Luis E. Díaz

Microbial degradation, cytotoxicity and antibacterial activity of polyurethanes based on modified castor oil and polycaprolactone

Abstract The objective of this study was to assess the effects of type of polyol and concentration of polycaprolactone (PCL) in polyurethanes (PUs) on microbial degradability, cytotoxicity, biological properties and antibacterial activity to establish whether these materials may have biomedical applications. Chemically modified and unmodified castor oil, PCL and isophorone diisocyanate in a 1:1 ratio of NCO/OH were used. PUs were characterized by stress/strain fracture tests and hardness (ASTM D 676-59). Hydrophilic character was determined by contact angle trials and morphology was evaluated by scanning electron microscopy. Degradability with Escherichia coli and Pseudomonas aeruginosa was evaluated by measuring variations in the weight of the polymers. Cytotoxicity was evaluated using the ISO 10993-5 (MTT) method with mouse embryonic fibroblasts L-929 (ATCC® CCL-1) in direct contact with the PUs and with NIH/3T3 cells (ATCC® CRL-1658) in indirect contact with the PUs. Antimicrobial activity against E. coli and P. aeruginosa was determined. PUs derived from castor oil modified (P0 and P1) have higher mechanical properties than PUs obtained from castor oil unmodified (CO). The viability of L-929 mouse fibroblasts in contact with polymers was greater than 70%. An assessment of NIH/3T3 cells in indirect contact with PUs revealed no-toxic degradation products. Finally, the antibacterial effect of the PUs decreased by 77% for E. coli and 56% for P. aeruginosa after 24 h. These results indicate that PUs synthesized with PCL have biocidal activity against Gram-negative bacteria and do not induce cytotoxic responses, indicating the potential use of these materials in the biomedical field.

Effect of the incorporation of chitosan on the physico-chemical, mechanical properties and biological activity on a mixture of polycaprolactone and polyurethanes obtained from castor oil.

In the present study, polyurethane materials were obtained from castor oil, polycaprolactone and isophorone diisocyanate by incorporating different concentrations of chitosan (0.5, 1.0 and 2.0% w/w) as an additive to improve the mechanical properties and the biological activity of polyurethanes. The polyurethanes were characterized by Fourier transform infrared spectroscopy, thermogravimetric analysis, scanning electron microscopy, stress/strain fracture tests and swelling analysis, and the hydrophilic character of the surface was determined by contact angle trials. The objectives of the study were to evaluate the effect of the incorporation of chitosan on the changes of the physico-chemical and mechanical properties and the in vitro biological activity of the polyurethanes. It was found that the incorporation of chitosan enhances the ultimate tensile strength of the polyurethanes and does not affect the strain at fracture in polyurethanes with 5% w/w of polycaprolactone and concentrations of chitosan ranging from 0 to 2% w/w. In addition, PCL5-Q-PU formulations and their degradation products did not affect cell viability of L929 mouse fibroblast and 3T3, respectively. Polyurethane formulations showed antibacterial activities against Staphylococcus aureus and Escherichia coli bacteria. The results of this study have highlighted the potential biomedical application of this polyurethanes related to soft and cardiovascular tissues.

Use of a mixed culture strategy to isolate halophilic bacteria with antibacterial and cytotoxic activity from the Manaure solar saltern in Colombia

