Stroke rehabilitation: strategies to enhance motor recovery.

Recent evidence indicates that the brain can remodel after stroke, primarily through synaptogenesis. Task-specific and repetitive exercise appear to be key factors in promoting synaptogenesis and are central elements in rehabilitation of motor weakness following stroke. Expert medical management ensures a patient is well enough to participate in rehabilitation with minimal distractions due to pain or depression. Contraint-induced motor therapy and body-weight-supported ambulation are forms of exercise that "force use" of an impaired upper extremity. Technologies now in common use include robotics, functional electrical stimulation, and, to a lesser degree, transcranial magnetic stimulation and virtual reality. The data on pharmacological interventions are mixed but encouraging; it is hoped such treatments will directly stimulate brain tissue to recovery. Mitigation of factors preventing movement, such as spasticity, might also play a role. Research evaluating these motor recovery strategies finds them generally good at the movement level but somewhat less robust when looking at functional performance. It remains unclear whether inconsistent evidence for functional improvement is a matter of poor treatment efficacy or insensitive outcome measures.

A novel γ-PGA composite gellan membrane containing glycerol for guided bone regeneration.

An ideal barrier membrane design should incorporate the function of a delivery vehicle for transporting drugs and osteoinductive factors to where the body is under inflammation. In the present study, a functional hydrogel-based barrier membrane is fabricated using calcium-form poly-γ-glutamic acid (γ-PGA) and glycerol blending into gellan gum. The concentration of the calcium-form poly-γ-glutamic acid (γ-PGA) and the glycerol ratio are studied for improving practicability in easy-handling and expanding the coverage area. Gellan gum-based membranes with uniformly distributed calcium aggregates are not only successfully manufactured but also providing excellent characteristics for protein adsorption, bioactivity, and bone cell maturation. Our composite gellan gum-based membranes were tested including to their morphology, mechanical properties, swelling behavior, protein adsorption, drug diffusion, and lysozyme degradation. The biocompatibility, proliferation, and osteoblastic response of membranes were examined by osteoblast-like (MG63) cells. Our results indicate that adequate physical cross-linking with γ-PGA improves the original mechanical properties and delays degradation. Growing glycerol ratio not only enhances the elongation at break and diffusion rate, but it also changes the tensile strength and the remaining weight. In vitro biocompatibility tests, an adequate ratio of γ-PGA modification significantly enhances the proliferation, the secretion of alkaline phosphatase (ALP) and mineralization. However, worth noting is the glycerol-modified membrane cannot bear a close resemblance with the non-glycerol group in the high level of osteoblastic response. In general, these tunable materials with biocompatibility, biodegradability, and positive osteoblastic responses were poised to be possible candidates for bone defect repair.

Dual RGD-immobilized poly(L-lactic acid) by atmospheric pressure plasma jet for bone tissue engineering.

Surface treatment on PLA substrates by atmospheric pressure plasma jet (APPJ) for polymerization of dual RGD-peptides were investigated. Peptide-modified surfaces have been highlighted as the most promising approach to improve the integration of implants into surrounding bones. By varying the RF power, PLA substrates treated by APPJ process have a tendency to form a hydrophobic surface. The effects on the proliferation and differentiation of MG63 cells were evaluated and osteocalcin (OCN) expression was analyzed using RT-PCR. The water contact angle of the W/APPJ process PLA was approximately 54% of that of the W/O APPJ process PLA substrates. W/APPJ process significantly increased cell proliferation, improved the functionality of the material without using a complicated procedure. We believe that pretreatment using the APPJ processes and dual RGD grafting can be more appropriate than traditional surface modification methods, with more potential for application to bone materials.

Tracking the Reflective Light Particles Spreading on the Cornea: An Emerging Assessment for Tear Film Homeostasis.

