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A facile one-step sonochemical method was employed for the first time for gentamicin nanoparticles (GNPs) fabrication and embedding into the surface of parylene C implant coating. The developed system was thoroughly characterized in terms of particle size (NTA, STEM/EDX), surface dispersion (IR-image) and drug release kinetics (UV-Vis). It was revealed that the optimization of the applied ultrasound conditions resulted in the formation of GNPs with an average size in the narrow range of 30-70 nm and their docking into the parylene C nanopores, while the molecular structure of the antibiotic was preserved as confirmed by the FTIR spectra. The obtained surface morphology resulted in controlled elution of the drug up to 7 days, and the kinetics followed the Korsmeyer-Peppas model. The apparent benefits of the proposed sonochemical approach (short preparation time, direct drug accessibility, lack of chemical wastes) are pointed out.
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Materiais Revestidos Biocompatíveis , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Gentamicinas/metabolismo , Nanopartículas/química , Polímeros/química , Próteses e Implantes , Xilenos/química , Antibacterianos/química , Antibacterianos/metabolismo , Preparações de Ação Retardada , Gentamicinas/químicaRESUMO
Polyurethanes (PUs) are versatile polymers used in medical applications due to their high flexibility and fatigue resistance. PUs are widely used for synthetic blood vessels, wound dressings, cannulas, and urinary and cardiovascular catheters. Many scientific reports indicate that surface wettability is crucial for biocompatibility and bacterial adhesion. The use of oxygen plasma to modify PUs is advantageous because of its effectiveness in introducing oxygen-containing functional groups, thereby altering surface wettability. The purpose of this study was to investigate the effect of the modification of the oxygen plasma of polyurethane on its biocompatibility with lung tissue (A549 cell line) and the adhesion of Gram-positive bacteria (S. aureus and S. epidermidis). The results showed that the modification of polyurethane by oxygen plasma allowed the introduction of functional groups containing oxygen (-OH and -COOH), which significantly increased its hydrophilicity (change from 105° ± 2° to 9° ± 2°) of PUs. Surface analysis by atomic force microscopy (AFM) showed changes in PU topography (change in maximum height from â¼110.3 nm to â¼32.1 nm). Moreover, biocompatibility studies on A549 cells showed that on the PU-modified surface, the cells exhibited altered morphology (increases in cell surface area and length, and thus reduced circularity) without concomitant effects on cell viability. However, serial dilution and plate count and microscopic methods confirmed that plasma modification significantly increased the adhesion of S. aureus and S. epidermidis bacteria. This study indicate the important role of surface hydrophilicity in biocompatibility and bacterial adhesion, which is important in the design of new medical biomaterials.
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At the time when pathogens are developing robust resistance to antibiotics, the demand for implant surfaces with microbe-killing capabilities has significantly risen. To achieve this goal, profound understanding of the underlying mechanisms is crucial. Our study demonstrates that graphene oxide (GO) nano films deposited on stainless steel (SS316L) exhibit superior antibacterial features. The physicochemical properties of GO itself play a pivotal role in influencing biological events and their diversity may account for the contradictory results reported elsewhere. However, essential properties of GO coatings, such as oxygen content and the resulting electrical conductivity, have been overlooked so far. We hypothesize that the surface potential and electrical resistance of the oxygen content in the GO-nano films may induce bacteria-killing events on conductive metallic substrates. In our study, the GO applied contains 52 wt% of oxygen, and thus exhibits insulating properties. When deposited as a nano film on an electrically conducting steel substrate, GO flakes generate a Schottky barrier at the interface. This barrier, consequently, impedes the transfer of electrons to the underlying conductive substrate. As a result, this creates reactive oxygen species (ROS), leading to bacterial death. We confirmed the presence of GO coatings and their hydrolytic stability by using X-ray photoelectron spectroscopy (XPS), µRaman spectroscopy, scanning electron microscopy (SEM), and Kelvin probe force microscopy (KPFM) measurements. The biological evaluation was performed on the MG63 osteoblast-like cell line and two selected bacteria species: S. aureus and P. aeruginosa, demonstrating both the cytocompatibility and antibacterial behavior of GO-coated SS316L substrates. We propose a two-step bactericidal mechanism: electron transfer from the bacteria membrane to the substrate, followed by ROS generation. This mechanism finds support in changes observed in contact angle, surface potential, and work function, identified as decisive factors. By addressing overlooked factors and effectively bridging the gap between understanding and practicality, we present a transformative approach for implant surfaces, combating microbial resistance, and offering new application possibilitie.
