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Biomedical subjects

Huiguang Zhu

Publications and source records attributed to Huiguang Zhu.

17 recordsLinked to original sources

Poly(D,L-lactic acid)-block-(ligand-tethered poly(ethylene glycol)) copolymers as surface additives for promoting chondrocyte attachment and growth.

The poly(D,L-lactic acid)-block-(ligand-tethered poly(ethylene glycol)) copolymer was explored to engineer poly(D,L-lactic acid) (PLA) material to promote chondrocyte attachment and growth. The poly(D,L-lactic acid)-block-poly(ethylene glycol) copolymer (PLE) was synthesized by a coupling reaction between PLA and poly(ethylene glycol) (PEG) (M(n) 1000, 2000, and 4000 respectively), with the use of 4,4'-methylenediphenyl diisocyanate (MDI). Then the PLE was activated by methyl sulfonyl chloride and the amino acids or arginine-glycine-aspartic acid tripeptide (RGD) was attached, which was verified by the ninhydrin-UV method. The modified PLA films were simply prepared by blending PLA with PLE derivatives. ATR-FTIR, XPS, contact angle, and AFM results clearly showed that the PEG chain stably enriched on the surface of PLE-modified PLA films. The chondrocyte cytocompatibility test showed the modified PLA films could significantly improve chondrocyte attachment and proliferation.

Cell Adhesion↗

Enzymatic fluorescent microsphere glucose sensors:evaluation of response under dynamic conditions.

Most previous attempts at the development of a "smart tattoo" for glucose monitoring in diabetes--implantable fluorescent microspheres that can be implanted intradermally and interrogated transdermally using light--have focused on the encapsulation of a competitive binding assay for glucose within hydrogel microspheres or polyelectrolyte microcapsules. We recently reported on the development of a microsphere sensor based on an enzymatic scheme, combined with an oxygen-quenched fluorescent reporter element. A novel feature of this design is the use of polyelectrolyte multilayer nanofilms, which are assembled stepwise on the surface of the microspheres, for modulation of glucose and oxygen mass transport, allowing them to be designed for sensitive and safe operation within the "oxygen deficit" present in the skin. In this work, a flow-through sensor testing apparatus was used to demonstrate the reversible response of the sensors under controlled, dynamic conditions. The sensors were shown to be sensitive over at least 0-140 mg/dL glucose, with a response time of less than 2 min. This report also identifies some current problems with the approach, and gives potential solutions towards the development of a practical implantable sensor for glucose monitoring in diabetes.

Biosensing Techniques↗

Loading of hydrophobic materials into polymer particles: implications for fluorescent nanosensors and drug delivery.

A straightforward method for loading hydrophobic materials into commercially available polymer nano- or microparticles is described. PMMA and PS nano/microparticles were swelled by an organic solvent with an ionic surfactant (SDS) to stabilize the particles in aqueous solution. FITC and Ru(dpp)3Cl2 were loaded into those particles based on the principle of "like dissolves like". Further surface modification of the loaded particles was achieved via layer-by-layer (LbL) self-assembly. Culture of fibroblasts with the dye-doped, coated particles showed that the cells internalized the fluorescent particles with no apparent toxic effects. The findings suggest the facile process could be useful in a wide range of applications for fluorescent micro/nanosensors and drug delivery.

Drug Delivery Systems↗

Stable encapsulation of active enzyme by application of multilayer nanofilm coatings to alginate microspheres.

In an effort to improve the stability for long-term biosensor use, layer-by-layer self-assembly was explored as a potential technique to provide a diffusion barrier to encapsulated glucose oxidase inside alginate microspheres (<5 microm), fabricated using an emulsification technique. The total loss of encapsulated enzyme was reduced to less than 25 and 15% with the application of single PAH/PSS and crosslinked PAH/PAA coatings, respectively, in comparison to at least a 45% loss observed with uncoated and PDDA/PSS-coated microspheres. Furthermore, it was found that enzyme within PAH/PSS- and crosslinked PAH/PAA-coated spheres retained more than 84 and 60% of initial activity, respectively, after three months, whereas uncoated and PDDA/PSS-coated microspheres retained less than 20%.

