Multicolor diagnosis of salivary alkaline phosphatase triggered by silver-coated gold nanobipyramids.

Alkaline phosphatase (ALP) is one of the most versatile biomarkers for early detection of several diseases, such as oral carcinomas and periodontitis; therefore, great efforts have been dedicated for developing an ALP biosensor. Multicolor detection of ALP in saliva is ideal for a point-of-care diagnosis; however, this approach is very challenging since spectral responses over wavelengths of several tens of nanometers have thus far remained difficult to achieve. In this work, a colorimetric biosensor for ALP assay has been developed based on ALP affinity to dephosphorylate glucose phosphate into glucose, which has the affinity to deposit Ag nanoshells onto Au nanobipyramids with a multicolor response. This approach provides a blue shift of localized surface plasmon resonance (LSPR) as large as 190 nm corresponding to distinctive color changes, from yellowish brown to red based on the thickness of the formed Ag shell around the Au nanobipyramids. The change in the LSPR has been conducted for highly sensitive quantitative bioassay of ALP with a detectable multicolor change with linear dynamic range of 0.1-20 U/L and low limit of detection (LOD) of 0.085 U/L. Furthermore, the developed multicolor ALP biosensor exhibits high selectivity with high recovery of 98.6% demonstrating  its reliability and suitability for a point-of-care diagnosis.

Smartphone-assisted point-of-care colorimetric biosensor for the detection of urea via pH-mediated AgNPs growth.

Smartphone-assisted point-of-care (POC) bioassay has brought a giant leap in personal healthcare system and environmental monitoring advancements. In this study, we developed a rapid and reliable colorimetric urea biosensor assisted by a smartphone. We employed hydrolysis of urea into NH3 by urease, which activates the reduction power of tannic acid, to generate silver nanoparticles for a dramatic colorimetric response. The proposed urea biosensor was validated in a solution to provide high selectivity against various interferents in human urine. It had high sensitivity, with a limit of detection as low as 0.0036 mM, and a high reliability of 99% ± 2.9% via the standard addition method. The urea biosensor was successfully implanted on a paper to facilitate smartphone-assisted POC readout with a limit of detection of 0.58 mM and wide detection range of 500 mM, whereby direct diagnosis of human urine without dilution was realized. Our smartphone-assisted POC colorimetric urea biosensor will pave the way for daily monitoring systems of renal and hepatic dysfunction diseases.

A colorimetric alkaline phosphatase biosensor based on p-aminophenol-mediated growth of silver nanoparticles.

Alkaline phosphatase (ALP) is an enzyme that catalyzes the dephosphorylation of proteins, nucleic acids, and biomolecules. It is a potential biomarker for diverse diseases such as breast cancer, osteopenia, and hepatobiliary. Herein, we developed a colorimetric sensor for the ALP assay based on its enzymatic activity to dephosphorylate the p-aminophenol phosphate (pAPP) into pAP. In a solution containing silver nanoparticles (AgNPs) and Ag+ ions prepared using a low concentration of NaBH4, pAP mediates the growth of AgNPs by reducing the concentration of Ag+ ions to enhance the intensity of localized surface plasmon resonance as the pAPP cannot induce a reduction of the remaining Ag+ due to the masking of the hydroxyl with phosphate. The quantitative assay of the ALP was demonstrated via the colorimetric detection of the pAP-mediated growth of AgNPs in the presence of an ALP. The highly sensitive enzymatic growth of AgNPs provided a wider dynamic linear range of 0.5-225 U/L with a lower limit of detection of 0.24 U/L than that previously reported. The use of pAP resulted in excellent selectivity of the sensor for the ALP assay in human serum, yielding a high recovery rate and a high precision of 99.2 ± 1.5 % for the standard addition method.

Solid-state colorimetric polydiacetylene liposome biosensor sensitized by gold nanoparticles.

Polydiacetylene (PDA), a conjugated polymer, has attracted attention for realization of a label-free real-time colorimetric biosensor because it exhibits large and rapid colorimetric responses upon the binding of biomolecules. This is due to the conformational distortion of its conjugated backbone. However, solid-state PDA biosensors for point-of-care diagnosis remain unexplored. We describe a highly sensitive solid-state biosensor based on PDA liposomes. We employed gold nanoparticles (AuNPs) on PDA liposomes as the molecular-binding-signal sensitizer, which provides additional conformational distortion in the backbone structure of PDA by exerting steric repulsion to the attached biomolecules. To prove the concept, AuNPs and a thrombin-binding-aptamer were individually functionalized on PDA liposomes, which were attached to a substrate for the detection of thrombin. We found that the sensitivity was enhanced 2.5 times in the presence of AuNPs compared with the case without AuNPs. Because the steric repulsion of the AuNPs is target-independent, we believe that our solid-state biosensor provides a path toward advanced solid-state biosensors.

Polyurethane foam containing rhEGF as a dressing material for healing diabetic wounds: Synthesis, characterization, in vitro and in vivo studies.

Diabetic wounds are a major health issue associated with diabetes mellitus. To surmount this issue, we developed polyurethane foams (PUFs) incorporating varying amounts of recombinant human epidermal growth factor (rhEGF) (rhEGF-PUFs) as a wound dressing for diabetic wounds. From electron microscopy images, it was found that the pore size of the rhEGF-PUFs surface (the wound contact layer) was less than 100µm, regardless of rhEGF content. The release of rhEGF from the PUFs was evaluated using an enzyme-linked immunosorbent assay. The result showed that the release of rhEGF was time and concentration dependent, i.e., the amount of released rhEGF significantly increased as the immersion time and the rhEGF content of the PUFs increased. In vitro cytotoxicity testing showed that rhEGF-PUFs increased the viability of HaCaT human keratinocytes and CCD986-sk human fibroblasts, which indicated that the incorporated rhEGF maintained its biological activity. In an in vitro scratch wound healing assay, the wound closure rate was faster in CCD986-sk human fibroblasts than in HaCaT human keratinocytes. Finally, the rhEGF-PUFs were evaluated as an in vivo treatment in a full-thickness wound model in diabetized Sprague-Dawley rats. The result indicated that compared with PUFs, rhEGF-PUFs accelerated wound healing by promoting wound contraction, re-epithelialization, collagen deposition and the formation of a skin appendage. These findings demonstrate that rhEGF-PUFs are a promising dressing for diabetic wounds.

Evaluation of AgHAP-containing polyurethane foam dressing for wound healing: synthesis, characterization, in vitro and in vivo studies.

Silver-containing dressings have been widely used to control wound infection. In this study, we developed various amounts of silver-hydroxyapatite (AgHAP)-containing polyurethane foams (PUFs) (AgHAP-PUFs), and their biological properties including biocompatibility, antibacterial activities, and in vivo wound healing properties were evaluated in the Sprague-Dawley rat model. From electron microscopy imaging, it was found that AgHAP particles are uniformly dispersed inside PUFs. The release of Ag from PUFs was dependent on both time and concentration, i.e., the amount of released Ag was significantly higher with increasing immersion time and Ag content in the PUFs. From the cytotoxicity test, AgHAP-PUFs exhibited high antibacterial efficacy against four pathogenic bacteria, and they were not cytotoxic against L-929 fibroblast cells. AgHAP-PUF treated groups exhibited scar-free wound healing by promoting re-epithelialization and collagen deposition in the infected excision wound model. Overall, it is evident that AgHAP-PUFs may be considered as a good antibacterial wound dressing for infected wounds due to their good antibacterial activity, biocompatibility, and wound healing rate.