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Study of novel chitosan-gelatin artificial skin in vitro.

A novel absorbable scaffold composed of chitosan and gelatin was fabricated by freezing and lyophilizing methods, resulting in an asymmetric structure. This bilaminar texture is suitable for preparing a bilayer skin substitute. The methods employed to confirm the applicability of this chitosan-gelatin scaffold as an ideal skin substitute were a water uptake ability test, in vitro fibroblast proliferation, and scaffold tests in which fibroblasts were co-cultured with keratinocytes. The chitosan-gelatin scaffolds were more wettable and adsorbed more water than did chitosan alone. In static cell culture the thinner scaffold is better than the thicker one, and because of diffusion limitations in the scaffold, culture time must be within 3 weeks before transplantation to living tissues. Keratinocytes were co-cultured with fibroblasts in chitosan-gelatin scaffolds to construct an artificial bilayer skin in vitro. The artificial skin obtained was flexible and had good mechanical properties. Moreover, there was no contraction observed in the in vitro cell culture tests. The data from this study suggest that chitosan-gelatin scaffolds are suitable for skin tissue engineering goals.

Absorption↗

Control of cell adhesion and detachment using temperature and thermoresponsive copolymer grafted culture surfaces.

The hydrophobic monomer, n-butyl methacrylate (BMA) has been incorporated into thermoresponsive poly(N-isopropylacrylamide) (PIPAAm) to lower PIPAAm phase transition temperatures necessary for systematically regulating cell adhesion on and detachment from culture dishes at controlled temperatures. Poly(IPAAm-co-BMA)-grafted dishes were prepared by electron beam irradiation methods, systematically changing BMA content in the feed. Copolymer-grafted surfaces decreased grafted polymer transition temperatures with increasing BMA content as shown by water wettabilities compared to homopolymer PIPAAm-grafted surfaces. Bovine endothelial cells readily adhered and proliferated on copolymer-grafted surfaces above collapse temperature at 37 degrees C, finally reaching confluence. Cell sheet detachment behavior from copolymer-grafted surfaces depended on the culture temperature and BMA content. In conclusion, cell attachment/detachment can be controlled to an arbitrary temperature by varying the content of hydrophobic monomer incorporated into PIPAAm grafted to culture surfaces.

Animals↗

Rat bone marrow stromal cell osteogenic differentiation and fibronectin adsorption on chitosan membranes: the effect of the degree of acetylation.

Cell adhesion, migration, and proliferation of a few anchorage-dependent cells cultured on chitosan (Ch) matrices are influenced by the degree of N-acetylation (DA) of Ch. In the present work, we examined the influence of the DA on the attachment, spreading, proliferation, and osteogenic differentiation of rat bone marrow stromal cells (rBMSCs). Ch membranes were characterized in terms of surface morphology, roughness, and wettability, and in terms of adsorption of an adhesive protein, fibronectin (Fn). Chs with DAs in the range of 4 to 49% were used. Among the Ch samples, the DA of 4% led to the highest Fn surface concentration, both from single protein solution and from diluted serum. Furthermore, the levels of Fn adsorbed from serum found for this DA were threefold higher than for the tissue culture polystyrene control, indicating that in the presence of competitive proteins Ch is more specific toward Fn adsorption than tissue culture polystyrene. rBMSCs cultured on Ch carrying a DA of 4% were able to spread, proliferate, and differentiate, reaching a higher level of osteogenic differentiation than on the control, despite the lower cell attachment observed for all Ch samples. Because the Ch sample with a DA of 4% showed the highest Fn adsorption from serum, we suggest that cell adhesion, spreading, and osteogenic differentiation of rBMSCs on Ch may be mediated by the adsorbed layer of Fn.

Acetylation↗

Attachment, growth, and activity of rat osteoblasts on polylactide membranes treated with various low-temperature radiofrequency plasmas.

Nonporous and porous membranes from poly(L/DL-lactide) 80/20% were treated with low-temperature oxygen, ammonia, or sulphur dioxide-hydrogen plasmas and the late effects of plasma treatment on physicochemical characteristics of the membranes' surface were analyzed. The plasma treatment resulted in the permanent attachment of sulphur and nitrogen functionalities to the membrane's surface, and increased the surface concentration of oxygen, thereby increasing the surface wettability. To assess whether the plasma treatment affects the cellular response, primary rat osteoblasts were cultured on nontreated and plasma-treated nonporous and microporous membranes, and attachment, growth, and activity of cells were investigated. It was found that attachment and growth of osteoblasts on all the plasma-treated membranes were greater compared with nontreated controls. The treatment with ammonia plasma was most efficacious. The beneficial effects of plasma treatment on cells were most pronounced for microporous polylactide membranes irrespective of the plasma used. The results of the study suggest that the treatment of porous polylactide structures with plasma can be an effective means of enhancing their suitability for tissue engineering. Plasma exposure may also have an advantageous effect on bone healing when polylactide membranes are used to treat bone defects.

