Margarida Figueiredo

Microencapsulation of hemoglobin in chitosan-coated alginate microspheres prepared by emulsification/internal gelation

Chitosan-coated alginate microspheres prepared by emulsification/internal gelation were chosen as carriers for a model protein, hemoglobin (Hb), owing to nontoxicity of the polymers and mild conditions of the method. The influence of process variables related to the emulsification step and microsphere recovering and formulation variables, such as alginate gelation and chitosan coating, on the size distribution and encapsulation efficiency was studied. The effect of microsphere coating as well its drying procedure on the Hb release profile was also evaluated. Chitosan coating was applied by either a continuous microencapsulation procedure or a 2-stage coating process. Microspheres with a mean diameter of less than 30 μm and an encapsulation efficiency above 90% were obtained. Calcium alginate cross-linking was optimized by using an acid/CaCO3 molar ratio of 2.5, and microsphere-recovery with acetate buffer led to higher encapsulation efficiency. Hb release in gastric fluid was minimal for air-dried microspheres. Coating effect revealed a total release of 27% for 2-stage coated wet microspheres, while other formulations showed an Hb release above 50%. Lyophilized microspheres behaved similar to wet microspheres, although a higher total protein release was obtained with 2-stage coating. At pH 6.8, uncoated microspheres dissolved in less than 1 hour; however, Hb release from air-dried microspheres was incomplete. Chitosan coating decreased the release rate of Hb, but an incomplete release was obtained. The 2-stage coated microspheres showed no burst effect, whereas the 1-stage coated microspheres permitted a higher protein release.

In Situ Forming Chitosan Hydrogels Prepared via Ionic/Covalent Co-Cross-Linking

In situ forming chitosan hydrogels have been prepared via coupled ionic and covalent cross-linking. Thus, different amounts of genipin (0.05, 0.10, 0.15, and 0.20% (w/w)), used as a chemical cross-linker, were added to a solution of chitosan that was previously neutralized with a glycerol-phosphate complex (ionic cross-linker). In this way, it was possible to overcome the pH barrier of the chitosan solution, to preserve its thermosensitive character, and to enhance the extent of cross-linking in the matrix simultaneously. To investigate the contributions of the ionic cross-linking and the chemical cross-linking, separately, we prepared the hydrogels without the addition of either genipin or the glycerol-phosphate complex. The addition of genipin to the neutralized solution disturbs the ionic cross-linking process and the chemical cross-linking becomes the dominant process. Moreover, the genipin concentration was used to modulate the network structure and performance. The more promising formulations were fully characterized, in a hydrated state, with respect to any equilibrium swelling, the development of internal structure, the occurrence of in vitro degradability and cytotoxicity, and the creation of in vivo injectability. Each of the hydrogel systems exhibited a notably high equilibrium water content, arising from the fact that their internal structure (examined by conventional SEM, and environmental SEM) was highly porous with interconnecting pores. The porosity and the pore size distribution were quantified by mercury intrusion porosimetry. Although all gels became degraded in the presence of lysozyme, their degradation rate greatly depended on the genipin load. Through in vitro viability tests, the hydrogel-based formulations were shown to be nontoxic. The in vivo injection of a co-cross-linking formulation revealed that the gel was rapidly formed and localized at the injection site, remaining in position for at least 1 week.

Physicochemical characterization of biomaterials commonly used in dentistry as bone substitutes--comparison with human bone

