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Diêgo dos Santos Ferreira,1,2 Samilla Dornelas Faria,1 Sávia Caldeira de Araújo Lopes,1 Cláudia Salviano Teixeira,1 Angelo Malachias,3 Rogério Magalhães-Paniago,3 José Dias de Souza Filho,4 Bruno Luis de Jesus Pinto Oliveira,2 Alexander Ramos Guimarães,2 Peter Caravan,2 Lucas Antônio Miranda Ferreira,1 Ricardo José Alves,1 Mônica Cristina Oliveira1 1Department of Pharmaceutical Products, Faculty of Pharmacy,Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil; 2Athinoula A Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA; 3Department of Physics, 4Department of Chemistry, Institute of Exact Sciences, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil Background: Despite recent advances in cancer therapy, the treatment of bone tumors remains a major challenge. A possible underlying hypothesis, limitation, and unmet need may be the inability of therapeutics to penetrate into dense bone mineral, which can lead to poor efficacy and high toxicity, due to drug uptake in healthy organs. The development of nanostructured formulations with high affinity for bone could be an interesting approach to overcome these challenges.Purpose: To develop a liposomal formulation with high affinity for hydroxyapatite and the ability to release doxorubicin (DOX) in an acidic environment for future application as a tool for treatment of bone metastases.Materials and methods: Liposomes were prepared by thin-film lipid hydration, followed by extrusion and the sulfate gradient-encapsulation method. Liposomes were characterized by average diameter, ζ-potential, encapsulation percentage, X-ray diffraction, and differential scanning calorimetry. Release studies in buffer (pH 7.4 or 5), plasma, and serum, as well as hydroxyapatite-affinity in vitro analysis were performed. Cytotoxicity was evaluated by MTT assay against the MDA-MB-231 cell line, and biodistribution was assessed in bone metastasis-bearing animals.Results: Liposomes presented suitable diameter (~170 nm), DOX encapsulation (~2 mg/mL), controlled release, and good plasma and serum stability. The existence of interactions between DOX and the lipid bilayer was proved through differential scanning calorimetry and small-angle X-ray scattering. DOX release was faster when the pH was in the range of a tumor than at physiological pH. The bone-targeted formulation showed a strong affinity for hydroxyapatite. The encapsulation of DOX did not interfere in its intrinsic cytotoxicity against the MDA-MB-231 cell line. Biodistribution studies demonstrated high affinity of this formulation for tumors and reduction of uptake in the heart.Conclusion: These results suggest that bone-targeted pH-sensitive liposomes containing DOX can be an interesting strategy for selectively delivering this drug into bone-tumor sites, increasing its activity, and reducing DOX-related toxicity. Keywords: hydroxyapatite-targeted formulations, bisphosphonates, pH-responsive nanostructures, bone-tumor treatment

Juliane S Lanza,1 Flaviana R Fernandes,1 José D Corrêa-Júnior,2 José MC Vilela,3 Rogério Magalhães-Paniago,4 Lucas AM Ferreira,5 Margareth S Andrade,3 Cynthia Demicheli,6 Maria N Melo,7 Frédéric Frézard1 1Department of Physiology and Biophysics, 2Department of Morphology, Instituto de Ciências Biológicas (ICB), Universidade Federal de Minas Gerais (UFMG), 3Innovation and Technology Center SENAI FIEMG – Campus CETEC, 4Department of Physics, Instituto de Ciências Exatas (ICEX), 5Department of Pharmaceutical Products, Faculty of Pharmacy, Universidade Federal de Minas Gerais (UFMG), 6Department of Chemistry, Instituto de Ciências Exatas (ICEX), 7Department of Parasitology, Instituto de Ciências Biológicas (ICB), Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, Brazil Abstract: There is a great need for orally active drugs for the treatment of the neglected tropical disease leishmaniasis. Amphiphilic Sb(V) complexes, such as 1:3 Sb–N-octanoyl-N-methylglucamide complex (SbL8), are promising drug candidates. It has been previously reported that SbL8 forms kinetically stabilized nanoassemblies in water and that this simple dispersion exhibits antileishmanial activity when given by oral route to a murine model of visceral leishmaniasis. The main objective of the present work was to interfere in the structural organization of these nanoassemblies so as to investigate their influence on the oral bioavailability of Sb, and ultimately, optimize an oral formulation of SbL8 for the treatment of cutaneous leishmaniasis. The structural organization of SbL8 nanoassemblies was manipulated through addition of propylene glycol (PG) to the aqueous dispersion of SbL8. The presence of 50% (v/v) PG resulted in the loss of hydrophobic microenvironment, as evidenced by fluorescence probing. However, nanostructures were still present, as demonstrated by dynamic light scattering, small-angle X-ray scattering, and atomic force microscopy (AFM). A remarkable property of these nanoassemblies, as revealed by AFM analysis, is the flexibility of their supramolecular organization, which showed changes as a function of the solvent and substrate polarities. The formulation of SbL8 in 1:1 water:PG given orally to mice promoted significantly higher and more sustained serum levels of Sb, when compared to SbL8 in water. The new formulation, when given as repeated doses (200 mg Sb/kg/day) to BALB/c mice infected with Leishmania amazonensis, was significantly more effective in reducing the lesion parasite burden, compared to SbL8 in water, and even, the conventional drug Glucantime® given intraperitoneally at the same dose. In conclusion, this work introduces a new concept of polarity-sensitive nanocarrier that was successfully applied to optimize an oral formulation of Sb(V) for treating cutaneous leishmaniasis. Keywords: propylene glycol, antimony, Leishmania amazonensis, AFM, amphiphilic complex, SAXS