BackgroundWater evaporation in solar salterns creates salinity gradients that promote the adaptation of microbial species to different salinities. This competitive habitat challenges the metabolic capabilities of microorganisms and promotes alterations in their production of secondary metabolites. Thus, solar salterns are a potentially important source of new natural products. In Colombia, the most important and representative solar saltern is located in Manaure (La Guajira) in the north of Colombia. The aim of this study was to develop an alternative screening strategy to select halophilic bacteria as producers of bioactive compounds from mixed microbial cultures rather than individual environmental isolates. Brine and sediment samples from different ponds (across a salinity gradient) were inoculated in seven different culture media to grow bacteria and archaea, allowing for a total of 40 different mixed cultures. An organic extract from each mixed culture was obtained and tested against multidrug resistant pathogens, including Klebsiella pneumoniae, vancomycin-resistant Enterococcus faecium, methicillin-resistant Staphylococcus aureus and Bacillus subtilis. In addition, the extracts were tested against two human cancer cell lines, cervical adenocarcinoma (SiHa) and lung carcinoma (A-549).ResultsTwenty-four of the forty extracts from mixed cultures obtained from brine and sediment samples from the Manaure solar saltern showed antibacterial activity against Bacillus subtilis. Two extracts, referred to as A1SM3–29 and A1SM3–36, were also active against a methicillin-resistant Staphylococcus aureus, with the latter extract also showing slight cytotoxic activity against the assayed human lung cancer cell line. From this mixed culture, nine isolates were cultivated, and their extracts were tested against the same pathogens, resulting in the identification of a Vibrio sp. strain (A1SM3–36-8) with antimicrobial activity that was similar to that observed for the mixed culture extract. The extract of this strain was subjected to a bioautography assay, and 3 different fractions exhibited antibacterial activity against methicillin-resistant Staphylococcus aureus. Based on the amount obtained for each fraction, F3 was selected to isolate and identify its metabolites. The major compound was identified by NMR and HRMS as 13-cis-docosenamide, an amide that has been previously reported to be an antimicrobial and cytotoxic compound.ConclusionsOur results shows the utility of our strategy in detecting bioactive molecules in initial mixed cultures by biological assays, resulting in the isolation and characterization of Vibrio sp. A1SM3–36-8, a halophilic strain with great antibacterial and cytotoxic potential.

Polyurethanes from modified castor oil and chitosan: Synthesis, characterization, in vitro degradation, and cytotoxicity

Polyurethanes (PUs) from castor oil (CO), modified CO (MCO) by transesterification reaction, isophorone diisocyanate (IPDI) in an NCO/OH ratio equal to 1, and chitosan (CS) were synthesized to assess their potential as biomaterials. PUs were characterized by Fourier transform infrared spectroscopy, hydroxyl value (ASTM D1957), thermogravimetric analysis, Shore A hardness (ASTM D2240), and scanning electronic microscopy (SEM). Also, contact angle, water retention and in vitro degradation in PBS, and cell viability on fibroblast were performed. The hydroxyl value confirms CO modification, and IR analysis confirms urethane bond formation. The thermal assay does not show new degradation stages and polyol with a high functionality had better hardness performance due to the increase in cross-linking. The micrograph shows micro-phase separation of both polymers. The contact angle shows the hydrophobic surface with an angle over 65°, and the CS and polyol type do not affect swelling and in vitro degradation due to phase separation between both polymers. The cell viability was over 70% in all cases, and solid polymers and degradation products involve non-cytotoxic effects on the samples. The results suggest a potential for these formulations in the biomedical field.

Poliuretanos obtenidos a partir de aceite de higuerilla modificado y poli-isocianatos de lisina: síntesis, propiedades mecánicas y térmicas y degradación in vitro

Biodegradable polyurethanes (PUR) were prepared from polyols derived from castor oil by transesterification of pentaerythritol-modified castor oil and lysine polyisocyanates (LDI and LTI). The polyurethanes obtained were characterized by Fourier transform infrared spectroscopy, thermogravimetric analysis (TGA), and dynamic mechanical thermal analysis (DMTA). The mechanical behavior of the polyurethanes was measured by Shore A hardness and tensile testing (stress-strain curves). The biodegradable nature of the material was determined by contact angle, water absorption tests, and in vitro degradation in PBS solution. This study aims to examine the effect of the structure and functionality of diisocyanate on the mechanical properties and in vitro degradation of the material. The results were compared with homologous materials obtained from isophorone diisocyanate (IPDI) used in previous works. The objective was to evaluate candidate materials that can be potentially used in tissue engineering.

