In vitro detection of allergen sensitized basophils by HSA-DNP antigen-anchored liquid crystal microdroplets.

A simple and easy to handle biosensing technique for in vitro detection of HSA-DNP antigen induced allergen reactions in patients has been developed through the detection of sensitized basophils expressed with anti-IgE receptor (FcεRI) by using human serum albumin-dinitrophenol (HSA-DNP) antigen-anchored liquid crystal (LC) microdroplets emulsion. The radial to bipolar transition in nematic 4-cyano-4'-pentyl biphenyl liquid crystal molecules (5CB) confined in HSA-DNP antigen anchored LC microdroplets (8.5 pg HSA-DNP/LC microdroplet) is found to be sensitive in PBS solution in detection of allergen sensitized basophils expressed with a minimum amount of anti HSA-DNP (anti-IgE) receptor (≥4.5 pg/basophil). The detection of allergen sensitized basophils was possible within a contact time of 30 min in presence of control cells and with 10% solution of human blood plasma. The HSA-DNP antigen anchored LC microdroplets in presence of macrophages or non-sensitized basophils did not show radial to bipolar transition in 5CB molecules in PBS or solution with 10 wt% human blood plasma. Thus HSA-DNP antigen anchored LC microdroplets biosensor may be used for in vivo detection of stage I allergen reaction basophils in blood samples.

Photoinduced development of antibacterial materials derived from isosorbide moiety.

A straightforward method for immobilizing in situ generated silver nanoparticles on the surface of a photoactivable isosorbide-derived monomer is developed with the objective to design a functional material having antibacterial properties. The photoinduced thiol-ene mechanism involved in these syntheses is described by the electron spin resonance/spin trapping technique. The resulting materials with or without silver nanoparticles (Ag NPs) were used as films or as coatings on glass substrate. The surface of the synthesized materials was characterized by X-ray photoelectron spectroscopy and scanning electron microscopy, and their thermal and mechanical properties were evaluated by dynamic-mechanical thermal tests, differential scanning calorimetry, thermogravimetric analyses, along with pencil hardness, nanoindentation, and scratch resistance tests. The photoinduced formation of Ag NPs is also confirmed by UV spectrophotometry. Finally, a primary investigation demonstrates the antibacterial properties of the isosorbide-derived material against Staphylococcus aureus and Escherichia coli, as well as its cytocompatibility toward NIH 3T3 fibroblastic cells.

Poly(acrylic acid)-grafted graphene oxide as an intracellular protein carrier.

A pH-sensitive poly(acrylic acid)-grafted graphene oxide (GO-PAA) nanocarrier was synthesized by in situ atom transfer radical polymerization to allow the oral delivery of hydrophilic macromolecular proteins in their active forms to specific cells or organs. The synthesis, morphology, and physiochemical properties of GO-PAA were examined. A model protein, bovine serum albumin (BSA) labeled with fluorescein isothiocyanate (FITC) (BSAFITC), was loaded onto GO-PAA through noncovalent interactions and its release was arrested at acidic pH similar to stomach, whereas at pH similar to intestine it was reduced, which paves way for site specific delivery without its degradation in the gastrointestinal tract. Confocal laser microscopy showed that the BSAFITC-loaded GO-PAA was internalized by KB cells by endocytosis and released into cytoplasm. Thus the GO-PAA as a transmembrane transporter is a new class of drug transporters with potential protein delivery applications.

Folate ligand anchored liquid crystal microdroplets emulsion for in vitro detection of KB cancer cells.

A KB cancer cell-selective, liquid crystal microdroplets emulsion is prepared using folic acid-conjugated block copolymers (PS-b-PAA-FA) and sodium dodecyl sulfate (SDS) as a mediator to induce configurational transitions in 4-cyano-4'-pentylbiphenyl (5CB) liquid crystal microdroplets emulsion. The prepared liquid crystal microdroplets emulsion has shown a configurational transition from radial to bipolar on interacting with KB cancer cells, but no transition from radial to bipolar configuration is observed when liquid crystal microdroplets emulsion was allowed to interact with other normal cells such as fibroblast and osteoblast. The KB cancer cell selectivity of liquid crystal microdroplets emulsion has been considered due to the presence of KB cancer cell folate receptor-specific ligand (FA) at the surface of liquid crystal microdroplets, which allowed liquid crystal microdroplets to interact specifically with KB cancer cells. The ligand-receptor interactions have been considered responsible for triggering the configurational transitions from radial to bipolar in liquid crystal microdroplets emulsion. Thus, folate ligand anchored liquid crystal microdroplets emulsion has shown a potential to be used for in vitro detection of KB cancer cells in the early stage of tumor development.