An in situ combination of atomic force microscopy and micro x-ray diffraction was developed to study the elastic behavior of nanosized objects. This technique offers the means to locally access the Young elastic moduli and Poisson ratios of individual nanostructures. Here, we investigated the elastic behavior of a single self-assembled 450 nm high SiGe island. As pressure was applied on the island, the resonance frequency of the atomic force microscope tuning fork was tracked together with the x-ray diffraction stemming from this individual crystal. The change in the tip-island contact stiffness could be derived from the variation in the resonance frequency of the tuning fork, whereas the island mean lattice parameter was inferred from the center of mass of the island’s Bragg scattering. From this information, the reduced elastic modulus of the tip-island system could be directly determined, which is in very good agreement with literature values. The pressure needed to compress the island lattice to the Si value amounts to about 3 GPa and is in good accordance with finite-element method simulations of the displacement field in the pressurized object. [ABSTRACT FROM AUTHOR]

We investigated structural and optical properties of type-II InP/GaAs quantum dots using reflection high energy electron diffraction, transmission electron microscopy, atomic force microscopy, grazing incidence x-ray diffraction, and photoluminescence techniques. The InP dots present an efficient optical emission even when they are uncapped, which is attributed to the low surface recombination velocity in InP. We compare the difference in the optical properties between surface free dots, which are not covered by any material, with dots covered by a GaAs capping layer. We observed a bimodal dispersion of the dot size distribution, giving rise to two distinct emission bands. The results also revealed that the strain accumulated in the InP islands is slightly relieved for samples with large InP amounts. An unexpected result is the relatively large blue shift of the emission band from uncapped samples as compared to capped dots. [ABSTRACT FROM AUTHOR]

We investigated the thermal evolution of the magnetic properties of MnAs epitaxial films grown on GaAs(001) during the coexistence of hexagonal/orthorhombic phases using polarized resonant (magnetic) soft x-ray scattering and magnetic force microscopy. The results of the diffuse satellite x-ray peaks were compared to those obtained by magnetic force microscopy and suggest a reorientation of ferromagnetic terraces as temperature rises. By measuring hysteresis loops at these peaks we show that this reorientation is common to all ferromagnetic terraces. The reorientation is explained by a simple model based on the shape anisotropy energy. Demagnetizing factors were calculated for different configurations suggested by the magnetic images. We noted that the magnetic moments flip from an in-plane monodomain orientation at lower temperatures to a three-domain out-of-plane configuration at higher temperatures. The transition was observed when the ferromagnetic stripe width L is equal to 2.9 times the film thickness d. This is in good agreement with the expected theoretical value of L=2.6d. [ABSTRACT FROM AUTHOR]

Si islands were grown on Ge (111) in Volmer-Weber growth mode with a 40-nm thick Ge0.85Si0.15 buffer. The state of strain and chemical composition of these islands were evaluate by grazing incidence anomalous x-ray diffraction. The results show evidence of lattice coherence and Ge-Si intermixing. A direct relationship between increase in substrate temperature and enhancement of alloying was found, evidencing the importance of atomic interdiffusion in this growth mode. [ABSTRACT FROM AUTHOR]

