Design and Characterization of Silicone and Surfactant Based Systems for Topical Drug Delivery

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Acção de extractos vegetais de camomila, macela e arnica do Brasil, no processo de modificação da cor dos cabelos, em diferentes veículos extractores

Apreocupação quanto ao aspecto sensorial faz com que vários formuladores opte pela utilização de diferentes tipos de silicones em cosméticos, uma vez que, além de formarem uma película protetora no cabelo não oclusiva, melhoram o aspecto final do produto. Plantas como Matricaria chamomilla (camomila), Achyrocline satureoides (macela) e Solidago microglossa (arnica de Brasil) são ricas em flavonóides e vêm sendo usadas em fórmulas destinadas ao clareamento dos cabelos. Deste modo, este trabalho tem como objetivo verificar a efetividade dos extractos dessas plantas, obtida por veículos extractores diferentes (hidroglicólico e hidrosilicônico), no clareamento capilar. Os extractos, obtidos por percolação e por digestão seguida de maceração, foram aplicados em mechas de cabelos castanhos claros, as quais posteriormente foram avaliadas por colorímetro de reflectância com o objetivo de avaliar a qualidade da extração em termos de material extraído. Os resultados obtidos indicam que dentre os diferentes veículos extractores, os melhores resultados foram obtidos com a mistura de água: DC 193 (PEG-12 dimeticone). Além disso, das três plantas pesquisadas, a camomila foi a que apresentou alterações estatisticamente significativas de cor nas mechas de cabelos, evidenciando a ação clareadora deste extracto.

Effect of surfactants and polyethylene glycol on the activity and stability of a lipase from oilseeds of Pachira aquatica

Lipases from oilseeds have a great potential for commercial exploration as industrial enzymes. Lipases are used mixed with surfactants in cleaning and other formulated products, and accordingly, both components must be compatible with each other. This work presents the results of the effects of anionic, cationic and nonionic surfactants, polyethylene glycol and urea on the activity and stability of a lipase extracted of oilseeds from Pachira aquatica. The enzyme was purified and the spectrophotometric assays were done using p-nitrophenyl acetate (p-NPA) as substrate pH 7.5 and 25 degrees C. The activity was significantly enhanced by the cationic surfactant CTAB. Bile salts increased the lipase activity in the tested concentration range, whereas anionic and nonionic surfactants showed an inhibitory effect. Aqueous solutions of PEG activated the lipase and maximum activation (161%) occurred in PEG 12,000. This effect on lipase that can be due to exposition of some hydrophobic residues located in the vicinity of the active site or aggregation.

Caracterização bioquímica de lipase extraída de sementes oleaginosas de Pachira aquatica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fabrication using a rapid prototyping system and in vitro characterization of PEG-PCL-PLA scaffolds for tissue engineering

n the field of tissue engineering new polymers are needed to fabricate scaffolds with specific properties depending on the targeted tissue. This work aimed at designing and developing a 3D scaffold with variable mechanical strength, fully interconnected porous network, controllable hydrophilicity and degradability. For this, a desktop-robot-based melt-extrusion rapid prototyping technique was applied to a novel tri-block co-polymer, namely poly(ethylene glycol)-block-poly(epsi-caprolactone)-block-poly(DL-lactide), PEG-PCL-P(DL)LA. This co-polymer was melted by electrical heating and directly extruded out using computer-controlled rapid prototyping by means of compressed purified air to build porous scaffolds. Various lay-down patterns (0/30/60/90/120/150°, 0/45/90/135°, 0/60/120° and 0/90°) were produced by using appropriate positioning of the robotic control system. Scanning electron microscopy and micro-computed tomography were used to show that 3D scaffold architectures were honeycomb-like with completely interconnected and controlled channel characteristics. Compression tests were performed and the data obtained agreed well with the typical behavior of a porous material undergoing deformation. Preliminary cell response to the as-fabricated scaffolds has been studied with primary human fibroblasts. The results demonstrated the suitability of the process and the cell biocompatibility of the polymer, two important properties among the many required for effective clinical use and efficient tissue-engineering scaffolding.