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Antibacterianos , Grafite , Staphylococcus aureus , Espécies Reativas de Oxigênio/metabolismo , Antibacterianos/farmacologia , Antibacterianos/química , Metais/farmacologia , Oxigênio/farmacologiaRESUMO
BACKGROUND: The use of nanotechnology in the production of medical equipment has opened new possibilities to fight bacterial biofilm developing on their surfaces, which can cause infectious complications. In this study, we decided to use gentamicin nanoparticles. An ultrasonic technique was used for their synthesis and immediate deposition onto the surface of tracheostomy tubes, and their effect on bacterial biofilm formation was evaluated. METHODS: Polyvinyl chloride was functionalized using oxygen plasma followed by sonochemical formation and the embedment of gentamicin nanoparticles. The resulting surfaces were characterized with the use of AFM, WCA, NTA, FTIR and evaluated for cytotoxicity with the use of A549 cell line and for bacterial adhesion using reference strains of S. aureus (ATCC® 25923™) and E. coli (ATCC® 25922™). RESULTS: The use of gentamicin nanoparticles significantly reduced the adhesion of bacterial colonies on the surface of the tracheostomy tube for S. aureus from 6 × 105 CFU/mL to 5 × 103 CFU/mL and for E. coli from 1.655 × 105 CFU/mL to 2 × 101 CFU/mL, and the functionalized surfaces did not show a cytotoxic effect on A549 cells (ATTC CCL 185). CONCLUSIONS: The use of gentamicin nanoparticles on the polyvinyl chloride surface may be an additional supporting method for patients after tracheostomy in order to prevent the colonization of the biomaterial by potentially pathogenic microorganisms.
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The solid-aqueous boundary formed upon biomaterial implantation provides a playground for most biochemical reactions and physiological processes involved in implant-host interactions. Therefore, for biomaterial development, optimization, and application, it is essential to understand the biomaterial-water interface in depth. In this study, oxygen plasma-functionalized polyurethane surfaces that can be successfully utilized in contact with the tissue of the respiratory system were prepared and investigated. Through experiments, the influence of plasma treatment on the physicochemical properties of polyurethane was investigated by atomic force microscopy, attenuated total reflection infrared spectroscopy, differential thermal analysis, X-ray photoelectron spectroscopy, secondary ion mass spectrometry, and contact angle measurements, supplemented with biological tests using the A549 cell line and two bacteria strains (Staphylococcus aureus and Pseudomonas aeruginosa). The molecular interpretation of the experimental findings was achieved by molecular dynamics simulations employing newly developed, fully atomistic models of unmodified and plasma-functionalized polyurethane materials to characterize the polyurethane-water interfaces at the nanoscale in detail. The experimentally obtained polar and dispersive surface free energies were consistent with the calculated free energies, verifying the adequacy of the developed models. A 20% substitution of the polymeric chain termini by their oxidized variants was observed in the experimentally obtained plasma-modified polyurethane surface, indicating the surface saturation with oxygen-containing functional groups.