We explore the donor/acceptor interaction between distinct polymers at sequential bilayer interfaces to understand if their emission is influenced by the solvent. Resonant soft X-ray reflectivity and near-edge X-ray absorption fine structure measurements were used to investigate optical and morphological properties of polymer stacks, which were made using polymer chain structures diluted with different or equal solvents. We identified coupled and uncoupled bilayer systems using as a probe the exciton energy transfer effect inferred from the donor/acceptor interdiffusion region at the heterojunction interfaces. We also show that the overall emission is dominated by the affinity of the chemical structures at the interface, regardless of the solvent used for the deposition of each layer. [ABSTRACT FROM AUTHOR]

Melting and premelting phenomena in self-organized organic systems have been extensively explored in the literature, exploring distinct behaviors of different molecule lengths and morphologies. Nevertheless, the influence of the supramolecular assembly configuration on the occurrence of premelting remains poorly explored. Here we use phosphonic acids as model systems for self-organized molecular assemblies. These molecules exhibit long-range order on different types of substrates. The balance between chain-to-chain and head-to-head interactions leads to distinct types of stackings. Although their structural configurations are well understood, very little is known about their behavior near the melting transition. We show here that premelting occurs in lamellar structures and that its behavior depends directly on the ordered configuration assumed in the studied multilayers. Two molecules with different chain lengths were investigated: octadecyl phosphonic and octyl phosphonic acids. Although almost no dependence on the molecule length was observed, the occurrence of premelting is strongly influenced by their lamellar packing configuration. For tilted packings premelting is unfavored while in straight configurations, where alkyl chain interactions are weakened with respect to head-to-head interactions, strong premelting is observed. We find that the onset of premelting occurs at the domain boundaries with straight lamellar configurations and the domain sizes exhibit power law temperature dependences. [ABSTRACT FROM AUTHOR]

Iron silicide samples were grown on Si (111) substrates by solid phase epitaxy and reactive deposition epitaxy. The different iron silicide phases and their correlations with the growth parameters were analyzed by x-ray photoelectron spectroscopy, conversion electron Mössbauer spectroscopy, x-ray diffraction, atomic force microscopy, and magnetic force microscopy. The authors investigated the potential of each technique for identifying and quantifying of the phases. In particular, the authors used a semiquantitative analysis of magnetic force microscopy images to spatially resolve the semiconductor β-FeSi2 phase. [ABSTRACT FROM AUTHOR]

Atomic force microscopy (AFM) and micro-x-ray diffraction are combined to investigate nanostructures during in situ indentation. This technique allows the determination of elastic properties of individual nanoscale objects, particularly here SiGe/Si(001) self-assembled islands. Using this novel technique it was possible to select a specific island, align it in the microfocused beam, and apply a pressure onto it, using the AFM tip. Simultaneously, the x-ray diffuse scattering map from the island and the surrounding substrate was recorded in order to probe the lattice parameter change during indentation. An elastic reduction of the island lattice parameter of up to 0.6% was achieved. [ABSTRACT FROM AUTHOR]

Lattice parameter profiles and the chemical structure of InP self-assembled islands grown on GaAs(001) were determined with x-ray resonant scattering. By accessing four different photon energies, near x-ray absorption edges of two of the atomic species present on the samples, composition maps of all four atomic constituents of these islands were obtained. This experiment was performed for samples grown at two different temperatures and the effect of temperature was associated to Ga-interdiffusion and strain relief in the dots. [ABSTRACT FROM AUTHOR]

The authors have investigated strain relaxation in InAs/InGaAs/InP nanowires (NW’s). Transmission electron microscopy images show an additional stress field attributed to compositional modulation in the ternary layer, which disrupts NW formation and drives Ga interdiffusion into InAs, according to grazing incidence x-Ray diffraction under anomalous scattering conditions. The strain profile along the NW, however, is not significantly affected when interdiffusion is considered. Results show that the InAs NW energetic stability is preserved with the introduction of ternary buffer layer in the structure. [ABSTRACT FROM AUTHOR]

