Interactions between Damaged Hair Keratin and Juglone as a Possible Restoring Agent: A Vibrational and Scanning Electron Microscopy Study.

Juglone, a quinonic compound present in walnut extracts, was proposed as a restoring agent for hair keratin treated with permanent or discoloration processes. The proposed mechanism of restoration by juglone involves the formation of a Michael adduct between the quinone and the thiol moieties of cysteine residues. To this purpose, the first part of the present paper involved the spectroscopic study of the product of the reaction between juglone and N-acetyl-L-cysteine as a model compound. IR spectroscopy and Scanning Electron Microscopy (SEM) monitored the chemical and morphological variations induced by applying juglone to hair keratin. In order to simulate the most common hair treatments (i.e., permanent and discoloration), juglone was applied to hair that had been previously treated with a reducing agent, i.e., methyl thioglycolate (MT) or with bleaching agents (based on hydrogen peroxide and persulfates) followed by sodium hydrogen sulfite. IR spectroscopy allowed us to monitor the formation of Michael adducts between juglone and cysteine residues: the Michael adducts' content was related to the cysteine content of the samples. In fact, MT and sodium hydrogen sulfite favored the reduction of the disulfide bonds and increased the content of free cysteine residues, which can react with juglone. SEM analyses confirmed the trend observed by IR spectroscopy since hair samples treated with juglone adopted a more regular hair surface and more imbricated scales, thus supporting the possible use of juglone as a restoring agent for damaged hair keratins.

Guttapercha Improves In Vitro Bioactivity and Dentin Remineralization Ability of a Bioglass Containing Polydimethylsiloxane-Based Root Canal Sealer.

Guttapercha (GP, trans-1,4-polyisoprene) is the most used tooth root filling material, and it must be used with an appropriate cement (typically a polydimethylsiloxane (PDMS)-based sealer) to ensure an adequate canal obturation. This study aimed to assess the bioactivity and dentin remineralization ability of a bioglass containing PDMS commercial endodontic sealer, BG-PDMS (GuttaFlow Bioseal), and to evaluate the possible influence of a GP cone (Roeko GP point) on the mineralization process. To this end, BG-PDMS disks were aged alone or in the presence of a GP cone in Hank's Balanced Salt Solution (28 d, 37 °C). Dentin remineralization experiments were carried out under the same conditions. Micro-Raman and IR analyses demonstrated that BG-PDMS is bioactive, thanks to the formation of a silica-rich layer with nucleation sites for B-type carbonated apatite deposition. This phase was thicker when BG-PDMS was aged in the presence of GP. The two materials influenced each other because GP, which alone did not show any bioactivity, nucleated a calcium phosphate phase under these conditions. Analogously, dentin remineralization experiments showed that BG-PDMS is able to remineralize dentin, especially in the presence of GP. Under the experimental conditions, GP acted as a templating agent for calcium phosphate deposition.

Proteolytically Resistant Bioactive Peptide-Grafted Sr/Mg-Doped Hardystonite Foams: Comparison of Two Covalent Functionalization Strategies.

Hardystonite-based (HT) bioceramic foams were easily obtained via thermal treatment of silicone resins and reactive oxide fillers in air. By using a commercial silicone, incorporating strontium oxide and magnesium oxide precursors (as well as CaO and ZnO), and treating it at 1100 °C, a complex solid solution (Ca1.4Sr0.6Zn0.85Mg0.15Si2O7) that has superior biocompatibility and bioactivity properties compared to pure hardystonite (Ca2ZnSi2O7) can be obtained. Proteolytic-resistant adhesive peptide mapped on vitronectin (D2HVP), was selectively grafted to Sr/Mg-doped HT foams using two different strategies. Unfortunately, the first method (via protected peptide) was unsuitable for acid-sensitive materials such as Sr/Mg-doped HT, resulting in the release of cytotoxic levels of Zinc over time, with consequent negative cellular response. To overcome this unexpected result, a novel functionalization strategy requiring aqueous solution and mild conditions was designed. Sr/Mg-doped HT functionalized with this second strategy (via aldehyde peptide) showed a dramatic increase in human osteoblast proliferation at 6 days compared to only silanized or non-functionalized samples. Furthermore, we demonstrated that the functionalization treatment does not induce any cytotoxicity. Functionalized foams enhanced mRNA-specific transcript levels coding IBSP, VTN, RUNX2, and SPP1 at 2 days post-seeding. In conclusion, the second functionalization strategy proved to be appropriate for this specific biomaterial and was effective at enhancing the material's bioactivity.