Bilayer dressing based on aerogel/electrospun mats with self-catalytic hydrogen sulfide generation and enhanced antioxidant ability.

Hydrogen sulfide (H2S) releasing wound dressings have attracted much attention for their ability to promote cell proliferation, stimulate angiogenesis, and resist inflammation. Mimicking the skin structure, a bilayer wound dressing based on aerogel/mats with H2S release capability was designed and fabricated. A bio-macromolecular H2S donor based on a keratin-TA conjugate (KTC) was first synthesized through a thiol-disulfide exchange reaction. As an inner layer, KTC was then loaded into a gelatin hydrogel with large pores to absorb the wound exudates and generate H2S self-catalytically. Subsequently, polyurethane was electrospun with glutathione (GSH) to be used as an outer layer with small pores, which provided mechanical support, supplied GSH, and prevented bacterial invasion. The bilayer dressing was capable of generating H2S self-catalytically, achieving a controlled and sustained release. The dressing could also promote cell proliferation and migration. In addition, the dress possessed enhanced antioxidant ability and reactive oxygen species (ROS) scavenging capability. The bilayer dressing on promoting wound healing was investigated in a full-thickness excisional cutaneous wound model in rats. The results demonstrated that it could reduce inflammation, promote vascularization, and facilitate hair follicle regeneration, thereby accelerating wound healing. Overall, the bilayer dressing has great potential applications in the field of the wound dressing.

Immobilization of Collagen on the Surface of a PEEK Implant with Monolayer Nanopores.

Polyetheretherketone (PEEK) is the only polymer material that can replace titanium implants in the field of orthopedics. This is because the mechanical properties of PEEK are similar to those of bone, and PEEK has natural radiolucency, chemical stability, and sterilization resistance. Despite these advantages, PEEK has a disadvantage-that it is bio-inert. Therefore, many studies have attempted to change the bio-inertness of PEEK into bioactivity. Among them, a method of forming pores by acid treatment is attracting attention. In this study, an attempt was made to form pores on the surface of PEEK implant using a mixed acid of sulfuric acid and nitric acid. As a result, it was found that the condition when the PEEK surface is in contact with the acid is very important. That is, it was possible to form single-layered nanopores on the surface by contacting PEEK with a mixed acid under ultrasound. Additionally, by immobilizing type I collagen on the porous PEEK surface through dopamine coating, it was possible to obtain collagen-immobilized porous PEEK (P-PEEK-Col) with high compatibility with osteoblasts. This P-PEEK-Col has high potential for use as a bone substitute that promotes bone formation.

In vitro anti-bacterial and cytotoxic properties of silver-containing poly(L-lactide-co-glycolide) nanofibrous scaffolds.

Electrospinning has recently emerged as a leading technique for the formation of nanofibrous structures made of organic and inorganic components. In this study, nanofibrous scaffolds were prepared by electrospining a bend solution of poly(L-lactide-co-glycolide) (PLGA) and silver nanoparticles in 1,1,1,3,3,3,-hexafluoro-2-propanol (HFIP). The resulting fibers ranged from 420 to 590 nm in diameter. To evaluate the possibility of using silver-containing PLGA as a tissue engineering scaffold, experiments on cell viability and antibacterial activity were carried out. As a result, PLGA nanofibrous scaffolds having silver nanoparticles of more than 0.5 wt% showed antibacterial effect against Staphylococcus aureus and Klebsiella pneumonia. Furthermore, silver-containing PLGA nanofibrous scaffolds showed viability, indicating their possible application in the field of tissue engineering.

Nitric oxide-releasing poly(ε-caprolactone)/S-nitrosylated keratin biocomposite scaffolds for potential small-diameter vascular grafts.