Alginates↗

Stabilization of glucose oxidase in alginate microspheres with photoreactive diazoresin nanofilm coatings.

The nanoassembly and photo-crosslinking of diazo-resin (DAR) coatings on small alginate microspheres for stable enzyme entrapment is described. Multilayer nanofilms of DAR with poly(styrene sulfonate) (PSS) were used in an effort to stabilize the encapsulation of glucose oxidase enzyme for biosensor applications. The activity and physical encapsulation of the trapped enzyme were measured over 24 weeks to compare the effectiveness of nanofilm coatings and crosslinking for stabilization. Uncoated spheres exhibited rapid loss of activity, retaining only 20% of initial activity after one week, and a dramatic reduction in effective activity over 24 weeks, whereas the uncrosslinked and crosslinked {DAR/PSS}-coated spheres retained more than 50% of their initial activity after 4 weeks, which remained stable even after 24 weeks for the two and three bilayer films. Nanofilms comprising more polyelectrolyte layers maintained higher overall activity compared to films of the same composition but fewer layers, and crosslinking the films increased retention of activity over uncrosslinked films after 24 weeks. These findings demonstrate that enzyme immobilization and stabilization can be achieved by using simple modifications to the layer-by-layer self-assembly technique.

Adsorption↗

Macromolecule encapsulation in diazoresin-based hollow polyelectrolyte microcapsules.

A stable enzyme encapsulation technique based on the conversion of weak interactions between diazo resin/poly(styrene sulfonate) to covalent bonds was explored. Photosensitive diazoresin-based polyelectrolyte microcapsules were prepared via layer-by-layer electrostatic self-assembly of poly(styrene sulfonate) and diazoresin on MnCO(3) templates. UV-vis and zeta-potential measurements confirmed the alternate deposition of {PSS/DAR} multilayers on the micrometer-sized dissolvable templates. The DAR-based microcapsules were demonstrated to be permeable to enzymes prior to UV irradiation, while the permeability of the multilayer wall was changed substantially after photo-cross-linking. Encapsulated molecules were stably entrapped after UV irradiation, as shown by confocal microscopy and atomic force microscopy images. Activity assays revealed that encapsulated glucose oxidase possessed 52.8% of the catalytic activity exhibited by the same amount of free enzyme, proving the preservation of native conformation and accessibility of substrate. This encapsulation technique is promising for many biomedical and biotechnological applications, particularly enzyme biosensors, which require stable immobilization of functional components while allowing sufficient transport rates for substrate molecules.

Azo Compounds↗

Osteoblast growth promotion by protein electrostatic self-assembly on biodegradable poly(lactide).

Extracellular matrix (ECM)-like coating was developed on biodegradable biomaterials based on the electrostatic self-assembly (ESA) technique to promote osteoblast growth. Poly(ethylenimine) (PEI) was first employed to obtain a stable positively charged surface on poly (DL-lactide) (PDL-LA) substrate. Gelatin was selected as ECM-like biomacromolecule to deposit on the activated PDL-LA substrate using the ESA technique. zeta-Potential results showed alternating charge of polyelectrolytes (PEI/gelatin) layering on PDL-LA microspheres. Quartz crystal microbalance (QCM) measurement further verified the gradual deposition of PEI/gelatin on PDL-LA thin film. Osteoblast cells (MC3T3) were chosen to test the cell behavior on modified PDL-LA substrates. The osteoblast test about cell activity, intracellular total DNA content, total protein content and cell morphology by SEM investigation on ECM-like multilayer-modified PDL-LA substrate showed to promote osteoblast growth. Comparing conventional coating methods, polyelectrolyte multilayers are easy and stable to prepare. It may be a good choice for the surface modification of complex biomedical devices. These very flexible systems allow broad medical applications for drug delivery and tissue engineering.

3T3 Cells↗

Protein electrostatic self-assembly on poly(DL-lactide) scaffold to promote osteoblast growth.

