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. 2023 Jun 15;12(6):1053.
doi: 10.3390/antibiotics12061053.

Cranberry/Chondroitin Sulfate Co-precipitate as a New Method for Controlling Urinary Tract Infections

Concetta Caglioti  1   2 , Rossana Iannitti  3 , Giada Ceccarelli  3 , Laura Selan  4 , Marco Artini  4 , Rosanna Papa  4 , Antonio Malvasi  5 , Rosaria Gentile  1 , Diletta Del Bianco  1 , Florinda Apone  1 , Paola Angelini  1 , Federico Palazzetti  1 , Bernard Fioretti  1
Affiliations

Cranberry/Chondroitin Sulfate Co-precipitate as a New Method for Controlling Urinary Tract Infections

Concetta Caglioti et al. Antibiotics (Basel). .

Abstract

Urinary tract infections (UTI), which are among the most frequent cases of infectious diseases, mainly affect women. The most common treatment approach involves the use of antibiotics, although this solution is not always the most suitable, mainly because of the resistance that bacterial strains develop. Proanthocyanidins are a class of polyphenols, abundantly contained in cranberry extracts, which have shown beneficial effects in the treatment of urinary tract infections, due to their anti-adhesive properties toward bacteria, with respect to the membranes of the cells of the urothelium and intestine, thus reducing their virulence. In this work, we demonstrate via microscopy and scattering measurements how a mixture of cranberry and chondroitin sulfate can form a crosslinked structure with barrier properties. By using a design of experiment (DOE), we optimized the mass ratio to obtain a precipitate between cranberry extract and chondroitin sulfate in the presence of N-acetylcysteine and hyaluronic acid. By using transepithelial electrical resistance (TEER) chambers, we confirmed the barrier properties of the best mixture obtained with the DOE. Lastly, the antibiofilm action was investigated against five strains of Escherichia coli with different antibiotic sensitivity. The precipitate displayed a variable inhibitory effect in biofilm formation with major effects in UTI with an antibiotic resistance profile.

Keywords: N-acetylcysteine; bacterial biofilm; chondroitin sulfate; cranberry; hyaluronic acid; mucosal protector; physical barrier; urinary tract infections.

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Conflict of interest statement

R.I., F.P. and B.F. are inventors of the co-precipitate, with a patent pending.

Figures

Figure 1
Figure 1
Cranberry/chondroitin sulfate complex formation. (A,B) Exemplificative microscopic imaging of solutions A and B reported in Table 1. (C) Cranberry/chondroitin sulfate complex precipitate obtained by microscopic analysis after 5 min mixture of solutions A and B in 1:1 volume ratio as displayed in Table 2.
Figure 2
Figure 2
Scatter profile of mixture solutions. (A) Schematic view of scatter process observed during spectrophotometric recording. (B) Absorption/scattering values for mixtures reported in Table 2 (see Section 2 for description).
Figure 3
Figure 3
DOE of the precipitation process in (A) hydro-alcoholic solution and (B) aqueous solution (see Table 2 and Table 3, respectively) obtained by evaluating the scattering process. The double circle represents the best ratio to obtain the scatter process. Colors represent the predicted scattering: blue for low values, green for intermediate values, yellow and orange for intermediate–high values, and red for high values.
Figure 4
Figure 4
Cranberry v/v solution ratio impact on precipitation process evaluated by scattering and TEER analysis. (A) Optimization of the v/v ratio of the cranberry solution evaluated with scattering methods for some solutions reported in in Table 3. (B) Experimental scheme of the apparatus used in the measurement of the TEER (transepithelial electrical resistance) for the estimation of the electrical resistance of the precipitates. (C) Determination of the electrical resistance (Ω/cm2) of the precipitate obtained 1 h after mixing the v/v ratios for the mixtures indicated in Table 2 and Table 3. For TBS, see Section 4 “Materials and methods”.
Figure 5
Figure 5
Effect of on biofilm formation of different clinical strains, as well as the ATCC25922 and ATCC8739 reference strains. Cranberry/chondroitin/NAC/hyaluronic acid precipitate was added to the culture medium at time zero (0 h, pre-adhesion period) diluted 1:300, and the biofilm was analyzed after overnight incubation. On the ordinate axis, the bars indicate the percentage of bacterial biofilm production. Data are expressed as the percentage of biofilm formed in the presence of cranberry/chondroitin/NAC/hyaluronic acid precipitate compared with the untreated bacteria. Each data point is composed of three independent experiments, each performed in at least three replicates.
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