Rapid endothelialization and regulation of smooth muscle cell proliferation are crucial for small-diameter vascular grafts to address poor compliance, thromboembolism, and intimal hyperplasia, and achieve revascularization. As a gaseous signaling molecule, nitric oxide (NO) regulates cardiovascular homeostasis, inhibits blood clotting and intimal hyperplasia, and promotes the growth of endothelial cells. Due to the instability and burst release of small molecular NO donors, a novel biomacromolecular donor has generated increasing interest. In the study, a low toxic NO donor of S-nitrosated keratin (KSNO) was first synthesized and then coelectrospun with poly(ε-caprolactone) to afford NO-releasing small-diameter vascular graft. PCL/KSNO graft was capable to generate NO under the catalysis of ascorbic acid (Asc), so the graft selectively elevated adhesion and growth of human umbilical vein endothelial cells (HUVECs), while inhibited the proliferation of human aortic smooth muscle cells (HASMCs) in the presence of Asc. In addition, the graft displayed significant antibacterial properties and good blood compatibility. Animal experiments showed that the biocomposite graft could inhibit thrombus formation and preserve normal blood flow via single rabbit carotid artery replacement for 1 month. More importantly, a complete endothelium was observed on the lumen surface. Taken together, PCL/KSNO small-diameter vascular graft has potential applications in vascular tissue engineering with rapid endothelialization and vascular remolding.

In vitro assessment of antibacterial activity and cytocompatibility of silver-containing PHBV nanofibrous scaffolds for tissue engineering.

Infections with bacteria have become a serious problem in joint arthroplasty. This study reports about in vitro antibacterial activity and in vitro cell compatibility of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) nanofibers loaded with metallic silver particles of a size of 5-13 nm. In vitro antibacterial activity against Staphylococcus aureus and Klebsiella pneumoniae was studied by microplate proliferation tests. The adhesion, viability, and proliferation properties of fibroblasts (NIH 3T3) and differentiation of osteoblasts (MC3T3-E1) were done to study in vitro cell compatibility of the scaffolds. As the results, only silver-containing PHBV nanofibrous scaffolds showed a high antibacterial activity and an inhibitory effect on the growth of both Staphylococcus aureus and Klebsiella pneumoniae bacteria. The nanofibrous scaffolds having silver nanoparticles <1.0% were free of in vitro cytotoxicity. To sum up, the PHBV nanofibrous scaffolds having nanoparticles <1.0 wt % showed not only good antibacterial activity but also good in vitro cell compatibility. It is considered that the PHBV nanofibrous scaffolds with silver nanoparticles <1.0 wt % have a potential to be used in joint arthroplasty.

Immobilization of biomolecules on the surface of inorganic nanoparticles for biomedical applications.

Various inorganic nanoparticles have been used for drug delivery, magnetic resonance and fluorescence imaging, and cell targeting owing to their unique properties, such as large surface area and efficient contrasting effect. In this review, we focus on the surface functionalization of inorganic nanoparticles via immobilization of biomolecules and the corresponding surface interactions with biocomponents. Applications of surface-modified inorganic nanoparticles in biomedical fields are also outlined.

Immobilization of lactobionic acid on the surface of cadmium sulfide nanoparticles and their interaction with hepatocytes.

In the current study, beta-galactose-carrying lactobionic acid (LA) was conjugated on the surface of mercaptoacetic acid-coated cadmium sulfide nanoparticles (CSNPs) to ensure specific recognition of liver cells (hepatocytes) and to enhance biocompatibility. Maltotrionic acid-coated CSNPs (MCSNPs) were also prepared for use as a control. The results showed that LA-immobilized CSNPs (LCSNPs) were selectively and rapidly internalized into hepatocytes and emitted more intense fluorescence images as well as demonstrated increased biocompatible behavior in vitro than those of CSNPs and MCSNPs. Furthermore, the uptake amount of LCSNPs into hepatocytes was higher than that of CSNPs and MCSNPs. All these results indicate that LCSNPs may find ever-growing applications in biological labels and detection or contrast agents in life science and medical diagnostics.

