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Low Concentration Fe-Doped Alumina Catalysts Using Sol-Gel and Impregnation Methods: The Synthesis, Characterization and Catalytic Performance during the Combustion of Trichloroethylene

A Technology Platform to Test the Efficacy of Purification of Alginate

Department of Pathology and Medical Biology, Section of Immunoendocrinology, University Medical Center Groningen, University of Groningen, Hanzeplein 1, EA11, 9700 RB Groningen, The Netherlands
Author to whom correspondence should be addressed.
Materials 2014, 7(3), 2087-2103;
Received: 10 January 2014 / Revised: 12 February 2014 / Accepted: 5 March 2014 / Published: 12 March 2014
Alginates are widely used in tissue engineering technologies, e.g., in cell encapsulation, in drug delivery and various immobilization procedures. The success rates of these studies are highly variable due to different degrees of tissue response. A cause for this variation in success is, among other factors, its content of inflammatory components. There is an urgent need for a technology to test the inflammatory capacity of alginates. Recently, it has been shown that pathogen-associated molecular patterns (PAMPs) in alginate are potent immunostimulatories. In this article, we present the design and evaluation of a technology platform to assess (i) the immunostimulatory capacity of alginate or its contaminants, (ii) where in the purification process PAMPs are removed, and (iii) which Toll-like receptors (TLRs) and ligands are involved. A THP1 cell-line expressing pattern recognition receptors (PRRs) and the co-signaling molecules CD14 and MD2 was used to assess immune activation of alginates during the different steps of purification of alginate. To determine if this activation was mediated by TLRs, a THP1-defMyD88 cell-line was applied. This cell-line possesses a non-functional MyD88 coupling protein, necessary for activating NF-κB via TLRs. To identify the specific TLRs being activated by the PAMPs, we use different human embryonic kidney (HEK) cell-line that expresses only one specific TLR. Finally, specific enzyme-linked immunosorbent assays (ELISAs) were applied to identify the specific PAMP. By applying this three-step procedure, we can screen alginate in a manner, which is both labor and cost efficient. The efficacy of the platform was evaluated with an alginate that did not pass our quality control. We demonstrate that this alginate was immunostimulatory, even after purification due to reintroduction of the TLR5 activating flagellin. In addition, we tested two commercially available purified alginates. Our experiments show that these commercial alginates contained peptidoglycan, lipoteichoic acid, flagellin, and even lipopolysaccharides (LPS). The platform presented here can be used to evaluate the efficacy of purification procedures in removing PAMPs from alginates in a cost-efficient manner. View Full-Text
Keywords: alginate purification; pathogen-associated molecular patterns (PAMPs); microencapsulation; immunology alginate purification; pathogen-associated molecular patterns (PAMPs); microencapsulation; immunology
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MDPI and ACS Style

Paredes-Juarez, G.A.; De Haan, B.J.; Faas, M.M.; De Vos, P. A Technology Platform to Test the Efficacy of Purification of Alginate. Materials 2014, 7, 2087-2103.

AMA Style

Paredes-Juarez GA, De Haan BJ, Faas MM, De Vos P. A Technology Platform to Test the Efficacy of Purification of Alginate. Materials. 2014; 7(3):2087-2103.

Chicago/Turabian Style

Paredes-Juarez, Genaro A., Bart J. De Haan, Marijke M. Faas, and Paul De Vos. 2014. "A Technology Platform to Test the Efficacy of Purification of Alginate" Materials 7, no. 3: 2087-2103.

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