This letter shows that aligned zinc oxide (ZnO) nanowires growth on sapphire substrates is epitaxial and demonstrates the crystallographic relation between the two using grazing incidence synchrotron x-ray diffraction (XRD). The in-plane lattice match between the sapphire and the nanowires was directly probed by using XRD at grazing angles of incidence, where the lattice match between the (0001) plane of the sapphire and the (11-20) plane of the ZnO were observed simultaneously. It will also be shown that gold acts as a catalyst to initiate ZnO nanowire growth, but it does not interfere with the epitaxial mechanism between the nanowires and the sapphire substrate. [ABSTRACT FROM AUTHOR]

This work presents a systematic study of the ratio between the integrated intensities of the disorder-induced D and G Raman bands (ID/IG) in nanographite samples with different crystallite sizes (La) and using different excitation laser energies. The crystallite size La of the nanographite samples was obtained both by x-ray diffraction using synchrotron radiation and directly from scanning tunneling microscopy images. A general equation for the determination of La using any laser energy in the visible range is obtained. Moreover, it is shown that ID/IG is inversely proportional to the fourth power of the laser energy used in the experiment. [ABSTRACT FROM AUTHOR]

MnAs films grown on GaAs (001) exhibit a progressive transition between hexagonal (ferromagnetic) and orthorhombic (paramagnetic) phases at wide temperature range instead of abrupt transition during the first-order phase transition. The coexistence of two phases is favored by the anisotropic strain arising from the constraint on the MnAs films imposed by the substrate. This phase coexistence occurs in ordered arrangement alternating periodic terrace steps. We present here a method to study the surface morphology throughout this transition by means of specular and diffuse scattering of soft x rays, tuning the photon energy at the Mn 2p resonance. The results show the long-range arrangement of the periodic stripe-like structure during the phase coexistence and its period remains constant, in agreement with previous results using other techniques. [ABSTRACT FROM AUTHOR]

Three-dimensional strain mapping of InAs self-assembled nanowires on an InP substrate using grazing incidence x-ray diffraction is reported. A remarkable anisotropy was observed for the strain components, parallel [-220] and perpendicular [220] to the wire axis. The highest strain relaxation was measured along the [220] direction. The relationship between the interatomic distances along the [-220] and [220] directions, for each z position (height) in the nanostructure, was obtained by angular scans in the vicinity of the (040) reciprocal lattice point. [ABSTRACT FROM AUTHOR]

Multilayer epitaxial graphene was obtained from a 6H-SiC(001) substrate subjected to a temperature gradient from 1250 to 1450 °C. Scanning tunneling microscopy and x-ray diffraction were used to identify the structure and morphology of the surface, from which the formation of a metastable phase was inferred. By a comparison between microscopy and diffraction data, we report the appearance of misoriented Si-doped graphene in cold regions (1250 °C) of the substrate. This metastable phase occurs in domains where silicon sublimation is incomplete and it coexists with small domains of epitaxial graphene. At 1350 °C this phase disappears and one observes complete graphene-like layers (although misoriented), where rotational registry between the underlying epitaxial graphene and additional layers is absent. At 1450 °C the stacking among layers is established and the formation of highly oriented single crystalline graphite is complete. The stability of this Si-rich metastable phase at 1250 °C was confirmed by first-principles calculations based on the density functional theory. [ABSTRACT FROM AUTHOR]

Structural and chemical properties of self-assembled InAs islands grown on GaAs(001) were studied using x-ray scattering. Two measurements performed under grazing incidence geometry were correlated to obtain the three-dimensional strain and chemical status of InGaAs coherent islands. Grazing incidence diffraction was employed to reveal the in-plane strain-size interplay. Mapping out the reciprocal space near the GaAs(022) reflection and correlating the in-plane and out-of-plane strain information, we have been able to quantify the tetragonal distortion of the unit cells at any position inside the islands. Simple theory of elasticity of alloys enabled us to analyse the elastic deformation of the unit cells. Any variation in the expected tetragonal distortion of the unit cell was associated to the presence of Ga atoms inside the islands. Using this method, the Ga content in our islands was shown to vary linearly from 25% (island bottom) to 8% (island top). [ABSTRACT FROM AUTHOR]

In this paper we present a method to determine the occupation of Mn ions in epitaxial (In,Mn)As nanostructures by resonant x-ray diffraction near the Mn K-absorption edge, exploiting the dependency of the intensity of both (200) superstructure and the (400) fundamental reflections. The concentrations of Mn atoms on substitutional In sites, as well as on In- and As-interstitial sites, were unambiguously determined. A threshold concentration for the interstitial sites, which are occupied first for low nominal Mn deposition content, was found. Calculations using density-functional theory indicate that a higher occupancy of such sites can be favorable with respect to the occupation of substitutional sites, depending on surface potentials and growth kinetic effects. [ABSTRACT FROM AUTHOR]

