Object
Title: Functionalization of the poly(vinyl chloride) tracheostomy tube surface to prevent the growth of bacterial biofilm for use in patients undergoing tracheostomy procedure
Abstract:
Introduction Patients who require continuous ventilator therapy or who have upper airway obstruction most frequently undergo tracheostomies, which involve opening the front wall of the trachea. Most frequently, a poly(vinyl chloride) tracheostomy tube is utilized to protect the airway. In the medical field, it is used to make items like drains, catheters, blood and infusion containers, endotracheal tubes, and tracheostomy tubes in a plasticized form PVC. This polymer is a crucial component of scientific research because of how widely it is used in healthcare settings. Functionalization of the polymer surface with drug nanoparticles is one of the intriguing study topics. It has been demonstrated that the use of antibiotic nanoparticles during the placement of orthopedic or maxillofacial surgery implants lowers the incidence of postoperative infection problems. One method for reducing systemic toxicity in local medication administration is the inclusion of nanoparticles into a polymer. Tracheostomy tubes receive less attention in the literature that details studies on medical devices. This opens up a new area for investigation, particularly with regard to functionalizing the polymeric material with antibiotic nanoparticles. When utilized in patients, biomaterial antibacterial qualities are crucial for minimizing local and systemic issues brought on by the growth of bacterial b ; iofilm on their surfaces. Pathogenic bacteria are stored in biofilms, which are frequently to blame for patient infections. This doctoral dissertation examined the microbiological and physicochemical composition of poly(vinyl chloride) tracheostomy tubes used by patients. Then, using "green chemistry" - compliant chemical techniques, an innovative type of functionalized polymer surface was created. This surface might one day be employed to create a new form of antibacterial tracheostomy tube. Objectives The major objective of this PhD thesis was to create a functionalized poly(vinyl chloride) surface with gentamicin nanoparticles, which can be used for producing tracheostomy tubes. With a focus on biocompatibility in contact with patient tissues, such functionalization intended to slow down or inhibit the formation of bacterial biofilms while retaining reliable performance metrics. The doctoral dissertation's precise goals have been divided up into three research strands: 1) evaluation of the drug resistance of isolated microorganisms to antibiotics as well as the profile of bacteria that form a biofilm on the surface of tracheostomy tubes used by tracheostomy patients. 2) Two-stage functionalization of the poly(vinyl chloride) surface: introduction of oxygen functional groups (using oxygen plasma), gentamicin nanoparticles (using the sonochemical method), and comprehensiv ; e physicochemical characterization of the tested systems. 3) Evaluation of the effectiveness of surface functionalization and its impact on the growth of bacterial biofilm using reference strains of Staphylococcus aureus and Escherichia coli. Evaluation of the cytotoxicity of obtained biomaterials using the 549-cell line. Materials and Methods The period of collecting clinical material lasted from June 2018 to March 2020. The study was approved by Jagiellonian University Bioethical Committee no. 1072.1612.153.2019. Patients for the study were recruited at the 5th Military Hospital with Polyclinic in Krakow. The study involved 10 women and 35 men. All patients were older than 18 years with a mean age of 67.9 years (range 36-89 years) and had used a tracheostomy tube for at least 24 hours. Conventional microbiological cultures were made from the collected material, along with the assessment of drug resistance of bacterial strains. Material for experimental research in the field of physiochemistry consisted of sterile packed discs with dimensions of 15 mm in diameter and 2 mm in thickness, cut from the same material as tracheostomy tubes made of poly(vinyl chloride). The surface of the discs was functionalized using oxygen plasma, and then formed and immediately sonochemically deposited gentamicin nanoparticles. The resulting surfaces were characterized by AFM, W ; CA, NTA, FTIR and evaluated for cytotoxicity using the A549 cell line (ATCC CCL-185) and bacterial adhesion using reference strains of S. aureus (ATCC® 25923™) and E. coli (ATCC ® 25922™). The evaluation of gentamicin cytotoxicity was performed using the MTT protocol. Optical microscopy, SEM and the LIVE/DEAD method using fluorescence microscopy were used to image the cytotoxic effect. Results Every patient-provided tracheostomy tube was culture-positive. In total, 58 isolates (10 species) were isolated and identified. In this study, bacteria that are typically considered to be part of the physiological microbiota were removed, while bacteria from the PPM (potentially pathogenic microorganisms) group were included in further investigation. 