PURPOSE: To implement an emerging noninvasive approach for assessing the dynamic tear film (TF) homeostasis. METHODS: The video records of dynamic TF from 12 healthy orthokeratology lens wearers were obtained by a clinically available TF analyzer and decomposed as image sequences. The trajectories of TF particles were analyzed by two tracking models, the full-span model (FSM) and the fixed-duration model (FDM). FSM tracked a particle for a complete opening blink cycle, while FDM tracked 1 second of the same cycle. A power-law fitting operation [Formula: see text] was used to extract homeostasis markers based on the tracking model for each subject. RESULTS: Comparing two tracking models (N = 6), only one subject had statistical difference in averaged momentary moving speed (MMS; P = 0.0488), while none had significant difference in averaged momentary moving direction (MMD). However, both models showed good correlations in average MMS (ρ = 0.94, P = 0.0048) and MMD (ρ = 1.00, P < 0.0001) and all extracted homeostasis markers [α, ß, MMS(0.1), and MMS(2.0)]. Assessing interblink reliability in these markers under FDM tracking (N = 12), only one subject in the MMS (0.1) and another subject in the MMS (2.0) were outside 95% limits of agreement, respectively. CONCLUSIONS: FDM is a good alternative to FSM and has tracking properties of higher efficiency and easier implementation. The homeostasis markers under FDM tracking showed a good interblink consistence; therefore this approach will be a promising method for analyzing dynamic TF homeostasis in future practice. TRANSLATIONAL RELEVANCE: FDM analytical architecture can practice the past experimental platform on a TF analyzer to obtain homeostasis markers of TF.

A new protein A assay based on Raman reporter labeled immunogold nanoparticles.

A unique, sensitive, highly specific, and photobleaching-resistant immunoassay system utilizing gold nanoparticles and surface-enhanced Raman scattering (SERS) is described. This new system, featuring a capability of bifunctional analysis, is manufactured by chemisorption of antibody immunoglobulin G (IgG) on gold nanoparticles (AuNP), followed by coupling the Raman-active reporter molecule, 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) to the surface of IgG-AuNP. The adsorbed DTNB molecules exhibit strong Raman signals via both electromagnetic and chemical enhancement. The narrow spectral widths and high photostability assure the system to be an excellent detection label. This SERS-based immunoassay is applied to the detection of protein A, which is a specific surface antigen of Staphylococcus aureus. A working curve is obtained by plotting the intensity of the SERS signal of symmetric NO(2) stretching of DTNB at 1,333 cm(-1) versus the concentration of the analyte (antigen). A dynamic range of two to three orders of magnitude and a detection limit of 1 pg/mL of protein A are achieved.

Novel electrospun polycaprolactone/graphene oxide/Fe3O4 nanocomposites for biomedical applications.

In this study, one of the most promising methods of tailoring a composite scaffold material in nano sized diameters, electrospinning method were used to produce Polycaprolactone (PCL)/Graphene Oxide (GO)/Iron(II, III) Oxide (Fe3O4) nanocomposite fibers as biocompatible scaffolds for biomedical applications. Products were analyzed by scanning electron microscopy (SEM) for morphological analysis of the electrospun nanocomposites and Fourier Transform Infrared Spectroscopy (FTIR) was used to determine functional groups of the PCL, GO, and Fe3O4 materials in the electrospun nanocomposites. For physical properties, viscosity, density, permittivity, dielectric loss and liquid and solid state alternating current conductivity, measurements were done for each nanocomposite fibers. Effects of concentration percentage of GO on permittivity, dielectric loss and AC conductivity have been analyzed by using measured and calculated data. Trend lines have been drawn for permittivity, dielectric loss and conductivity via concentration percentage of GO. The relation between ac conductivity and frequency have been studied for each concentration percentage of GO and interpretations have been done by using the obtained results.

Characterization of pegylated copolymeric micelles and in vivo pharmacokinetics and biodistribution studies.