The development of protein coating on 3D biodegradable scaffold based on electrostatic self-assembly (ESA) to promote osteoblast growth is reported. Poly (ethylenimine) (PEI) was employed to obtain a stable positively charged surface on poly(DL-lactide) (PDL-LA) substrate. An extracellular-matrix (ECM)-like biomacromolecule, gelatin, was chosen as the polyelectrolyte to deposit on the activated PDL-LA substrate via ESA technique. Osteoblast (MC3T3) was then cultured on unmodified and gelatin-modified PDL-LA scaffolds. Osteoblast testing regarding total intracellular protein content, total DNA content, cell activity, and cell morphology on the ECM-like multilayer-modified PDL-LA scaffold showed that osteoblast growth was promoted. It will be easy to replace the gelatin with osteoinductive proteins or other polyelectrolytes to promote specific osteoblast functions. In comparison with conventional coating methods, polyelectrolyte multilayers are easy and stable to prepare. They may be a good choice for the surface modification of complex biomedical devices, especially for the 3D tissue-engineering scaffold. These very flexible systems allow broad medical applications for drug delivery and tissue engineering.

3T3 Cells↗

Surface tailoring of poly(DL-lactic acid) by ligand-tethered amphiphilic polymer for promoting chondrocyte attachment and growth.

The ligand-tethered poly(ethylene oxide-propylene oxide-ethylene oxide) (PEO-PPO-PEO) triblock copolymer was explored to engineer poly(DL-lactic acid) (PDL-LA) material to promote cell attachment and growth. The PEO-PPO-PEO was activated by methyl sulfonyl chloride and the amino acid, and peptide were attached. By blending the PDL-LA with the ligand-tethered PEO-PPO-PEO derivatives, the surface of modified PDL-LA film was investigated by ATR-FTIR, XPS and contact angle. The chondrocytes test on different PDL-LA films indicated that the PEO-PPO-PEO amino acid and RGD derivatives modified PDL-LA films could promote chondrocyte attachment and growth. This simple surface treatment method may have potentials for tissue engineering and other biomedical applications.

Adsorption↗

Construction of multilayer coating onto poly-(DL-lactide) to promote cytocompatibility.

Thin polymer films were formed on poly-(DL-lactide) (PDL-LA) using polyelectrolyte multilayer technique to promote the chondrocyte cytocompatibility. PDL-LA substrates were activated by poly-(ethylenimine) to obtain stable positively charged surface. The polyelectrolytes such as alginate and poly-(L-lysine) were alternatively deposited onto the activated PDL-LA substrates. The multilayer-modified PDL-LA films were investigated by X-ray photoelectron spectroscopy, attenuated total reflection FTIR, contact angle and atomic force microscopy. The in vitro chondrocyte test indicated that the multilayer-modified PDL-LA substrates promoted chondrocyte attachment and growth. In comparison to conventional coating methods, polyelectrolyte multilayers are easy to prepare and the procedure is valid whatever the shape of the solid. It allows broad medical applications for drug delivery and tissue engineering.

Animals↗

Surface engineering of poly(D,L-lactic acid) by entrapment of chitosan-based derivatives for the promotion of chondrogenesis.

Chitosan and chitosan-amino acid derivatives were explored to engineer poly(D,L-lactic acid) (PDL-LA) as an extracellular matrix-like surface to promote cell adhesion and growth. Four kinds of chitosan-amino acid derivatives were prepared to mimic the carbohydrate moieties of cell matrix glycoprotein. The chitosan-amino acid derivatives were characterized by using Fourier transform infrared and ultraviolet spectra. The amino acid content on chitosan-amino acid derivatives was determined by using a ninhydrin-ultraviolet method. A new strategy, entrapment, was therefore used to modify the PDL-LA membrane with chitosan and chitosan-amino acid derivatives. The results of X-ray photoelectron spectroscopy, attenuated total reflectance-Fourier transform infrared, and contact angle confirmed that a stable thin film of chitosan and its derivatives can be entrapped on the surface of the PDL-LA membrane. From the results of chondrocyte cytocompatibility, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assays, and cell morphology, the chitosan-amino acid derivative modified PDL-LA membranes were shown to promote chondrogenesis. The novel surface treatment method combines the good mechanical property of PDL-LA with the good cytocompatibility of chitosan derivatives, which may have potential for tissue engineering.