Vibrational Study on the Structure, Bioactivity, and Silver Adsorption of Silk Fibroin Fibers Grafted with Methacrylonitrile.

Natural fibers have received increasing attention as starting materials for innovative applications in many research fields, from biomedicine to engineering. Bombyx mori silk fibroin has become a material of choice in the development of many biomedical devices. Grafting represents a good strategy to improve the material properties according to the desired function. In the present study, Bombyx mori silk fibroin fibers were grafted with methacrylonitrile (MAN) with different weight gains. The potential interest in biomedical applications of MAN functionalization relies on the presence of the nitrile group, which is an acceptor of H bonds and can bind metals. IR and Raman spectroscopy were used to characterize the grafted samples and the possible structural changes induced by grafting. Afterward, the same techniques were used to study the bioactivity (i.e., the calcium phosphate nucleation ability) of MAN-grafted silk fibroins after ageing in simulated body fluid (SBF) for possible application in bone tissue engineering, and their interaction with Ag+ ions, for the development of biomaterials with enhanced anti-microbial properties. MAN was found to efficiently polymerize on silk fibroin through polar amino acids (i.e., serine and tryptophan), inducing an enrichment in silk fibroin-ordered domains. IR spectroscopy allowed us to detect the nucleation of a thin calcium phosphate layer and the uptake of Ag+ ions through the nitrile group, which may foster the application of these grafted materials in biomedical applications.

Chemical-Physical Properties and Bioactivity of New Premixed Calcium Silicate-Bioceramic Root Canal Sealers.

The aim of the study was to analyze the chemical−physical properties and bioactivity (apatite-forming ability) of three recently introduced premixed bioceramic root canal sealers containing varied amounts of different calcium silicates (CaSi): a dicalcium and tricalcium silicate (1−10% and 20−30%)-containing sealer with zirconium dioxide and tricalcium aluminate (CERASEAL); a tricalcium silicate (5−15%)-containing sealer with zirconium dioxide, dimethyl sulfoxide and lithium carbonate (AH PLUS BIOCERAMIC) and a dicalcium and tricalcium silicate (10% and 25%)-containing sealer with calcium aluminate, tricalcium aluminate and tantalite (NEOSEALER FLO). An epoxy resin-based sealer (AH PLUS) was used as control. The initial and final setting times, radiopacity, flowability, film thickness, open pore volume, water absorption, solubility, calcium release and alkalizing activity were tested. The nucleation of calcium phosphates and/or apatite after 28 days aging in Hanks balanced salt solution (HBSS) was evaluated by ESEM-EDX, vibrational IR and micro-Raman spectroscopy. The analyses showed for NeoSealer Flo and AH Plus the longest final setting times (1344 ± 60 and 1300 ± 60 min, respectively), while shorter times for AH Plus Bioceramic and Ceraseal (660 ± 60 and 720 ± 60 min, respectively). Radiopacity, flowability and film thickness complied with ISO 6876/12 for all tested materials. A significantly higher open pore volume was observed for NeoSealer Flo, AH Plus Bioceramic and Ceraseal when compared to AH Plus (p < 0.05), significantly higher values were observed for NeoSealer Flo and AH Plus Bioceramic (p < 0.05). Ceraseal and AH Plus revealed the lowest solubility. All CaSi-containing sealers released calcium and alkalized the soaking water. After 28 days immersion in HBSS, ESEM-EDX analyses revealed the formation of a mineral layer that covered the surface of all bioceramic sealers, with a lower detection of radiopacifiers (Zirconium for Ceraseal and AH Plus Bioceramic, Tantalum for NeoSealer Flo) and an increase in calcium, phosphorous and carbon. The calcium phosphate (CaP) layer was more evident on NeoSealer Flo and AH Plus Bioceramic. IR and micro-Raman revealed the formation of calcium carbonate on the surface of all set materials. A thin layer of a CaP phase was detected only on AH Plus Bioceramic and NeoSealer Flo. Ceraseal did not show CaP deposit despite its highest calcium release among all the tested CaSi-containing sealers. In conclusion, CaSi-containing sealers met the required chemical and physical standards and released biologically relevant ions. Slight/limited apatite nucleation was observed in relation to the high carbonation processes.

