Alexandre António Antunes Barros

Bioresorbable ureteral stents from natural origin polymers

In this work, stents were produced from natural origin polysaccharides. Alginate, gellan gum, and a blend of these with gelatin were used to produce hollow tube (stents) following a combination of templated gelation and critical point carbon dioxide drying. Morphological analysis of the surface of the stents was carried out by scanning electron microscopy. Indwelling time, encrustation, and stability of the stents in artificial urine solution was carried out up to 60 days of immersion. In vitro studies carried out with simulated urine demonstrated that the tubes present a high fluid uptake ability, about 1000%. Despite this, the materials are able to maintain their shape and do not present an extensive swelling behavior. The bioresorption profile was observed to be highly dependent on the composition of the stent and it can be tuned. Complete dissolution of the materials may occur between 14 and 60 days. Additionally, no encrustation was observed within the tested timeframe. The ability to resist bacterial adherence was evaluated with Gram-positive Staphylococcus aureus and two Gram-negatives Escherichia coli DH5 alpha and Klebsiella oxytoca. For K. oxytoca, no differences were observed in comparison with a commercial stent (Biosoft(®) duo, Porges), although, for S. aureus all tested compositions had a higher inhibition of bacterial adhesion compared to the commercial stents. In case of E. coli, the addition of gelatin to the formulations reduced the bacterial adhesion in a highly significant manner compared to the commercial stents. The stents produced by the developed technology fulfill the requirements for ureteral stents and will contribute in the development of biocompatible and bioresorbable urinary stents.

Carboxymethylation of ulvan and chitosan and their use as polymeric components of bone cements.

Ulvan, extracted from the green algae Ulva lactuca, and chitosan, extracted from Loligo forbesis squid-pen, were carboxymethylated, yielding polysaccharides with an average degree of substitution of ∼98% (carboxymethyl ulvan, CMU) and ∼87% (carboxymethyl chitosan, N,O-CMC). The carboxymethylation was confirmed by Fourier transform infrared spectroscopy and quantified by conductimetric titration and 1H nuclear magnetic resonance. The average molecular weight increased with the carboxymethylation (chitosan, Mn 145→296 kDa and Mw 227→416 kDa; ulvan, Mn 139→261 kDa and Mw 368→640 kDa), indicating successful chemical modifications. Mixtures of the modified polysaccharides were tested in the formulation of polyacrylic acid-free glass-ionomer bone cements. Mechanical and in vitro bioactivity tests indicate that the inclusion of CMU in the cement formulation, i.e. 0.50:0.50 N,O-CMC:CMU, enhances its mechanical performance (compressive strength 52.4±8.0 MPa and modulus 2.3±0.3 GPa), generates non-cytotoxic cements and induces the diffusion of Ca and/or P-based moieties from the surface to the bulk of the cements.

Ketoprofen-eluting biodegradable ureteral stents by CO2 impregnation: In vitro study

Ureteral stents are indispensable tools in urologic practice. The main complications associated with ureteral stents are dislocation, infection, pain and encrustation. Biodegradable ureteral stents are one of the most attractive designs with the potential to eliminate several complications associated with the stenting procedure. In this work we hypothesize the impregnation of ketoprofen, by CO2-impregnation in a patented biodegradable ureteral stent previously developed in our group. The biodegradable ureteral stents with each formulation: alginate-based, gellan gum-based were impregnated with ketoprofen and the impregnation conditions tested were 100 bar, 2 h and three different temperatures (35 °C, 40 °C and 50 °C). The impregnation was confirmed by FTIR and DSC demonstrated the amorphization of the drug upon impregnation. The in vitro elution profile in artificial urine solution (AUS) during degradation of a biodegradable ureteral stent loaded with ketoprofen was evaluated. According to the kinetics results these systems have shown to be very promising for the release ketoprofen in the first 72 h, which is the necessary time for anti-inflammatory delivery after the surgical procedure. The in vitro release studied revealed an influence of the temperature on the impregnation yield, with a higher impregnation yield at 40 °C. Higher yields were also obtained for gellan gum-based stents. The non-cytotoxicity characteristic of the developed ketoprofen-eluting biodegradable ureteral stents was evaluated in L929 cell line by MTS assay which demonstrated the feasibility of this product as a medical device.

Drug-eluting biodegradable ureteral stent: new approach for urothelial tumors of upper urinary tract cancer

Upper urinary tract urothelial carcinoma (UTUC) accounts for 5-10% of urothelial carcinomas and is a disease that has not been widely studied as carcinoma of the bladder. To avoid the problems of conventional therapies, such as the need for frequent drug instillation due to poor drug retention, we developed a biodegradable ureteral stent (BUS) impregnated by supercritical fluid CO2 (scCO2) with the most commonly used anti-cancer drugs, namely paclitaxel, epirubicin, doxorubicin, and gemcitabine. The release kinetics of anti-cancer therapeutics from drug-eluting stents was measured in artificial urine solution (AUS). The in vitro release showed a faster release in the first 72h for the four anti-cancer drugs, after this time a plateau was achieved and finally the stent degraded after 9days. Regarding the amount of impregnated drugs by scCO2, gemcitabine showed the highest amount of loading (19.57μg drug/mg polymer: 2% loaded), while the lowest amount was obtained for paclitaxel (0.067μg drug/mg polymer: 0.01% loaded). A cancer cell line (T24) was exposed to graded concentrations (0.01-2000ng/ml) of each drugs for 4 and 72h to determine the sensitivities of the cells to each drug (IC50). The direct and indirect contact study of the anti-cancer biodegradable ureteral stents with the T24 and HUVEC cell lines confirmed the anti-tumoral effect of the BUS impregnated with the four anti-cancer drugs tested, reducing around 75% of the viability of the T24 cell line after 72h and demonstrating minimal cytotoxic effect on HUVECs.