Amino Acids↗

Surface engineering of poly(DL-lactic acid) by entrapment of alginate-amino acid derivatives for promotion of chondrogenesis.

Alginate-amino acid derivatives were explored to engineer poly(DL-lactic acid)(PDL-LA) as glycocalyx-like surface to promote cell adhesion and growth. Four different kinds of alginate-amino acid derivatives were synthesized to mimic the glycocalyx of cell membrane to promote chondrogenesis. The alginate-amino acid derivatives were characterized by FT-IR, 1H NMR and UV spectra and the amino acid content on alginate-amino acid derivatives was given by ninhydrin-UV method. A new strategy, entrapment, was then employed to modify PDL-LA membranes with alginate and its amino acid derivatives. The results of XPS, ATR-FTIR and contact angle confirmed that a stable thin film of alginate and its amino acid derivatives can be entrapped on the surface of PDL-LA membrane. The chondrocyte cytocompatibility test and MTT assays indicated that the alginate-amino acid derivatives modified PDL-LA membranes could promote chondrogenesis. The novel surface treatment method may have potentials for tissue engineering and other biomedical applications.

Alginates↗

Combined physical and chemical immobilization of glucose oxidase in alginate microspheres improves stability of encapsulation and activity.

Chemical sensors utilizing immobilized enzymes and proteins are important for monitoring chemical processes and biological systems. In this study, calcium-cross-linked alginate hydrogel microspheres were fabricated as enzyme carriers by an emulsification technique. Glucose oxidase (GOx) was encapsulated in alginate microspheres using three different methods: physical entrapment (emulsion), chemical conjugation (conjugation), and a combination of physical entrapment and chemical conjugation (emulsion-conjugation). Nano-organized coatings were applied on alginate/GOx microspheres using the layer-by-layer self-assembly technique in order to stabilize the hydrogel/enzyme system under biological environment. The encapsulation of GOx and formation of nanofilm coating on alginate microspheres were verified with FTIR spectral analysis, zeta-potential analysis, and confocal laser scanning microscopy. To compare both the immobilization properties of enzyme encapsulation techniques and the influence of nanofilms with uncoated microspheres, the relationship between enzyme loading, release, and effective GOx activity (enzyme activity per unit protein loading) were studied over a period of four weeks. The results produced four key findings: (1) the emulsion-conjugation technique improved the stability of GOx in alginate microspheres compared to the emulsion technique, reducing the GOx leaching from microsphere from 50% to 17%; (2) the polyelectrolyte nanofilm coatings increased the GOx stability over time, but also reduced the effective GOx activity; (3) the effective GOx activity for the emulsion-conjugation technique (about 3.5 x 10(-)(5) AU microg(-)(1) s(-)(1)) was higher than that for other methods, and did not change significantly over four weeks; and (4) the GOx concentration, when compared after one week for microspheres with three bilayers of poly(allylamine hydrochloride)/sodium poly(styrene sulfonate) ({PAH/PSS}) coating, was highest for the emulsion-conjugation technique. As a result, the comparison of these three techniques showed the emulsion-conjugation technique to be a potentially effective and practical way to fabricate alginate/GOx microspheres for implantable glucose biosensor application.

Alginates↗

Surface engineering of poly(DL-lactide) via electrostatic self-assembly of extracellular matrix-like molecules.