Characterization of Actinobacterial Communities from Arauca River Sediments (Colombia) Reveals Antimicrobial Potential Presented in Low Abundant Isolates

Introduction: New strategies have been arisen to set a rapid and effective screening for selection of microorganism with bioactive potential. This study suggests that combination of physicochemical pretreatments and taxonomic dereplication of microbial collections through MALDI-TOF MS, facilitates the detection of low abundance actinobacteria with potential as a source of antimicrobial agents. Material and Methods: An unstudied microbial community from a tropical river sediment in Colombian Orinoquía is described, applying an extended cultivation strategy using physicochemical pretreatments, biological screenings and taxonomic dereplication through MALDI-TOF MS approach. Results: Actinobacteria-like isolates (790) were growth and their antimicrobial activity was assessed against methicillin-resistant Staphylococcus aureus, Vancomycin-resistant Enterococcus faecium, extended-spectrum β-lactamase Klebsiella pnumoniae, and clinical isolates of Cladosporium cladosporioides and Epicoccum nigrum. Seventy-eight isolates, belonging to the Streptomycetaceae family according to 16S rDNA analysis were found to have antimicrobial activity and were categorized as low abundance actinobacteria by MALDI-TOF MS. Conclusion: The results suggest that combination of physicochemical pretreatments and taxonomic dereplication of microbial collections through MALDI-TOF MS, facilitates the detection of low abundance actinobacteria with potential as a source of antimicrobial agents.

Optimization of the Cytotoxic Activity of Three Streptomyces Strains Isolated from Guaviare River Sediments (Colombia, South America)

Three Streptomyces strains isolated from Guaviare sediments (Colombia, South America) with cytotoxic activity against prostate cancer (PC3), breast cancer (MDA-MB-231), and lung cancer (A549) line cells were studied. The present investigation reveals the enhancement of the cytotoxic activity evaluating different values of pH, carbon sources (sucrose, glucose, xylose, maltose, and starch), and nitrogen sources (malt extract, yeast extract, meat extract, peptone, and potassium nitrate). Based on the response surface methodology, the isolates Streptomyces aburaviensis (73) had the maximum activity for lung cancer (IC50= 25.00 ± 1.86 ppm) with 4% of yeast extract, 3% of starch, and a pH value of 7. Streptomyces gramineus (386) had the maximum activity against prostate cancer (IC50= 6.14 ± 2.07 ppm) with 5% of malt extract, 3% of glucose, and a pH value of 6. Finally, Streptomyces psammoticus (519) had the maximum activity against breast cancer (IC50= 35.53 ± 2.71 ppm) with 1% of yeast extract, 4% of starch, and a pH 8. The results suggest that the ethyl acetate extracts from isolates Streptomyces aburaviensis (73), Streptomyces gramineus (386), and Streptomyces psammoticus (519) have a potential for use in pharmaceuticals as cytotoxic agents.

Actividad antibacteriana, degradación hidrolítica y microbiana y citotoxicidad in vitro de poliuretanos sintetizados con polioles derivados del aceite de higuerilla

Biological activities from polyurethanes (PUs) synthesized from polyols (derived from castor oil) and isophorone diisocyanate were evaluated. In vitro degradability was evaluated by phosphate buffered saline (PBS) and antibacterial degradation with Escherichia coli and Pseudomonas aeruginosa. The biocompatibility was analyzed by: i) the antimicrobial activity against E. coli, S. aureus and P. aeruginosa, and ii) in vitro cytotoxicity assays using mouse embryonic fibroblast cell line L-929 in direct contact with the PUs and with NIH/3T3 cells in indirect contact with the PUs degradation products. PUs with polyol P1 (2,64% molar relation of pentaerythritol and castor oil) was the material with better biodegradation properties: 1.125±0.110% in 72 h by PBS at 100 oC. In general, all materials were degraded up to 1.000±0.223% and 2.251±0.010% in 72 h by E. coli and P. aeruginosa, respectively. On the other hand, it was determined a close relationship between functionality of polyol and bacterial inhibition. The antibacterial effect of the PUs decreased by 67% for E. coli, 55% for S. aureus and 56% for P. aeruginosa after 24 h. Chemical modification of castor oil did not generate a cytotoxic effect on the tested cell lines. The evaluated materials are suggested as candidates to obtain biomaterials due to their mechanical properties and biocompatibility presented in untransformed cells.