The effect of composition, electron irradiation and quenching on ionic conductivity in a new solid polymer electrolyte: (PEG) (x) NH4I

We have prepared, characterized and investigated a new PEG-2000 based solid polymer electrolyte (PEG) x NH4I. Ionic conductivity measurements have been made as a function of salt concentration as well as temperature in the range 265–330 K. Selected compositions of the electrolyte were exposed to a beam of 8 MeV electrons to an accumulated dose of 10 kGy to study the effect on ionic conductivity. The electrolyte samples were also quenched at liquid nitrogen temperature and conductivity measurements were made. The ionic conductivity at room temperature exhibits a characteristic double peak for the composition x = 20 and 70. Both electron beam irradiation and quenching at low temperature have resulted in an increase in conductivity by 1–2 orders of magnitude. The enhancement of conductivity upon irradiation and quenching is interpreted as due to an increase in amorphous region and decrease in crystallinity of the electrolyte. DSC and proton NMR measurements also support this conclusion.

Investigation of the (PEG)xLiCl system using conductivity, DSC and NMR techniques

Electrolytes based on polyethylene glycol (PEG, mol.wt.8000) and LiCl of compositions, (PEG)(x)LiCl, x=4, 6, 8, 10, 12, 40, 60, where x is the O/Li ratio, were prepared by solution casting from methanol solutions. FTIR studies indicate that the ether oxygens of the polymer chain participate in Li+ ion conduction. The presence of a salt-polymer complex that melts around 190 degrees C was evidenced by DSC measurements for the electrolytes with compositions x<12. The highest conductivity was obtained at the composition x=10 which was attributed to the presence of a mostly amorphous compound. NMR measurements indicated two regions of motional narrowing, one attributable to the glass transition and another to translational diffusion.

Enhanced lithium-ion transport in PEG-based composite polymer electrolyte with Mn0.03Zn0.97Al2O4 nanoparticles

The ion conduction and thermal properties of composite solid polymer electrolyte (SPE) comprising Poly(ethylene) Glycol (PEG, mol wt. 2000), lithium perchlorate (LiClO4) and insulating Mn0.03Zn0.97Al2O4 nanoparticle fillers were studied by complex impedance analysis and DSC techniques. The average size of the nanoparticles was determined by powder X-ray diffraction (XRD) using Scherrer's equation and was found to be similar to 8 nm. The same was also determined by TEM imaging and found to be similar to 12 nm. The glass transition temperature T, as measured by differential scanning calorimeter (DSC), showed a minimum at 5 mol% of narroparticles. Fractional crystallinity was determined using DSC. NMR was used to deter-mine crystallinity of a pure PEG sample, which was then used as the standard. Fractional crystallinity X. was the lowest for 5 mol% and beyond. The ionic conductivity of the composite polymer electrolyte containing 5 mol% Mn0.03Zn0.97Al2O4 nanoparticles was found to be 1.82 x 10(-5) S/cm, while for the pristine one, it was 7.27 x 10(-7) S/cm at room temperature. As a function of nanoparticle content, conductivity was observed to go through two maxima, one at around 5 mol% and another shallower one at around 12 mol%. The temperature dependence of conductivity could be divided into two regions, one consistent with Arrhenius behaviour and the other with VTF. We conclude that the enhancement of ionic conductivity on the addition of Mn0.03Zn0.97Al2O4 nanoparticles is a result of reduction in both the T, and the crystallinity. (C) 2002 Elsevier Science B.V. All rights reserved.