The Influence of the Matrix on the Apatite-Forming Ability of Calcium Containing Polydimethylsiloxane-Based Cements for Endodontics.

This study aimed to characterize the chemical properties and bioactivity of an endodontic sealer (GuttaFlow Bioseal) based on polydimethylsiloxane (PDMS) and containing a calcium bioglass as a doping agent. Commercial PDMS-based cement free from calcium bioglass (GuttaFlow 2 and RoekoSeal) were characterized for comparison as well as GuttaFlow 2 doped with dicalcium phosphate dihydrate, hydroxyapatite, or a tricalcium silicate-based cement. IR and Raman analyses were performed on fresh materials as well as after aging tests in Hank's Balanced Salt Solution (28 d, 37 °C). Under these conditions, the strengthening of the 970 cm−1 Raman band and the appearance of the IR components at 1455−1414, 1015, 868, and 600−559 cm−1 revealed the deposition of B-type carbonated apatite. The Raman I970/I638 and IR A1010/A1258 ratios (markers of apatite-forming ability) showed that bioactivity decreased along with the series: GuttaFlow Bioseal > GuttaFlow 2 > RoekoSeal. The PDMS matrix played a relevant role in bioactivity; in GuttaFlow 2, the crosslinking degree was favorable for Ca2+ adsorption/complexation and the formation of a thin calcium phosphate layer. In the less crosslinked RoekoSeal, such processes did not occur. The doped cements showed bioactivity higher than GuttaFlow 2, suggesting that the particles of the mineralizing agents are spontaneously exposed on the cement surface, although the hydrophobicity of the PDMS matrix slowed down apatite deposition. Relevant properties in the endodontic practice (i.e., setting time, radiopacity, apatite-forming ability) were related to material composition and the crosslinking degree.

Synthesis of thia-Michael-Type Adducts between Naphthoquinones and N-Acetyl-L-Cysteine and Their Biological Activity.

A series of naphthoquinones, namely, 1,4-naphthoquinone, menadione, plumbagin, juglone, naphthazarin, and lawsone, were reacted with N-acetyl-L-cysteine, and except for lawsone, which did not react, the related adducts were obtained. After the tuning of the solvent and reaction conditions, the reaction products were isolated as almost pure from the complex reaction mixture via simple filtration and were fully characterized. Therefore, the aim of this work was to evaluate whether the antitumor activity of new compounds of 1,4-naphthoquinone derivatives leads to an increase in ROS in tumor cell lines of cervical carcinoma (HeLa), neuroblastoma (SH-SY5Y), and osteosarcoma (SaOS2, U2OS) and in normal dermal fibroblast (HDFa). The MTT assay was used to assay cell viability, the DCF-DA fluorescent probe to evaluate ROS induction, and cell-cycle analysis to measure the antiproliferative effect. Compounds 8, 9, and 12 showed a certain degree of cytotoxicity towards all the malignant cell lines tested, while compound 11 showed biological activity at higher IC50 values. Compounds 8 and 11 induced increases in ROS generation after 1 h of exposure, while after 48 h of treatment, only 8 induced an increase in ROS formation in HeLa cells. Cell-cycle analysis showed that compound 8 caused an increase in the number of G0/G1-phase cells in the HeLa experiment, while for the U2OS and SH-SY5Y cell lines, it led to an accumulation of S-phase cells. Therefore, these novel 1,4-naphthoquinone derivatives may be useful as antitumoral agents in the treatment of different cancers.

Mn-Containing Bioactive Glass-Ceramics: BMP-2-Mimetic Peptide Covalent Grafting Boosts Human-Osteoblast Proliferation and Mineral Deposition.