The production of controlled doping in two-dimensional semiconductor materials is a challenging issue when introducing these systems into current and future technology. In some compounds, the coexistence of distinct crystallographic phases for a fixed composition introduces an additional degree of complexity for synthesis, chemical stability, and potential applications. In this work, we demonstrate that a multiphase In 2 Se 3 layered semiconductor system, synthesized with three distinct structures─rhombohedral α and β-In 2 Se 3 and trigonal δ-In 2 Se 3 ─exhibits chemical stability and well-behaved n-type doping. Scanning tunneling spectroscopy measurements reveal variations in the local electronic density of states among the In 2 Se 3 structures, resulting in a compound system with electronic bandgaps that range from infrared to visible light. These characteristics make the layered In 2 Se 3 system a promising candidate for multigap or broad spectral optical devices, such as detectors and solar cells. The ability to tune the electronic properties of In 2 Se 3 through structural phase manipulation makes it ideal for integration into flexible electronics and the development of heterostructures with other materials.

A long-circulating and pH-sensitive liposome containing paclitaxel (SpHL-PTX) was recently developed by our group. Once in an acidic environment, for example, tumors, these liposomes undergo destabilization, releasing the encapsulated drug. In this way, the aim of this study was to evaluate the molecular and supramolecular interactions between the lipid bilayer and PTX in similar biological environment conditions. High-sensitivity analyses of SpHL-PTX structures were obtained by the small-angle X-ray scattering technique combined with other techniques such as dynamic light scattering, asymmetric flow field-flow fractionation, transmission electron microscopy, and high-performance liquid chromatography. The results showed that PTX incorporation in the liposomal bilayer clearly leads to changes in supramolecular organization of dioleoylphosphatidylethanolamine (DOPE) molecules, inducing the formation of more ordered structures. Changes in supramolecular organization were observed at lower pH, indicating that pH sensitivity was preserved even in the presence of fetal bovine serum proteins. Furthermore, morphological and physicochemical characterization of SpHL-PTX evidenced the formation of nanosized dispersion suitable for intravenous administration. In conclusion, a stable nanosized dispersion of PTX was obtained at pH 7.4 with suitable parameters for intravenous administration. At lower pH conditions, the pH sensitivity of the system was clearly evidenced by changes in the supramolecular organization of DOPE molecules, which is crucial for the delivery of PTX into the cytoplasm of the targeted cells. In this way, the results obtained by different techniques confirm the feasibility of SpHL as a promising tool to PTX delivery in acidic environments, such as tumors.

The goal of this work is to study transformations that occur upon heating Bi 2 Se 3 to temperatures up to 623 K. X-ray diffraction (XRD) and scanning tunneling microscopy (STM) and spectroscopy (STS) techniques were used in our investigation. XRD was measured following the 00L and 01L truncation rods. These measurements revealed that upon heating there is a coexistence of a major Bi 2 Se 3 phase and other ones that present structures of quintuple-layers intercalated with Bismuth bilayers. STM measurements of the surface of this material showed the presence of large hexagonal Bi x Se y domains embedded in a Bi 2 Se 3 matrix. STS experiments were employed to map the local electronic density of states and characterize the modifications imposed by the presence of the additional phases. Finally, density functional theory (DFT) calculations were performed to support these findings.

