Urinary Biomaterials Research Articles

Application of Novel 3,4-Dihydroxyphenylalanine-Containing Antimicrobial Polymers for the Prevention of Uropathogen Attachment to Urinary Biomaterials.

Introduction and Objective: Urinary catheters and stents are frequently prone to catheter-associated urinary tract infections (CAUTI) through biofilm formation. Several strategies have been evaluated in search of a stent coating to reliably prevent adherence of bacteria and biofilm. Previous in vivo and in vitro research with methoxylated polyethylene glycol 3,4-dihydroxyphenylalanine (DOPA) copolymer as a candidate coating showed promising results to reduce the bacterial attachment. We aimed to further enhance this antimicrobial activity by adding antimicrobial agents to newly synthesized DOPA-based copolymers. Materials and Methods: Building on our previous experience, novel copolymers were engineered based on DOPA. Quaternary ammonium groups and silver particles were added by cross-linking to increase the antimicrobial activity through both kill-by-contact and planktonic killing. After coating polyurethane sheets and measuring contact angles, all candidate coatings were challenged in vitro with an Escherichia coli culture. The most promising coatings were then further evaluated against a panel of seven clinically relevant uropathogens and planktonic killing, and microbial attachment was determined. Results: Initially, seven coatings were developed, referred to as Surphys 093-099. The most significant increase in contact angle was identified in Surphys-095 and -098. Surphys coatings S-094, S-095, and S-098 were cross-linked with silver and exhibited profound antimicrobial properties when challenged with E. coli. Further testing demonstrated S-095 to have antimicrobial efficacy against gram-positive and gram-negative bacteria at different silver-loading concentrations. The final coating, consisting of a 2 mg/mL solution of S-095 cross-linked with 0.25 mg/mL AgNO3, appeared to be highly bactericidal showing a ≥99.9% bacterial killing effect while remaining below cytotoxicity levels. Conclusions: We were able to engineer DOPA-based copolymers and add quaternary ammonium and silver particles, thus increasing the bactericidal properties of the coating. These coatings have exhibited a biologically significant ability to prevent uropathogens from attaching to biomaterials and represent a realistic opportunity to combat CAUTI.

Minimal Device Encrustation on Vesair Intravesical Balloons in the Treatment of Stress Urinary Incontinence: Analysis of Balloons Removed from Women in the SOLECT Trial.

IntroductionEncrustation of urinary biomaterials is common; however, the incidence of surface deposition on the Vesair® intravesical pressure-attenuation balloon has not been previously reported. The purpose of this analysis is to determine the incidence and potential risk factors for encrustation of the Vesair intravesical balloon.MethodsThe SOLECT trial is a prospective randomized controlled trial conducted at several European centers to evaluate the safety and efficacy of the Vesair balloon for the treatment of female stress urinary incontinence (SUI). Women included in the study demonstrated SUI symptoms for more than 12 months without complicating factors, such as history of recurrent urinary tract infections or nephrolithiasis. All balloons removed from women enrolled in the SOLECT trial were analyzed for surface characteristics and encrustation. Surface deposition severity was quantified and composition analyzed with infrared spectroscopy and scanning electron microscopy. Incidence of surface deposition was tabulated and risk factors analyzed.ResultsOne hundred and five balloons removed from 75 women were included in this analysis. Measurable stone deposition of less than 1.5 mm was found on four balloons (3.8%), surface granules were noted on 42 (40.0%), surface film on 11 (10.5%), and both granules and film on two (1.9%). Analysis identified calcium oxalate both in measurable encrustation deposits as well as those with surface granulation. Pooled analysis found that dwell time was a risk factor for calcium deposition.ConclusionThe rate of encrustation on the Vesair intravesical balloon is low and does not appear to increase the rate of adverse outcomes or reduce clinical efficacy.FundingSolace Therapeutics, Inc.

An Infection-Responsive Approach To Reduce Bacterial Adhesion in Urinary Biomaterials.

Infection is an inevitable consequence of chronic urinary catheterization with associated problems of recurrent catheter encrustation and blockage experienced by approximately 50% of all long-term catheterized patients. In this work, we have exploited, for the first time, the reported pathogen-induced elevation of urine pH as a trigger for "intelligent" antimicrobial release from novel hydrogel drug delivery systems of 2-hydroxyethyl methacrylate and vinyl-functionalized nalidixic acid derivatives, developed as candidate infection-resistant urinary catheter coatings. Demonstrating up to 20-fold faster rates of drug release at pH 10, representing infected urine pH, than at pH 7 and achieving reductions of up to 96.5% in in vitro bacterial adherence, our paradigm of pH-responsive drug delivery, which requires no external manipulation, therefore represents a promising development toward the prevention of catheter-associated urinary tract infections in vivo.