7.41 Ureteral Stents Technology: Biodegradable and Drug-Eluting Perspective

An ureteral stent is a versatile and indispensable common medical device in the management of several urological diseases. Nonetheless, the actual stent technology is far from the ideal. The ureteral stents available in the market are associated with clinical complications including bacterial adhesion, infection, encrustation development, pain and discomfort for the patients. Innovative stent materials and more research of new ureteral stent designs are necessary in order to avoid these complications and pursue the ideal stent, ie, a stent that can maintain the adequate urine flow from the kidney to the bladder and mitigation of hydronephrosis. In past years different approaches have been explored, including novel stent coatings, drug-eluting stent and biodegradable ureteral stents. The common goal of these new technologies is to increase the biocompatibility of the biomaterials and avoid the stent-related symptoms. New developments in ureteral stent field, namely drug-eluting biodegradable ureteral stents, are also envisaged to address new urological clinical scenarios. In summary there is no perfect ureteral stent that avoids all ureteral stent associated complications but there have been significant advances in last years in stent technology and particularly technologies such biodegradable drug-eluting stents are seen as very promising.

In vitro bioactivity studies of ceramic structures isolated from marine sponges.

In this work, we focused on the potential of bioceramics from different marine sponges-namely Petrosia ficiformis, Agelas oroides and Chondrosia reniformis-for novel biomedical/industrial applications. The bioceramics from these sponges were obtained after calcination at 750 °C for 6 h in a furnace. The morphological characteristics were evaluated by scanning electron microscopy (SEM). The in vitro bioactivity of the bioceramics was evaluated in simulated body fluid (SBF) after 14 and 21 d. Observation of the bioceramics by SEM after immersion in SBF solution, coupled with spectroscopic elemental analysis (EDS), showed that the surface morphology was consistent with a calcium-phosphate (Ca/P) coating, similar to hydroxyapatite crystals (HA). Evaluation of the characteristic peaks of Ca/P crystals by Fourier transform infrared spectroscopy and x-ray diffraction further confirmed the existence of HA. Cytotoxicity studies were carried out with the different ceramics and these were compared with a commercially available Bioglass(®). In vitro tests demonstrated that marine bioceramics from these sponges are non-cytotoxic and have the potential to be used as substitutes for synthetic Bioglass(®).

Bioactive Composites Reinforced with Inorganic Glasses and Glass–Ceramics for Tissue Engineering Applications

Bioactive composites, prepared by the combination of glasses or glass–ceramics with natural or synthetic polymers or blends, have been extensively exploited in bone tissue engineering. Their bioactive character is usually derived from the glass or glass–ceramic phase and is one of the most relevant properties to generate bone bonding. Herein we focus on the development of bioactive composite structures that target tissue engineering applications, with special emphasis on bone regeneration. Some concepts, e.g., bioactivity and biocompatibility, are initially introduced, followed by a description of the synthetic approaches that have been reported for the preparation of bioactive inorganic glasses or glass–ceramics. Different strategies to compound these inorganic particles with polymeric phases are detailed, spanning from conventional methodologies and wet spinning to rapid prototyping. Finally, a series of systems that have been developed for bone tissue engineering are described (including injectable systems, 3D scaffolds, membranes, and biomimetic layer-by-layer structures), as well as their in vitro biological response.

Upper urinary tract urothelial tumours targeted with biodegradable drug-eluting stents

3bs Research Group, Dept. of Polymer Engineering, Barco, Portugal, University of Berkeley, Dept. of Bioengineering and Materials Science and Engineering, Berkeley, United States of America, Life and Health Sciences Research Institute (ICVS), Dep. of Health Sciences, Braga, Portugal, 3bs Research Group, Dept. of Polymer Engineering, Portugal, Life and Health Sciences Research Institute (ICVS), Braga, Portugal

Bioactive ceramics for tissue engineering and regenerative medicine derived from marine sponges

Introduction: Decellularized engineered extracellular matrices (ECM) are used in a variety of regenerative medicine applications. Existing decellularization strategies rely on cell lysis and generally result in a variable but significant impairment of the ECM structure/composition. As an alternative, we aimed at activating the apoptotic pathway in order to decellularize engineered matrices while preserving their osteo-inductive properties [1]. Materials and methods: We generated a death-inducible, immortalized human Mesenchymal Stromal Cell (hMSC) line [2]. Cells were seeded on ceramic scaffolds and cultured for 4 weeks in osteogenic medium in a 3D perfusion bioreactor system (U-cup, Cellec). The ECM was decellularized by direct supply of the apoptotic inducer in the 3D culture system. Grafts were implanted in a rat cranial defect model to assess their regenerative potential after 12 weeks. Results: Cells were successfully seeded and differentiated, leading to deposition of a dense ECM during 3D culture. The apoptosis induction allowed for efficient decellularization while preserving the secreted matrix. These “apoptized” cell-free ECM coated constructs induced superior bone regeneration than control materials (Fig. 2). Areas of de novo bone formation not connected with surrounding bone suggest osteoinductive properties of the grafts.