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Ten downward portions in the large oscillatory force-distance curve reported earlier are analyzed to understand a nanoscale water meniscus confined between a sharp probe and a flat substrate in air. The sigmoidal shape of each portion leads to the assumption that the meniscus is made up of

The need for hollow microneedle arrays is important for both drug delivery and wearable sensor applications; however, their fabrication poses many challenges. Hollow metal microneedle arrays residing on a flexible metal foil substrate were created by combining additive manufacturing, micromolding, and electroplating approaches in a process we refer to as electromolding. A solid microneedle with inward facing ledge was fabricated with a two photon polymerization (2PP) system utilizing laser direct write (LDW) and then molded with polydimethylsiloxane. These molds were then coated with a seed layer of Ti/Au and subsequently electroplated with pulsed deposition to create hollow microneedles. An inward facing ledge provided a physical blocking platform to restrict deposition of the metal seed layer for creation of the microneedle bore. Various ledge sizes were tested and showed that the resulting seed layer void could be controlled via the ledge length. Mechanical properties of the PDMS mold was adjusted via the precursor ratio to create a more ductile mold that eliminated tip damage to the microneedles upon removal from the molds. Master structures were capable of being molded numerous times and molds were able to be reused. SEM/EDX analysis showed that trace amounts of the PDMS mold were transferred to the metal microneedle upon removal. The microneedle substrate showed a degree of flexibility that withstood over 100 cycles of bending from side to side without damaging. Microneedles were tested for their fracture strength and were capable of puncturing porcine skin and injecting a dye.

Suturing has been the gold standard approach to close wounds for many decades. However, suturing causes tissue damage, which is accompanied by foreign body reaction, entry of pathogens, complications, infection, or death. In addition, the procedure is usually time-consuming, requiring manual dexterity and free moving space. Other adhesive approaches have been proposed and demonstrated with great potential, including laser-assisted tissue closure with either photothermal or photochemical reactions, application of nanoparticles, glues, constructs based on extracellular matrix (ECM), microbarbs, bio-inspired structures, and tape. The quality of closure has been evaluated by histological methods, indexing, morphology, tensile testing, patency rate, leakage pressure, and burst pressure. All the novel tissue joining methods aim to provide an adhesive with appropriate strength, non-cytotoxicity, and minimal damage. The capability for rapid attachment and release may further reduce surgical procedure time. More research is needed to prove the feasibility of new tissue joining techniques based on the type of tissue, surface chemistry, and working environment.

We explored the potential of biomimetic thin films fabricated by means of matrix-assisted pulsed laser evaporation (MAPLE) for releasing combinations of active substances represented by flavonoids (quercetin dihydrate and resveratrol) and antifungal compounds (amphotericin B and voriconazole) embedded in a polyvinylpyrrolidone biopolymer, the antifungal activity of the film components was evaluated using in vitro microbiological assays. Thin films were deposited using a pulsed KrF* excimer laser source which were structurally characterized using atomic force microscopy (AFM) and Fourier transform infrared spectroscopy (FTIR). High-quality thin films with chemical structures similar to dropcast ones were created using an optimum laser fluence of ~80 mJ/cm2. Bioactive substances were included within the polymer thin films using the MAPLE technique. The results of the in vitro microbiology assay, which utilized a modified disk diffusion approach and were performed using two fungal strains (Candida albicans American Type Culture Collection (ATCC) 90028 and Candida parapsilosis American Type Culture Collection (ATCC) 22019), revealed that voriconazole was released in an active form from the polyvinylpyrrolidone matrix. The results of this study show that the MAPLE-deposited bioactive thin films have a promising potential for use in designing combination devices, such as drug delivery devices, and medical device surfaces with antifungal activity.