Osteoblast behaviours on nanorod hydroxyapatite-grafted glass surfaces.

BACKGROUND: The goal of this study is to obtain basic information to improve the bone adhesion of silica components, which are used as the main ingredient in glass ionomer cement (GIC). To achieve this, nanorod hydroxyapatite (nHA) was grafted to the surface of silica cover glass. Surface analysis confirmed nHA was joined to the glass surface and biocompatibility with osteoblasts was investigated. RESULTS: The grafting of nHA on the surface of slide cover glass (Glass) was confirmed by X-ray photoelectron spectroscopy (XPS) and contact angle (θ) measurement. MC3T3-E1 cells were more stretched out on the nHA-grafted cover glass (Glass-nHA) in comparison to the Glass. In addition, the Glass-nHA was more bioactive in supporting the proliferation of MC3T3-E1 cells in comparison to cells seeded on the Glass. CONCLUSION: The Glass-nHA was to be highly bioactive and this might be useful information for property modification of GIC.

Biocompatible Polymers and their Potential Biomedical Applications: A Review.

BACKGROUND: Biocompatible polymers are gaining great interest in the field of biomedical applications. The term biocompatibility refers to the suitability of a polymer to body and body fluids exposure. Biocompatible polymers are both synthetic (man-made) and natural and aid in the close vicinity of a living system or work in intimacy with living cells. These are used to gauge, treat, boost, or substitute any tissue, organ or function of the body. A biocompatible polymer improves body functions without altering its normal functioning and triggering allergies or other side effects. It encompasses advances in tissue culture, tissue scaffolds, implantation, artificial grafts, wound fabrication, controlled drug delivery, bone filler material, etc. OBJECTIVES: This review provides an insight into the remarkable contribution made by some well-known biopolymers such as polylactic-co-glycolic acid, poly(ε-caprolactone) (PCL), polyLactic Acid, poly(3- hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), Chitosan and Cellulose in the therapeutic measure for many biomedical applications. METHODS: Various techniques and methods have made biopolymers more significant in the biomedical fields such as augmentation (replaced petroleum based polymers), film processing, injection modeling, blow molding techniques, controlled / implantable drug delivery devices, biological grafting, nano technology, tissue engineering etc. RESULTS: The fore mentioned techniques and other advanced techniques have resulted in improved biocompatibility, nontoxicity, renewability, mild processing conditions, health condition, reduced immunological reactions and minimized side effects that would occur if synthetic polymers are used in a host cell. CONCLUSION: Biopolymers have brought effective and attainable targets in pharmaceutics and therapeutics. There are huge numbers of biopolymers reported in the literature that has been used effectively and extensively.

Hydroxyapatite Nanorod-Modified Sand Blasted Titanium Disk for Endosseous Dental Implant Applications.

BACKGROUND: Sand blasted titanium (Ti) is commonly used in designing endosseous dental implants due to its biocompatibility and ability to form bonds with bone tissues. However, titanium implants do not induce strong interactions with teeth bones. To increase strong interactions between Ti disk implants and teeth bones, the l-glutamic acid grafted hydroxyapatite nanorods (nHA) were immobilized on albumin modified Ti disk implants (Ti-Alb). METHODS: For modification of Ti disk implants by nHA, the l-glutamic acid grafted nHA was synthesized and then immobilized on albumin modified Ti disk implants. Fourier transformed infrared spectroscopy, X-ray photoelectron spectroscopy, scanning electron microscope; energy dispersive spectroscopy and confocal laser scanning microscopy were used to confirm the modification of Ti disk implants. The bioactivity of nHA-modified Ti disk implants was evaluated by seeding MC3T3-E1 cells on Ti-nHA implants. RESULTS: Characterization techniques have confirmed the successful modification of Ti disk implants by l-glutamic acid grafted nHA. The nHA-modified Ti disk implants have shown enhanced adhesion, proliferation and cytotoxicity of MC3T3-E1 cells in comparison to pristine Ti implants. CONCLUSIONS: The modification of Ti implants by l-glutamic acid grafted nHA has produced highly osteogenic Ti disk plants in comparison to pristine Ti disk implants due to the formation of bioactive surfaces by hydroxyapatite nano rods on Ti disk implants. Ti-nHA disk implants showed enhanced adhesion, proliferation, and MC3T3-E1 cells viability in comparison to pristine Ti disk implants. Thus nHA might be to be useful to enhance the osseointegration of Ti implants with teeth bones.