双结晶性PEG-PCL两嵌段共聚物的合成，结晶行为和形态

聚乙二醇－聚ε-己内酯两嵌段共聚物（PEG-PCL）由于其在生物医用材料中的潜在应用而受到广泛的关注。然而，研究表明这类嵌段共聚物的许多性质，如药物渗透性，降解性能和机械性质等，都要受到它们的结晶行为与聚集态结构的显著影响。而在本课题开始之前，还没有关于PEG-PCL的结晶行为与形态的系统研究报道。因此，本文希望通过对PEG-PCL两嵌段共聚物结晶行为与形态的研究，能为这类生物降解高分子材料的工业应用提供一定的科学依据。本文使用辛酸亚锡为催化剂，甲氧基聚乙二醇（mPEG）为大分子引发剂，合成了一系列分子量分布比较窄，PCL质量百分含量为0.16-0.93的PEG-PCL两嵌段共聚物。两嵌段共聚物中的PEG段分子量固定为5000，共聚物的组成通过改变PCL链段的长度来调节。本文使用DSC，WAXD，常温或变温FTIR详细研究了PEG-PCL的结晶和熔融行为，使用偏光显微镜（POM）观察了PEG-PCL的结晶形态及结晶生长行为，利用SAXS研究了PEG-PCL的微观形态，得出了如下结果：（1）WAXD与FTIR的结果表明，两嵌段共聚物中的PEG与PCL形成微相分离的结晶微区，不存在两者的共晶或混晶。PCL含量为0.23-0.87的两嵌段共聚物中都能观察到的PEG与PCL的结晶。变温FTIR结果显示，当PCL含量低于或等于0.36，两嵌段共聚物中的PEG先从熔体中结晶；反之，当PCL含量等于或大于0.43，则熔体中PCL结晶先出现。（2）DSC结果表明，随着PEG-PCL中PCL段长度的增加，PCL段的结晶和熔融温度显著增加；相反，PEG段的结晶和熔融温度则显著降低。当PCL的质量分数由0增加至0.93，PEG的结晶度从79%降低至0，然而PCL的结晶度却不是单调变化，而是出现一个最大值。（3）在POM下观察PEG-PCL的36 oC等温熔体结晶过程，当PCL质量分数不超过0.36时，在偏光显微镜下只能观察到PEG球晶；而当PCL质量分数大于或等于0.56时，只能观察到PCL球晶；PCL含量为0.43和0.50的两种两嵌段共聚物中观察到了一种独特的同心球晶，同心球晶的中心部分形态类似于PCL球晶，而外部则类似于PEG球晶。PEG球晶与PCL球晶生长速率受PCL含量的影响显著：当PCL质量分数从0增加至0.50，PEG球晶的生长速率大大降低；然而，PCL球晶的生长速率却不是单调变化，而是在PCL质量分数为0.62时达到最大值。（4）SAXS结果表明，结晶后的PEG-PCL的微区结构是由交替的PEG与PCL的层状微区组成。共聚物的长周期在PCL质量分数为0.50时达到最大值。当PCL质量分数由0增加至0.50时，由于PCL层厚度的显著增加，共聚物的长周期显著增加；而当PCL含量由0.50继续增加至0.87，由于PEG层厚度的急剧降低，又使得共聚物的长周期迅速降低。（5）首次利用POM和微区红外光谱详细研究了PEG-PCL50/50(w/w)同心球晶的形成过程，发现同心球晶的形成是由于一种独特的结晶动力学造成的。另外，即使同心球晶的中心和外部的形态差别巨大，但是红外显微镜结果显示，两部分的组成却是相同的。

Isothermal crystallization behavior of the poly(L-lactide) block in poly(L-lactide)-poly(ethylene glycol) diblock copolymers: Influence of the PEG block as a diluted solvent

Isothermal crystallization kinetics and morphology of the poly(L-lactide) block in poly(L-lactide)poly(ethylene glycol) diblock copolymers were studied by differential scanning calorimetry (DSC) and polarized optical microscopy (POM), respectively. The results were compared with that of the PLLA homopolymer. The introduction of the PEG block accelerated the crystallization rate of the PLLA block and promoted to form ring-banded spherulites. The analysis of isothermal crystallization kinetics has shown that the PLLA homopolymer accorded with the Avrami equation. But the PLLA block of the diblock copolymers deviated from the Avrami equation, which resulted from increasing of the crystallization rate and occurring of the second crystallization process. The equilibrium melting temperature (T,,) of the PLLA block fell with its molecular weight decreasing. The conditions to obtain more regular ring-banded spherulites were below: the sample was the PLLA block of LA(5) EG(5); the crystallization temperature was about from 95 degrees C to 100 degrees C, which almost corresponded to regime II.