The purpose of this study was to investigate the interactions between microorganisms, including the planktonic and adherent organisms, and biopolymer (polyvinylpyrrolidone), flavonoid (quercetin dihydrate and resveratrol)-biopolymer, and silver nanoparticles-biopolymer composite thin films that were deposited using matrix assisted pulsed laser evaporation (MAPLE). A pulsed KrF * excimer laser source was used to deposit the aforementioned composite thin films, which were characterized using Fourier transform infrared spectroscopy (FT-IR), infrared microscopy (IRM), scanning electron microscopy (SEM), Grazing incidence X-ray diffraction (GIXRD) and atomic force microscopy (AFM). The antimicrobial activity of thin films was quantified using an adapted disk diffusion assay against Gram-positive and Gram-negative bacteria strains. FT-IR, AFM and SEM studies confirmed that MAPLE may be used to fabricate thin films with chemical properties corresponding to the input materials as well as surface properties that are appropriate for medical use. The silver nanoparticles and flavonoid-containing films exhibited an antimicrobial activity both against Gram-positive and Gram-negative bacterial strains demonstrating the potential use of these hybrid systems for the development of novel antimicrobial strategies.

Itraconazole is a triazole agent that is routinely used for treatment of nail infections and other fungal infections. Recent studies indicate that itraconazole can also inhibit the growth of basal cell carcinoma (BCC) through suppression of the Sonic Hedgehog (SHH) signaling pathway. In this study, polyglycolic acid microneedle arrays and stainless steel microneedle arrays were used for transdermal delivery of itraconazole to a human BCC model which was regenerated on mice. One-by-four arrays of 642-μm-long polyglycolic acid microneedles with sharp tips were prepared using injection molding and drawing lithography. Arrays of 85 stainless steel 800-μm-tall microneedles attached to syringes were obtained for comparison purposes. Skin grafts containing devitalized split-thickness human dermis that had been seeded with human keratinocytes transduced to express human SHH protein were sutured to the skin of immunodeficient mice. Mice with this human BCC model were treated daily for 2 weeks with itraconazole dissolved in 60% dimethylsulfoxane and 40% polyethylene glycol-400 solution; transdermal administration of the itraconazole solution was facilitated by either four 1 × 4 polyglycolic acid microneedle arrays or stainless steel microneedle arrays. The epidermal tissues treated with polyglycolic acid microneedles or stainless steel microneedles were markedly thinner than that of the control (untreated) graft tissue. These preliminary results indicate that microneedles may be used to facilitate transdermal delivery of itraconazole for localized treatment of BCC.

Electroplated iron was investigated as a novel material for microneedle fabrication due to its recent success as a biocompatible metal in other medical device applications. Hollow polymer microneedles were made using a laser direct write process that involved two-photon polymerization of a commercially available Class 2a biocompatible polymer and subsequent electroplating of this structure with iron. Electroplating bath and deposition conditions were shown to affect the mechanical properties of both iron plated microneedles and iron plated on planar polymer substrates. Conditions for depositing the iron coatings were investigated in terms of grain size, residual strain, and elemental composition for planar iron samples. Fracture strength and puncture mechanics into ex vivo porcine skin for iron coated hollow microneedles were examined. Biocompatibility testing was performed using the MTT assay against human epidermal keratinocytes with several concentrations of iron extract to investigate iron as a material used for transdermal applications. Iron coatings proved to significantly improve the strength of the hollow polymer microneedles and sustained structural integrity up to 7 insertions into porcine skin without bending. A commercially available device (Medtronic MiniMed Quick-Serter®) was used for controlled application of microneedles into porcine skin and estimations of insertion forces for the device were made. Plating conditions were optimized such that an adherent, uniform, and high purity iron coating was deposited onto polymer substrates and polymer microneedles without delamination or fracturing of the microneedles upon ex vivo insertion into porcine skin.

Although a great number of antibiotics are currently available, they are often rendered ineffective by the ability of microbial strains to develop genetic resistance and to grow in biofilms. Since many antimicrobial agents poorly penetrate biofilms, biofilm-associated infections often require high concentrations of antimicrobial agents for effective treatment. Among the various strategies that may be used to inhibit microbial biofilms, one strategy that has generated significant interest involves the use of bioactive surfaces that are resistant to microbial colonization. In this respect, we used matrix assisted pulsed laser evaporation (MAPLE) involving a pulsed KrF* excimer laser source ( λ = 248 nm, τ = 25 ns, ν = 10 Hz) to obtain thin composite biopolymeric films containing natural (flavonoid) or synthetic (antibiotic) compounds as bioactive substances. Chemical composition and film structures were investigated by Fourier transform infrared spectroscopy and X-ray diffraction. Films morphology was studied by scanning electron microscopy and transmission electron microscopy. The antimicrobial assay of the microbial biofilms formed on these films was assessed by the viable cell counts method. The flavonoid-containing thin films showed increased resistance to microbial colonization, highlighting their potential to be used for the design of anti-biofilm surfaces.

