Ioanna Sandvig

L‐DOPA‐Coated Manganese Oxide Nanoparticles as Dual MRI Contrast Agents and Drug‐Delivery Vehicles

Manganese oxide nanoparticles (MONPs) are capable of time-dependent magnetic resonance imaging contrast switching as well as releasing a surface-bound drug. MONPs give T2/T2* contrast, but dissolve and release T1-active Mn(2+) and L-3,4-dihydroxyphenylalanine. Complementary images are acquired with a single contrast agent, and applications toward Parkinsons disease are suggested.

RGD-peptide modified alginate by a chemoenzymatic strategy for tissue engineering applications

One of the main challenges in tissue engineering and regenerative medicine is the ability to maintain optimal cell function and survival post-transplantation. Biomaterials such as alginates are commonly used for immunoisolation, while they may also provide structural support to the cell transplants by mimicking the extracellular matrix. In this study, arginine-glycine-aspartate (RGD)-peptide-coupled alginates of tailored composition were produced by adopting a unique chemoenzymatic strategy for substituting the nongelling mannuronic acid on the alginate. Alginates with and without RGD were produced with high and low content of G. Using carbodiimide chemistry 0.1-0.2% of the sugar units were substituted by peptide. Furthermore, the characterization by (1)H-nuclear magnetic resonance (NMR) revealed by-products from the coupling reaction that partly could be removed by coal filtration. Olfactory ensheathing cells (OECs) and myoblasts were grown in two-dimensional (2D) and 3D cultures of RGD-peptide modified or unmodified alginates obtained by the chemoenzymatically strategy and compared to native alginate. Both OECs and myoblasts adhered to the RGD-peptide modified alginates in 2D cultures, forming bipolar protrusions. OEC encapsulation resulted in cell survival for up to 9 days, thus demonstrating the potential for short-term 3D culture. Myoblasts showed long-term survival in 3D cultures, that is, up to 41 days post encapsulation. The RGD modifications did not result in marked changes in cell viability in 3D cultures. We demonstrate herein a unique technique for tailoring peptide substituted alginates with a precise and flexible composition, conserving the gel forming properties relevant for the use of alginate in tissue engineering.

Injectable Extracellular Matrix Hydrogels as Scaffolds for Spinal Cord Injury Repair.

Restoration of lost neuronal function after spinal cord injury (SCI) still remains a big challenge for current medicine. One important repair strategy is bridging the SCI lesion with a supportive and stimulatory milieu that would enable axonal rewiring. Injectable extracellular matrix (ECM)-derived hydrogels have been recently reported to have neurotrophic potential in vitro. In this study, we evaluated the presumed neuroregenerative properties of ECM hydrogels in vivo in the acute model of SCI. ECM hydrogels were prepared by decellularization of porcine spinal cord (SC) or porcine urinary bladder (UB), and injected into a spinal cord hemisection cavity. Histological analysis and real-time qPCR were performed at 2, 4, and 8 weeks postinjection. Both types of hydrogels integrated into the lesion and stimulated neovascularization and axonal ingrowth into the lesion. On the other hand, massive infiltration of macrophages into the lesion and rapid hydrogel degradation did not prevent cyst formation, which progressively developed over 8 weeks. No significant differences were found between SC-ECM and UB-ECM. Gene expression analysis revealed significant downregulation of genes related to immune response and inflammation in both hydrogel types at 2 weeks post SCI. A combination of human mesenchymal stem cells with SC-ECM did not further promote ingrowth of axons and blood vessels into the lesion, when compared with the SC-ECM hydrogel alone. In conclusion, both ECM hydrogels bridged the lesion cavity, modulated the innate immune response, and provided the benefit of a stimulatory substrate for in vivo neural tissue regeneration. However, fast hydrogel degradation might be a limiting factor for the use of native ECM hydrogels in the treatment of acute SCI.

In vivo MRI of olfactory ensheathing cell grafts and regenerating axons in transplant mediated repair of the adult rat optic nerve

The purpose of the present study was to use magnetic resonance imaging (MRI) as a tool for monitoring transplant‐mediated repair of the adult rat visual pathway. We labelled rat olfactory ensheathing cells (OECs) using micron‐sized particles of iron oxide (MPIO) and transplanted them by: i) intravitreal injection (ivit) and ii) intra‐optic nerve (ON) injection (iON) in adult rats with ON crush (ONC) injury. We applied T2‐weighted MRI and manganese‐enhanced MRI (MEMRI) to visualise transplanted cells and ON axons at specific times after injury and cell engraftment. Our findings demonstrate that ivit MPIO‐labelled OECs are unequivocally detected by T2‐weighted MRI in vivo and that the T1‐weighted 3D FLASH sequence applied for MEMRI facilitates simultaneous visualisation of Mn2+−enhanced regenerating retinal ganglion cell (RGC) axons and MPIO‐labelled OEC grafts. Furthermore, analysis of MRI data and ultrastructural findings supports the hypothesis that iON OEC transplants mediate regeneration and remyelination of RGC axons post injury. Copyright

Current developments in cell- and biomaterial-based approaches for stroke repair.

