Two-step nanoprecipitation for the production of protein-loaded PLGA nanospheres

نویسندگان

  • Moraima Morales-Cruz
  • Giselle M. Flores-Fernández
  • Myreisa Morales-Cruz
  • Elsie A. Orellano
  • José A. Rodriguez-Martinez
  • Mercedes Ruiz
  • Kai Griebenow
چکیده

One of the first methods to encapsulate drugs within polymer nanospheres was developed by Fessi and coworkers in 1989 and consisted of one-step nanoprecipitation based on solvent displacement. However, proteins are poorly encapsulated within polymer nanoparticles using this method because of their limited solubility in organic solvents. To overcome this limitation, we developed a two-step nanoprecipitation method and encapsulated various proteins with high efficiency into poly(lactic-co-glycolic)acid (PLGA) nanospheres (NP). In this method, a protein nanoprecipitation step is used first followed by a second polymer nanoprecipitation step. Two model enzymes, lysozyme and α-chymotrypsin, were used for the optimization of the method. We obtained encapsulation efficiencies of >70%, an amount of buffer-insoluble protein aggregates of typically <2%, and a high residual activity of typically >90%. The optimum conditions identified for lysozyme were used to successfully encapsulate cytochrome c(Cyt-c), an apoptosis-initiating basic protein of similar size, to verify reproducibility of the encapsulation procedure. The size of the Cyt-c loaded-PLGA nanospheres was around 300-400 nm indicating the potential of the delivery system to passively target tumors. Cell viability studies, using a human cervical cancer cell line (HeLa), demonstrate excellent biocompatibility of the PLGA nanoparticles. PLGA nanoparticles carrying encapsulated Cyt-c were not efficient in causing apoptosis presumably because PLGA nanoparticles are not efficiently taken up by the cells. Future systems will have to be optimized to ascertain efficient cellular uptake of the nanoparticles by, e.g., surface modification with receptor ligands.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Preparation of protein-loaded PLGA-PVP blend nanoparticles by nanoprecipitation method: entrapment, Initial burst and drug release kinetic studies

Objective(s):Despite of wide range applications of polymeric nanoparticles in protein delivery, there are some problems for the field of protein entrapment, initial burst and controlled release profile.   Materials and Methods: In this study, we investigated the influence of some changes in PLGA nanoparticles formulation to improve the initial and controlled release profile. Selected parameters ...

متن کامل

PLGA Nanospheres Loaded with Autoclaved Leishmania Major (ALM) and CpG-ODN: Preparation and in vitro Characterization

Objective(s) Several antigens, adjuvants and delivery systems have been evaluated for induction of protective immune responses against Leishmaniasis, but most of them have been inefficient. In this study, PLGA nanospheres as antigen delivery system CpG-ODN as an immunoadjuvant for increasing the immune responses against Autoclaved Leishmania major (ALM) were prepared and characterized. Materi...

متن کامل

Preparation and characterization of PLGA nanospheres loaded with inactivated influenza virus, CpG-ODN and Quillaja saponin

  Objective(s): The purpose of this study was preparation and evaluation of PLGA nanospheres containing the influenza virus and different adjuvants, Quillaja saponin (QS) and CpG-ODN.   Materials and Methods: Nanospheres were prepared using the double emulsion-solvent evaporation method. The morphological and physicochemical properties were studied by scanning electron microscopy...

متن کامل

Preparation of Methotrexate loaded PLGA nanoparticles coated with PVA and Poloxamer188

Objective(s): Nanoparticles offer an attractive platform for drug delivery through a wide variety of the body's physiological barriers. Furthermore, modification of nanoparticle surface with moeites such as Poloxamer188 can enhance their accumulation and localization at disease site. In this work, we investigated the physiochemical effect of a scavenger receptor (SR-BI) interac...

متن کامل

Controlled Antibiotic Delivery by Gelatin Nanospheres: Optimization, Characterization and Antibacterial Evaluation

The present work focuses on preparation and characterization of erythromycin loaded gelatin nanoparticles through nanoprecipitation method. The procedure consists of the addition of the aqueous gelatin solution to the non-solvent phase containing Lutherol F127. Three different measures of cross-linker and polymer concentration were also examined, and the optimum concentration was found. The mor...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره 2  شماره 

صفحات  -

تاریخ انتشار 2012