The addition of Mn in bioceramic formulation is gaining interest in the field of bone implants. Mn activates human osteoblast (h-osteoblast) integrins, enhancing cell proliferation with a dose-dependent effect, whereas Mn-enriched glasses induce inhibition of Gram-negative or Gram-positive bacteria and fungi. In an effort to further optimize Mn-containing scaffolds' beneficial interaction with h-osteoblasts, a selective and specific covalent functionalization with a bioactive peptide was carried out. The anchoring of a peptide, mapped on the BMP-2 wrist epitope, to the scaffold was performed by a reaction between an aldehyde group of the peptide and the aminic groups of silanized Mn-containing bioceramic. SEM-EDX, FT-IR, and Raman studies confirmed the presence of the peptide grafted onto the scaffold. In in vitro assays, a significant improvement in h-osteoblast proliferation, gene expression, and calcium salt deposition after 7 days was detected in the functionalized Mn-containing bioceramic compared to the controls.

Vibrational Study on Structure and Bioactivity of Protein Fibers Grafted with Phosphorylated Methacrylates.

In the last decades, silk fibroin and wool keratin have been considered functional materials for biomedical applications. In this study, fabrics containing silk fibers from Bombyx mori and Tussah silk fibers from Antheraea pernyi, as well as wool keratin fabrics, were grafted with phosmer CL and phosmer M (commercial names, i.e., methacrylate monomers containing phosphate groups in the molecular side chain) with different weight gains. Both phosmers were recently proposed as flame retarding agents, and their chemical composition suggested a possible application in bone tissue engineering. IR and Raman spectroscopy were used to disclose the possible structural changes induced by grafting and identify the most reactive amino acids towards the phosmers. The same techniques were used to investigate the nucleation of a calcium phosphate phase on the surface of the samples (i.e., bioactivity) after ageing in simulated body fluid (SBF). The phosmers were found to polymerize onto the biopolymers efficiently, and tyrosine and serine underwent phosphorylation (monitored through the strengthening of the Raman band at 1600 cm-1 and the weakening of the Raman band at 1400 cm-1, respectively). In grafted wool keratin, cysteic acid and other oxidation products of disulphide bridges were detected together with sulphated residues. Only slight conformational changes were observed upon grafting, generally towards an enrichment in ordered domains, suggesting that the amorphous regions were more prone to react (and, sometimes, degrade). All samples were shown to be bioactive, with a weight gain of up to 8%. The most bioactive samples contained the highest phosmers amounts, i.e., the highest amounts of phosphate nucleating sites. The sulphate/sulphonate groups present in grafted wool samples appeared to increase bioactivity, as shown by the five-fold increase of the IR phosphate band at 1040 cm-1.

Vibrational Raman and IR data on brown hair subjected to bleaching.

Brown human hair was bleached three times (45 min × 3) and four times (45 min × 3 + 15 min) with commercial formulations containing persulfate salts and hydrogen peroxide. The hair samples were characterized by Raman and IR spectroscopy in the Attenuated Total Reflectance (ATR) mode to gain more insights into the possible secondary structure and Cα-Cß-S-S-Cß-Cα conformational changes induced by bleaching. The latter were evaluated through band-fitting procedures; the relative content of the disulfide bridges and oxidized sulfur species (cysteic acid, Bunte salt, cystine oxides) was assessed. The observed conformational changes could be significant in developing restoring agents to be used after hair decoloration. The use of two different spectroscopic techniques allowed to discriminate the information coming from the cortical region of hair (Raman) and the cuticle (ATR/IR). This article refers to "Structural investigation on damaged hair keratin treated with α,ß-unsaturated Michael acceptors used as repairing agents" (Di Foggia et al., Int. J. Biol. Macromol. 167 (2021) 620-632 https://doi.org/10.1016/j.ijbiomac.2020.11.194).

SERS Investigation on Oligopeptides Used as Biomimetic Coatings for Medical Devices.

The surface-enhanced Raman scattering (SERS) spectra of three amphiphilic oligopeptides derived from EAK16 (AEAEAKAK)2 were examined to study systematic amino acid substitution effects on the corresponding interaction with Ag colloidal nanoparticles. Such self-assembling molecular systems, known as "molecular Lego", are of particular interest for their uses in tissue engineering and as biomimetic coatings for medical devices because they can form insoluble macroscopic membranes under physiological conditions. Spectra were collected for both native and gamma-irradiated samples. Quantum mechanical data on two of the examined oligopeptides were also obtained to clarify the assignment of the prominent significative bands observed in the spectra. In general, the peptide-nanoparticles interaction occurs through the COO- groups, with the amide bond and the aliphatic chain close to the colloid surface. After gamma irradiation, mimicking a free oxidative radical attack, the SERS spectra of the biomaterials show that COO- groups still provide the main peptide-nanoparticle interactions. However, the spatial arrangement of the peptides is different, exhibiting a systematic decrease in the distance between aliphatic chains and colloid nanoparticles.