Paclitaxel is a potent antimicrotubule chemotherapeutic agent widely used for clinical treatment of a variety of solid tumors. However, the low solubility of the drug in aqueous medium and the toxic effects of the commercially available formulation, Taxol(®), has hindered its clinical application. To overcome these paclitaxel-related disadvantages, several drug delivery approaches have been thoroughly investigated. In this context, our research group has developed long-circulating and pHsensitive liposomes containing paclitaxel composed of dioleylphosphatidylethanolamine, cholesterylhemisuccinate and distearoylphosphatidylethanolamine-polyethylene glycol2000, which have shown to be very promising carriers for this taxane. For the destabilization of pH-sensitive liposomal systems and the release of the encapsulated drug in the cytoplasm of tumor cells, the occurrence of a phase transition from a lamellar to a non-lamellar phase of dioleylphosphatidylethanolamine molecules is essential. Two techniques, differential scanning calorimetry and small angle X-ray scattering, were used to investigate the influence of the liposomal components and paclitaxel in the phase transition process of dioleylphosphatidylethanolamine molecules and to evaluate the pH-sensitivity of the formulation under low hydration conditions. The findings clearly evidence the phase transition of dioleylphosphatidylethanolamine molecules in the presence and absence of PTX indicating that the introduction of the drug in the system does not bring damage to the pH-sensitivity of the system, which resulting in liposome destabilization at low pH regions and encapsulated paclitaxel release preferentially in a desired target tissue., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Mucopolysaccharidosis type I (MPS I) is an autosomal disease caused by alpha-L-iduronidase deficiency. This study proposed the use of cationic nanoemulsions as non-viral vectors for a plasmid (pIDUA) containing the gene that codes for alpha-L-iduronidase. Nanoemulsions composed of medium chain triglycerides (MCT)/1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)/1,2-dioleoyl-sn-glycero-3-trimethylammonium propane (DOTAP)/1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG) were prepared by high pressure homogenization. Formulations were prepared by the adsorption or encapsulation of preformed pIDUA-DOTAP complexes into the oil core of nanoemulsions at different charge ratios. pIDUA complexed was protected from enzymatic degradation by DNase I. The physicochemical characteristics of complexes in protein-containing medium were mainly influenced by the presence of DSPE-PEG. Bragg reflections corresponding to a lamellar organization were identified for blank formulations by energy dispersive X-ray diffraction, which could not be detected after pIDUA complexation. The intravenous injection of these formulations in MPS I knockout mice led to a significant increase in IDUA activity (fluorescence assay) and expression (RT-qPCR) in different organs, especially the lungs and liver. These findings were more significant for formulations prepared at higher charge ratios (+4/-), suggesting a correlation between charge ratio and transfection efficiency. The present preclinical results demonstrated that these nanocomplexes represent a potential therapeutic option for the treatment of MPS I., (Copyright © 2015. Published by Elsevier B.V.)

Ursolic acid (UA) is a triterpene found in different plant species that has been shown to possess significant antitumor activity. However, UA presents a low water solubility, which limits its biological applications. In this context, our research group has proposed the incorporation of UA in long-circulating and pH-sensitive liposomes (SpHL-UA).These liposomes, composed of dioleylphosphatidylethanolamine (DOPE), cholesteryl hemisuccinate (CHEMS), and distearoylphosphatidylethanolamine-polyethylene glycol2000 (DSPE-PEG2000), were shown to be very promising carriers for UA. Considering that the release of UA from SpHL-UA and its antitumor activity depend upon the occurrence of the lamellar to non-lamellar phase transition of DOPE, in the present work, the interactions of UA with the components of the liposomes were evaluated, aiming to clarify their role in the structural organization of DOPE. The study was carried out by differential scanning calorimetry (DSC) and small-angle X-ray scattering (SAXS) under low hydration conditions. DSC studies revealed that DOPE phase transition temperatures did not shift significantly upon UA addition. On the other hand, in SAXS studies, a different pattern of DOPE phase organization was observed in the presence of UA, with the occurrence of the cubic phase Im3m at 20 °C and the cubic phase Pn3m at 60 °C. These findings suggest that UA interacts with the lipids and changes their self-assembly. However, these interactions between the lipids and UA were unable to eliminate the lamellar to non-lamellar phase transition, which is essential for the cytoplasmic delivery of UA molecules from SpHL-UA.

The determination of the molecular structure of a porphyrin is achieved by using nuclear magnetic resonance (NMR) and scanning tunneling microscopy (STM) techniques. Since macroscopic crystals cannot be obtained in this system, this combination of techniques is crucial to solve the molecular structure without the need for X-ray crystallography. For this purpose, previous knowledge of the flatness of the reagent molecules (a porphyrin and its functionalizing group, a naphthalimide) and the resulting molecular structure obtained by a force-field simulation are used. The exponents of the I-V curves obtained by scanning tunneling spectroscopy (STS) allow us to check whether the thickness of the film of molecules is greater than a monolayer, even when there is no direct access to the exposed surface of the metal substrate. Photoluminescence (PL), optical absorption, infrared (IR) reflectance and solubility tests are used to confirm the results obtained here with this NMR/STM/STS combination.