The C-terminal fragment of prostate-specific antigen, a 2331 Da peptide, as a new urinary pathognomonic biomarker candidate for diagnosing prostate cancer.

Background and ObjectivesProstate cancer (PCa) is one of the most common cancers and leading cause of cancer-related deaths in men. Mass screening has been carried out since the 1990s using prostate-specific antigen (PSA) levels in the serum as a PCa biomarker. However, although PSA is an excellent organ-specific marker, it is not a cancer-specific marker. Therefore, the aim of this study was to discover new biomarkers for the diagnosis of PCa.Materials and MethodsWe focused on urine samples voided following prostate massage (digital rectal examination [DRE]) and conducted a peptidomic analysis of these samples using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MSn). Urinary biomaterials were concentrated and desalted using CM-Sepharose prior to the following analyses being performed by MALDI-TOF/MSn: 1) differential analyses of mass spectra; 2) determination of amino acid sequences; and 3) quantitative analyses using a stable isotope-labeled internal standard.ResultsMultivariate analysis of the MALDI-TOF/MS mass spectra of urinary extracts revealed a 2331 Da peptide in urine samples following DRE. This peptide was identified as a C-terminal PSA fragment composed of 19 amino acid residues. Moreover, quantitative analysis of the relationship between isotope-labeled synthetic and intact peptides using MALDI-TOF/MS revealed that this peptide may be a new pathognomonic biomarker candidate that can differentiate PCa patients from non-cancer subjects.ConclusionThe results of the present study indicate that the 2331 Da peptide fragment of PSA may become a new pathognomonic biomarker for the diagnosis of PCa. A further large-scale investigation is currently underway to assess the possibility of using this peptide in the early detection of PCa.

Ureteral Stents and Foley Catheters-Associated Urinary Tract Infections: The Role of Coatings and Materials in Infection Prevention.

Urinary tract infections affect many patients, especially those who are admitted to hospital and receive a bladder catheter for drainage. Catheter associated urinary tract infections are some of the most common hospital infections and cost the health care system billions of dollars. Early removal is one of the mainstays of prevention as 100% of catheters become colonized. Patients with ureteral stents are also affected by infection and antibiotic therapy alone may not be the answer. We will review the current evidence on how to prevent infections of urinary biomaterials by using different coatings, new materials, and drug eluting technologies to decrease infection rates of ureteral stents and catheters.

Synthesis and release kinetics of polymerisable ester drug conjugates: towards pH-responsive infection-resistant urinary biomaterials

Herein we report the synthesis, characterisation and hydrolytic release kinetics of a suite of novel, polymerisable ester quinolone conjugates with varying alkenyl chain lengths. Hydrolysis was shown to proceed up to 17-fold faster upon elevation of pH from neutral to pH 9.29, making these conjugates attractive for the development of ‘designer’ infection-resistant urinary biomaterials exploiting the increase in urine pH reported at the onset of catheter-associated infection to trigger drug release.

Development of sustained-release antibacterial urinary biomaterials through using an antimicrobial as an organic modifier in polyurethane nanocomposites

Urinary catheters are among the most frequently used medical devices in clinical practice. However, their use is associated with high rates of nosocomial infection. This study investigates the use of polyurethane nanocomposites (PUNCs) incorporating an antimicrobial agent, chlorhexidine diacetate (CHX), behaving as nanoparticle dispersant and model drug/active agent, as sustained-release antibacterial biomaterials in urinary devices. A range of PUNCs incorporating organically modified silicate (OMS) nanoparticles with CHX was fabricated using a solution-cast method. PUNCs with free CHX added into the bulk polymer were also made. Materials were assessed for antibacterial activity in an in vitro urinary tract (UT) model and release kinetics of CHX was studied. PUNCs demonstrated sustained antibacterial activity against Staphylococcus epidermidis in the UT model, reaching ~50 days infection-free in materials with 2 wt % free CHX loading. Drug-release profiles demonstrated that, compared with microcomposite and unfilled polyurethane, the initial burst effect was significantly reduced in PUNCs. Prolonged drug release was achieved through incorporation of OMS, hypothesized to be due to a combination of barrier properties created by the nanoinclusions and strong interactions between CHX and MMT within the PUNCs. Use of PUNCs for sustained drug release in long-term urinary applications shows promise in addressing catheter-related nosocomial infections.