The aim of this study was to evaluate the influence of pegylated copolymeric micelle carrier on the biodistribution of drug in rats. The copolymers were synthesized via a modified ring-opening copolymerization of lactone monomers (epsilon-caprolactone, delta-valerolactone, L-lactide) and poly(ethylene glycol) (PEG(10,000) and PEG(4000)). The molecular weights and the polydispersities of synthesized copolymers were in the range of 15,000-31,000 g/mol and 1.7-2.7, respectively. All of the pegylated amphiphilic copolymers were micelles formed with low CMC values in the range of 10(-7)-10(-8)M. The drug-loaded micelles were prepared via a dialysis method. The average particle size of micelles was around 150-200 nm. The cytotoxicity in terms of cell viability after treated with PCL-PEG, PVL-PEG, and PLA-PEG micelles was insignificant. PCL-PEG and PVL-PEG micelles without branch side chain in structures had higher drug loading than PLA-PEG micelles. In vitro release profiles indicated the release of indomethacin from these micelles exhibited a sustained release behavior. The similar phenomenon was also observed in vivo in rats. The pegylated copolymeric micelles not only decreased drug uptake by the liver and kidney, but also prolonged drug retention in the blood.

Osteogenesis of human adipose-derived stem cells on poly(dopamine)-coated electrospun poly(lactic acid) fiber mats.

Electrospinning is a versatile technique to generate large quantities of micro- or nano-fibers from a wide variety of shapes and sizes of polymer. The aim of this study is to develop functionalized electrospun nano-fibers and use a mussel-inspired surface coating to regulate adhesion, proliferation and differentiation of human adipose-derived stem cells (hADSCs). We prepared poly(lactic acid) (PLA) fibers coated with polydopamine (PDA). The morphology, chemical composition, and surface properties of PDA/PLA were characterized by SEM and XPS. PDA/PLA modulated hADSCs' responses in several ways. Firstly, adhesion and proliferation of hADSCs cultured on PDA/PLA were significantly enhanced relative to those on PLA. Increased focal adhesion kinase (FAK) and collagen I levels and enhanced cell attachment and cell cycle progression were observed upon an increase in PDA content. In addition, the ALP activity and osteocalcin of hADSCs cultured on PDA/PLA were significantly higher than seen in those cultured on a pure PLA mat. Moreover, hADSCs cultured on PDA/PLA showed up-regulation of the ang-1 and vWF proteins associated with angiogenesis differentiation. Our results demonstrate that the bio-inspired coating synthetic degradable PLA polymer can be used as a simple technique to render the surfaces of synthetic biodegradable fibers, thus enabling them to direct the specific responses of hADSCs.

Poly(dopamine) coating of 3D printed poly(lactic acid) scaffolds for bone tissue engineering.

3D printing is a versatile technique to generate large quantities of a wide variety of shapes and sizes of polymer. The aim of this study is to develop functionalized 3D printed poly(lactic acid) (PLA) scaffolds and use a mussel-inspired surface coating to regulate cell adhesion, proliferation and differentiation of human adipose-derived stem cells (hADSCs). We prepared PLA 3D scaffolds coated with polydopamine (PDA). The chemical composition and surface properties of PDA/PLA were characterized by XPS. PDA/PLA modulated hADSCs' responses in several ways. Firstly, adhesion and proliferation, and cell cycle of hADSCs cultured on PDA/PLA were significantly enhanced relative to those on PLA. In addition, the collagen I secreted from cells was increased and promoted cell attachment and cell cycle progression were depended on the PDA content. In osteogenesis assay, the ALP activity and osteocalcin of hADSCs cultured on PDA/PLA were significantly higher than seen in those cultured on pure PLA scaffolds. Moreover, hADSCs cultured on PDA/PLA showed up-regulation of the ang-1 and vWF proteins associated with angiogenic differentiation. Our results demonstrate that the bio-inspired coating synthetic PLA polymer can be used as a simple technique to render the surfaces of synthetic scaffolds active, thus enabling them to direct the specific responses of hADSCs.

Clinical outcome and radiographic change of ipsilateral scapular neck and clavicular shaft fracture: comparison of operation and conservative treatment.

