Angel Contreras-García

Stimuli–responsive networks grafted onto polypropylene for the sustained delivery of NSAIDs

Co-polymers of N-isopropyl acrylamide (NIPAAm) and N-(3-aminopropyl) methacrylamide hydrochloride (APMA) were grafted on polypropylene (PP) films by means of a γ-ray pre-irradiation method, with the aim of developing medical devices able to load non-steroidal anti-inflammatory drugs (NSAIDs) and to control their release under physiological conditions. The NIPAAm/APMA molar ratios in the grafts, estimated by Fourier transform infrared attenuated total reflection spectroscopy and X-ray photoelectron spectroscopy analysis, were 4.76 and 1.23 for PP-g-(1M NIPAAm-r-0.5M APMA) and PP-g-(1M NIPAAm-r-1M APMA), respectively. By varying the reaction time, different degrees of grafting were achieved, while the monomer ratio was kept constant. PP-g-(NIPAAm-r-APMA) films showed temperature-responsive swelling, smaller friction coefficients, hemolysis and thrombogenicity and higher cell compatibility, did not elicit secretion of cytokines, and took up remarkable amounts of diclofenac and ibuprofen and sustained delivery for several hours in phosphate buffer, pH 7.4. Coating with carboxymethyl dextran of diclofenac-loaded PP-g-(NIPAAm-r-APMA) films caused a minor discharge of the drug but did not alter the drug release rate.

Biofilm inhibition and drug-eluting properties of novel DMAEMA-modified polyethylene and silicone rubber surfaces

Poly(2-(dimethylaminoethyl) methacrylate) (pDMAEMA) was grafted to low density polyethylene (LDPE) and silicone rubber (SR) in order to make them less susceptible to microbial biofilm formation. The direct grafting of DMAEMA using γ-rays was an efficient and fast procedure for obtaining modified materials, which could be quaternized in a second step using methyl iodide. Raman spectroscopy showed that the grafting occurred only at the surface of the LDPE, but both at the surface and in the bulk of the SR. Consequently, the grafted chains caused changes in the surface-related features of the LDPE (water contact angle and viscoelastic behavior in the dry state) and in the bulk-related properties of the SR (swelling and viscoelasticity in the swollen state). The microbiological assays revealed that the grafted DMAEMA reduced Candida albicans biofilm formation (almost no biofilm on SR), while the quaternized surfaces inhibited C. albicans and Staphylococcus aureus biofilm by more than 99% compared to pristine materials. Modified LDPE and SR were capable of holding considerable amounts of nalidixic acid, an anionic antimicrobial drug, and sustained the release for several hours. In addition, the grafted materials were cytocompatible (fibroblast cell survival > 70%). In conclusion, these materials have the ability to inhibit microbial biofilm formation and at the same time act as drug-eluting systems, and for that reason may hold great promise for anti-biofouling applications.

Low-Pressure Plasma Polymerization of Acetylene–Ammonia Mixtures for Biomedical Applications

Past research in this laboratory has focused on the deposition of nitrogen- (N)-rich thin organic coatings for biomedical applications; among usual fabrication methods are plasma polymerization (PP) at low- (“L”) or atmospheric- (high-, “H”)-pressure. In the “L” case, ethylene (“E”, C2H4)/ammonia (NH3) feed-gas mixtures with different flow ratios, R, are used, by which the nitrogen- and primary amine concentrations, [N] and [–NH2], respectively, can be reproducibly controlled. The generic symbol we use for that family of deposits is L-PPE:N. In the present research, we used acetylene (“A”, C2H2) as the hydrocarbon feed, because our earlier experience with “H”-type materials (H-PPE:N and H-PPA:N) revealed striking differences in physico-chemical (e.g. [N] and [–NH2], and solubility) characteristics, which are important for applications. We now find that such differences also exist between the L-PPA:N and L-PPE:N families of coatings. This is attributed to the fundamentally different bonding structures of “A” and “E”, namely CH≡CH and CH2=CH2; the former leads to more highly cross-linked, [NH2]-leaner deposits, as was also noted for the “H”-type deposits mentioned above.

Surface functionalization of polypropylene devices with hemocompatible DMAAm and NIPAAm grafts for norfloxacin sustained release

To improve the hemocompatibility and elution of antimicrobial agents for medical devices, N,N′-dimethylacrylamide (DMAAm) and N-isopropylacrylamide (NIPAAm) were sequentially grafted onto polypropylene (PP) films. Various (PP-g-DMAAm)-g-NIPAAm grafts were prepared using different time periods of irradiation while keeping the radiation dose constant. The hydrogel layer that formed on the surface of the PP was temperature-responsive (shifted from 32°C for NIPAAm to 37°C for the copolymer). The (PP-g-DMAAm)-g-NIPAAm films adsorbed serum albumin but not fibrinogen and had significantly lower hemolytic and thrombogenic activity. The DMAAm promoted the loading of norfloxacin (13.3 μg cm—2) when the hydrogel layer was swollen; as the NIPAAm shrank, a sustained delivery (∼6 h) occurred at body temperature. These functionalized PP films have potential as hemo- and cyto-compatible materials for medical devices and drug delivery products.