Infection-Responsive Drug Delivery from Urinary Biomaterials Controlled by a Novel Kinetic and Thermodynamic Approach

The pH-dependent physicochemical properties of the antimicrobial quinolone, nalidixic acid, were exploited to achieve 'intelligent' drug release from a potential urinary catheter coating, poly(2-hydroxyethylmethacrylate) (p(HEMA)), in direct response to the elevated pH which occurs at the onset of catheter infection. p(HEMA) hydrogels, and reduced-hydrophilicity copolymers incorporating methyl methacrylate, were loaded with nalidixic acid by a novel, surface particulate localization method, and characterized in terms of pH-dependent drug release and microbiological activity against the common urease-producing urinary pathogen Proteus mirabilis. The pH-dependent release kinetics of surface-localized nalidixic acid were 50- and 10-fold faster at pH 9, representing the alkaline conditions induced by urease-producing urinary pathogens, compared to release at pH 5 and pH 7 respectively. Furthermore, microbiological activity against P. mirabilis was significantly enhanced after loading surface particulate nalidixic acid in comparison to p(HEMA) hydrogels conventionally loaded with dispersed drug. The more hydrophobic methyl methacrylate-containing copolymers also demonstrated this pH-responsive behavior, but additionally exhibited a sustained period of zero-order release. The paradigm presented here provides a system with latent, immediate infection-responsive drug release followed by prolonged zero-order antimicrobial delivery, and represents an 'intelligent', infection-responsive, self-sterilizing biomaterial.

Development of a lubricity test for urinary biomaterials

Development of a lubricity test for urinary biomaterials

Effects of hyaluronan and SPARC on fibroproliferative events assessed in an in vitro bladder acellular matrix model

Bladder acellular matrix (BAM) is a promising candidate for urinary biomaterials development. In the current work we have modified the BAM construct to include two biologically active components; hyaluronan (HA) and a peptide (SP 4.2) derived from secreted protein, acidic, rich in cysteine (SPARC), a matricellular glycoprotein. In order to assess the potential of an HA/SP 4.2 modified BAM to influence cellular functions associated with bladder healing, experiments were conducted to evaluate the individual and combined effects of these molecules on in vitro fibroproliferative endpoints within a co-culture model. Thiol-modified HA (246 kDa, 15 mg/ml)±SP 4.2 (200 μ m) was incorporated and cross-linked into BAM disks through disulfide bond formation. The following scaffolds compositions were then evaluated in a bladder smooth muscle cell (SMC)–urothelial (UEC) cell co-culture model: BAM unmodified; BAM+HA, BAM+SP 4.2 (media addition); BAM+HA+SP 4.2 (media addition); BAM+HA+SP 4.2 (matrix incorporated). At 3, 7 and 14 days post-seeding, SMC-mediated matrix contraction and gelatinolytic activity were evaluated. HA-modified BAM exhibited a significantly higher degree of contraction and gelatinase activity compared to unmodified BAM. In contrast, addition of SP 4.2 to BAM produced a negligible effect on contraction, while significantly reducing gelatinase activity. Matrices containing both molecules displayed significant increases in contraction, while gelatinase activity was dependent upon the method of peptide delivery. These results demonstrate that both HA and SP 4.2 have significant, yet distinct effects on the contractile and proteolytic activity of bladder SMCs and suggest that a modified BAM may be capable of modulating processes associated with post-surgical graft contracture and scar formation.

Physicochemical characterisation and biological evaluation of hydrogel-poly(ε-caprolactone) interpenetrating polymer networks as novel urinary biomaterials