We report the development of new biomacromolecule coatings on biodegradable biomaterials based on electrostatic assembly of extracellular matrix-like molecules. Poly(ethylene imine) (PEI) was employed to engineer poly(dl-lactide) (PDL-LA) substrate to obtain a stable positively charged surface. An extracellular matrix- (ECM-) like biomacromolecule, gelatin, was selected as the polyelectrolyte to deposit on the activated PDL-LA substrate via the electrostatic assemble technique. The extracellular matrix-like multilayer on the PDL-LA substrate was investigated by attenuated total reflection (ATR-FTIR), X-ray photoelectron spectrscopy (XPS), contact angle, and atomic force microscopy (AFM). The gradual buildup of the protein layer was investigated by UV-vis spectra, and it was further given a quantitative analysis of the protein layer on the PDL-LA substrate via the radioiodination technique. The stability of the protein layer under aqueous condition was also tested by the radiolabeling method. Chondrocyte was selected as the model system for testing the cell behavior and morphology on modified PDL-LA substrates. The chondrocyte test about cell attachment, proliferation, cell activity and cell morphology by SEM, and confocal laser scanning microscopy (CLSM) investigation on extracellular matrix-like multilayer modified PDL-LA substrate was shown to promote chondrocyte attachment and growth. Comparing conventional coating methods, polyelectrolyte multiplayers are easy and stable to prepare. It may be a good choice for the modification of 3-D scaffolds used in tissue engineering. These very flexible systems allow broad medical applications for drug delivery and tissue engineering.

Animals↗

Biomacromolecules electrostatic self-assembly on 3-dimensional tissue engineering scaffold.

A poly(ethylenimine) (PEI) was employed to obtain a stable positively charged surface on a poly(D,L-lactide) (PDL-LA) tissue engineering scaffold. An extracellular matrix (ECM)-like biomacromolecule, gelatin, was selected as polyelectrolyte and deposit alternately with PEI on the activated PDL-LA scaffold via ESA technique. The zeta-potential result showed alternating charge of polyelectrolytes (PEI/gelatin) layering on PDL-LA microspheres. Quartz crystal microbalance (QCM) measurement further verified the gradual deposition of PEI/gelatin on the PDL-LA thin film. The combination of PEI aminolysis and the layer-by-layer technique was then explored to construct gelatin coating onto the 3-D porous PDL-LA scaffold. Scanning electronic microscopy showed that there is no notable difference between modified and unmodified PLA scaffolds, with regard to the porosity, pore diameter, and scaffold integration. The dual-tunnel confocal laser scanning microscopy indicated uniform gelatin distribution on the inner surface of the 3-D porous scaffold. The gradual build-up of protein layer on scaffold was investigated by radioiodination technique. Chondrocyte was chosen to test the cell behavior on modified and unmodified PDL-LA scaffolds. The results of the cell viability, total intracellular protein content, and cell morphology on the PEI/gelatin multilayers modified PDL-LA scaffold showed to promote chondrocyte growth. Comparing conventional coating methods, polyelectrolyte multilayers are easy and stable to prepare. It may be a promising choice for the surface modification of complex biomedical devices. These very flexible systems allow broad medical applications for drug delivery and tissue engineering.

Animals↗

Spontaneous loading of positively charged macromolecules into alginate-templated polyelectrolyte multilayer microcapsules.

A simple and high-efficiency approach to loading macromolecules into microscale carriers is presented. Calcium-cross-linked alginate hydrogel microspheres were fabricated by an emulsification technique and then used as negatively charged templates to form polyelectrolyte multilayer coatings. A calcium ion chelator, EDTA, was used to free the Ca(2+)-cross-linked alginate hydrogel within {poly(allylamine hydrochloride)/poly (styrene sulfonate)}(4) ({PAH/PSS}(4)) coating, allowing partial release of alginate. The retention of alginate in {PAH/PSS}(4) microcapsule was confirmed by FTIR spectroscopy and confocal microscopy. Real-time confocal microscopy was used to investigate the loading process of positively charged macromolecules (dextran-amino, and peroxidase) into alginate-templated microcapsules, which showed the loading occurred in <2 min for dextran-amino and <10 min for peroxidase, respectively. A high loading efficiency of 25 mug peroxidase in approximately 1.0 x 10(7) microcapsules (2.5 pg POx/capsule) was achieved with a low concentration of peroxidase loading solution (10 mug/mL). This spontaneous loading technique for encapsulating positively charged molecules in alginate-templated polyelectrolyte microcapsules shows strong potential for biosensor and drug delivery applications.

Alginates↗