Microneedles are minimally invasive transdermal medical devices that are utilized for various applications, including drug delivery, fluid sampling, micro-dialysis, and electrochemical sensing. These devices are associated with less pain and tissue damage as compared with conventional hypodermic needle-based devices. In this study, we demonstrate fabrication of titanium alloy microneedle arrays with nitrogen-incorporated ultrananocrystalline diamond (N-UNCD) coatings. Microneedles were micromachined from ASTM F136 ELI Ti –6Al–4V alloy, a widely used medical-grade titanium alloy. N-UNCD coatings were deposited on the microneedles using microwave plasma enhanced chemical vapor deposition to enhance mechanical strength, increase hardness, improve biocompatibility, and provide an electrochemically stable surface. The structural and chemical properties of the N-UNCD titanium alloy microneedle arrays were evaluated using scanning electron microscopy and Raman spectroscopy. The mechanical robustness and skin penetration capability of the devices were demonstrated using cadaveric porcine skin. Finally, the electrochemical properties of the N-UNCD electrodes were evaluated; in vitro electrochemical detection of uric acid and dopamine was demonstrated using unmodified N-UNCD electrodes. These results demonstrate the application potential of N-UNCD-coated titanium alloy microneedles for transdermal electrochemical biosensing applications.

Miconazole is an imidazole used for treatment of fungal infections that exhibits poor solubility in polar solvents (e.g., aqueous solutions). Microneedles, small-scale lancet-shaped devices that are commonly used for delivery of pharmacologic agents and vaccines, were made out of an acid anhydride copolymer using visible light dynamic mask micro-stereolithography/micromolding and loaded with miconazole using a piezoelectric inkjet printer. The miconazole-coated microneedles showed biodegradation and antifungal activity against the organism Candida albicans (ATCC 90028) on Sabouraud dextrose agar using an in vitro agar plating method. The results of this study demonstrate that piezoelectric inkjet printing may be used load microneedles and other drug delivery devices with pharmacologic agents. Miconazole-loaded microneedles prepared by the visible light dynamic mask micro-stereolithography–micromolding–piezoelectric inkjet printing approach have potential use in transdermal treatment of cutaneous fungal infections.

The delivery of amphotericin B, a pharmacologic agent with activity against a broad spectrum of fungi as well as against parasitic protozoa, has been complicated by the fact that amphotericin B exhibits poor solubility in aqueous solutions at physiologic pH levels. In this study, piezoelectric inkjet printing was used to modify the surfaces of Gantrez 169 BF microneedles (Ashland, Covington, KY). These amphotericin B-loaded microneedles demonstrated activity against Candida parapsilosis in a radial diffusion assay. The results of this study suggest that a combination of visible light dynamic mask microstereolithography, micromolding, and piezoelectric inkjet printing may be used to prepare amphotericin B-loaded microneedles with antifungal properties. It is envisioned that microneedles containing amphotericin B may be used for transdermal delivery of pharmacologic agents for the treatment of cutaneous fungal infections as well as cutaneous leishmaniasis.

We designed and developed a high-speed atomic force microscope (HSAFM) utilizing a force-feedback scheme for imaging large biological samples. The system collects three simultaneous images: a deflection image, a topographic image, and a force image. We demonstrated that this force-feedback HSAFM is capable of acquiring large topographic images of Escherichia coli biofilms at approximately one frame per second in air. We discuss how the self-actuating cantilever and the piezo tube follow those larger biological topographic features during the HSAFM imaging process.