Hydrogels are frequently employed as medical device biomaterials due to their advantageous biological properties, e.g. resistance to infection and encrustation, biocompatibility; however, their poor mechanical properties generally limit the scope of application to coatings of medical devices. To address this limitation, this study described the formulation of sequential interpenetrating polymer networks (IPN) of poly(ε-caprolactone) (PCL) and poly(hydroxyethylmethacrylate) (p(HEMA)). IPN containing 20% w/w PCL, p(HEMA), both in the presence or absence of ethyleneglycol dimethacrylate (EGDMA 1% w/w), were prepared by free radical polymerisation. Following preparation the degradation and the mechanical and surface properties of the biomaterials and, in addition, the resistances to microbial adherence and encrustation in vitro were examined. In comparison to p(HEMA) the various IPN exhibited substantially greater tensile properties (ultimate tensile strength, % elongation, Young's modulus) that were accredited to the discrete distribution of PCL within the hydrogel network. The IPN exhibited two glass transition temperatures that were statistically similar to those of the individual components, thereby providing evidence of the immiscible nature of the two polymers. The IPN possessed higher receding contact angles and lower equilibrium water contents in comparison to p(HEMA), whereas the limited degradation of the IPN at both pH 7 and 9 was deemed suitable for clinical usage for periods of at least 4 weeks. The resistances of the various IPN to bacterial adherence and urinary encrustation were examined using in vitro models. Importantly the resistance of the IPN to encrustation was, in general, similar to that of p(HEMA) but greater than that of PCL whereas, the resistance of the IPN to bacterial adherence was frequently greater than that of p(HEMA) and PCL. Therefore, this study has shown that the mechanical properties of p(HEMA) may be substantially increased by the formation of IPN with PCL whilst maintaining other appropriate physicochemical properties and resistances to urinary encrustation and bacterial adherence. It is suggested that these IPN may be suitable for device fabrication thereby expanding the manufacturing application of hydrogels without compromising their potential clinical efficacy.

Immobilization of an oxalate-degrading enzyme on silicone elastomer.

Urinary biomaterials are compromised by device-related urinary tract infections, bacterial biofilm formation, and biomineral encrustation. In the absence of urinary infection, calcium oxalate is the prevalent encrustation mineral formed. Considering this, a novel approach was taken in the study reported here, in that an oxalate-degrading enzyme, oxalate oxidase (OXO), was immobilized on the surface of silicone elastomer (PDMS), a common urological biomaterial. It was hypothesized that the enzymatic action of OXO could lower urinary oxalate levels near the device surface, thereby preventing calcium oxalate crystal formation. The PDMS surface was functionalized with the use of radio-frequency plasma discharge (RFPD) in the presence of water vapor, then coated with 3-aminopropyltriethoxysilane (AMEO). The resulting aminated surface was covalently coupled with OXO via glutaraldehyde bioconjugation. The ability of the OXO-coated PDMS to prevent calcium oxalate encrustation was evaluated with the use of a modified Robbins device (MRD) encrustation model. RFPD performed on PDMS resulted in an increase in the hydrophilicity of treated surfaces, as measured by contact angle. X-ray photoelectron spectroscopy studies showed increases in elemental oxygen, after water-vapor plasma, and in nitrogen after AMEO derivatization. The immobilized enzyme was shown to retain 47.5% of its specific enzymatic activity as compared to free enzyme. In vitro experiments for 6 days, with the use of a MRD, showed 53% less encrustation deposits on discs coated with OXO than control discs. The results from the current study suggest that PDMS-immobilized oxalate-degrading enzymes are active against calcium oxalate encrustation.

Cultured differentiated human urothelial cells in the biomaterials field

To review the use of normal cultured differentiated human urothelial cells in the biomaterials field, we checked the literature for human urothelial cells in culture (HUC) both for their use in biocompatibility assessment and as bioartificial devices. The in vitro culture of differentiated human urothelium is now a simple and reliable procedure. These techniques provide new tools for biocompatibility assessment of urinary biomaterials, because for the rational design of a testing procedure, it is preferable that the particular cell culture models selected should be closely related to the end-use application. The emerging use of HUC culture should lead to the development of bioartificial tissue for urinary tract reconstruction. Tissue engineering techniques require urothelial cells and cell delivery matrices. The cytocompatibility of novel artificial delivery matrices should be assessed in vitro before implantation using cultured HUC to find the best material available.

41] Biofilm and biofilm-related encrustation of urinary tract devices

This chapter focuses on the development of artificial urine models that simulate in vivo encrustation resulting from device-related infection and can be used to assess biomaterial encrustation and the ability of biomaterials to resist intraluminal blockage. The extent to which different urinary biomaterials will encrust in vivo is a problem that needs to be addressed if new, less encrusting biomaterials are to be discovered. The optimal level of agitation within a model is determined by the portion of the urinary tract in which the test biomaterial is destined to reside. Artificial urine models prove useful for testing novel biodegradable biomaterials for use as urinary prostheses. In practice, urine flow may encourage surface layers of the degradable polymer to be shed, thereby assisting the removal of encrusting deposits and adherent microorganisms. Such biomimetic biomaterials may have a role to play in the development of improved urinary tract devices.

Characeristationi of polyurethane/polymethylmethacrylate (PU/PMMA) networks as urinary biomaterials

Characeristationi of polyurethane/polymethylmethacrylate (PU/PMMA) networks as urinary biomaterials