OBJECTIVE: The purpose of this study is to compare glenopolar angle (GPA) and the functional outcomes of fixation of both the clavicle and the scapular neck, fixation of the clavicle alone, and conservative treatment for floating-shoulder injuries. METHODS: A prospective stratified randomized study was performed in 39 adult patients who suffered floating-shoulder injuries and underwent fixation of both the clavicle and the scapular neck (group A), or fixation of the clavicle alone (group B), or conservative treatment (group C) between January 2005 and September 2011. The GPA, Disabilities of the Arm, Shoulder and Hand (DASH) score, and Constant-Murley Shoulder Outcome (Constant) score were compared between the three groups. RESULTS: All 39 patients were followed up for more than 2 years. GPA after bony consolidation was significantly better in group A than in groups B and C (p = 0.015). Functional outcomes measured by DASH and Constant scores were significantly better in group A at final follow-up (p = 0.008 and 0.002, respectively). Both DASH and Constant scores were highly correlated with GPA after consolidation (p < 0.001, respectively). The receiver operating characteristic (ROC) analysis showed that of the two randomly selected DASH scores, the smaller DASH score would have a larger GPA than the larger DASH score. Similarly, the larger Constant score would have a larger GPA than the smaller Constant score. CONCLUSIONS: Fixation of both the clavicle and the scapular neck may correct GPA and improve functional outcomes for the treatment of floating-shoulder injuries. GPA after fracture consolidation is a useful prognostic indicator of a satisfactory clinical outcome as defined by either DASH score or Constant score.

Urea permeation and hydrolysis through hollow fiber dialyzer immobilized with urease: storage and operation properties.

The surface of polyacrylonitrile hollow fibers was hydrolyzed and covalently bonded with urease via glutaraldehyde. Immobilized urease retained higher relative activity than native urease when storing at various pHs. The stabilities of immobilized urease to pH were higher than those of native enzyme. Immobilized urease retained 86% of initial activity after reusing 15 times at pH 7. After storing for 42d at 4 degrees C and pH 7, the immobilized urease can hydrolyze 15% of initial concentration of urea at pH 7 and 37 degrees C after 4h, while native urease lost almost its catalytic ability. The removal of urea using urease-immobilized dialyzer was demonstrated with in vitro dialysis and showed faster removing rate of urea than a regular dialyzer by 2 times. Furthermore, the improvement in the urea clearance by the urease immobilization to a dialyzer increased with the dialysate velocity.

Cholesterol oxidation using hollow fiber dialyzer immobilized with cholesterol oxidase: effect of storage and reuse.

Cholesterol oxidase (COD) was covalently bonded onto the surface of polyacrylonitrile (PAN) hollow fiber via glutaraldehyde. The optimum conditions were found to be at pH 7, -20 degrees C and in the dry state. Immobilized COD retained relative activity above 42% after a 30-day period, when storing at 25 degrees C. After reusing for 30 times, the activity of dry-stored immobilized COD retained 58% of activity. By rinsing with PBS, the decrease in the activity can be greatly reduced. At a flow speed of 1.09cm/s, the immobilized COD can decrease the concentration of cholesterol by about 40% in a 4-h run.

Cholesterol oxidation using hollow fiber dialyzer immobilized with cholesterol oxidase: preparation and properties.

The surface of polyacrylonitrile hollow fibers were hydrolyzed and covalently bonded with cholesterol oxidase (COD) via glutaraldehyde. The immobilized amount of the COD increased with the concentration of glutaraldehyde. However, COD immobilized with 10% glutaraldehyde had higher activity than with other concentrations. The stabilities of immobilized COD to pH and temperature were higher than those of native enzyme. The immobilized enzyme retained 80% of initial activity after 15 days when stored at 4 degrees C, which was longer than native COD. After being reused six times, the COD-immobilized hollow fiber retained more than 80% of the activity.

The effect of pH on the decomposition of hydrophenols in aqueous solutions by ultraviolet direct photolysis and the ultraviolet-hydrogen peroxide process.