Poly(glycolic acid) microneedle arrays were fabricated using a drawing lithography process; these arrays were modified with a drug release agent and an antifungal agent by piezoelectric inkjet printing. Coatings containing poly(methyl vinyl ether-co-maleic anhydride), a water-soluble drug release layer, and itraconazole (an antifungal agent), were applied to the microneedles by piezoelectric inkjet printing. Microscopic evaluation of the microneedles indicated that the modified microneedles contained the piezoelectric inkjet printing-deposited agents and that the surface coatings were released in porcine skin. Energy dispersive x-ray spectrometry aided in confirmation that the piezoelectric inkjet printing-deposited agents were successfully applied to the desired target areas of the microneedle surface. Fourier transform infrared spectroscopy was used to confirm the presence of the component materials in the piezoelectric inkjet printing-deposited material. Itraconazole-modified microneedle arrays incubated with agar plates containing Candida albicans cultures showed zones of growth inhibition.

A custom-designed microneedle sampling system was prepared using dynamic mask microstereolithography; this sampling system was used for determination of histamine content in fresh, histamine-spiked, and spoiled tuna flesh. Lateral flow (test strip) assays were successfully utilized in the microneedle sampling system to assess histamine content. Good agreement was noted between data obtained from the microneedle sampling system and a commercially available histamine detection kit. A discrepancy was noted in the results from the microneedle sampling system and the commercially available histamine detection kit at low (negative) levels of histamine. There was an improvement in the agreement between the microneedle sampling system and the commercially available histamine detection kit at higher histamine levels. The results, which showed an improvement in the test duration and the amount of reagent needed for histamine detection, indicate the promise of printed microneedle sampling systems for histamine detection in seafood samples and other types of food testing.

Tissue barbs are small-scale structures that may be used for sutureless joining of tissues. In this study, several types of tissue barbs were fabricated using two-photon polymerization/micromolding, including two-pronged tissue barbs, eight-pronged tissue barbs, 10-pronged tissue barbs, and 16-pronged tissue barbs. Tissue barb penetration in porcine tissue was observed using confocal laser scanning microscopy. Constructs containing medical tape and tissue barbs were created by applying tissue barbs in a parallel arrangement to Transpore™ medical tape. These results suggest that two-photon polymerization/micromolding is an indirect rapid prototyping approach that may be used for high-throughput replication of tissue barbs and other microstructured solid wound sealants.

Summary Cantilever-based optical interfacial force microscopy (COIFM) was applied to the investigation of the mechanical properties of soft materials to avoid the double-spring effect and snap-to-contact problem associated with atomic force microscopy (AFM). When a force was measured as a function of distance between an oxidized silicon probe and the surface of a soft hydrocarbon film, it increases nonlinearly in the lower force region below ∼10 nN, following the Herzian model with the elastic modulus of ∼50 MPa. Above ∼10 nN, it increases linearly with a small oscillatory sawtooth pattern with amplitude 1–2 nN. The pattern suggests the possible existence of the layered structure within the film. When its internal part of the film was exposed to the probe, the force depends on the distance linearly with an adhesive force of −20 nN. This linear dependence suggests that the adhesive internal material behaved like a linear spring with a spring constant of ∼1 N/m. Constant-force images taken in the repulsive and attractive contact regimes revealed additional features that were not observed in the images taken in the noncontact regime. At some locations, however, contrast inversions were observed between the two contact regimes while the average roughness remained constant. The result suggests that some embedded materials had spring constants different from those of the surrounding material. This study demonstrated that the COIFM is capable of imaging mechanical properties of local structures such as small impurities and domains at the nanometer scale, which is a formidable challenge with conventional AFM methods. SCANNING 35:59-67, 2013. © 2012 Wiley Periodicals, Inc.

Microneedles are small-scale needle-like projections that may be used for transdermal delivery of pharmacologic agents, including protein-containing and nucleic acid-containing agents. Commercial translation of polymeric microneedles would benefit from the use of facile and cost effective fabrication methods. In this study, visible light dynamic mask microstereolithography, a rapid prototyping technique that utilizes digital light projection for selective polymerization of a liquid resin, was used for fabrication of solid microneedle array structures out of an acrylate-based polymer. Pulsed laser deposition was used to deposit silver and zinc oxide coatings on the surfaces of the visible light dynamic mask microstereolithography-fabricated microneedle array structures. Agar diffusion studies were used to demonstrate the antimicrobial activity of the coated microneedle array structures. This study indicates that light-based technologies, including visible light dynamic mask microstereolithography and pulsed laser deposition, may be used to fabricate microneedles with antimicrobial properties for treatment of local skin infections.