The present work focuses on the physicochemical characterization of selected mineral-based biomaterials that are frequently used in dental applications. The selected materials are commercially available as granules from different biological origins: bovine, porcine, and coralline. Natural and calcined human bone were used for comparison purposes. Besides a classical rationalization of chemical composition and crystallinity, a major emphasis was placed on the measurement of various morphostructural properties such as particle size, porosity, density, and specific surface area. Such properties are crucial to acquiring a full interpretation of the in vivo performance. The studied samples exhibited distinct particle sizes (between 200 and 1000 microm) and shapes. Mercury intrusion revealed not only that the total sample porosity varied considerably (33% for OsteoBiol, 50% for PepGen P-15, and 60% for BioOss) but also that a significant percentage of that porosity corresponded to submicron pores. Biocoral was not analyzed by this technique as it possesses larger pores than those of the porosimeter upper limit. The density values determined for the calcined samples were close to the theoretical values of hydroxyapatite. However, the values for the collagenated samples were lower, in accordance with their lower mineral content. The specific surface areas ranged from less than 1 m(2)/g (Biocoral) up to 60 m(2)/g (BioOss). The chemical and phase composition of most of the samples, the exception being Biocoral (aragonite), were hydroxyapatite based. Nonetheless, the samples exhibited different organic material content as a consequence of the distinct heat treatments that each had received.

Comparative study of cellulose fragmentation by enzymes and ultrasound

Abstract The stability in aqueous suspensions of two particulate celluloses, Sigmacell type 100 and Avicel PH101, was analyzed. The effect of the presence of a cellulase from Trichoderma reesei , ionic strength, and ultrasonic agitation on the fragmentation/aggregation phenomena was studied. Particle size distributions of the powders were obtained with three different particle sizers: the Galai CIS 100, the Coulter Multisizer II, and the Malvern 2600c. The differences in the obtained absolute values are discussed according to the measuring principles of each technique; however, the overal conclusions are independent of the particle sizer used. The enzyme breaks up the Avicel aggregates more effectively than ultrasound while the Sigmacell particles are stable under the present experimental conditions. The stabilizing effect of cellulases was tentatively explained using the DLVO (Derjaguin, Landau, Verweye, and Overbeek) theory. The adsorbed enzyme did not change significantly the zeta potential of the fibers; hence, the stabilizing effect was attributed to a reduction in the attractive van der Waals forces and hydration effects.

Tailoring the properties of gelatin films for drug delivery applications: influence of the chemical cross-linking method.

Two types of chemically cross-linked gelatin films were prepared and characterized. The first type of films was cross-linked with 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide hydrochloride (EDC) under heterogeneous conditions and are named Gel-E. In the second type of films, gelatin was previously functionalized with methacrylamide side groups by the reaction with methacrylic anhydride and for that is named Gel-MA. The modified gelatin was subsequently cross-linked by a photoinitiated radical polymerization. These films were characterized relatively to their degree of cross-linking, buffer uptake capacity, resistance to hydrolytic and proteolytic degradation, and mechanical and thermal properties. Results show that the employed cross-linking method, together with the degree cross-linking, dictate the final properties of the films. Gel-E films have significant lower buffer uptake capacities and higher resistance to collagenase digestion when compared to Gel-MA films. Additionally, Gel-E films exhibit higher values of stress at break and lower strains at break. Moreover, the films properties could be modified by varying the extent of the chemical cross-linking, which in turn could be controlled by varying the concentration of EDC, for the first type of films (Gel-E), or by using gelatins with different degrees of functionalization, in the case of the second type of films (Gel-MA).

Shrinkage Characteristics and Porosity of Pears during Drying

The high moisture content of pears (about 80%) results in a high degree of shrinkage when drying takes place, which strongly affects the fruit structure and quality. The aim of the present work was to study the shrinkage characteristics of pears during air drying by evaluating their dimensions and volume along the drying process and by predicting particle and bulk densities as well as porosity as a function of material moisture content. The latter properties were estimated using a mathematical model available from the literature whose parameters were either directly measured or determined by fitting some of the equations of the proposed model to the experimental data. The results predicted were subsequently validated by independent measurements.

