal regeneration without increasing inflammation. These results support the incorporation of biofunctionalized electroconducting microfibers in neuro-electronic interfaces and lesion-bridging systems for the treatment of SCI. © 2016 Elsevier Ltd.

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