The decomposition of catechol (o-hydrophenol) and resorcinol (m-hydrophenol) in aqueous solution by 254-nm UV direct photolysis and the UV-hydrogen peroxide (H2O2) process under various solution pH values was studied. The light absorbance and photolytic properties of catechol and resorcinol were found to be highly dependent on solution pH and can be adequately described with a three-species distribution model. For the UV-H2O2 process, the individual contribution to the decomposition of pollutants by direct photolysis and indirect hydroxyl radical destruction was differentiated by studying the linear addition of UV light absorbance of various reactant species. The contribution to the decomposition of the two hydrophenols by hydroxyl radical destruction was more than 95% in acidic and neural solutions for treatment with the UV-H2O2 process.

AuNPs@mesoSiO2 composites for SERS detection of DTNB molecule.

A surface enhanced Raman spectroscopy (SERS) substrate made of gold nanoparticles-embedded mesoporous silica (AuNPs@mesoSiO2) varying with size and gold concentrations was applied for SERS applications. In this study, the AuNPs@mesoSiO2 substrate produced notable intensity and more distinguishable peaks when compared with the spectra collected directly from an Au/Cr-coated substrate with regard to the detection of a lower concentration of chemicals or environmental contamination. Both aqueous and dried coffee rings of 5-5'-Dithio-bis (2-nitrobenzoic acid) (DTNB) solutions were examined. SERS spectra obtained from the substrate showed more fingerprint peaks with significant enhancement on the spectra signals. The correlation between the SERS signal and the DTNB concentration was found to be linear within a range of 10(-2) to 10(-12) M. SERS enhancement between 25 and 8 times greater can be achieved for DTNB detection using AuNPs@mesoSiO2 compared with the normal Raman spectra obtained from the aqueous solution and the contact line of the dried coffee ring, respectively.

Osteogenesis of human adipose-derived stem cells on hydroxyapatite-mineralized poly(lactic acid) nanofiber sheets.

Electrospun fiber sheets with various orientations (random, partially aligned, and aligned) and smooth and roughened casted membranes were prepared. Hydroxyapatite (HA) crystals were in situ formed on these material surfaces via immersion in 10× simulated body fluid solution. The size and morphology of the resulting fibers were examined using scanning electron microscopy. The average diameter of the fibers ranged from 225±25 to 1050±150 nm depending on the electrospinning parameters. Biological experiment results show that human adipose-derived stem cells exhibit different adhesion and osteogenic differentiation on the three types of fiber. The cell proliferation and osteogenic differentiation were best on the aligned fibers. Similar results were found for phosphorylated focal adhesion kinase expression. Electrospun poly(lactic acid) aligned fibers mineralized with HA crystals provide a good environment for cell growth and osteogenic differentiation and thus have great potential in the tissue engineering field.

Chitosan-coated electrospun PLA fibers for rapid mineralization of calcium phosphate.

In this work, hydroxyapatite (HA) mineralized on chitosan (CS)-coated poly(lactic acid) (PLA) nanofiber mat was prepared and compared in terms of mineralization characteristics. Significant calcium phosphate crystals formed on various concentrations of CS-coated PLA fiber mat with better uniformity after 2h of incubation in 10 times simulated body fluid (10× SBF). X-ray diffraction results further indicated that the composition of the deposited mineral was a mixture of dicalcium phosphate dehydrates and apatite. Chitosan, a cationic polysaccharide, can promote more nucleation and growth of calcium phosphate under conditions of 0.4% chitosan concentrations. These results indicated that HA-mineralized on CS-coated PLA fiber mat can be prepared directly via simply using CS coating followed by SBF immersion, and the results also suggest that this composite can mimic structural, compositional, and biological functions of native bone and can serve as a good candidate for bone tissue engineering (BTE).

The role of integrin αv in proliferation and differentiation of human dental pulp cell response to calcium silicate cement.

