Qiuchen Dong

A Biocompatible and Biodegradable Protein Hydrogel with Green and Red Autofluorescence: Preparation, Characterization and In Vivo Biodegradation Tracking and Modeling

Because of its good biocompatibility and biodegradability, albumins such as bovine serum albumin (BSA) and human serum albumin (HSA) have found a wide range of biomedical applications. Herein, we report that glutaraldehyde cross-linked BSA (or HSA) forms a novel fluorescent biological hydrogel, exhibiting new green and red autofluorescence in vitro and in vivo without the use of any additional fluorescent labels. UV-vis spectra studies, in conjunction with the fluorescence spectra studies including emission, excitation and synchronous scans, indicated that three classes of fluorescent compounds are presumably formed during the gelation process. SEM, FTIR and mechanical tests were further employed to investigate the morphology, the specific chemical structures and the mechanical strength of the as-prepared autofluorescent hydrogel, respectively. Its biocompatibility and biodegradability were also demonstrated through extensive in vitro and in vivo studies. More interestingly, the strong red autofluorescence of the as-prepared hydrogel allows for conveniently and non-invasively tracking and modeling its in vivo degradation based on the time-dependent fluorescent images of mice. A mathematical model was proposed and was in good agreement with the experimental results. The developed facile strategy to prepare novel biocompatible and biodegradable autofluorescent protein hydrogels could significantly expand the scope of protein hydrogels in biomedical applications.

In situ microfluidic fabrication of SERS nanostructures for highly sensitive fingerprint microfluidic-SERS sensing

A microfluidic device with integrated silver nanoparticles (AgNPs) was fabricated via in situ galvanic replacement of a pre-patterned copper substrate in a microfluidic channel. The integrated microfluidic device with AgNPs serves as a highly active Raman substrate which can be applied for in-channel surface-enhanced Raman scattering (SERS). Preparation of the SERS active substrate and subsequent SERS experiments are all completed within the microfluidic device allowing for easy integration and application. In conjunction with the high sensitivity, easy fabrication and mobility of the microfluidic device, the developed microfluidic-SERS system provides an excellent sensing platform for sensitive, real-time fingerprint detection of target molecules. Crystal violet is first used as a model compound to demonstrate the effectiveness of the microfluidic-SERS system. Specifically, the in situ fabricated SERS active substrate demonstrates high sensitivity and exhibits an apparent enhancement factor of 2.2 × 107, high robustness and reusability, making it a perfect fit for the real time detection of pesticides. Finally the detection of pesticides and herbicides such as Carbofuran and Alachlor as low as 5 ppb was demonstrated using the as-developed microfluidic-SERS system. This study opens a new avenue to fabricate an integrated microfluidic-SERS sensing system with high performance.

ELP-OPH/BSA/TiO2 nanofibers/c-MWCNTs based biosensor for sensitive and selective determination of p-nitrophenyl substituted organophosphate pesticides in aqueous system

A novel biosensor for rapid, sensitive and selective monitoring of p-nitrophenyl substituted organophosphate pesticides (OPs) in aqueous system was developed using a functional nanocomposite which consists of elastin-like-polypeptide-organophosphate hydrolase (ELP-OPH), bovine serum albumin (BSA), titanium dioxide nanofibers (TiO2NFs) and carboxylic acid functionalized multi-walled carbon nanotubes (c-MWCNTs). ELP-OPH was simply purified from genetically engineered Escherichia coli based on the unique phase transition of ELP and thus served as biocatalyst for OPs, while BSA was used to stabilize OPH activity in the nanocomposite. TiO2NFs was employed to enrich organophosphates in the nanocomposite due to its strong affinity with phosphoric group in OPs, while c-MWCNTs was used to enhance the electron transfer in the amperometric detection as well as for covalent immobilization of ELP-OPH. ELP-OPH/BSA/TiO2NFs/c-MWCNTs nanocomposite were systematically characterized using field emission scanning electron microscopy (SEM), Raman spectra, Fourier Transform infrared spectroscopy (FTIR) and X-ray Diffraction (XRD). Under the optimized operating conditions, the ELP-OPH/BSA/TiO2NFs/c-MWCNTs based biosensor for OPs shows a wide linear range, a fast response (less than 5s) and limits of detection (S/N=3) as low as 12nM and 10nM for methyl parathion and parathion, respectively. Such excellent sensing performance can be attributed to the synergistic effects of the individual components in the nanocomposite. Its further application for selectively monitoring OPs compounds spiked in lake water samples was also demonstrated with good accuracy. These features indicate that the developed nanocomposite offers an excellent biosensing platform for rapid, sensitive and selective detection of organophosphates compounds.

