Investigation of the Impact of Poly(Ethylene Glycol)-Modulation of Poly(B-Hydroxybutyrate) Syntheses on Cell Interactions of the Resulting Polymers

Abstract

Poly(-hydroxybutyrate), PHB is a bacterial polyester known for its excellent bone compatibility, however, the material lacks blood and tissue compatibility. Poly(ethylene glycol), PEG,-modulated fermentation of Alcaligenes latus and Azotobacter vinelandii UWD was employed to yield copolymers consisting of PHB and PEG that exhibit diminished cell-adhesion surface properties. PEGs with molecular weights of 3400, 2000, and 400 as well as diethylene glycol, DEG, and pentaerythritol ethoxylate, PEE, were used in a concentration of 2% (w/v) for amending the fermentation broths. This modulation of the fermentation conditions did not influence polymer yields. However, the resulting copolymers had drastically reduced molecular weights, 82% less for the DEG-amended fermentation of A. latus. The reduction in molecular weight was attributed to an end-capping reaction of the nascent PHB-chain with PEG and/or early chain termination by water facilitated by the presence of the highly hydrophilic PEG-molecules. The formation of a covalent linkage was proven unambiguously by H-NMR-spectroscopic methods only for the copolymers obtained in the DEG-modified fermentations of both strains. Cell growth experiments using SK-MEL 28 and MDA-MB 231 cells were used for the evaluation of polymer-cell interaction. Copolymer films obtained from PEG-modulated syntheses showed significantly less cell adhesion with reductions in cell adhesions; up to 74% less in the two-day experiments (MDA-MB 231 on the copolymer obtained in DEG-modified fermentation of A. latus) and 48% less in the seven-day experiments (SK-MEL 28 on the copolymer obtained in PEG 400-modified fermentation of A. vinelandii UWD). In the two-day experiments, no differences in the cellinteraction was observed between the polymers obtained from two different bacterial sources, the polymers differed in their long-term, seven-day, cell interaction with copolymers obtained from A. vinelandii UWD maintaining more effective cell repulsion.