Changes in organic compounds secreted by roots in two Poaceae species (Hordeum vulgare and Polypogon monspenliensis) subjected to iron deficiency.

Despite their economic and ecological interests, Poaceae are affected by the low availability of iron in calcareous soils. Several studies focused on the capacity of this family to secrete phytosiderophores and organic acids as a mechanism of tolerance to iron deficiency. This work aimed at studying the physiological responses of two Poaceae species; Hordeum vulgare (cultivated barley) and Polypogon monspenliensis (spontaneous species) to iron deficiency, and evaluate especially the release of phytosiderophores and organic acids. For this purpose, seedlings of these two species were cultivated in complete nutrient solution with or without iron. The biomass production, iron status, phytosiderophores and organic acids release by roots were studied. The results demonstrated that Polypogon monspenliensis was relatively more tolerant to iron deficiency than Hordeum vulgare. Polypogon monspenliensis had the ability to secrete a higher amount of phytosiderophores and organic acids, especially citric, acetic, oxalic and malic acids, compared to Hordeum vulgare. We propose this spontaneous species as a forage plant in calcareous soils and in intercropping systems with fruit trees to prevent iron chlorosis.

Structural investigation on damaged hair keratin treated with α,ß-unsaturated Michael acceptors used as repairing agents.

Many restoring formulations for damaged hair keratin have been developed. Some patents claim that the hair repair occurs through the reconstruction of disulfide bridges of keratin, through α,ß-unsaturated Michael acceptors, such as shikimic acid and bis-aminopropyl diglycol dimaleate. To gain more insights into the possible repairing mechanism, this study is aimed at assessing, by IR and Raman spectroscopies coupled to scanning electron microscopy (SEM), the structural changes induced in keratin from bleached hair by the treatment with commercial reconstructive agents as well as shikimic acid and dimethyl maleate, chosen as model compounds. Vibrational spectroscopy revealed that shikimic acid- and maleate-based restoring agents interacted with hair fibers modifying both their cortex and cuticle regions. None of the investigated treatments induced an increase in the SS disulfide bridges content of the hair cortex, although it cannot be excluded that this phenomenon could have occurred in the cuticle. SS rearrangements were found to occur. None of our results can be interpreted as direct evidence of the sulfa-Michael reaction/cross-linking. From a morphological point of view, beneficial effects of the restoring agents were observed by SEM analyses, in terms of a more regular hair surface and more imbricated scales.

Preliminary Study on the Activity of Phycobiliproteins against Botrytis cinerea.

Phycobiliproteins (PBPs) are proteins of cyanobacteria and some algae such as rhodophytes. They have antimicrobial, antiviral, antitumor, antioxidative, and anti-inflammatory activity at the human level, but there is a lack of knowledge on their antifungal activity against plant pathogens. We studied the activity of PBPs extracted from Arthrospiraplatensis and Hydropuntiacornea against Botrytiscinerea, one of the most important worldwide plant-pathogenic fungi. PBPs were characterized by using FT-IR and FT-Raman in order to investigate their structures. Their spectra differed in the relative composition in the amide bands, which were particularly strong in A. platensis. PBP activity was tested on tomato fruits against gray mold disease, fungal growth, and spore germination at different concentrations (0.3, 0.6, 1.2, 2.4, and 4.8 mg/mL). Both PBPs reduced fruit gray mold disease. A linear dose-response relationship was observed for both PBPs against disease incidence and H. cornea against disease severity. Pathogen mycelial growth and spore germination were reduced significantly by both PBPs. In conclusion, PBPs have the potential for being also considered as natural compounds for the control of fungal plant pathogens in sustainable agriculture.

Degradative Ability of Mushrooms Cultivated on Corn Silage Digestate.

