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Increased antiparkinson efficacy of the combined administration of VEGF- and GDNF-loaded nanospheres in a partial lesion model of Parkinson's disease.

Herrán E, Requejo C, Ruiz-Ortega JA, Aristieta A, Igartua M, Bengoetxea H, Ugedo L, Pedraz JL, Lafuente JV, Hernández RM - Int J Nanomedicine (2014)

Bottom Line: The NS particle size was about 200 nm and the simultaneous addition of VEGF NS and GDNF NS resulted in significant protection of the PC-12 cell line against 6-OHDA in vitro.The results showed that VEGF NS and GDNF NS significantly decreased the number of amphetamine-induced rotations at the end of the study.In addition, tyrosine hydroxylase immunohistochemical analysis in the striatum and the external substantia nigra confirmed a significant enhancement of neurons in the VEGF NS and GDNF NS treatment group.

View Article: PubMed Central - PubMed

Affiliation: NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque Country (UPV/EHU), School of Pharmacy, Vitoria, Spain ; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Vitoria, Spain.

ABSTRACT
Current research efforts are focused on the application of growth factors, such as glial cell line-derived neurotrophic factor (GDNF) and vascular endothelial growth factor (VEGF), as neuroregenerative approaches that will prevent the neurodegenerative process in Parkinson's disease. Continuing a previous work published by our research group, and with the aim to overcome different limitations related to growth factor administration, VEGF and GDNF were encapsulated in poly(lactic-co-glycolic acid) nanospheres (NS). This strategy facilitates the combined administration of the VEGF and GDNF into the brain of 6-hydroxydopamine (6-OHDA) partially lesioned rats, resulting in a continuous and simultaneous drug release. The NS particle size was about 200 nm and the simultaneous addition of VEGF NS and GDNF NS resulted in significant protection of the PC-12 cell line against 6-OHDA in vitro. Once the poly(lactic-co-glycolic acid) NS were implanted into the striatum of 6-OHDA partially lesioned rats, the amphetamine rotation behavior test was carried out over 10 weeks, in order to check for in vivo efficacy. The results showed that VEGF NS and GDNF NS significantly decreased the number of amphetamine-induced rotations at the end of the study. In addition, tyrosine hydroxylase immunohistochemical analysis in the striatum and the external substantia nigra confirmed a significant enhancement of neurons in the VEGF NS and GDNF NS treatment group. The synergistic effect of VEGF NS and GDNF NS allows for a reduction of the dose by half, and may be a valuable neurogenerative/neuroreparative approach for treating Parkinson's disease.

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In vitro neurotoxicity assay.Notes: (A) DAPI immunostaining after different 6-OHDA concentration treatments to establish the neurotoxic cell model. (B) PC-12 cell viability evaluation after 6-OHDA induced toxicity in vitro. *P<0.05, V 10 + G 10 (+6-OHDA) versus C−, and C− (+6-OHDA) groups. ***P<0.001, V 5 + G 5 (+6-OHDA) versus C−, and C− (+6-OHDA) groups.Abbreviations: 6-OHDA, 6-hydroxydopamine; C−, negative control; DAPI, 4′,6-diamidino-2-phenylindole; G 5, GDNF 5 ng/mL; G 10, GDNF 10 ng/mL; GDNF, glial cell line-derived neurotrophic factor; OD, optical density; V 5, VEGF 5 ng/mL; V 10, VEGF 10 ng/mL; VEGF, vascular endothelial growth factor.
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f3-ijn-9-2677: In vitro neurotoxicity assay.Notes: (A) DAPI immunostaining after different 6-OHDA concentration treatments to establish the neurotoxic cell model. (B) PC-12 cell viability evaluation after 6-OHDA induced toxicity in vitro. *P<0.05, V 10 + G 10 (+6-OHDA) versus C−, and C− (+6-OHDA) groups. ***P<0.001, V 5 + G 5 (+6-OHDA) versus C−, and C− (+6-OHDA) groups.Abbreviations: 6-OHDA, 6-hydroxydopamine; C−, negative control; DAPI, 4′,6-diamidino-2-phenylindole; G 5, GDNF 5 ng/mL; G 10, GDNF 10 ng/mL; GDNF, glial cell line-derived neurotrophic factor; OD, optical density; V 5, VEGF 5 ng/mL; V 10, VEGF 10 ng/mL; VEGF, vascular endothelial growth factor.

Mentions: In this assay, we tested whether VEGF and GDNF released from PLGA NS were able to protect PC-12 cells from 6-OHDA toxicity. As shown in Figure 3A, when the PC-12 cell line was exposed to different 6-OHDA concentrations, the damage to cells was evident under fluorescence microscopy when 0.1 mM and 0.2 mM concentrations were added. These results have been convincingly corroborated by cell viability assay, giving rise to a selection of 0.1 mM 6-OHDA as the ideal dose with which to damage half of the cells (data not shown).