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Materiais Biocompatíveis , Poliuretanos , Poliuretanos/química , Propriedades de Superfície , Água , OxigênioRESUMO
<b>Introduction:</b> In hospitalized patients, tracheostomy tubes (TTs) are susceptible to colonization by biofilm- producing potentially pathogenic microorganisms (PPMs). Contact with TTs, which are situated in a critical region of the body with enormous microbial exposure, may lead to the emer-gence of resistant respiratory infections.</br></br> <b>Objective:</b> Our study aimed to isolate and identify Gram-positive and Gram-negative PPMs, mark their antibiotic resistance and determine the bacteriological pattern of the biofilm colonizing the TTs. </br></br> <b>Methods:</b> The study was conducted on 45 tracheostomy tubes obtained from 45 hospitalized adult patients with tracheostomy with intubation periods ranging from 1 to 28 days. Tracheal aspirates (TA) obtained from polyvinyl chloride (PVC) TTs were used for the analysis. Bacteria in biofilms were identified by standard microbiological techniques, tested for antibiotic resistance and phenotypic resistance according to the EUCAST guidelines and visualized by SEM.</br></br> <b>Results:</b> Out of 45 TTs, 100% were found to be positive in bacterial cultures with 58 PPM isolates (10 spe-cies) correlating well with the SEM findings. Overall, 72% of isolates were Gram-negative bacilli, followed by Gram-positive cocci (28%). Staphylococcus aureus was the predominant bacterium (identified in 35.5% of patients), followed by Klebsiella pneumoniae (identified in 23.8%). Among the Gram-negative PPMs, 50% of isolates were identified as multidrug-resistant (MDR), 8.6% as extremely drug-resistant (XDR) and 5.2% were pandrug-resistant (PDR).</br></br><b>Conclusions:</b> Our study showed a rapid colonization of the TT surface by biofilm- producing PPMs. Patients with tracheosto- mies, also those with non-infectious conditions, were mainly colonized with highly re-sistant bacteria.
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Bactérias Gram-Negativas , Traqueostomia , Adulto , Humanos , Staphylococcus aureus , Farmacorresistência Bacteriana Múltipla , BiofilmesRESUMO
Background: Bacterial biofilm on the surface of tracheostomy tubes (TTs) is a potential reservoir of potentially pathogenic bacteria, including S. aureus. For this reason, our study aimed to investigate biofilm production in vitro and the presence of icaAD and MSCRAMM genes in clinical S. aureus strains derived from TTs, with respect to antibiotic resistance and genetic variability. Methods: The clonality of the S. aureus strains was analyzed by the PFGE method. The assessment of drug resistance was based on the EUCAST recommendations. The isolates were evaluated for biofilm production by the microtiter plate method and the slime-forming ability was tested on Congo red agar (CRA). The presence of icaAD genes was investigated by PCR and MSCRAMM genes were detected by multiplex PCR. Results: A total of 60 patients were enrolled in the study. One TT was obtained from each patient (n = 60). Twenty-one TTs (35%) were colonized with S. aureus. A total of 24 strains were isolated as 3 patients showed colonization with 2 SA clones (as confirmed by PFGE). PFGE showed twenty-two unique molecular profiles. Two isolates (8%) turned out to be MRSA, but 50% were resistant to chloramphenicol, 25% to erythromycin and 8% to clindamycin (two cMLSB and four iMLSB phenotypes were detected). The microtiter plate method with crystal violet confirmed that 96% of the strains were biofilm formers. Representative strains were visualized by SEM. All isolates had clfAB, fnbA, ebpS and icaAD. Different MSCRAMM gene combinations were observed. Conclusions: the present study showed that the S. aureus isolated from the TTs has a high diversity of genotypes, a high level of antibiotic resistance and ability to produce biofilm.
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Group B streptococcus (GBS) is one of the uropathogens that causes urinary tract infections (UTIs). The aims of this article were molecular characterization, an analysis of antimicrobial susceptibility profiles, adherence to bladder endothelial cells, and the detection of immunoreactive proteins of 94 clinical strains of GBS isolated from adult Polish patients with UTI. Antibiotic susceptibilities were determined by disk diffusion. Serotyping and Alp family genes detection were studied using multiplex PCR. Genetic profiles were determined by pulsed-field gel electrophoresis. The adherence ability of the studied strains was estimated by incubation on human bladder microvascular endothelial cell line. Immunoreactive proteins were studied by immunoblotting. Antibiotic susceptibility investigation revealed that 22% of GBS strains were resistant to erythromycin, whereas 18% demonstrated resistance to clindamycin. cMLSB was present in 76% of the resistant strains, M phenotype was detected in 14%, whereas iMLSB was present for 10%. The most common serotype was serotype III (31%), followed by serotype V (27%), and serotype Ia (17%). The genes that dominated among other Alp genes were: epsilon (29%), alp2 (27%), and rib (23%). The most common co-occurring serotypes and Alp genes were: Ia and epsilon, III and rib, III and alp2, V and alp2, and V and alp3 (p < 0.001). The PFGE method showed high clonality for serotype V and cMLSB (p < 001). The PFGE method showed high clonality for serotype V. Furthermore, this serotype was significantly associated with the cMLSB phenotype (p < 0.001). The most common immunoreactive proteins demonstrated masses of 50 kDa and 45-47 kDa. Although examined GBS isolates showed high genetic diversity, immunoreactive proteins were common for most of the studied GBS isolates, which may indicate their conservation, and allows to consider them as potential immunodiagnostic markers. Although the examined GBS isolates showed high genetic diversity, immunoreactive proteins were shared by most of the studied GBS isolates. It may indicate their conservation, thus allowing to consider them as potential immunodiagnostic markers.
