Preparation and Characterization of Chitosan Microspheres for Controlled Release of Rh-BMP-2
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MRS Advances © 2019 Materials Research Society DOI: 10.1557/adv.2019.426
Preparation and Characterization of Chitosan Microspheres for Controlled Release of Rh-BMP-2 Cindy Sinaí Velázquez-González1, Ena Athenea Aguilar-Reyes1, Rosa Elvira Núñez-Anita2, Carlos Alberto León-Patiño1 1
Instituto de Investigación en Metalurgia y Materiales, Universidad Michoacana de San Nicolás de Hidalgo, Morelia, México.
2 Facultad de Medicina Veterinaria y Zootecnia, Unidad de Proteómica y Bioingeniería Celular, Universidad Michoacana de San Nicolás de Hidalgo, Tarímbaro, México.
ABSTRACT
The implantation and controlled release of growth factors can enhance the proliferation and differentiation of cells that promote new bone formation at defect sites. Therefore, chitosan polymer microspheres were prepared by the water-in-oil emulsion (W/O) method and solvent freeze-drying, using glutaraldehyde as an ionic crosslinker, along with the lyophilization of solvents, to microencapsulate growth factors, preventing denaturation. The microspheres were loaded with recombinant bone morphogenetic protein 2 (Rh-BMP-2). They were spherical in shape, with a rough surface ranging in particle size from 0.4 to 1.6 Pm. The yield percentage with respect to the polymer was 70% and the BMP-2 load was regulated by the initial protein dose. BMP-2 release experiments were performed for 7 days in PBS solutions at pH 4 and 7.4. The results showed that the protein release rate was only 2% lower at pH 7.4. BMP-2/chitosan microspheres were compatible with the MG-63 cell line (ATCC£ CRL1427¥ Homo sapiens bone osteosarcoma) and could be considered drug delivery vehicles in bone tissue engineering applications. *Corresponding author: Dr. Ena Athenea Aguilar Reyes (Email:
3259
[email protected])
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INTRODUCTION Alternative vehicles that can be easily manipulated for delivering drugs, growth factors, or therapeutic proteins and peptides in specific areas and preserving their nature, have been explored in several studies [1-3]. Microencapsulation is a technique whereby small liquid drops containing the active substance are coated with a porous polymeric wall. The microcapsules are then administered or implanted in certain regions. The polymeric barrier delays the chemical reactions between the active compound and the surrounding environment, thus increasing the useful lifetime of the substance and achieving its sustained release [4,5]. Chitosan (1-4-β-2 amino, 2-deoxy-D-glucose or D-glucosamine) is a natural polymer with excellent antimicrobial properties, good adhesion, coagulation capacity, and immunostimulating activity. It is a linear cationic polymer, with a high molecular weight, that can be found naturally in the cell walls of plants and fungi. It can also be obtained through the chemical or enzymatic deacetylation of chitin. Chitosan i
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