Animals↗

Biomechanical evaluation of the interfacial strength of a chemically modified sandblasted and acid-etched titanium surface.

The functional capacity of osseointegrated dental implants to bear load is largely dependent on the quality of the interface between the bone and implant. Sandblasted and acid-etched (SLA) surfaces have been previously shown to enhance bone apposition. In this study, the SLA has been compared with a chemically modified SLA (modSLA) surface. The increased wettability of the modSLA surface in a protein solution was verified by dynamic contact angle analysis. Using a well-established animal model with a split-mouth experimental design, implant removal torque testing was performed to determine the biomechanical properties of the bone-implant interface. All implants had an identical cylindrical shape with a standard thread configuration. Removal torque testing was performed after 2, 4, and 8 weeks of bone healing (n = 9 animals per healing period, three implants per surface type per animal) to evaluate the interfacial shear strength of each surface type. Results showed that the modSLA surface was more effective in enhancing the interfacial shear strength of implants in comparison with the conventional SLA surface during early stages of bone healing. Removal torque values of the modSLA-surfaced implants were 8-21% higher than those of the SLA implants (p = 0.003). The mean removal torque values for the modSLA implants were 1.485 N m at 2 weeks, 1.709 N m at 4 weeks, and 1.345 N m at 8 weeks; and correspondingly, 1.231 N m, 1.585 N m, and 1.143 N m for the SLA implants. The bone-implant interfacial stiffness calculated from the torque-rotation curve was on average 9-14% higher for the modSLA implants when compared with the SLA implants (p = 0.038). It can be concluded that the modSLA surface achieves a better bone anchorage during early stages of bone healing than the SLA surface; chemical modification of the standard SLA surface likely enhances bone apposition and this has a beneficial effect on the interfacial shear strength.

Biocompatible Materials↗

Characterization of nano hydroxyapatite/collagen surfaces and cellular behaviors.

Osseointegration at the bone-implant interface is a prerequisite for endosseous implants to succeed in achieving and maintaining their long-term stability in bone tissue. The achievement of osseointegration is significantly affected by surface nature of implants. To optimize osseointegration, this study presents the characterization of synthesized nanocrystalline hydroxyapatite (nano HA) and in vitro studies on nano HA, nano-HA/collagen, and titanium surfaces. Voids were found within the grain of nano HA, which consisted of the shell and the core. The finding assists the clarification of microstructures of nano HA. By low-temperature mixing nano-HA sol with collagen gel (nano-HA/collagen 80:20), nano HA, and nano-HA/collagen coated on pure titanium or porous anodic titanium oxides resulted in higher wettability and lower roughness. The in vitro studies showed that porous structures produced by anodic oxides on titanium served as positive anchorage sites for cell filopodia to connect, and nano HA decreased cell attachment of osteoblasts and induced well-developed long filopodia and broad lamellipodia, thereby enhancing cellular motility. Collagen involvement enhanced cell adhesion to nano HA. Cell reactions to nano HA, nano-HA/collagen, native, and porous titanium surfaces provide some guidance for an optimal osseointegration by their application in surface modifications for implants.

Cell Line↗

A poly(lactide-co-glycolide)/hydroxyapatite composite scaffold with enhanced osteoconductivity.

Biodegradable polymer/ceramic scaffolds can overcome the limitations of conventional ceramic bone substitutes. However, the conventional methods of polymer/ceramic scaffold fabrication often use organic solvents, which might be harmful to cells or tissues. Moreover, scaffolds fabricated with the conventional methods have limited ceramic exposure on the scaffold surface since the polymer solution envelopes the ceramic particles during the fabrication process. In this study, we developed a novel fabrication method for the efficient exposure of ceramic onto the scaffold surface, which would enhance the osteoconductivity and wettability of the scaffold. Poly(D,L-lactide-co-glycolide)/nanohydroxyapatite (PLGA/HA) scaffolds were fabricated by the gas foaming and particulate leaching (GF/PL) method without the use of organic solvents. Selective staining of ceramic particles indicated that HA nanoparticles exposed to the scaffold surface were observed more abundantly in the GF/PL scaffold than in the conventional solvent casting and particulate leaching (SC/PL) scaffold. Both types of scaffolds were implanted to critical size defects in rat skulls for 8 weeks. The GF/PL scaffolds exhibited significantly enhanced bone regeneration when compared with the SC/PL scaffolds. Histological analyses and microcomputed tomography of the regenerated tissues showed that bone formation was more extensive on the GF/PL scaffolds than on the SC/PL scaffolds. Compared with the SC/PL scaffolds, the enhanced bone formation on the GF/PL scaffolds may result from the higher exposure of HA nanoparticles to the scaffold surface. These results show that the biodegradable polymer/ceramic composite scaffolds fabricated with the novel GF/PL method can enhance bone regeneration compared with those fabricated with the conventional SC/PL method.