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Partially covered self-expandable metallic esophageal stent (SEMS) placement is the most frequently applied palliative treatment in esophageal cancer. Structural characterization of explanted 16 nitinol-polyurethane SEMS (the group of 6 females, 10 males, age 40-80) was performed after their removal due to dysfunction. The adverse bulk changes in the polymer structure were identified using differential scanning calorimetry (DSC), differential mechanical thermal analysis (DMTA), and attenuated total reflectance infrared spectroscopy (ATR-IR) and discussed in terms of melting point shift (9 °C), glass-transition shift (4 °C), differences in viscoelastic behavior, and systematic decrease of peaks intensities corresponding to C-H, CâO, and C-N polyurethane structural bonds. The scanning electron and confocal microscopic observations revealed all major types of surface degradation, i.e., surface cracks, peeling off of the polymer material, and surface etching. The changes in the hydrophobic polyurethane surfaces were also revealed by a significant decrease in wettability (74°) and the corresponding increase of the surface free energy (31 mJ/m2). To understand the in vivo degradation, the in vitro tests in simulated salivary and gastric fluids were performed, which mimic the environments of proximal and distal ends, respectively. It was concluded that the differences in the degradation of the proximal and distal ends of prostheses strongly depend on the physiological environment, in particular stomach content. Finally, the necessity of the in vivo tests for SEMS degradation is pointed out.
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Neoplasias Esofágicas , Stents Metálicos Autoexpansíveis , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias Esofágicas/complicações , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Cuidados Paliativos , Resultado do TratamentoRESUMO
In this work, polymeric and bioactive glass (BG)-modified composite films were successfully loaded with polyphenols (PPh) extracted from sage. It was hypothesized that PPh, alone and in combination with BGs particles, would affect physicochemical and biological properties of the films. Furthermore, sol-gel-derived BG particles would serve as an agent for control the release of the polyphenolic compounds, and other important properties related to the presence of PPh. The results showed that polyphenolic compounds significantly modified numerous material properties and also acted as biologically active substances. On the one hand, PPh can be considered as plasticizers for PCL, on the other hand, they can act as coupling agent in composite materials, improving their mechanical performance. The presence of PPh in materials improved their hydrophilicity and apatite-forming ability, and also provided antioxidant activity. What is important is that the aforementioned properties and kinetics of PPh release can be modulated by the use of various concentrations of PPh, and by the modification of PCL matrix with sol-gel-derived BG particles, capable of binding PPh. The films containing the lowest concentration of PPh exhibited cytocompatibility, significantly increased alkaline phosphatase activity and the expression of bone extracellular matrix proteins (osteocalcin and osteopontin) in human normal osteoblasts, while they reduced intracellular reactive oxygen species production in macrophages. Furthermore, materials loaded with PPh showed antibiofilm properties against Gram positive and Gram negative bacteria. The results suggest that obtained materials represent potential multifunctional biomaterials for bone tissue engineering with a wide range of tunable properties.