Increased antiparkinson efficacy of the combined administration of VEGF- and GDNF-loaded nanospheres in a partial lesion model of Parkinson's disease.

Herrán E, Requejo C, Ruiz-Ortega JA, Aristieta A, Igartua M, Bengoetxea H, Ugedo L, Pedraz JL, Lafuente JV, Hernández RM - Int J Nanomedicine (2014)

In vitro neurotoxicity assay.Notes: (A) DAPI immunostaining after different 6-OHDA concentration treatments to establish the neurotoxic cell model. (B) PC-12 cell viability evaluation after 6-OHDA induced toxicity in vitro. *P<0.05, V 10 + G 10 (+6-OHDA) versus C−, and C− (+6-OHDA) groups. ***P<0.001, V 5 + G 5 (+6-OHDA) versus C−, and C− (+6-OHDA) groups.Abbreviations: 6-OHDA, 6-hydroxydopamine; C−, negative control; DAPI, 4′,6-diamidino-2-phenylindole; G 5, GDNF 5 ng/mL; G 10, GDNF 10 ng/mL; GDNF, glial cell line-derived neurotrophic factor; OD, optical density; V 5, VEGF 5 ng/mL; V 10, VEGF 10 ng/mL; VEGF, vascular endothelial growth factor.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4043720&req=5

f3-ijn-9-2677: In vitro neurotoxicity assay.Notes: (A) DAPI immunostaining after different 6-OHDA concentration treatments to establish the neurotoxic cell model. (B) PC-12 cell viability evaluation after 6-OHDA induced toxicity in vitro. *P<0.05, V 10 + G 10 (+6-OHDA) versus C−, and C− (+6-OHDA) groups. ***P<0.001, V 5 + G 5 (+6-OHDA) versus C−, and C− (+6-OHDA) groups.Abbreviations: 6-OHDA, 6-hydroxydopamine; C−, negative control; DAPI, 4′,6-diamidino-2-phenylindole; G 5, GDNF 5 ng/mL; G 10, GDNF 10 ng/mL; GDNF, glial cell line-derived neurotrophic factor; OD, optical density; V 5, VEGF 5 ng/mL; V 10, VEGF 10 ng/mL; VEGF, vascular endothelial growth factor.
Mentions: In this assay, we tested whether VEGF and GDNF released from PLGA NS were able to protect PC-12 cells from 6-OHDA toxicity. As shown in Figure 3A, when the PC-12 cell line was exposed to different 6-OHDA concentrations, the damage to cells was evident under fluorescence microscopy when 0.1 mM and 0.2 mM concentrations were added. These results have been convincingly corroborated by cell viability assay, giving rise to a selection of 0.1 mM 6-OHDA as the ideal dose with which to damage half of the cells (data not shown).

Bottom Line: The NS particle size was about 200 nm and the simultaneous addition of VEGF NS and GDNF NS resulted in significant protection of the PC-12 cell line against 6-OHDA in vitro.The results showed that VEGF NS and GDNF NS significantly decreased the number of amphetamine-induced rotations at the end of the study.In addition, tyrosine hydroxylase immunohistochemical analysis in the striatum and the external substantia nigra confirmed a significant enhancement of neurons in the VEGF NS and GDNF NS treatment group.

View Article: PubMed Central - PubMed

Affiliation: NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque Country (UPV/EHU), School of Pharmacy, Vitoria, Spain ; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Vitoria, Spain.

ABSTRACT
Current research efforts are focused on the application of growth factors, such as glial cell line-derived neurotrophic factor (GDNF) and vascular endothelial growth factor (VEGF), as neuroregenerative approaches that will prevent the neurodegenerative process in Parkinson's disease. Continuing a previous work published by our research group, and with the aim to overcome different limitations related to growth factor administration, VEGF and GDNF were encapsulated in poly(lactic-co-glycolic acid) nanospheres (NS). This strategy facilitates the combined administration of the VEGF and GDNF into the brain of 6-hydroxydopamine (6-OHDA) partially lesioned rats, resulting in a continuous and simultaneous drug release. The NS particle size was about 200 nm and the simultaneous addition of VEGF NS and GDNF NS resulted in significant protection of the PC-12 cell line against 6-OHDA in vitro. Once the poly(lactic-co-glycolic acid) NS were implanted into the striatum of 6-OHDA partially lesioned rats, the amphetamine rotation behavior test was carried out over 10 weeks, in order to check for in vivo efficacy. The results showed that VEGF NS and GDNF NS significantly decreased the number of amphetamine-induced rotations at the end of the study. In addition, tyrosine hydroxylase immunohistochemical analysis in the striatum and the external substantia nigra confirmed a significant enhancement of neurons in the VEGF NS and GDNF NS treatment group. The synergistic effect of VEGF NS and GDNF NS allows for a reduction of the dose by half, and may be a valuable neurogenerative/neuroreparative approach for treating Parkinson's disease.

Show MeSH
Related in: MedlinePlus