Diamond has been considered for use in several medical applications due to its unique mechanical, chemical, optical, and biological properties. In this paper, methods for preparing synthetic diamond surfaces and particles are described. In addition, recent developments involving the use of diamond in prostheses, sensing, imaging, and drug delivery applications are reviewed. These developments suggest that diamond-containing structures will provide significant improvements in the diagnosis and treatment of medical conditions over the coming years.

Piezoelectric inkjet printing is a noncontact process that enables microscale processing of biological materials. In this research summary, the use of piezoelectric inkjet printing for patterning medical adhesives and sealants, including a two-component polyethylene glycol hydrogel-based medical sealant, an N-butyl cyanoacrylate tissue adhesive, and a mussel adhesive protein biological adhesive, is described The effect of Fe(III) on mussel adhesive protein structure was evaluated by means of atomic force microscopy. The ability to process microscale patterns of medical sealants and adhesives will provide an improvement in tissue joining, including enhanced tissue integrity, reduced bond lines, and decreased adhesive toxicity. Piezoelectric inkjet deposition of medical adhesives and sealants may be used in wound closure, fracture fixation, and microscale vascular surgery.

Designing, fabricating, and evaluating stretchable electronics is a growing area of materials research. Electronic devices have traditionally been fabricated using rigid, inorganic substrates (e.g., silicon) with metallic components and interconnections. Conventional electronic devices may face limitations when placed in environments that are dominated by stretchable or three-dimensional structures, including those within the human body. This paper describes the use of pulsed laser deposition to create diamond-like carbon microstructures on polydimethylsiloxane. The viability of human epidermal keratinocyte cells on polydimethylsiloxane surfaces coated with arrays of diamond-like carbon islands was similar to that on unmodified polydimethylsiloxane surfaces, which are commonly used in medical devices. It is anticipated that stretchable electronic devices may be incorporated within novel medical devices and prostheses that interface with stretchable or three-dimensional structures in the human body.

Biomimetic surfaces and materials may be utilized in biosensing applications, harnessing material properties that mimic the natural environment of a biomolecule in order to maintain its functionality, artificially create a complex that takes on the form of a biomolecular structure, or modify an environment to promote cellular affinity. Throughout the scientific literature, there are numerous mechanisms by which this concept may be accomplished. Synthetic lipid membranes, meant to mimic a cellular membrane, have been deposited onto sensor platforms for analyte detection with immobilized functional biomolecules. Natural biomolecules may also be immobilized on transducers, with care taken to protect their functionality (e.g., through polymer linkages), allowing them to operate as sensing units. Synthetic molecular constructs have been developed to mimic the activity of biomolecules. Molecularly imprinted polymers have been created, operating as artificial bioaffinity recognition sites for target molecules. Furthermore, whole cells may be immobilized onto sensing surfaces, acting as sensing units or mimics of larger tissue systems. In this chapter, the relevant literature examples are discussed, highlighting the means by which these biomimetic sensing approaches are accomplished.

We present the development of an electrochemical biosensor based on modified glassy carbon (GC) electrodes using hydrogel-based molecularly imprinted polymers (MIPs) has been fabricated for protein detection. The coupling of pattern recognition techniques via principal component analysis (PCA) has resulted in unique protein fingerprints for corresponding protein templates, allowing for MIP-based protein profiling. Polyacrylamide MIPs for memory imprinting of bovine haemoglobin (BHb), equine myoglobin (EMb), cytochrome C (Cyt C), and bovine serum albumin (BSA), alongside a non-imprinted polymer (NIP) control, were spectrophotometrically, and electrochemically characterised using modified GC electrodes. Rebinding capacities (Q) were revealed to be higher for larger proteins (BHb and BSA, Q ≈ 4.5) while (EMb and Cyt C, Q ≈ 2.5). Electrochemical results show that due to the selective nature of MIPs, protein arrival at the electrode via diffusion is delayed, in comparison to a NIP, by attractive selective interactions with exposed MIP cavities. However, at lower concentrations such discriminations are difficult due to low levels of MIP rebinding. PCA loading plots revealed 5 variables responsible for the separation of the proteins; Ep, Ip, E1/2 , Iat −0.8 V, �Idecay peak current to −0.8 V. Statistical symmetric measures of agreement using Cohen’s kappa coefficient (K) were revealed to be 63% for bare GC, 96% for NIP and 100% for MIP. Therefore, our results show that with the use of PCA such discriminations are achievable, also with the advantage of faster detection rates. The possibilities for this MIP technology once fully developed are vast, including uses in bio-sample clean-up or selective extraction, replacement of biological antibodies in immunoassays, as well as biosensors for medicine, food and the environment.