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Interactions at the solid-body fluid interfaces play a vital role in bone tissue formation at the implant surface. In this study, fully atomistic molecular dynamics (MD) simulations were performed to investigate interactions between the physiological components of body fluids (Ca2+, HPO42-, H2PO4-, Na+, Cl-, and H2O) and functionalized parylene C surface. In comparison to the native parylene C (-Cl surface groups), the introduction of -OH, -CHO, and -COOH surface groups significantly enhances the interactions between body fluid ions and the polymeric surface. The experimentally observed formation of calcium phosphate nanocrystals is discussed in terms of MD simulations of the calcium phosphate clustering. Surface functional groups promote the clustering of calcium and phosphate ions in the following order: -OH > -CHO > -Cl (parent parylene C) ≈ -COO-. This promoting role of surface functional groups is explained as stimulating the number of Ca2+ and HPO42- surface contacts as well as ion chemisorption. The molecular mechanism of calcium phosphate cluster formation at the functionalized parylene C surface is proposed.
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This paper reports on the plasma electrolytic oxidation (PEO) of titanium alloy Ti-15Mo in baths containing zinc to obtain biomaterials with bacteriostatic and antibacterial properties. The Ti-15Mo surface was oxidised in a 0.1â¯M Ca(H2PO2)2 bath containing zinc compound particles: ZnO or Zn3(PO4)2. During the PEO process, the applied voltage was 300â¯V, and the current density was 150â¯mAâcm-2. The surface morphology, roughness and wettability were determined. It has been noted that both roughness and wettability of Ti-15Mo alloy surface increased after PEO. EDX and XPS chemical composition analysis was carried out, and Raman spectroscopy was also performed indicating that Zn has been successfully incorporated into oxide layer. To investigate the antibacterial properties of the PEO oxide coatings, microbial tests were carried out. The bacterial adhesion test was performed using four different bacterial strains: reference Staphylococcus aureus (ATCC 25923), clinical Staphylococcus aureus (MRSA 1030), reference Staphylococcus epidermidis (ATCC 700296) and clinical Staphylococcus epidermidis (15560). Performed zinc-containing oxide coatings did not indicate the bacteria growth inducing effect. Additionally, the cytocompatibility of the formed oxide layers was characterised by MG-63 osteoblast-like live/dead tests. The surface bioactivity and cytocompatibility increased after the PEO process. The zinc was successfully incorporated into the titanium oxide layer. Based on the obtained results of the studies, it can be claimed that zinc-containing PEO layers can be an interesting course of bacteriostatic titanium biomaterials development.
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Ligas/farmacologia , Antibacterianos/farmacologia , Aderência Bacteriana/efeitos dos fármacos , Fosfatos/química , Compostos de Zinco/química , Óxido de Zinco/química , Ligas/química , Antibacterianos/química , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Humanos , Osteoblastos/classificação , Osteoblastos/efeitos dos fármacos , Análise Espectral Raman , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/crescimento & desenvolvimento , MolhabilidadeRESUMO
Multifunctional and biofunctional coatings for medical devices are an attractive strategy towards tailoring the interactions of the device with the body, thereby influencing the host response, and the susceptibility to microbial colonization. Here we describe the development of a coating process to yield amphiphilic, lubricious coatings, resistant to bacterial colonization, based on chitosan. Chitosan-fatty acid derivatives were obtained by simultaneous N,O-acylation of chitosan with either linoleic, α-linolenic, or dilinoleic acid. Chemical characterization of new materials was carried out using 1H NMR, FTIR, and XPS. Surface properties of coated polyester samples were studied using SEM and contact angle measurements, which indicated that the incorporation of hydrophobic constituents into chitosan macromolecules led to a decrease of both surface roughness and water contact angle. Importantly, tribological testing demonstrated that these new coatings decrease the coefficient of friction due to the self-organization of fatty acid (from 0.53 for the neat chitosan to 0.35 for chitosan-fatty acid derivative). Meanwhile, preliminary bacterial colonization tests indicated significant-over 80%-reduction in E. coli colonization following coating with chitosan-linoleic and chitosan-α-linolenic derivatives. Finally, cytotoxicity and hemocompatibility studies confirmed that all amphiphilic chitosan-fatty acid derivatives were non-toxic and non-hemolytic. Collectively, our results demonstrate the potential of the developed coating strategy, particularly the chitosan-linoleic and chitosan-α-linolenic acid derivatives, for applications as biofunctional catheter coatings.
