Category: Formulation and Quality
Purpose: Perinatal asphyxia caused due to hypoxia complicates and causes neonatal hypoxic-ischemic encephalopathy (HIE). A widely used treatment for HIE is therapeutic hypothermia. In recent times, various neuroprotective and anti-inflammatory agents are explored as adjuvant therapy with hypothermia in clinical settings. Previous reports suggest that lutein supplementation has neuroprotective properties in rats. However, lutein has poor oral bioavailability due to poor aqueous solubility. The aim of the present study is to encapsulate lutein into polymeric nanoparticles (NPs) (PLGA, PLGA-PEG and PLGA-PEG-FOL) and evaluate its uptake in neuroblastoma cells.
Methods: Lutein loaded PLGA, PLGA-PEG and PLGA-PEG-FOL NPs were prepared using oil in water (O/W) emulsion solvent evaporation method. Particle size, polydispersity index and zeta-potential of NPs were measured using dynamic light scattering (DLS) technique. Differential scanning calorimetry (DSC) was used to confirm encapsulation of lutein into NPs. Scanning electron microscopy (SEM) was used to determine surface morphology of NPs. Cellular uptake studies were conducted in human neuroblastoma cell line, SK-N-BE(2) (ATCC CRL-2271). Further characterization of NPs such as encapsulation efficiency (%EE), in-vitro release studies and stability studies were performed.
Results: Lutein was successfully encapsulated into NPs with particle size less than 200 nm, uniform distribution and high zeta potential. Particle size of lutein loaded PLGA, PLGA-PEG, and PLGA-PEG-FOL NPs were 132.6 ± 1.04 nm, 163.3 ± 2.86 nm, and 188.0 ± 4.06 nm, respectively. %EE of lutein was 48.82 ± 15.50, 63.45 ± 10.13 and 72.87 ± 7.22 for lutein PLGA, PLGA-PEG and PLGA-PEG-FOL NPs, respectively. DSC analysis confirmed encapsulation of lutein into NPs. Release profile had shown that NPs released lutein in a sustained manner with no initial burst release. Cumulative release of lutein was higher in PLGA NPs with 100% release within 24hours. In PLGA-PEG and PLGA-PEG-FOL NPs, cumulative release of lutein was ~69% and ~80% at 48h. Cellular uptake studies showed a significant increase uptake of lutein with PLGA-PEG-FOL NPs compared to PLGA-PEG and PLGA NPs (p< 0.0001). There was ~1.8 and ~6.5-fold enhanced uptake of lutein from PLGA-PEG-FOL NPs compared with PLGA-PEG and PLGA NPs.
Conclusion: In this study, we successfully encapsulated lutein into PLGA-PEG-FOL NPs and significantly enhanced the lutein uptake in neuroblastoma cells. Efficiency of lutein loaded PLGA-PEG-FOL NPs will be confirmed in vivo using HIE rat model.
Abhishek Arnipalli– Elpaso, Texas
Xiaoming Ging– El Paso, Texas
Venkata Kashyap Yellepeddi– Salt Lake City, Utah
Mahima Singh– Student, University of the Sciences in Philadelphia, Philadelphia, Pennsylvania
Devaraj Sambalingam– El Paso, Texas
Jwala Renukuntla– Assistant Professor, University of Texas at El Paso, El Paso, Texas