17 research outputs found
Intraarterial route increases the risk of cerebral lesions after mesenchymal cell administration in animal model of ischemia
Mesenchymal stem cells (MSCs) are a promising clinical therapy for ischemic stroke. However, critical parameters, such as the most effective administration route, remain unclear. Intravenous (i.v.) and intraarterial (i.a.) delivery routes have yielded varied outcomes across studies, potentially due to the unknown MSCs distribution. We investigated whether MSCs reached the brain following i.a. or i.v. administration after transient cerebral ischemia in rats, and evaluated the therapeutic effects of both routes. MSCs were labeled with dextran-coated superparamagnetic nanoparticles for magnetic resonance imaging (MRI) cell tracking, transmission electron microscopy and immunohistological analysis. MSCs were found in the brain following i.a. but not i.v. administration. However, the i.a. route increased the risk of cerebral lesions and did not improve functional recovery. The i.v. delivery is safe but MCS do not reach the brain tissue, implying that treatment benefits observed for this route are not attributable to brain MCS engrafting after stroke.This study has been partially supported by grants from Axencia Galega de Innovación (Xunta de Galicia), the Instituto de Salud Carlos III (PI13/00292; PI14/01879), the Spanish Research Network on Cerebrovascular Diseases RETICS-INVICTUS (RD12/0014), Xunta de Galicia (Consellería Educación GRC2014/027), the European Commission program FEDER and Promoting Active Ageing program: Functional Nanostructures For Alzheimer’s Disease At Ultra-Early Stages” (Pana_686009), a Research and Innovation Project, funded within the EU Horizon 2020 Programme”. Furthermore, this study was also co-funded within the POCTEP (Operational Programme for Cross-border Cooperation Spain-Portugal) program (0681_INVENNTA_1_E), co-financed by the ERDF (European Regional Development Fund). T. Sobrino (CP12/03121) and F. Campos (CP14/00154) are recipients of a research contract from Miguel Servet Program of Instituto de Salud Carlos III. Finally, P. Taboada thanks Mineco and Xunta de Galicia for funding through projects MAT2013-40971-R and EM2013-046, respectively. J Trekker is the recipient of an innovation grant from the IWT-VlaanderenS
Enzyme-Loaded Gel Core Nanostructured Lipid Carriers to Improve Treatment of Lysosomal Storage Diseases: Formulation and In Vitro Cellular Studies of Elosulfase Alfa-Loaded Systems
Mucopolysaccharidosis IVA (Morquio A) is a rare inherited metabolic disease caused by
deficiency of the lysosomal enzyme N-acetylgalatosamine-6-sulfate-sulfatase (GALNS). Until now,
treatments employed included hematopoietic stem cell transplantation and enzyme replacement
therapy (ERT); the latter being the most commonly used to treat mucopolysaccharidoses, but with
serious disadvantages due to rapid degradation and clearance. The purpose of this study was
to develop and evaluate the potential of nanostructured lipid carriers (NLCs) by encapsulating
elosulfase alfa and preserving its enzyme activity, leading to enhancement of its biological effect
in chondrocyte cells. A pegylated elosulfase alfa-loaded NLC was characterized in terms of size,
ζ potential, structural lipid composition (DSC and XRD), morphology (TEM microscopy), and
stability in human plasma. The final formulation was freeze-dried by selecting the appropriate
cryoprotective agent. Viability assays confirmed that NLCs were non-cytotoxic to human fibroblasts.
Imaging techniques (confocal and TEM) were used to assess the cellular uptake of NLCs loaded with
elosulfase alfa. This study provides evidence that the encapsulated drug exhibits enzyme activity
inside the cells. Overall, this study provides a new approach regarding NLCs as a promising delivery
system for the encapsulation of elosulfase alfa or other enzymes and the preservation of its activity
and stability to be used in enzymatic replacement therapy (ERT).This research was funded by Xunta de Galicia, grant number GRC2013/015 and GPC2017/015S
A new seipin-associated neurodegenerative syndrome
Background: Seipin/BSCL2 mutations can cause type
2 congenital generalised lipodystrophy (BSCL) or
dominant motor neurone diseases. Type 2 BSCL is
frequently associated with some degree of intellectual
impairment, but not to fatal neurodegeneration. In order
to unveil the aetiology and pathogenetic mechanisms of
a new neurodegenerative syndrome associated with a
novel BSCL2 mutation, six children, four of them
showing the BSCL features, were studied.
Methods: Mutational and splicing analyses of BSCL2
were performed. The brain of two of these children was
examined postmortem. Relative expression of BSCL2
transcripts was analysed by real-time reverse
transcription-polymerase chain reaction (RT-PCR) in
different tissues of the index case and controls.
Overexpressed mutated seipin in HeLa cells was analysed
by immunofluorescence and western blotting.
Results: Two patients carried a novel homozygous
c.985C>T mutation, which appeared in the other four
patients in compound heterozygosity. Splicing analysis
showed that the c.985C>T mutation causes an aberrant
splicing site leading to skipping of exon 7. Expression of
exon 7-skipping transcripts was very high with respect to
that of the non-skipped transcripts in all the analysed
tissues of the index case. Neuropathological studies
showed severe neurone loss, astrogliosis and intranuclear
ubiquitin(+) aggregates in neurones from multiple
cortical regions and in the caudate nucleus.
Conclusions: Our results suggest that exon 7 skipping
in the BSCL2 gene due to the c.985C>T mutation is
responsible for a novel early onset, fatal
neurodegenerative syndrome involving cerebral cortex
and basal ganglia.Instituto de Salud Carlos III (grant number PI 10/02873) and European
Regional Development Fund, FEDER (grant number 10PXIB208013PR) and
Consellería de Industria, Xunta de Galicia.S