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Quitosana/química , Materiais Revestidos Biocompatíveis/química , Ácidos Graxos/química , Animais , Antibacterianos/química , Escherichia coli/crescimento & desenvolvimento , Interações Hidrofóbicas e Hidrofílicas , Células L , Camundongos , Propriedades de SuperfícieRESUMO
In this paper, the preparation of a functional hybrid coating loaded with a drug (amoxicillin) on a promising titanium alloy - Ti-15Mo alloy is presented. The titanium alloy surface was anodized in solution with bioactive compounds to obtain a porous oxide layer favorable for MG-63 osteoblast-like cell adhesion. Then, a poly(lactide-co-glycolide) (PLGA) loaded with amoxicillin layer was formed using a dip-coating technique to cover the oxide layer, without filling in all of the pores. The morphology of the surface was evaluated using scanning electron microscopy supported by 3D Roughness Reconstruction software. The surface treatment of the Ti-15Mo alloy surface caused the surface roughness to increase up to 1.71⯵m. The anodization process caused the Ti-15Mo alloy surface to become slightly more hydrophilic; however, the formation of the PLGA layer loaded with drug increased the contact angle to 96.5°â¯±â¯2.2°, respectively. After 4â¯weeks of polymer layer degradation, the registered signals on the 1H NMR spectrum were identical to the signals registered for lactic acid (LAc), which confirms that the polymer layer was degraded within a short period of time. The concentration of drug released into the artificial saliva was investigated using high-performance liquid chromatography (HPLC) up to 12â¯h of coatings immersion. During the first hour of coating degradation in artificial saliva, and the concentration of the drug (13⯵g/ml) was enough to inhibit bacterial growth of S. aureus and S. epidermidis. These results were confirmed by agar plate diffusion method and evaluation of the minimal inhibitory concentration (MIC). The cytocompatibility of the materials was determined using the osteoblast-like cells MG-63, and the viability and cell morphology (live/dead staining) were also evaluated. The results showed that amoxicillin influences the osteoblast-like MG-63 cells' behavior during cell culture, especially for the first few hours. The influence on the type of surface treatment on MG-63 cell behavior during 7â¯days of culture is discussed in this paper. To the best of our knowledge, this is the first time that a fast-degrading layer with amoxicillin has been deposited on previously anodized Ti surface. The formation of functional coating may find application as a cytocompatible coating to prevent bacterial adhesion on long-term implant surfaces.
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Ligas/química , Amoxicilina/farmacologia , Implantes Dentários , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Titânio/química , Amoxicilina/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Materiais Revestidos Biocompatíveis/química , Eletrodos , Humanos , Testes de Sensibilidade Microbiana , Espectroscopia de Prótons por Ressonância Magnética , Saliva/química , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus epidermidis/efeitos dos fármacos , MolhabilidadeRESUMO
Solid-water interfaces play a vital role in biomaterials science because they provide a natural playground for most biochemical reactions and physiological processes. In the study, fully atomistic molecular dynamics simulations were performed to investigate interactions between water molecules and several surfaces modeling for unmodified and modified parylene C surfaces. The introduction of -OH, -CHO, and -COOH to the surface and alterations in their coverage significantly influence the energetics of interactions between water molecules and the polymer surface. The theoretical studies were complemented with experimental measurements of contact angle, surface free energy, and imaging of osteoblast cells adhesion. Both MD simulations and experiments demonstrate that the optimal interface, in terms of biocompatibility, is obtained when 60% of native -Cl groups of parylene C surface is exchanged for -OH groups. By exploring idealized models of bare and functionalized parylene C, we obtained a unique insight into molecular interactions at the water-polymer interface. The calculated values of interaction energy components (electrostatic and dispersive) correspond well with the experimentally determined values of surface free energy components (polar and dispersive), revealing their optimal ratio for cells adhesion. The results are discussed in the context of controllable tuning and functionalization of implant polymeric coating toward improved biocompatibility.