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Effects of BDNF-loaded poly (lactide-co-glycolide) nanoparticle on hBMEC cell viability and its effectiveness in penetrating the BBB model

Siti N. Kamarudin1*, Gabriele R A Froemming2, Minaketan Tripathy3, Igor Iezhitsa4, Renad N Alyautdin5 and Nafeeza Mohd Ismail1
  • 1 University Teknologi Mara (UiTM), Pharmacology/ Faculty of Medicine, Malaysia
  • 2 University Teknologi Mara (UiTM), IPPerForM / Faculty of Medicine, Malaysia
  • 3 University Teknologi Mara (UiTM), Fundamental of Pharmaceutics / Faculty of Pharmacy, Malaysia
  • 4 University Teknologi Mara (UiTM), Centre for Neuroscience Research, Faculty of Medicine, Malaysia
  • 5 Ministry of Health Russian Federation , Scientific Centre for Expertise of Medical Application Products, Russia

Poly lactic-co-glycolic acid (PLGA) based nanoparticles are considered to be a promising drug carrier due to their extensive bioavailability, better encapsulation, controlled release properties and low cytotoxicity. Brain Derived Neurotrophic Factor (BDNF) is beneficial to brain tissue especially after ischemic injury promoting the survival of mature neuron cells. The objectives of this research were: 1) to prepare BDNF-loaded PLGA NPs coated with Apolipoprotein E (ApoE) to enhance BBB crossing via LDL-receptor mediated endocytosis; 2) to evaluate in vitro intracellular uptake and possible cytotoxicity of NPs using hBMEC cell line. Cell viability was determined using MTS-assay and cellular uptake of the NPs by the hBMEC cells was investigated using coumarin 6 as a fluorescent probe. For cells toxicity experiments, 5×104 cells per well were cultured in 96-well plates. After 4 hrs incubation with PLGA particles, 20 µl CellTiter 96s Aqueous One Solution was added directly to the cells and the formation of formazan was measured. Percentage of viability has been calculated relative to untreated control. For penetration study, 1×105 cells per well were cultured in 2 well chamber slides. After 4 hrs incubation, the nuclei were stained with DAPI; the cells were fixed and then imaged by CLSM to visualize the internalization of NPs. Totally 50 cells were analyzed using Leica Qwin software. The fluorescence intensity of the coumarin was measured according to its wavelength. Cell viability increased by 10.4% and 10.1% compared to control after exposure of hBMEC to PLGA-BDNF and PLGA-BDNF-ApoE respectively (p<0.001). Cellular uptake of PLGA-BDNF-ApoE was found to be significantly higher than PLGA-BDNF. In conclusion, PLGA-BDNF nanoparticles does not cause toxicity to hBMEC and was able to penetrate the cells. The penetration was enhanced by addition of ApoE on the surface of PLGA nanoparticles.

Acknowledgements

We would like to acknowledge the financial support from grant 600-RMI/ERGS 5/3 (36/2013) by MOE, Malaysia.

Keywords: Blood-Brain Barrier, Vitamin E, Toxicity, in vitro, nanoparticle

Conference: 14th Meeting of the Asian-Pacific Society for Neurochemistry, Kuala Lumpur, Malaysia, 27 Aug - 30 Aug, 2016.

Presentation Type: Poster Presentation Session

Topic: 14th Meeting of the Asian-Pacific Society for Neurochemistry

Citation: Kamarudin SN, Froemming G, Tripathy M, Iezhitsa I, Alyautdin R and Mohd Ismail N (2016). Effects of BDNF-loaded poly (lactide-co-glycolide) nanoparticle on hBMEC cell viability and its effectiveness in penetrating the BBB model. Conference Abstract: 14th Meeting of the Asian-Pacific Society for Neurochemistry. doi: 10.3389/conf.fncel.2016.36.00153

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Received: 04 Aug 2016; Published Online: 11 Aug 2016.

* Correspondence: Dr. Siti N Kamarudin, University Teknologi Mara (UiTM), Pharmacology/ Faculty of Medicine, Sg Buloh, Selangor, Malaysia, syafika84@yahoo.com