Absorbable Implants↗

Properties and performance of sulfide-substituted plasticized poly(vinyl chloride) as a biomaterial.

Plasticized poly(vinyl chloride) (PVC) was surface modified by nucleophilic substitution of the chlorine atoms of PVC by sulfide ions in aqueous media in the presence of a phase-transfer catalyst (PTC) to make it migration resistant. The modified PVC was sterilized by steam autoclaving and gamma radiation and subjected to plasticizer extraction in an extraction medium such as hexane. Virtually no plasticizer migrated out of modified PVC into the medium even after prolonged incubation for 30 d. The properties of the modified surface were evaluated by contact angle measurements, scanning electron microscopy (SEM), cell-culture studies, hemolysis assay, and whole-blood clotting time measurements. The mechanical properties of modified sheets were evaluated by measurements of tensile strength and strain at break after subjecting the specimens to different modes of sterilization. The modified PVC surface showed enhanced wettability and was found to be non-hemolytic and non-cytotoxic. Whole-blood clotting profiles of unmodified and modified PVC were virtually the same. Modification imparted slight yellow color to the material. Surface modification resulted in an 8% decrease in the ultimate stress and 28% decrease in the strain at break values. The sulfide-substituted PVC was found to be blood compatible and would be useful in applications such as tubing for transfusion, dialysis etc., where migration resistance is important.

Animals↗

Electrospinning of gelatin fibers and gelatin/PCL composite fibrous scaffolds.

In this article, ultrafine gelatin (Gt) fibers were successfully produced with the use of the electrical spinning or electrospinning technique. A fluorinated alcohol of 2,2,2-trifluoroethanol (TFE) was used as the dissolving solvent. The morphology of the electrospun gelatin fibers was found to be dependent on the alteration of gelatin concentration ranging from 2.5% w/v to 12.5% w/v at 2.5% increment intervals. Based on the electrospun gelatin fibers obtained, 10% w/v gelatin/TFE solution was selected and mixed with 10% w/v poly(epsilon-caprolactone) (PCL) in TFE at a ratio of 50:50 and co-electrospun to produce gelatin/PCL composite membranes. Contact-angle measurement and tensile tests indicated that the gelatin/PCL complex fibrous membrane exhibited improved mechanical properties as well as more favorable wettability than that obtained from either gelatin or PCL alone. The gelatin/PCL fibrous membranes were further investigated as a promising scaffold for bone-marrow stromal cell (BMSC) culture. Scanning electron microscopy (SEM) and laser confocal microscopy observations showed that the cells could not only favorably attach and grow well on the surface of these scaffolds, but were also able to migrate inside the scaffold up to 114 microm within 1 week of culture. These results suggest the potential of using composite gelatin/PCL fibrous scaffolds for engineering three-dimensional tissues.

Animals↗

Scale microornamentation of uropeltid snakes.

Microornamentation was examined on the exposed oberhautchen surface of dorsal, lateral, and ventral scales from the midbody region of 20 species of the fossorial snake family Uropeltidae and seven species of fossorial scolecophidian and anilioid outgroups. No substantial variation was observed in microornamentation from the different areas around the midbody circumference within species. All oberhautchen cells were flat and exhibited no major surface features other than occasional posterior margin denticulations, small pores/pits, and narrow, low ridges. This is largely consistent with the hypothesis that friction reduction and dirt shedding are the main selective pressures on microornamentation, given that reducing shine is not of key importance in fossorial animals. Variations among taxa were observed in the shape and size of oberhautchen cells, in the presence of pores/pits, in the presence and size of denticulations on posterior cell margins, and in the level or imbricate nature of cell borders. Six microornamentation characters were formulated, scored, and plotted onto a selected phylogeny. Character evolution and phylogenetic signal were explored, accepting the incomplete understanding of intraspecific variation and of uropeltid interrelationships. There is evidence that all but one of these characters evolved homoplastically, probably by multiple independent origin. There is no clear evidence for character state reversal, but greater phylogenetic resolution is required to test this further. Phylogenetic signal appears to exist in some instances, including possible microornamentation synapomorphies for Uropeltidae and Melanophidium. These derived character states are found elsewhere within Squamata. A microornamentation of narrow, finely, and regularly spaced ridges is associated with scale iridescence. These ridges, and possibly pores/pits, are also associated with scales that are less wettable, and that therefore might be expected to be better at shedding dirt in moist conditions. Testable hypotheses are presented that might explain minor variations in the form of ridges and pits among uropeltids.