16 isolates of Staphylococcus aureus were examined from the Gram-positive PPM (found in 35.5% of patients). As many as 76.2% (32 isolates) of the identified bacteria in the case of Gram-negative PPM referred to the order Enterobacterales (family Enterobacteriaceae), followed by nonfermenting Gram-negative bacteria (NFGNB), which included Pseudomonas aeruginosa (n = 5, identified in 11.9%), Stenotrophomonas maltophilia (n = 4, in 9.5% of patients), and Acinetobacter baumani (n=1, 2,4% of patients). The most frequently found Enterobacterales bacterium was Klebsiella pneumoniae (n = 10, found in 23.8% of patients), followed by E ; nterobacter cloacae (n = 7, found in 16.7% of patients), Escherichia coli (n = 7, found in 16.7% of patients), Proteus spp. (n = 4, found in 9.5% of patients), and Serratia marcescens (n = 2, 4.8% of patients), and Serratia liquefaciens (n = 2, 4.8% of patients). Before modification, PVC had a contact angle of 86±2 degrees. Wettability is one of the key factors affecting bacterial adhesion Based on the conducted experiments, it was found that the optimal parameters for modifying the PVC surface with oxygen plasma are: 50 W, 0.2 mbar and 180 sec. The lowest contact angle with the selected parameters was 32 ±2 degrees. The one-stage synthesis of gentamicin nanoparticles and their deposition on the PVC surface were conducted according to known schemes developed by the Surface Chemistry Group of the Jagiellonian University. The presence of the antibiotic on the surface was confirmed using FTIR by locating binding characteristics for gentamicin, and its release profile from the surface was assessed (OPA method). The cytotoxicity of gentamicin nanoparticles was evaluated using the lung adenoma cell line A549 cell line. IC-50 was assessed in 2-time intervals (1 and 24 hours), setting its value at 1.25 mg/200 μl (6.25 mg/1 ml). The largest decrease in viable cells was observed after exceeding the concentration of 0.31 mg/200 μl (1.55 mg/1 ml) from 95% to 60% at the conc ; entration of 0.63 mg/200 μl (3.15 mg/1 ml). In addition, the cytotoxic effect was visualized by optical microscopy and SEM. . Using LIVE/DEAD double staining and fluorescence microscopy, the cytotoxicity of the functionalized surface was assessed. A549 cell line was used in a cytotoxicity investigation, and the functionalized surfaces had no discernible impact on cell death. Standard strains of Staphylococcus aureus ATCC 25923 (representative of Grampositive bacteria) and Escherichia coli ATCC 25922 (representative of Gram-negative bacteria) were used to check the surface for bacterial adhesion. The use of gentamicin nanoparticles significantly reduces the adhesion of bacterial colonies on the surface of the tracheostomy tube for S. aureus from 6E+05 CFU/mL to 5E+03 CFU/ml and for E. coli from 1.655+05 CFU/ml to 2E+01 CFU/ml. The result is statistically significant (p<0.001). Conclusions In order to enhance the qualities of PVC surfaces for tracheostomy patient applications, a series of surface chemistry and microbiology investigations were used in this PhD thesis. The chosen techniques enabled the creation of a novel functionalized PVC surface with tissuefriendly and antibacterial characteristics. According to the "green chemistry" tenets, the research was conducted using straightforward, more affordable, and ecologically benign approaches.
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Degree discipline:
mikrobiologia ; otorynolaryngologia
Degree grantor:
Rada Dyscypliny Nauki medyczne
Promoter:
Brzychczy-Włoch, Monika ; Kotarba, Andrzej
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Last modified:
Aug 13, 2024
In our library since:
Mar 8, 2024
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http://dl.cm-uj.krakow.pl:8080/publication/5053
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| Edition name | Date |
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| ZB-140220 | Aug 13, 2024 |
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