Fabrication of a Layered Microstructured Polymeric Microspheres as a Cell Carrier for Nucleus Pulposus Regeneration
ABSTRACT This study aimed to investigate the feasibility of nanostructured 3D poly(lactide-co-glycolide) (PLGA) constructs, which are loaded with dexamethasone (DEX) and growth factor embedded heparin/poly(l-lysine) nanoparticles by a layer-by-layer system, to serve as an effective scaffold for nucleus pulposus (NP) tissue engineering. Our results demonstrated that the microsphere constructs were capable of simultaneously releasing basic fibroblast growth factor and DEX with approximately zero-order kinetics. The dual bead microspheres showed no cytotoxicity, and promoted the proliferation of the rat mesenchymal stem cells (rMSCs) by lactate dehydrogenase assay and CCK-8 assay. After 4 weeks of culture in vitro, the rMSCs-scaffold hybrids contained significantly higher levels of sulfated GAG/DNA and type-II collagen than the control samples. Moreover, quantity real-time PCR analysis revealed that the expression of disc-matrix proteins, including type-II collagen, aggrecan and versican, in the rMSCs-scaffold hybrids was significantly higher than the control group, whereas the expression of osteogenic differentiation marker type-I collagen was decreased. Taken together, these data indicate that the heparin bound bFGF-coated and DEX-loaded PLGA microsphere constructs is an effective bioactive scaffold for the regeneration of NP tissue.
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ABSTRACT: We estimate the perturbative nuclear shadowing in the nuclear structure function F A 2 (x; Q 2 ), mainly at the kinematic region of HERA using the Glauber-Mueller approach. The contributions of the quark and gluon sectors to the nuclear shadowing are estimated. We predict that the nuclear shadowing corrections are important and that saturation of the ratio R 1 = F A 2 (x; Q 2 )=A F p 2 (x; Q 2 ) occurs once the shadowing in the gluon sector is considered. 11.80.La; 24.95.+p Typeset using REVT E X E-mail:firstname.lastname@example.org E-mail:email@example.com 1 The future ultrarelativistic heavy ion collider experiments at the BNL Relativistic Heavy Ion Collider (RHIC) and the CERN Large Hadron Collider (LHC) are expected to exhibit new phenomena associated with an ultradense environment that may be created in the central collision region of these reactions . The main conclusion emerging from the analysis of nucleus-nucleus collisions for RHIC energies and beyond, is that th...
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ABSTRACT: Growth, differentiation and migration factors facilitate the engineering of tissues but need to be administered with defined gradients over a prolonged period of time. In this study insulin as a growth factor for cartilage tissue engineering and a biodegradable PLGA delivery device were used. The aim was to investigate comparatively three different microencapsulation techniques, solid-in-oil-in-water (s/o/w), water-in-oil-in-water (w/o/w) and oil-in-oil-in-water (o/o/w), for the fabrication of insulin-loaded PLGA microspheres with regard to protein loading efficiency, release and degradation kinetics, biological activity of the released protein and phagocytosis of the microspheres. Insulin-loaded PLGA microspheres prepared by all three emulsification techniques had smooth and spherical surfaces with a negative zeta potential. The preparation technique did not affect particle degradation nor induce phagocytosis by human leukocytes. The delivery of structurally intact and biologically active insulin from the microspheres was shown using circular dichroism spectroscopy and a MCF7 cell-based proliferation assay. However, the insulin loading efficiency (w/o/w about 80%, s/o/w 60%, and o/o/w 25%) and the insulin release kinetics were influenced by the microencapsulation technique. The results demonstrate that the w/o/w microspheres are most appropriate, providing a high encapsulation efficiency and low initial burst release, and thus these were finally used for cartilage tissue engineering. Insulin released from w/o/w PLGA microspheres stimulated the formation of cartilage considerably in chondrocyte high density pellet cultures, as determined by increased secretion of proteoglycans and collagen type II. Our results should encourage further studies applying protein-loaded PLGA microspheres in combination with cell transplants or cell-free in situ tissue engineering implants to regenerate cartilage.Acta biomaterialia 12/2010; 7(4):1485-95. DOI:10.1016/j.actbio.2010.12.014 · 5.68 Impact Factor
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ABSTRACT: Poly(lactic-co-glycolic acid) (PLGA) microsphere has been a useful tool in delivering therapeutic drugs and biologically active proteins. In this study, a covered porous PLGA microsphere was manufactured using W(1)/O/W(2) double emulsion solvent evaporation method, utilizing hydrogen peroxide as a novel porogen. An enzymatic reaction between hydrogen peroxide and catalase produced oxygen bubbles and thus many internal pores within microsphere were naturally developed. When different molar ratios between lactide and glycolide in PLGA were examined, the ratio, 50:50 showed the most organized porous microstructure. Higher molecular weight of PLGA seemed to be favorable in creating a porous structure. By testing various concentrations of hydrogen peroxide, it was found that rather concentrated one was more efficient in developing a porous network in the microspheres. The source of the skin layer that covers the whole surface of the microsphere was found to be PLGA, not polyvinyl alcohol (PVA). The residual amount of hydrogen peroxide was negligible after a thorough evaporation of PLGA microsphere. When release profiles of dexamethasone (Dex) with morphologically different microspheres such as, nonporous, covered porous, and porous, were investigated for up to 28 days in vitro, their release patterns were found to be significantly different on a temporal basis. The present work demonstrated that the covered porous PLGA microspheres could be successfully fabricated using hydrogen peroxide and that the covered skin layer on the PLGA microsphere played an important role in determining the characteristic release profiles of Dex.Journal of Controlled Release 10/2008; 133(1):37-43. DOI:10.1016/j.jconrel.2008.09.006 · 7.26 Impact Factor