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Evaluation of bacterial uptake, antibacterial efficacy against Escherichia coli, and cytotoxic effects of moxifloxacin-loaded solid lipid nanoparticles

Merve Eylul Kiymaci
Merve Eylul Kiymaci
, 
Gizem Ruya Topal
Gizem Ruya Topal
, 
Ozgur Esim
Ozgur Esim
, 
Merve Bacanli
Merve Bacanli
, 
Cansel Kose Ozkan
Cansel Kose Ozkan
, 
Onur Erdem
Onur Erdem
, 
Ayhan Savaser
Ayhan Savaser
 and 
Yalcin Ozkan
Yalcin Ozkan
   | Dec 30, 2022
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Article Category: Original article
Published Online: Dec 30, 2022
Page range: 260 - 269
Received: Jul 01, 2022
Accepted: Nov 01, 2022
DOI: https://doi.org/10.2478/aiht-2022-73-3667
© 2022 Merve Eylul Kiymaci, Gizem Ruya Topal, Ozgur Esim, Merve Bacanli, Cansel Kose Ozkan, Onur Erdem, Ayhan Savaser, Yalcin Ozkan, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Figure 1

TEM images of SLN1 (a), SLN2 (b), NLC1 (c), and NLC2 (d)
TEM images of SLN1 (a), SLN2 (b), NLC1 (c), and NLC2 (d)

Figure 2

MOX release from nanoparticles
MOX release from nanoparticles

Figure 3

Flow cytometry results (a: blank NPS; b: fluorescein solution; c: SLN1; d: SLN2; e: NLC1; f: NLC2)
Flow cytometry results (a: blank NPS; b: fluorescein solution; c: SLN1; d: SLN2; e: NLC1; f: NLC2)

Figure 4

Photomicrographs show bacterial uptake of formulations (a: fluorescein solution; b: SLN1; c: SLN2; d: NLC1; e: NLC2)(40× magnification)
Photomicrographs show bacterial uptake of formulations (a: fluorescein solution; b: SLN1; c: SLN2; d: NLC1; e: NLC2)(40× magnification)

Cytotoxic effects of MOX, MOX-free, and MOX-loaded LNPs in RAW 264.7 cells

RAW 264.7 cell viability (%)
24 h 48 h 72 h
0.010 µg/mL 0.025 µg/mL 0.05 µg/mL 0.1 µg/mL 0.010 µg/mL 0.025 µg/mL 0.05 µg/mL 0.1 µg/mL 0.010 µg/mL 0.025 µg/mL 0.05 µg/mL 0.1 µg/mL
MOX 88.65±0.09 69.05±0.35 56.48±0.01 43.90±0.03 89.42±0.02 73.98±0.33 59.43±0.02 40.01±0.10 86.62±0.23 83.49±0.08 54.58±0.02 31.93±0.11
SLN1+MOX 81.56±0.26 75.43±0.29 75.92±0.23 74.55±0.23 94.62±0.02 81.95±0.11 71.44±0.03 58.43±0.06 92.13±0.08 91.46±0.34 89.06±0.28 73.70±0.22
SLN1 95.30±0.08 68.91±0.12 62.78±0.25 57.75±0.01 95.75±0.29 91.22±0.05 85.44±0.06 64.09±0.03 87.12±0.27 82.65±0.09 73.35±0.11 69.72±0.17
SLN2+MOX 93.14±0.08 80.42±0.19 66.66±0.46 62.10±0.21 88.73±0.06 77.09±0.12 63.16±0.12 56.05±0.04 92.13±0.08 91.46±0.34 89.06±0.28 73.71±0.22
SLN2 90.98±0.30 86.81±0.12 64.91±0.09 60.60±0.31 96.51±0.22 94.42±0.12 80.82±0.08 68.49±0.22 96.02±0.33 87.31±0.27 69.95±0.09 55.88±0.06
NLC1+MOX 80.46±0.09 81.65±0.18 85.84±0.26 81.47±0.09 88.72±0.09 72.27±0.04 62.23±0.02 51.10±0.15 85.24±0.09 76.56±0.30 73.78±0.09 56.03±0.32
NLC1 89.19±0.13 78.73±0.17 70.83±0.33 53.27±0.04 92.75±0.13 82.93±0.04 63.82±0.09 54.44±0.05 89.70±0.09 80.44±0.08 63.51±0.35 54.76±0.20
NLC2+MOX 78.43±0.31 65.99±0.05 56.58±0.29 50.72±0.04 76.23±0.27 59.40±0.03 53.59±0.06 53.05±0.08 94.10±0.27 79.09±0.03 73.90±0.06 51.68±0.17
NLC2 85.54±0.07 60.58±0.10 56.67±0.26 50.07±0.16 83.62±0.06 62.94±0.12 54.81±0.05 52.27±0.04 91.99±0.42 82.25±0.12 74.99±0.05 54.11±0.09

Lipid nanoparticle properties

LNP Particle size (nm) Polydispersity index Zeta potential (mV) Encapsulation efficiency (%)
SLN1 181.6±1.27 0.25±0.02 -14.6±0.32 68.61±0.20
SLN2 201.8±1.65 0.29±0.02 -17.5±0.14 74.53±0.34
NLC1 176.8±1.90 0.57±0.23 -11.6±0.61 77.82±0.21
NLC2 132.4±1.62 0.51±0.53 -6.3±0.38 77.17±0.21

Stability of lipid nanoparticles stored for one month at +4 °C

Particle size (nm) Polydispersity index Zeta potential (mV)
Type of LNPs Baseline value After one month storage Baseline value After one month storage Baseline value After one month storage
SLN1 181.6±1.27 186.1±2.89 0.25±0.02 0.36±0.25 -14.6±0.32 -10.0±0.51*
SLN2 201.8±1.65 200.5±3.56 0.29±0.02 0.39±0.12 -17.5±0.14 -11.5±0.45*
NLC1 176.8±1.90 180.3±1.64 0.57±0.23 0.60±0.46 -11.6±0.61 -8.30±0.78
NLC2 132.4±1.62 135.2±3.98 0.51±0.53 0.65±0.22 -6.3±0.38 -5.62±0.96

Composition of lipid nanoparticles

Formulation code Solid lipid (glyceryl dibehenate) Liquid lipid (docosahexaenoic acid, DHA) Liquid lipid (oleic acid) Tween 80 Poloxamer 407 Triethanolamine (TEA)
SLN1 150 mg - - 100 mg - -
SLN2 150 mg - - - 100 mg -
NLC1 110 mg 40 mg 10 mg - 100 mg 10 mg
NLC2 110 mg 40 mg - - 100 mg 10 mg

Antibacterial activity of MOX-loaded SLNs and NLCs

Formulations Inhibition diameter (mm) Minimal inhibition concentration (μg/mL)
Standard MOX solution for SLNs * (0.68 mg) 33±0.07 0.041
SLN1 34±0.10*** 0.020
SLN2 32±0.11 0.041
Standard MOX solution for NLCs** (0.78 mg) 35±0.08 0.023
NLC1 36±0.12 0.023
NLC2 35±0.05 0.023
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