Dissolution rate studies of pharmaceutical multisized powders : a practical approach using the Coulter method

Abstract The aim of this work is the study of the influence of particle size and related properties on the dissolution rate of a sparingly soluble drug indomethacin. Different size fractions were fully characterized concerning particle size distribution, specific surface area, density, degree of crystallinity and solubility. A particle size counter the Coulter Multisizer II — was used, not only to characterize the primary particle size distribution of the different fractions, but also to monitor the size and number of the suspended particles during the dissolution process. Although this information was applied to evaluate dissolution profiles (dissolution drug concentration vs. time) it can be further used to thoroughly study the dissolution phenomenon. The accuracy of this instrument to assess dissolution profiles was confirmed by comparing its results with those obtained by HPLC. As expected a strong influence of the fraction size on the dissolution rate was found. A correlation was established between the mean dissolution time (MDT) and the mean particle size of the various indomethacin fractions.

Modelling and simulation of pear drying

In the present work a diffusion based model was adopted to represent the drying behaviour of pears in a continuous convective drier, taking into consideration the variation of the properties of the pears along drying. The program developed simulated a significant number of situations resultant from the variation of some of the operating conditions. The temperatures tested were 30, 40 and 50^oC, the air velocities were 0.5, 1.0 and 1.5m/s, and the relative humidity of the drying air was 40%, 50% and 60%. From the situations analyzed, and considering the specifications required for the final product, it was concluded that the drying should be carried out at 40^oC, with a drying air velocity of 1.5m/s and containing 60% RH.

Development of UV cross-linked gelatin coated electrospun poly(caprolactone) fibrous scaffolds for tissue engineering

Cardiovascular disease is the leading cause of morbidity and mortality among industrialized countries. Vascular grafts are often required for the surgical treatments. Considering the limitations associated with the use of autografts and with the currently available synthetic materials, a growing demand in tissue engineered vascular grafts has been registered. During the work here described, electrospinning technique was used to prepared fibrous matrices to be applied as vascular implants. For that purpose, electrospun polycaprolactone (PCL) fibrous mats were produced and afterwards coated with different hydrogel formulations based in photocrosslinkable gelatin (GelMA) and the macromers poly(ethylene glycol) acrylate (PEGA) and poly(ethylene glycol) diacrylate (PEGDA). These were further photocrosslinked under UV irradiation using Irgacure® 2959 (by BASF) as the photoinitiator. The suitability of the coated scaffolds for the intended application, was evaluated by assessing their chemical/physical properties as well as their interaction with blood and endothelial cells.

Comparison of a xenogeneic and an alloplastic material used in dental implants in terms of physico-chemical characteristics and in vivo inflammatory response

Two commercial bone grafts used in dentistry (Osteobiol Gen-Os®, a xenograft of porcine origin, and Bonelike®, a hydroxyapatite based synthetic material), in the form of granules, were characterized and evaluated in vivo regarding the intensity of the tissue inflammatory response. These biomaterials were characterized in terms of morphology, particle size distribution, porosity and pore size, specific surface area and density. The chemical composition and structure of the materials were accessed by Fourier-Transform Infrared Spectroscopy (FTIR) and X-ray Diffraction (XRD). The graft materials were implanted in the gluteus maximus muscles of Wistar rats and the inflammatory response was evaluated through histological analysis, after one week of implantation. The results showed that the two grafts have quite different characteristics in practically all the evaluated properties. While Osteobiol® exhibits a structure and composition similar to the natural bone, Bonelike® is constituted by a main crystalline phase of hydroxyapatite and two secondary phases of α- and β-tricalcium phosphate. Osteobiol® granules, besides being larger, are irregular, and exhibit sharp-edged tips, while those of Bonelike® are approximately cylindrical, with round contours, and more uniform in size. The in vivo response evaluated from the inflammatory infiltrates revealed that although both implants did not cause severe inflammation, Bonelike® granules elicit a consistently more intense inflammatory reaction than that triggered by the granules of Osteobiol®, particularly in terms of collagen production and formation of fibrous capsule. This reaction was partly explained in terms of the characteristics evaluated for the granules of this material.