Preparation and in vivo evaluation of insulin-loaded biodegradable nanoparticles prepared from diblock copolymers of PLGA and PEG

The aim of this study was to design a controlled release vehicle for insulin to preserve its stability and biological activity during fabrication and release. A modified, double emulsion, solvent evaporation, technique using homogenisation force optimised entrapment efficiency of insulin into biodegradable nanoparticles (NP) prepared from poly (dl-lactic-co-glycolic acid) (PLGA) and its PEGylated diblock copolymers. Formulation parameters (type of polymer and its concentration, stabiliser concentration and volume of internal aqueous phase) and physicochemical characteristics (size, zeta potential, encapsulation efficiency, in vitro release profiles and in vitro stability) were investigated. In vivo insulin sensitivity was tested by dietinduced type II diabetic mice. Bioactivity of insulin was studied using Swiss TO mice with streptozotocin-induced type I diabetic profile. Insulin-loaded NP were spherical and negatively charged with an average diameter of 200–400 nm. Insulin encapsulation efficiency increased significantly with increasing ratio of co-polymeric PEG. The internal aqueous phase volume had a significant impact on encapsulation efficiency, initial burst release and NP size. Optimised insulin NP formulated from 10% PEG-PLGA retained insulin integrity in vitro, insulin sensitivity in vivo and induced a sustained hypoglycaemic effect from 3 hours to 6 days in type I diabetic mice.

Effect of Polymer Architecture on the Structural and Biophysical Properties of PEG-PLA Nanoparticles

Polymers made of poly(ethylene glycol) chains grafted to poly(lactic acid) chains (PEG-g-PLA) were used to produce stealth drug nanocarriers. A library of comb-like PEG-g-PLA polymers with different PEG grafting densities was prepared in order to obtain nanocarriers with dense PEG brushes at their surface, stability in suspension, and resistance to protein adsorption. The structural properties of nanoparticles (NPs) produced from these polymers by a surfactant-free method were assessed by DLS, zeta potential, and TEM and were found to be controlled by the amount of PEG present in the polymers. A critical transition from a solid NP structure to a soft particle with either a “micelle-like” or “polymer nano-aggregate” structure was observed when the PEG content was between 15 to 25% w/w. This structural transition was found to have a profound impact on the size of the NPs, their surface charge, their stability in suspension in presence of salts as well as on the binding of proteins to the surface of the NPs. The arrangement of the PEG-g-PLA chains at the surface of the NPs was investigated by 1H NMR and X-ray photoelectron spectroscopy (XPS). NMR results confirmed that the PEG chains were mostly segregated at the NP surface. Moreover, XPS and quantitative NMR allowed quantifying the PEG chain coverage density at the surface of the solid NPs. Concordance of the results between the two methods was found to be remarkable. Physical-chemical properties of the NPs such as resistance to aggregation in saline environment as well as anti-fouling efficacy were related to the PEG surface density and ultimately to polymer architecture. Resistance to protein adsorption was assessed by isothermal titration calorimetry (ITC) using lysozyme. The results indicate a correlation between PEG surface coverage and level of protein interactions. The results obtained lead us to propose such PEG-g-PLA polymers for nanomedecine development as an alternative to the predominant polyester-PEG diblock polymers.

Interactions of KLVFF-PEG peptide conjugate with fibrinogen in neutral aqueous solutions

In this work we report on the interaction of KLVFF-PEG with fibrinogen (Fbg) in neutral aqueous solutions at 20 degrees C, for particular ratios of KLVFF-PEG to Fbg concentration, Delta = CKLVFF-PEG/C-Fbg- Our results show the formation of Fbg/KLVFF-PEG complexes for Delta > 0, such that there is not an extended network of complexes throughout the solution. In addition, cleaved protein and Fbg dimers are identified in the solution for Delta >= 0. There is a dramatic change in the tertiary structure of the Fbg upon KLVFF-PEG binding, although the KLVFF-PEG binds to the Fbg without affecting the secondary structure elements of the glycoprotein.