Introduction: Stroke is one of the most devastating diseases and a leading cause of mortality worldwide. So far, clinical management of stroke involves surgical clot retrieval or thrombolytic treatment inducing reperfusion of the occluded vessels in the cerebral infarcted area, which is dependent on early intervention following insult. New treatment strategies involve the promotion of angiogenesis and neuroplasticity, stimulation of endogenous neurogenesis, remyelinization, and immunomodulation by means of cell transplantation and sustained drug delivery. Areas covered: This review describes different types of stem cells (endogenous and exogenous neural progenitors, pluripotent stem cell derivatives, mesenchymal stem cells [MSCs], olfactory ensheathing cells) and biomaterials, their routes of administration, means of noninvasive imaging, and the prerequisites and hurdles for the successful translation of the cell therapies to the clinic. Expert opinion: Neural precursors (NPs) derived from pluripotent stem cells, unlike MSCs, can not only remodel the CNS by promoting neuroplasticity, angiogenesis, and immunomodulation, but also replace damaged cells. To transfer NPs into the clinic, step by step guidelines for researchers are identified and discussed.

Axonal tracing of the normal and regenerating visual pathway of mouse, rat, frog, and fish using manganese-enhanced MRI (MEMRI)

To assess optic nerve (ON) regeneration after injury by applying manganese‐enhanced MRI (MEMRI) in a study of comparative physiology between nonregenerating rat and mouse species and regenerating frog and fish species.

Epigenetic Modulation of Stem Cells in Neurodevelopment: The Role of Methylation and Acetylation

The coordinated development of the nervous system requires fidelity in the expression of specific genes determining the different neural cell phenotypes. Stem cell fate decisions during neurodevelopment are strictly correlated with their epigenetic status. The epigenetic regulatory processes, such as DNA methylation and histone modifications discussed in this review article, may impact both neural stem cell (NSC) self-renewal and differentiation and thus play an important role in neurodevelopment. At the same time, stem cell decisions regarding fate commitment and differentiation are highly dependent on the temporospatial expression of specific genes contingent on the developmental stage of the nervous system. An interplay between the above, as well as basic cell processes, such as transcription regulation, DNA replication, cell cycle regulation and DNA repair therefore determine the accuracy and function of neuronal connections. This may significantly impact embryonic health and development as well as cognitive processes such as neuroplasticity and memory formation later in the adult.

Labelling of olfactory ensheathing cells with micron-sized particles of iron oxide and detection by MRI

A crucial issue in transplant-mediated repair of the damaged central nervous system (CNS) is serial non-invasive imaging of the transplanted cells, which has led to interest in the application of magnetic resonance imaging (MRI) combined with designated intracellular magnetic labels for cell tracking. Micron-sized particles of iron oxide (MPIO) have been successfully used to track cells by MRI, yet there is relatively little known about either their suitability for efficient labelling of specific cell types, or their effects on cell viability. The purpose of this study was to develop a suitable MPIO labelling protocol for olfactory ensheathing cells (OECs), a type of glia used to promote the regeneration of CNS axons after transplantation into the injured CNS. Here, we demonstrate an OEC labelling efficiency of >90% with an MPIO incubation time as short as 6 h, enabling intracellular particle uptake for single-cell detection by MRI without affecting cell proliferation, migration and viability. Moreover, MPIO are resolvable in OECs transplanted into the vitreous body of adult rat eyes, providing the first detailed protocol for efficient and safe MPIO labelling of OECs for non-invasive MRI tracking of transplanted OECs in real time for use in studies of CNS repair and axon regeneration.

Synthesis of gadolinium oxide nanodisks and gadolinium doped iron oxide nanoparticles for MR contrast agents

Herein, we report the synthesis of differently sized gadolinium oxide nanodisks and gadolinium doped iron oxide spherical and cubic nanoparticles through the thermal decomposition of an oleate precursor. We also demonstrate that these nanoparticles are promising candidates for MR contrast agents.

Using Manganese-Enhanced MRI to Assess Optic Nerve Regeneration

Experimental visual pathway lesion in the form of optic nerve (ON) crush or transection injury results in massive death of retinal ganglion cells (RGCs) and permanent loss of synaptic connections (Berkelaar et al., J Neurosci 14:4368-4374, 1994). Despite the fact that RGC axon regeneration is inhibited in a manner typical of other CNS lesions, the rodent ON injury model is one of the few models where robust axon regeneration has been achieved after therapeutic intervention (Berry et al., Restor Neurol Neurosci 26:147-174, 2008). However, assessment of the efficacy of therapeutic approaches in promoting ON regeneration has traditionally relied on histological methods, which necessitate the sacrifice of experimental animals and thus preclude longitudinal in vivo monitoring of individual subjects. Manganese-enhanced MRI (MEMRI) utilizes the paramagnetic properties and uptake and transport mechanisms of manganese ions (Mn(2+)) by neurons, thus enabling serial in vivo monitoring of the entire axonal projections (Sandvig et al., J Magn Reson Imaging 34:670-675, 2011; Thuen et al., J Magn Reson Imaging 4:492-500, 2005; Pautler et al., Magn Res Med 50:33-39, 2003; Saleem et al., Neurotechnique 34:685-700, 2000). The above properties of Mn(2+) render MEMRI a highly suitable technique for assessment of ON regeneration after injury, especially with a view to in vivo monitoring of neuronal connectivity and axon-regenerative responses to treatment. In this chapter, we provide a generic protocol for ON lesioning and MEMRI application for assessment of ON regeneration in rodents.