A novel use of cellulose based filter paper containing silver nanoparticles for its potential application as wound dressing agent.

The frequent use of antibiotics against microbial infections may lead to the emergence of antibiotic resistant microbial strains. To overcome these microbial strains, we need to fabricate alternative materials which can handle them. It is for this reason, we have fabricated cellulose (CE) based filter paper (FP) composite scaffolds comprising of adsorbed chitosan (CS) and sliver (Ag) nanoparticles (NPs). The AgNPs are incorporated in the CS layer of the composite scaffold. Prior to evaluate the efficacy of the scaffolds against gram positive and gram negative bacterial strains, the scaffolds were characterized for the presence of the Ag NPs with field emission scanning electron microscope (FE-SEM), fourier transform infrared (FTIR) spectroscopy and x-ray diffractometer (XRD). These techniques confirmed the presence of Ag NPs in the composite scaffold. The biocompatibility of the scaffolds was assessed by subjecting pristine FP, CS adsorbed FP (CS-FP) and Ag loaded CS-FP (Ag-CS-FP) composite scaffolds to in vitro studies. From the data obtained, it was observed that NIH3T3 fibroblastic cells adhered and proliferated onto all the scaffolds. Furthermore, the scaffolds exhibited good antibacterial activity against both strains of bacteria. It is, therefore, concluded that these scaffolds could find potential application in biomedical field, particularly as a wound dressing agent.

Surface modification of magnetite nanoparticles using lactobionic acid and their interaction with hepatocytes.

In the current study, superparamagnetic magnetite nanoparticles were surface-modified with lactobionic acid (LA) to improve their intracellular uptake and ability to target hepatocytes. Maltotrionic acid (MA)-modified nanoparticles were also synthesized as a control. Cell culture experiment showed that LA-modified nanoparticles were internalized into hepatocytes and atomic absorption spectrometer (AAS) measurement indicated that the uptake amount of LA-modified magnetite into hepatocytes was higher than that of unmodified and MA-modified nanoparticles. LA-modified nanoparticle solution was injected in rabbit and the magnetic resonance (MR) images obtained showed that LA-coated nanoparticles were selectively accumulated onto the hepatocytes. This result demonstrates that the LA-modified magnetite nanoparticles have a great potential to be used as contrast agent for liver diagnosis.

Effect of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) nanofiber matrices cocultured with hair follicular epithelial and dermal cells for biological wound dressing.

We tested the effects on the early-stage wound healing of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) nanofiber matrices cultured with hair follicular cells. PHBV only, PHBV/collagen, and PHBV/gelatin at a 7/3 weight ratio were produced by electrospinning, and their in vitro cell culture and in vivo wound healing as biological dressings were examined. In cell attachment and growth on matrices, dermal sheath (DS) cells attached to hydrophilic PHBV/collagen and PHBV/gelatin faster than hydrophobic PHBV at the early incubation stage (up to 6 h). From 6- to 24-h incubation, PHBV/collagen showed the best results in cell culture. Furthermore, PHBV/collagen cocultured for 3-5 days with DS and epithelial outer root sheath (ORS) cells expressed more extracellular materials, such as type I collagen, elastin, and alpha-smooth muscle actin, than cocultured PHBV with the same cells. However, there was no significant difference between PHBV and PHBV/collagen in the amounts of cytokeratin 8 expressed. Grafting of PHBV and PHBV/collagen matrices cocultured with ORS/DS cells for 3-5 days showed that PHBV promoted wound closure and re-epithelization more obviously than PHBV/collagen in both cocultured matrices and matrices alone. Cocultured matrices would heal wounds better than the corresponding matrices alone. Thus, PHBV cocultured with ORS/DS cells could be used as a cell-seeded biological dressing, thereby reducing preparation time as well as regenerating the epidermis efficiently during the early stage of wound healing.

Lamination of microfibrous PLGA fabric by electrospinning a layer of collagen-hydroxyapatite composite nanofibers for bone tissue engineering.

