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jctb4003-sup-0001-AppendixS1.docWord document66030K Appendix S1. Cytotoxicity and biocompatibility assays
jctb4003-sup-0002-FigureS1.docWord document136K Figure S1. Chemical structure of rhamnolipids (a) [Trummler et al. 2003]3 and surfactin with the heptapeptide ring (b) [Bonmatin et al. 1994]4
jctb4003-sup-0003-FigureS2.docWord document329K Figure S2. Kinetics profiles of emulsion polymerizations of polystyrene using rhamnolipid (a) and surfactin (b) with a stirring rate 350 rpm
jctb4003-sup-0004-FigureS3.docWord document7857K Figure S3. Conversion vs time plots for emulsion polymerizations of polystyrene using rhamnolipid (a) and surfactin (b) with a stirring rate 240 rpm
jctb4003-sup-0005-FigureS4.docWord document6964K Figure S4. Conversion vs time plots for emulsion polymerizations of polystyrene using rhamnolipid (a) and surfactin (b) with a stirring rate 500 rpm
jctb4003-sup-0006-FigureS5.docWord document48K Figure S5. TEM photographs of PS particles prepared by emulsion polymerization at the stirring rate 240 rpm (a) and 500 rpm (b)
jctb4003-sup-0007-FigureS6.docWord document280K Figure S6. SEM micrograph of polystyrene synthesized by rhamnolipid (above CMC, 0.2 mM; a) and surfactin (above CMC, 0.08 mM; b). Particle size: 180 nm. Emulsion polymerization was carried out at a stirring rate of 350 rpm
jctb4003-sup-0008-FigureS7.docWord document281K Figure S7. Particle size of polystyrene as a function of rhamnolipid (a) and surfactin (b) concentrations. Arrows indicate the CMC of the biosurfactants. Emulsion polymerization was carried out at the stirring rate of 350 rpm
jctb4003-sup-0009-FigureS8.docWord document319K Figure S8. Number of particles as a function of rhamnolipid (a) and surfactin (b) concentrations. * indicates the CMC of the biosurfactant. Emulsion polymerization was carried out at the stirring rate of 350 rpm
jctb4003-sup-0010-FigureS9.docWord document243K Figure S9. FTIR spectra of polystyrene synthesized using rhamnolipid
jctb4003-sup-0011-FigureS10.docWord document60K Figure S10. 1H NMR (a), DSC thermograms (b) and TGA profile (c and d) of PS/rhamnolipid nanoparticles prepared by emulsion polymerization at a stirring rate of 350 rpm
jctb4003-sup-0012-FigureS11.docWord document1555K Figure S11. HPLC analysis of biodegradation metabolites in in vitro biodegradation assay: (a) PSSDS, (c) PSrhl and (e) PSsur NPs; HPLC analysis of biodegradation metabolites in soil burial test: (b) PSSDS, (d) PSrhl and (f) PSsur NPs
jctb4003-sup-0013-FigureS12.docWord document13910K Figure S12. NRU assay of (a) PSSDS, (b) PSrhl and (c) PSsur NPs. Rat hepatocyte cells (1×106 mL−1) were plated into each well of 96-well flat-bottom plates with different concentration of PS NPs and incubated for 24 h and percent viability measured by MTT assay. Vertical bars represent mean ± S.E.M. (n=3). * Statistically significant difference as compared to the controls (p<0.05 for each)
jctb4003-sup-0014-FigureS13.docWord document9321K Figure S13. LDH leakage assay of (a) PSSDS, (b) PSrhl and (c) PSsur NPs. Rat hepatocyte cells (1×106 mL−1) were plated into each well of 96-well flat-bottom plates with different concentration of PS NPs and incubated for 24 h and percent viability measured by MTT assay. Vertical bars represent mean ± S.E.M. (n=3). * Statistically significant difference as compared to the controls (p<0.05 for each)
jctb4003-sup-0015-FigureS14.docWord document12282K Figure S14. Size- and dose dependent effects of PSSDS [a(i): 0-50 mg mL−1 and a(ii) 55-100 mg mL−1], PSrhl [b(i): 0-50 mg mL−1 and b(ii): 55-100 mg mL−1] and PSsur [c(i): 0-50 mg mL−1 and c(ii): 55-100 mg mL−1] NPs on GSH levels in rat hepatocytes. Cells were treated with different concentration of three types of bionanocomposites for 24 h. At the end of the exposure, cells were washed with PBS and GSH (control: 44.2 ± 2.6 nmol GSH mg protein−1) were measured as described in experimental section. Control cells cultured in PS NPs-free medium were run in parallel to the treated groups. Values are the mean ± S.E.M from three independent experiments. Significance indicated by: * p < 0.05 versus control cells.
jctb4003-sup-0016-FigureS15.docWord document12322K Figure S15. Cellular TBARS levels of rat hepatocytes after 24 h exposure to different sized PS NPs at 5-100 mg mL−1. a(i): PSSDS, 0-50 mg mL−1; a(ii): PSSDS, 55-100 mg mL−1; b(i): PSrhl, 0-50 mg mL−1; b(ii): PSrhl, 55-100 mg mL−1; c(i): PSsur, 0-50 mg mL−1; c(ii): PSsur, 55-100 mg mL−1. Control cells cultured in NPs-free medium were run in parallel to the treated groups (control: 6.2 ± 0.3 nmol TBARS mg protein−1). Values are mean ± S.E.M from three independent experiments. Significance indicated by: *p < 0.05 versus control cells
jctb4003-sup-0017-FigureS16.docWord document220K Figure S16. Histopathology of liver from rat treated with bionanocomposites prepared with SDS (a), rhamnolipid (b) and surfactin (c). Photomicrographs (×40) of liver sections (5 µm) stained with hematoxylin and eosin: Arrows show infiltration of hepatocyte cords by erythrocytes (a); asterisk, sloughing of hepatocyte remnants into congested sinusoids and microvesiculation (1 cm=25 µm).
jctb4003-sup-0018-TableS1.docWord document44K Table S1 The characteristics of surfactants used in this study
jctb4003-sup-0019-TableS2.docWord document285K Table S2 FTIR peak assignments of PSrhl and rhamnolipid

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