Fusion peptide P15‐CSP shows antibiofilm activity and pro‐osteogenic activity when deposited as a coating on hydrophilic but not hydrophobic surfaces

In the context of porous bone void filler for oral bone reconstruction, peptides that suppress microbial growth and promote osteoblast function could be used to enhance the performance of a porous bone void filler. We tested the hypothesis that P15-CSP, a novel fusion peptide containing collagen-mimetic osteogenic peptide P15, and competence-stimulating peptide (CSP), a cationic antimicrobial peptide, has emerging properties not shared by P15 or CSP alone. Peptide-coated surfaces were tested for antimicrobial activity toward Streptoccocus mutans, and their ability to promote human mesenchymal stem cell (MSC) attachment, spreading, metabolism, and osteogenesis. In the osteogenesis assay, peptides were coated on tissue culture plastic and on thin films generated by plasma-enhanced chemical vapor deposition to have hydrophilic or hydrophobic character (water contact angles 63°, 42°, and 92°, respectively). S. mutans planktonic growth was specifically inhibited by CSP, whereas biofilm formation was inhibited by P15-CSP. MSC adhesion and actin stress fiber formation was strongly enhanced by CSP, P15-CSP, and fibronectin coatings and modestly enhanced by P15 versus uncoated surfaces. Metabolic assays revealed that CSP was slightly cytotoxic to MSCs. MSCs developed alkaline phosphatase activity on all surfaces, with or without peptide coatings, and consistently deposited the most biomineralized matrix on hydrophilic surfaces coated with P15-CSP. Hydrophobic thin films completely suppressed MSC biomineralization, consistent with previous findings of suppressed osteogenesis on hydrophobic bioplastics. Collective data in this study provide new evidence that P15-CSP has unique dual capacity to suppress biofilm formation, and to enhance osteogenic activity as a coating on hydrophilic surfaces.

Wound debridement and antibiofilm properties of gamma-ray DMAEMA-grafted onto cotton gauzes

Cotton gauze fabric was functionalized with 2-(dimethylamino)ethyl methacrylate (DMAEMA) with the aim of developing wound dressings with antibiofilm activity and tunable debriding activity. Cotton-g-DMAEMA gauzes were prepared via one-step grafting (direct method) using 60Co γ-rays as source to initiate the polymerization process. The effects of absorbed dose, dose rate, and monomer concentration on the degree of grafting were evaluated in detail. Some cotton-g-DMAEMA gauzes were subsequently quaternized with methyl iodide. Grafting of DMAEMA and sequential quaternization were confirmed by FTIR-ATR spectroscopy; thermal properties were analyzed using TGA, and morphology by scanning electron microscopy. Grafting of DMAEMA gauzes enhanced blood absorption and collagenase activity, while further quaternization led to a remarkable inhibition of the proteinase activity. Their antimicrobial features were analyzed by evaluating their biofilm inhibiting and biofilm eradicating properties in an in vitro chronic wound model. Although the non-quaternized gauzes only displayed moderate biofilm inhibitory properties at best, the quaternized cotton-g-DMAEMA bearing the highest content of DMAEMA displayed strong biofilm inhibiting and biofilm eradicating properties. This indicates that quaternized cotton-g-DMAEMA gauzes are less prone to be colonized by bacteria and can notably reduce the number of colonies in an infected wound.

A Versatile Star PEG Grafting Method for the Generation of Nonfouling and Nonthrombogenic Surfaces

Polyethylene glycol (PEG) grafting has a great potential to create nonfouling and nonthrombogenic surfaces, but present techniques lack versatility and stability. The present work aimed to develop a versatile PEG grafting method applicable to most biomaterial surfaces, by taking advantage of novel primary amine-rich plasma-polymerized coatings. Star-shaped PEG covalent binding was studied using static contact angle, X-ray photoelectron spectroscopy (XPS), and quartz crystal microbalance with dissipation monitoring (QCM-D). Fluorescence and QCM-D both confirmed strong reduction of protein adsorption when compared to plasma-polymerized coatings and pristine poly(ethyleneterephthalate) (PET). Moreover, almost no platelet adhesion was observed after 15 min perfusion in whole blood. Altogether, our results suggest that primary amine-rich plasma-polymerized coatings offer a promising stable and versatile method for PEG grafting in order to create nonfouling and nonthrombogenic surfaces and micropatterns.