INTRODUCTION: It has been proved that integrin αv activity is related to cell proliferation, differentiation, migration, and organ development. However, the biological functions of integrin αv in human dental pulp cells (hDPCs) cultured on silicate-based materials have not been explored. The aim of this study was to investigate the role of integrin αv in the proliferation and odontogenic differentiation of hDPCs cultured with the effect of calcium silicate (CS) cement and ß-tricalcium phosphate (TCP) cement. METHODS: In this study, hDPCs were cultured on CS and TCP materials, and we evaluated fibronectin (FN) secretion and integrin αv expression during the cell attachment stage. After small interfering RNA transfection targeting integrin αv, the proliferation and odontogenesis differentiation behavior of hDPCs were analyzed. RESULTS: The results indicate that CS releases Si ion-increased FN secretion and adsorption, which promote cell attachment more effectively than TCP. The CS cement facilitates FN and αv subintegrin expression. However, the FN adsorption and integrin expression of TCP are similar to that observed in the control dish. Integrin αv small interfering RNA inhibited odontogenic differentiation of hDPCs with the decreased formation of mineralized nodules on CS. It also down-regulated the protein expression of multiple markers of odontogenesis and the expression of dentin sialophosphoprotein protein. CONCLUSIONS: These results establish composition-dependent differences in integrin binding and its effectiveness as a mechanism regulating cellular responses to biomaterial surface.

Osteogenesis and angiogenesis properties of dental pulp cell on novel injectable tricalcium phosphate cement by silica doped.

ß-Tricalcium phosphate (ß-TCP) is an osteoconductive material in clinical. In this study, we have doped silica (Si) into ß-TCP and enhanced its bioactive and osteostimulative properties. To check its effectiveness, a series of Si-doped with different ratios were prepared to make new bioactive and biodegradable biocomposites for bone repair. Formation of the diametral tensile strength, ions released and weight loss of cements was considered after immersion. In addition, we also examined the behavior of human dental pulp cells (hDPCs) cultured on Si-doped ß-TCP cements. The results showed that setting time and injectability of the Si-doped ß-TCP cements were decreased as the Si content was increased. At the end of the immersion point, weight losses of 30.1%, 36.9%, 48.1%, and 55.3% were observed for the cement doping 0%, 10%, 20%, and 30% Si into ß-TCP cements, respectively. In vitro cell experiments show that the Si-rich cements promote human dental pulp cell (hDPC) proliferation and differentiation. However, when the Si-doped in the cement is more than 20%, the amount of cells and osteogenesis protein of hDPCs was stimulated by Si released from Si-doped ß-TCP cements. The degradation of ß-TCP and osteogenesis of Si gives a strong reason to believe that these Si-doped ß-TCP cements may prove to be promising bone repair materials.

A filter-like AuNPs@MS SERS substrate for Staphylococcus aureus detection.

An accurate, highly sensitive and rapid identification assay of cells is extremely important in areas such as medical diagnosis, biological research, and environmental monitoring. Laboratory examinations of clinical isolates require time-consuming and complex processes to identify the colony count, with approximately 10(6)-10(8) cells needed for the characterization of strains. In the present study, a highly sensitive SERS filter-like substrate is prepared with AuNPs embedded in mesoporous silica (denoted as AuNPs@MS) synthesized by a simple one-spot method, and an example of its use for the filtration and concentration of analytes from aqueous samples is reported. In an application for Staphylococcus aureus SERS discrimination, the results show that the target cells can be concentrated on the filter-like AuNPs@MS substrates within a few seconds, with much better reproducibility with regard to the SERS spectra that are obtained. The experimental findings suggest that the AuNPs@MS substrate supports much higher intensity with more distinguishable peaks compared to Au/Cr-coated substrate, and the reproducibility is also significantly improved. The substrates investigated in this study generated 900 times more SERS signals at a concentration of 10(6)CFU/mL in the detection of S. aureus on mesoporous silica (Au wt%=0) when using AuNPs@MS with 16 wt% AuNPs. The limitation of this filter-like SERS substrate can be applicable for small volume samples (few to hundred microliter).