Self-healing of thermally-induced, biocompatible and biodegradable protein hydrogel

Serum albumin is the most abundant protein in the circulatory system to transport fatty acids, metabolites and drugs. In this study, a highly biocompatible protein hydrogel was prepared from bovine serum albumin (BSA) via thermal treatment. A circular dichroism study indicates that thermally-induced partial unfolding of the protein molecules exposes the buried hydrophobic groups in the core to the environment, thus leading to the formation of fine stranded 3-D networks. By controlling the heating temperature and protein concentration, the mechanical strength and structural stability of the as-prepared BSA hydrogel can be facilely manipulated. The moderate denaturation of the protein within the hydrogel system allows repetitive self-healing after damage when moderate heat was induced. The tensile strength and break strain of fully healed protein hydrogel were recovered to almost 100% of the original strength and elongation abilities. Additionally, the good biocompatibility of this hydrogel system was demonstrated through in vitro cytotoxicity analysis first. Furthermore, in vivo experiments using immunocompetent mice show that the subcutaneously injected hydrogel in mice can be fully degraded with negligible acute inflammatory response, indicating excellent in vivo biocompatibility. These features indicate that the as-developed self-healing protein hydrogel system with good biocompatibility and biodegradability holds great potential in the field of biomedical engineering.

Repetitive Biomimetic Self-healing of Ca(2+)-Induced Nanocomposite Protein Hydrogels.

Self-healing is a capacity observed in most biological systems in which the healing processes are autonomously triggered after the damage. Inspired by this natural behavior, researchers believed that a synthetic material possessing similar self-recovery capability could also be developed. Albeit various intrinsic self-healing systems have been developed over the past few decades, restriction on the biocompatibility due to the required synthetic conditions under extreme pH and with poisonous cross-linker significantly limits their application in biomedical field. In this study, a highly biocompatible nanocomposite protein hydrogel with excellent biomimetic self-healing property is presented. The self-healing protein gel is made by inducing calcium ions into the mixture of heat-induced BSA nano-aggregates and pristine BSA molecules at room temperature and under physiological pH due to the ion-mediated protein-protein association and the bridging effect of divalent Ca2+ ions. The as-prepared protein hydrogel shows excellent repetitive self-healing properties without using any external stimuli at ambient condition. Such outstanding self-recovery performance was quantitatively evaluated/validated by both dynamic and oscillatory rheological analysis. Moreover, with the presence of calcium ions, the self-healing behavior can be significantly facilitated/enhanced. Finally, the superior biocompatibility demonstrated by in vitro cytotoxicity analysis suggests that it is a promising self-healing material well-suited for biomedical applications.

Novel green and red autofluorescent protein nanoparticles for cell imaging and in vivo biodegradation imaging and modeling

Albumins are widely used in bioengineering due to their low-cost, good biocompatibility and biodegradability. Herein we report that cross-linked bovine serum albumin (BSA) forms a suspension of novel fluorescent nanoparticles with an average size of ∼40 nm, exhibiting strong green/red autofluorescence. UV-vis spectra, in conjunction with fluorescence emission spectra, suggest that three classes of fluorescent compounds presumably formed during the preparation. The size distribution and surface morphology of the autofluorescent BSA nanoparticles were characterized using various advanced techniques. After removal of excessive cross-linking agent through dialysis, the autofluorescent BSA nanoparticles were first demonstrated for cell bioimaging application using 293FT human kidney cell line. Its good biocompatibility and low cytotoxicity were further validated by an in vitro cytotoxicity assay and an in vivo histological study. The strong red autofluorescence of the BSA nanoparticles was further exploited in the realization of convenient and non-invasive tracking/modeling of its in vivo degradation based on real-time fluorescence imaging. A mathematical model was proposed and in good agreement with the experimental results. This study indicates that the as-prepared functional, biocompatible and biodegradable autofluorescent protein nanoparticles are suitable for a range of biomedical applications.

Sensitive and Selective NH3 Monitoring at Room Temperature Using ZnO Ceramic Nanofibers Decorated with Poly(styrene sulfonate)

Ammonia (NH3) gas is a prominent air pollutant that is frequently found in industrial and livestock production environments. Due to the importance in controlling pollution and protecting public health, the development of new platforms for sensing NH3 at room temperature has attracted great attention. In this study, a sensitive NH3 gas device with enhanced selectivity is developed based on zinc oxide nanofibers (ZnO NFs) decorated with poly(styrene sulfonate) (PSS) and operated at room temperature. ZnO NFs were prepared by electrospinning followed by calcination at 500 °C for 3 h. The electrospun ZnO NFs are characterized to evaluate the properties of the as-prepared sensing materials. The loading of PSS to prepare ZnO NFs/PSS composite is also optimized based on the best sensing performance. Under the optimal composition, ZnO NFs/PSS displays rapid, reversible, and sensitive response upon NH3 exposure at room temperature. The device shows a dynamic linear range up to 100 ppm and a limit of detection of 3.22 ppm and enhanced selectivity toward NH3 in synthetic air, against NO2 and CO, compared to pure ZnO NFs. Additionally, a sensing mechanism is proposed to illustrate the sensing performance using ZnO NFs/PSS composite. Therefore, this study provides a simple methodology to design a sensitive platform for NH3 monitoring at room temperature.