Animals↗

Immediate drug release from solid oral dosage forms.

Fast drug release from solid dosage forms requires a very fast contact of the vast majority of the drug particles with the solvent; this, however, is particularly delayed in tablets and granulations. Starch and cellulose substances favor the matrix disintegration during the starting phase and the generation of the effective dissolution surface of the drug substance, thereby. To investigate the very complex interrelation between the functionality of commonly used excipients and the structural effects of the production processes, wettability, porosity, water uptake, and drug release rates of several ketoprofen-excipient preparations (powder blends, granulations, tablets) were measured. Significant linear correlation between these parameters, however, was not achieved; only qualitative tendencies of the effects could be detected. In consequence, a general mathematical model describing the mechanistic steps of drug dissolution from solid dosage forms in a fully correct way was not realized. However, the time-dependent change of the effective dissolution surface follows stochastic models: a new dissolution equation is based on the differential Noyes-Whitney equation combined with a distribution function, e.g. the lognormal distribution, and numerically solved with the software system EASY-FIT by fitting to the observations. This new model coincides with the data to a considerably higher degree of accuracy than the Weibull function alone, particularly during the starting, matrix disintegration, and end phases. In combination with a procedure continuously quantifying the dissolved drug, this mathematical model is suitable for the characterization and optimization of immediate drug release by the choice and modification of excipients and unit operations. The interdependence of some characteristic effects of excipients and production methods is discussed.

Algorithms↗

Water adsorption kinetics and contact angles of pharmaceutical powders.

Water sorption kinetics and water contact angles have been characterized for a range of pharmaceutical powders: ambroxol hydrochloride, griseofulvin, N,n-octyl-D-gluconamide, paracetamol, sulfathiazole, and theophylline. The uptake of water by powder samples at saturated vapor pressure was modeled using a pseudo first-order kinetic relationship. Parameters from this model have been correlated with the concentration and reactivity of the active surface sites of the pharmaceutical powders and their contact angles. The study has shown that analysis of water adsorption kinetics can be a powerful technique for characterizing the surface chemistry and wettability of pharmaceutical powders, and is particularly sensitive to their surface modification through excipient adsorption: ethyl(hydroxyethyl)cellulose treatment of griseofulvin and butyryl chloride treatment of sulfathiazole are reported as case studies.

Adsorption↗

Physicochemical and biopharmaceutical characterization of endo-2-(8-methyl-8-azabicyclo[3.2.1]oct-3-yl)-2,3-dihydro-1H-benz[e]isoindol-1-one (CR3124) a novel potent 5-HT3 receptor antagonist.

The physicochemical and biopharmaceutical properties, such as pK(a), crystal habit, water solubility, logD, molecular structure and dynamics, and membrane permeability of CR3124 (endo-2-(8-methyl-8-azabicyclo[3.2.1]oct-3-yl)-2,3-dihydro-1H-benz[e]isoindol-1-one, a novel potent 5-HT(3) receptor antagonist) have been studied in order to obtain preformulation information. The study showed that CR3124 is a very rigid molecule in which conformational freedom due to the presence of a rotatable bond is restricted by the interaction between an activated hydrogen and the amide oxygen and the conformation of the tropane piperidine ring is regulated by the environment in such a manner as to optimize the intermolecular interactions with the solvent. This chameleon behavior appears to be capable of explaining the biopharmaceutical properties showed by CR3124, such as low wettability, relatively good solubility, and very high membrane permeability.

1-Octanol↗

Hot-melt extrusion for enhanced delivery of drug particles.