We studied the stability of force-feedback high-speed atomic force microscopy (HSAFM) by imaging soft, hard, and biological sample surfaces at various applied forces. The HSAFM images showed sudden topographic variations of streaky fringes with a negative applied force when collected on a soft hydrocarbon film grown on a grating sample, whereas they showed stable topographic features with positive applied forces. The instability of HSAFM images with the negative applied force was explained by the transition between contact and noncontact regimes in the force–distance curve. When the grating surface was cleaned, and thus hydrophilic by removing the hydrocarbon film, enhanced imaging stability was observed at both positive and negative applied forces. The higher adhesive interaction between the tip and the surface explains the improved imaging stability. The effects of imaging rate on the imaging stability were tested on an even softer adhesive Escherichia coli biofilm deposited onto the grating structure. The biofilm and planktonic cell structures in HSAFM images were reproducible within the force deviation less than ∼0.5 nN at the imaging rate up to 0.2 s per frame, suggesting that the force-feedback HSAFM was stable for various imaging speeds in imaging softer adhesive biological samples.

Due to their ability to serve as fluorophores and drug delivery vehicles, quantum dots are a powerful tool for theranostics-based clinical applications. In this study, microneedle devices for transdermal drug delivery were fabricated by means of two-photon polymerization of an acrylate-based polymer. We examined proliferation of cells on this polymer using neonatal human epidermal keratinocytes and human dermal fibroblasts. The microneedle device was used to inject quantum dots into porcine skin; imaging of the quantum dots was performed using multiphoton microscopy.

Diamond has been considered for use in several medical applications due to its unique mechanical, chemical, optical, and biological properties. In this chapter, methods for preparing synthetic diamond surfaces and particles are described. In addition, recent developments involving the use of diamond in prostheses, sensing, imaging, and drug delivery applications are reviewed. These developments suggest that diamond-containing structures will provide significant improvements in the diagnosis and treatment of medical conditions over the coming years.

Microneedles are needle-like projections with microscale features that may be used for transdermal delivery of a variety of pharmacologic agents, including antibacterial agents. In the study described in this paper, an indirect rapid prototyping approach involving a combination of visible light dynamic mask micro-stereolithography and micromolding was used to prepare microneedle arrays out of a biodegradable acid anhydride copolymer, Gantrez(®) AN 169 BF. Fourier transform infrared spectroscopy, energy dispersive x-ray spectrometry and nanoindentation studies were performed to evaluate the chemical and mechanical properties of the Gantrez(®) AN 169 BF material. Agar plating studies were used to evaluate the in vitro antimicrobial performance of these arrays against Bacillus subtilis, Candida albicans, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. Large zones of growth inhibition were noted for Escherichia coli, S. aureus, Enterococcus faecalis and B. subtilis. The performance of Gantrez(®) AN 169 BF against several bacteria suggests that biodegradable acid anhydride copolymer microneedle arrays prepared using visible light dynamic mask micro-stereolithography micromolding may be useful for treating a variety of skin infections.

In this study, biodegradable acid anhydride copolymer microneedles containing quantum dots were fabricated by means of visible light dynamic mask micro-stereolithography-micromolding and inkjet printing. Nanoindentation was performed to obtain the hardness and the Young's modulus of the biodegradable acid anhydride copolymer. Imaging of quantum dots within porcine skin was accomplished by means of multiphoton microscopy. Our results suggest that the combination of visible light dynamic mask micro-stereolithography-micromolding and inkjet printing enables fabrication of solid biodegradable microneedles with a wide range of geometries as well as a wide range of pharmacologic agent compositions.