Célia F. Cruz

Keratins and lipids in ethnic hair

Human hair has an important and undeniable relevance in society due to its important role in visual appearance and social communication. Hair is mainly composed of structural proteins, mainly keratin and keratin associated proteins and lipids. Herein, we report a comprehensive study of the content and distribution of the lipids among ethnic hair, African, Asian and Caucasian hair. More interestingly, we also report the study of the interaction between those two main components of hair, specifically, the influence of the hair internal lipids in the structure of the hair keratin. This was achieved by the use of a complete set of analytical tools, such as thin layer chromatography‐flame ionization detector, X‐ray analysis, molecular dynamics simulation and confocal microscopy. The experimental results indicated different amounts of lipids on ethnic hair compositions and higher percentage of hair internal lipids in African hair. In this type of hair, the axial diffraction of keratin was not observed in X‐ray analysis, but after hair lipids removal, the keratin returned to its typical packing arrangement. In molecular dynamic simulation, lipids were shown to intercalate dimers of keratin, changing its structure. From those results, we assume that keratin structure may be influenced by higher concentration of lipids in African hair.

Potential of human γD-crystallin for hair damage repair : insights into the mechanical properties and biocompatibility

The objective of this work was to develop a new strategy to physically ‘repair’ chemically damaged hair. Hence the human eye γD‐crystallin, a protein from the superfamily characterized structurally by the Greek key motif, was studied. The human γD‐crystallin was chosen based on the ability of proteins belonging to this superfamily to be involved in the coating of specific structures. Two crystallins were used on the study, the wild type (Protein Data Bank ID: 1HK0) and the mutant protein. The mutant form was intended to induce a strong and quick protein polymerization as well to have new possible points of anchorage to hair.

The effects of solvent composition on the affinity of a peptide towards hair keratin: experimental and molecular dynamics data

The study of the interaction between hair filaments and formulations or peptides is of utmost importance in fields like cosmetic research. Keratin intermediate filament structure is not fully described, limiting the molecular dynamics (MD) studies in this field despite its high potential to improve the area. We developed a computational model of a truncated protofibril, simulated its behavior in alcoholic based formulations and with one peptide. The simulations showed a strong interaction between the benzyl alcohol molecules of the formulations and the model, leading to the disorganization of the keratin chains, which regress with the removal of the alcohol molecules. This behavior can explain the increase of peptide uptake in hair shafts evidenced in fluorescence microscopy pictures. The model developed is valid to computationally reproduce the interaction between hair and alcoholic formulations and provide a robust base for new MD studies about hair properties. It is shown that the MD simulations can improve hair cosmetic research, improving the uptake of a compound of interest.

Insights on the mechanical behavior of keratin fibrils

A computational molecular model of a truncated keratin protofibril (8 chains of hair keratin, PDB provided in Supplementary material) was used, to run a series of steered molecular dynamics simulations obtaining strain-stress curves. These results were compared with experimental mechanical data on hair fibers. Our data demonstrate that the molecular dynamics simulations can model hair mechanical properties. Simulations done in vacuum showed a better agreement with experimental Youngs Modulus (YM) values. The role of hydrogen bonds and the secondary structure of keratin on the mechanical properties was evaluated in detail. The incubation with a fragment of one surfactant protein, the SPD-2 peptide (QAAFSQ), showed the improvement of YM of the hair keratin either by simulations and experimental data. For the first, our research provides mechanistic insights on mechanical microscopic properties of keratin protofibrils through molecular dynamics simulations.

Effect of a peptide in cosmetic formulations for hair volume control

The capacity of hair to absorb water causes changes in its physical and cosmetic properties under different environmental conditions. Hence, the control of hair volume in variable relative humidity settings is an important topic in cosmetics. The behaviour of two types of hair, Caucasian and Asian, was studied regarding their volume change in different relative humidity conditions. The ability of a peptide as a hair volume treatment was evaluated in two climate control formulations.

Peptide—protein interactions within human hair keratins

We selected 1235 decapeptides from human hair proteins encoded by human genes of keratins and keratin associated proteins. The peptides were linked to glass arrays and screened for their affinity towards a solution of human hair extracted keratin fraction. Based on the physicochemical properties of the peptides, ten variables were studied: content of different types of amino acid side chains (cysteine, hydrophobic, polar, basic, acidic, aromatic rings, amide, alcohol side chains), isoelectric point, and net charge. We found differences statistically significant on the binding affinity of peptides based on their content of cysteine, hydrophobic and polar amino acids, mainly containing alcohols. These results point to the formation of hydrophobic interactions and disulfide bonds between small peptides and human hair keratins as the main driving forces for the interaction of possible cosmetic peptides, namely designed to strength human hair. As so, our results enlighten the nature of the interaction of keratin based materials with human hair, which are claimed to enhance hair fiber strength, and enable a more directed and sustained hair care peptide design.

Changing the shape of hair with keratin peptides

Chemical straightening of curly human hair fibres involves the use of strong reducing agents at alkaline pH. Human hair is made of keratin, and the fixation of fibre shape involves the reduction and reformation of new disulphide bonds between keratin molecules. Here, we propose an alternative and green methodology using keratin peptide sequences (10–13 residues) derived from the human genome. In a previous study, we analysed 1235 cysteine-containing peptides encoded by all human genes of hair keratin and keratin-associated proteins. These peptide fragments have been designed by nature to interact with keratin. Here we tested eight peptides, which were select based on their affinity for human hair keratin solution as shown by Matrix-Assisted Laser-Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF/TOF) and by molecular dynamics simulation. The peptides were characterized in detail regarding their ability to act as hair straightening modulators and to improve the tensile strength and elasticity of hair. Of the eight tested peptides, PepE, PepG and KP showed the highest ability to interact with a keratin peptide model, and to improve hair mechanical properties and straightening efficiency. The proposed solutions presented here will replace harsh reducing agents at alkaline pH by peptide formulations acting at neutral pH to change hair shape through the re-conformation of disulphide bonds. Here, we provide experimental evidence which explains at a molecular level how keratin decapeptides can interact with large keratin molecules in human hair, opening an innovative green approach to changing the shape of hair fibre.

Assessment of liposome disruption to quantify drug delivery in vitro

Efficient liposome disruption inside the cells is a key for success with any type of drug delivery system. The efficacy of drug delivery is currently evaluated by direct visualization of labeled liposomes internalized by cells, not addressing objectively the release and distribution of the drug. Here, we propose a novel method to easily assess liposome disruption and drug release into the cytoplasm. We propose the encapsulation of the cationic dye Hoechst 34580 to detect an increase in blue fluorescence due to its specific binding to negatively charged DNA. For that, the dye needs to be released inside the cell and translocated to the nucleus. The present approach correlates the intensity of detected fluorescent dye with liposome disruption and consequently assesses drug delivery within the cells.