With the recent advent of nanotechnology for pharmaceutical applications, drug particle engineering is the focus of increasing interest as a viable approach for overcoming solubility limitations of poorly water-soluble drugs. Although these particle engineering techniques have been proven successful for enhancing the dissolution properties of many poorly water-soluble drugs, there are limitations associated with them such as particle aggregation, morphological instability, and poor wettability. The aim of this study was to demonstrate a processing technique in which hot-melt extrusion (HME) is utilized to overcome these limitations. Micronized particles of amorphous itraconazole (ITZ) stabilized with PVP or HPMC were produced and subsequently melt extruded with poloxamer 407 and PEO 200 M to deaggregate and disperse the particles into the hydrophilic polymer matrix. Differential scanning calorimetry, X-ray diffraction, and scanning electron microscopy were used to demonstrate that the HME process did not alter the properties of the micronized particles. Dissolution testing conducted at sink conditions revealed that the dissolution rate of the micronized particles was improved by HME due to particle deaggregation and enhanced wetting. Supersaturation dissolution testing demonstrated that the ITZ-HPMC micronized particle extrudates provided superior supersaturation of ITZ compared to the ITZ-PVP micronized particle extrudates. Supersaturation dissolution testing incorporating a pH change (from pH 1.2 to 6.8 at 2 h) revealed that neither micronized particle extrudate formulation significantly reduced the rate of ITZ precipitation from supersaturated solution once pH was increased. Moreover, the two extrudate formulations performed very similarly when only considering dissolution testing from just before pH adjustment through the duration of testing at neutral pH. From oral dosing of rats, it was determined that the two extrudate formulations performed similarly in vivo as confirmed by their statistically equivalent AUC values. By correlating the results of supersaturation dissolution testing with pH change to the in vivo AUC, it appears that rapid precipitation of ITZ occurs upon entrance into the more neutral pH environment of the small intestine resulting in a brief opportunity for absorption. This suggests that perhaps the optimum formulation approach for ITZ is to control drug release so as to retard precipitation as pH is increased and extend the absorption window in the small intestine.

Animals↗

Effect of phthalate plasticizer on blood compatibility of polyvinyl chloride.

The amount of a phthalate plasticizer on the surface of a sheet of polyvinyl chloride used in the fabrication of blood storage bags was quantified using attenuated total reflectance spectroscopy, water contact angle measurement, and weight loss due to methanol extraction. Water wettability increased as the amount of surface phthalate extracted by methanol increased, which indicates that the accumulation of phthalate on the surface increases hydrophobicity. The extraction of phthalate by methanol consists of two steps: (a) methanol first dissolves surface phthalate, and (b) phthalate in the bulk then diffuses through the surface. The adsorption of plasma proteins was investigated to determine the initial events as blood contacts the surface. The composition of adsorbed proteins on the methanol-cleansed surface differs from that on the uncleansed polyvinyl chloride surface. Albumin adsorption onto phthalate-contaminated surfaces is less than on cleansed surfaces while adsorption of gamma-globulin and fibrinogen is greater on phthalate-contaminated surfaces. Protein adsorption can be related to surface thrombus formation. Increases in platelet numbers appeared on phthalate-contaminated surfaces as compared with methanol-treated surfaces. A phthalate may enhance platelet adhesion and aggregation when it exists on a polymer surface.

Adsorption↗

Dissolution rates of corticosteroids utilizing sugar glass dispersions.

A method of increasing the dissolution rates of some orally administered corticosteroids was investigated. This method involved glass dispersions using dextrose, galactose, and sucrose as the carriers. These dispersions were prepared by the fusion process and were subjected to a modified NF XIII dissolution rate determination. The results revealed a marked increase in the dissolution rate of the corticosteroids contained in the solid dispersions when compared to the dissolution rate of the plan corticosteroid powder. The increase in dissolution rates was attributed to the presence of the corticosteroid in a very fine state of subdivision and to the increased wettability of the corticosteroid powder.

Betamethasone↗

Effect of primidone concentration on glass transition temperature and dissolution of solid dispersion systems containing primidone and citric acid.

The glass transition temperatures of glasses containing various concentrations of primidone in citric acid were measured and found to increase as the primidone concentration increased. Dissolution studies of these systems and particle-size measurements of primidone precipitated during dissolution of devitrified glasses suggest that the increase in the dissolution rate of the devitrified systems is due to both the small size of the precipitated crystals and the excellent wettability of these systems.

Chemistry, Pharmaceutical↗

Direct preparation of spherically agglomerated salicylic acid crystals during crystallization.

Needle-like salicylic acid crystals were transformed into a spherically shaped dense form during crystallization by the spherical crystallization technique. Agitation of a mixture of ethanol-water-chloroform containing salicylic acid yielded spherically agglomerated salicylic acid crystals. The crystallinity of the agglomerated salicylic acid the amount of ethanol in the solvent mixture was decreased. The wettability of the agglomerated crystals increased when the amount of ethanol in the solvent mixture was decreased, and this enhanced the dissolution rate of the crystals. The remarkable improvements in the flow and packing of the agglomerated crystals enabled the direct compression of the crystals.

Chemical Phenomena↗