BACKGROUND: To mimic the muscle inspired cells adhesion through proteins secretion, the lamination of collagen-hydroxyapatite nanorod (nHA) composite nanofibers has been carried out successfully on polydopamine (PDA)-coated microfibrous polylactide-co-glycolide (PLGA) fabrics. The lamination of collagen-hydroxyapatite composite nanofibers on polydopamine-coated microfibrous PLGA fabrics was carried through electrospinning the solution of collagen containing L-glutamic acid-grafted hydroxyapatite nanorods (nHA-GA) at a flow rate of 1.5 mL/h and an applied voltage of 15 kV. RESULTS: In comparison to pristine PLGA, dopamine-coated PLGA and collagen-hydroxyapatite composite nanofiber lamination has produced more wettable surfaces and surface wettability is found to higher with dopamine-coated PLGA fabrics then pristine PLGA. The SEM micrographs have clearly indicated that the lamination of polydopamine-coated PLGA fabric with collagen-hydroxyapatite composite nanofibers has shown increased adhesion of MC3T3E1 cells in comparison to pristine PLGA fabrics. CONCLUSION: The results of these studies have clearly demonstrated that collagen-nHA composites fibers may be used to create bioactive 3D scaffolds using PLGA as an architectural support agent.

Preparation and characterization of anodized titanium surfaces and their effect on osteoblast responses.

In this study, titanium (Ti) surface was modified by anodizing with a mixture of beta-glycerophosphate sodium and calcium (Ca) acetate, and the anodized surfaces were characterized by scanning electron microscopy, X-ray diffraction, and electron probe microanalysis. In vitro osteoblast response to anodized oxide was also evaluated. The anodic oxide produced was observed to have interconnected pores (0.5-2 microm in diameter) and intermediate roughness (0.60-1.00 microm). In addition, anodic oxide was observed to have amorphous and anatase oxide. Calcium and phosphorus ions were deposited on the Ti oxide during anodization. Osteoblast differentiation, as indicated by alkaline phosphatase production, was enhanced on anodized surfaces. It was thus concluded from this study that Ca phosphate can be deposited on Ti surfaces by anodization. It was also concluded that the phenotypic expression of osteoblast was enhanced by the presence of Ca phosphate and higher roughness on anodized Ti surfaces.

Micro/Nano Multilayered Scaffolds of PLGA and Collagen by Alternately Electrospinning for Bone Tissue Engineering.

The dual extrusion electrospinning technique was used to fabricate multilayered 3D scaffolds by stacking microfibrous meshes of poly(lactic acid-co-glycolic acid) (PLGA) in alternate fashion to micro/nano mixed fibrous meshes of PLGA and collagen. To fabricate the multilayered scaffold, 35 wt% solution of PLGA in THF-DMF binary solvent (3:1) and 5 wt% solution of collagen in hexafluoroisopropanol (HFIP) with and without hydroxyapatite nanorods (nHA) were used. The dual and individual electrospinning of PLGA and collagen were carried out at flow rates of 1.0 and 0.5 mL/h, respectively, at an applied voltage of 20 kV. The density of collagen fibers in multilayered scaffolds has controlled the adhesion, proliferation, and osteogenic differentiation of MC3T3-E1 cells. The homogeneous dispersion of glutamic acid-modified hydroxyapatite nanorods (nHA-GA) in collagen solution has improved the osteogenic properties of fabricated multilayered scaffolds. The fabricated multilayered scaffolds were characterized using FT-IR, X-ray photoelectron spectroscopy, and transmission electron microscopy (TEM). The scanning electron microscopy (FE-SEM) was used to evaluate the adhesion and spreads of MC3T3-E1 cells on multilayered scaffolds. The activity of MC3T3-E1 cells on the multilayered scaffolds was evaluated by applying MTT, alkaline phosphatase, Alizarin Red, von Kossa, and cytoskeleton F-actin assaying protocols. The micro/nano fibrous PLGA-Col-HA scaffolds were found to be highly bioactive in comparison to pristine microfibrous PLGA and micro/nano mixed fibrous PLGA and Col scaffolds.