A high-performance electrochemical sensor for biologically meaningful l-cysteine based on a new nanostructured l-cysteine electrocatalyst

As a new class of l-cysteine electrocatalyst explored in this study, Au/CeO2 composite nanofibers (CNFs) were employed to modify the screen printed carbon electrode (SPCE) to fabricate a novel l-cysteine (CySH) electrochemical sensor with high performance. Its electrochemical behavior and the roles of Au and CeO2 in the composite toward electro-oxidation of CySH were elucidated and demonstrated using cyclic voltammetry and amperometry techniques for the first time through the comparison with pure CeO2 NFs. More specifically, the Au/CeO2 CNFs modified SPCE possessed greatly enhanced electrocatalytic activity toward CySH oxidation. An ultra high sensitivity of 321 μA mM-1cm-2 was obtained, which is almost 2.7 times higher than that of pure CeO2 NFs, revealing that the presence of Au imposed an important influence on the electrocatalytic activity toward CySH. The detailed reasons on such high performance were also discussed. In addition, the as-prepared sensor showed a low detection limit of 10 nM (signal to noise ratio of 3), a wide linear range up to 200 μM for the determination of CySH, an outstanding reproducibility and good long-term stability, as well as an excellent selectivity against common interferents such as tryptophan, tyrosine, methionine, ascorbic acid and uric acid. All these features indicate that the Au/CeO2 composite nanofiber is a promising candidate as a new class of l-cysteine electrocatalyst in the development of highly sensitive and selective CySH electrochemical sensor.

A Simple SERS-Based Trace Sensing Platform Enabled by AuNPs-Analyte/AuNPs Double-Decker Structure on Wax-Coated Hydrophobic Surface

In this work, a simple and versatile SERS sensing platform enabled by AuNPs-analyte/AuNPs double-decker structure on wax-coated hydrophobic surface was developed using a portable Raman spectrometer. Wax-coated silicon wafer served as a hydrophobic surface to induce both aggregation and concentration of aqueous phase AuNPs mixed with analyte of interest. After drying, another layer of AuNPs was drop-cast onto the layer of AuNPs-analyte on the substrate to form double-decker structure, thus introducing more “hot spots” to further enhance the Raman signal. To validate the sensing platform, methyl parathion (pesticide), and melamine (a nitrogen-enrich compound illegally added to food products to increase their apparent protein content) were employed as two model compounds for trace sensing demonstration. The as-fabricated sensor showed high reproducibility and sensitivity toward both methyl parathion and melamine detection with the limit of detection at the nanomolar and sub-nanomolar concentration level, respectively. In addition, remarkable recoveries for methyl parathion spiked into lake water samples were obtained, while reasonably good recoveries for melamine spiked into milk samples were achieved. These results demonstrate that the as-developed SERS sensing platform holds great promise in detecting trace amount of hazardous chemicals for food safety and environment protection.

Dual functional rhodium oxide nanocorals enabled sensor for both non-enzymatic glucose and solid-state pH sensing

Both pH-sensitive and glucose-responsive rhodium oxide nanocorals (Rh2O3 NCs) were synthesized through electrospinning followed by high-temperature calcination. The as-prepared Rh2O3 NCs were systematically characterized using various advanced techniques including scanning electron microscopy, X-ray powder diffraction and Raman spectroscopy, and then employed as a dual functional nanomaterial to fabricate a dual sensor for both non-enzymatic glucose sensing and solid-state pH monitoring. The sensing performance of the Rh2O3 NCs based dual sensor toward pH and glucose was evaluated using open circuit potential, cyclic voltammetry and amperometric techniques, respectively. The results show that the as-prepared Rh2O3 NCs not only maintain accurate and reversible pH sensitivity of Rh2O3, but also demonstrate a good electrocatalytic activity toward glucose oxidation in alkaline medium with a sensitivity of 11.46 μA mM-1 cm-2, a limit of detection of 3.1 μM (S/N = 3), and a reasonable selectivity against various interferents in non-enzymatic glucose detection. Its accuracy in determining glucose in human serum samples was further demonstrated. These features indicate that the as-prepared Rh2O3 NCs hold great promise as a dual-functional sensing material in the development of a high-performance sensor forManjakkal both solid-state pH and non-enzymatic glucose sensing.