The current management practice of digestate from biogas plants involves its use for land application as a fertilizer. Nevertheless, the inadequate handling of digestate may cause environmental risks due to losses of ammonia, methane and nitrous oxide. Therefore, the key goals of digestate management are to maximize its value by developing new digestate products, reducing its dependency on soil application and the consequent air pollution. The high nitrogen and lignin content in solid digestate make it a suitable substrate for edible and medicinal mushroom cultivation. To this aim, the mycelial growth rate and degradation capacity of the lignocellulosic component from corn silage digestate, undigested wheat straw and their mixture were investigated on Cyclocybe aegerita, Coprinus comatus, Morchella importuna, Pleurotus cornucopiae and Pleurotus ostreatus. The structural modification of the substrates was performed by using attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy. Preliminary in vitro results demonstrated the ability of P. ostreatus, P. cornucopiae and M. importuna to grow and decay hemicellulose and lignin of digestate. Cultivation trials were carried out on C. aegerita, P. cornucopiae and P. ostreatus. Pleurotus ostreatus showed the highest biological efficiency and fruiting body production in the presence of the digestate; moreover, P. ostreatus and P. cornucopiae were able to degrade the lignin. These results provide attractive perspectives both for more sustainable digestate management and for the improvement of mushroom cultivation efficiency.

Aureobasidium pullulans volatile organic compounds as alternative postharvest method to control brown rot of stone fruits.

Volatile compounds produced by L1 and L8 strains were assayed against mycelia and conidia growth of Monilinia laxa, M. fructicola, M. polystroma, and M. fructigena of stone fruits. Results showed that volatile metabolites inhibited significantly pathogens growth, in particular M. fructigena mycelium growth (70% by L1 and 50% by L8) and M. fructicola conidia germination (85% by L1 and 70% by L8) compared to the control. Moreover, the antagonistic activity was enhanced by the addition of asparagine (120 mg L-1) in the culture media composition. Synthetic pure compounds were tested in vitro on pathogens mycelial and conidia growth and their EC50 values were estimated, confirming 2-phenethyl as the most active compound. For this reason 2-phenethyl and VOCs of both yeast strains were assayed in vivo on cherry, peach, and apricot fruits. Regarding peach fruit, both treatments, yeasts and pure compounds, displayed the best inhibiting action against all the pathogens especially against M. laxa (100% by L1, 84% by L8 and 2-phenethyl). ATR/IR spectroscopy analysis showed how VOCs produced by both strains increase the fruit waxes complexity reducing the pathogens attack so playing an essential role in the antagonistic activity of both yeast strains and on fruit structural composition.

Spectroscopic and morphological data assessing the apatite forming ability of calcium hydroxide-releasing materials for pulp capping.

A pulp capping material must perform as a barrier and protect the dental pulpal complex by inducing the formation of a new dentin bridge or dentin-like tissue. Being a favorable condition for the healing process, the apatite forming ability of TheraCal (light-curable Portland-dimethacrylate cement) and Dycal (calcium hydroxide-based) pulp capping materials was studied in two simulated body fluids, i.e. Dulbecco's Phosphate Buffered Saline (DPBS) and Hank's Balanced Salt Solution (HBSS). The cements were analyzed before and after soaking in these media for different times (1-28 days) by ESEM-EDX, micro-Raman and IR spectroscopy. This data article refers to "An in vitro study on dentin demineralization and remineralization: collagen rearrangements and influence on the enucleated phase" (Di Foggia et al., 2019).

An in vitro study on dentin demineralization and remineralization: Collagen rearrangements and influence on the enucleated phase.

Dentin remineralization is of clinical relevance in the therapy of caries and dentin hypersensitivity. This study is aimed at gaining more insights on a molecular scale, through IR spectroscopy, into dentin demineralization and remineralization. The dentin demineralization by ethylenediaminetetraacetic acid, EDTA (17%, 2 h) significantly altered the secondary structure distribution of collagen, upon loss of interaction with calcium ions. To investigate dentin remineralization, previously demineralized human dentin slices were soaked in Dulbecco's Phosphate Buffered Saline (DPBS) or Hank's Balanced Salt Solution HBSS, in close contact with three commercial cements used as sustained releasing sources of Ca2+ and OH- ions (i.e. calcium hydroxide- and calcium silicate-based cements). IR spectroscopy showed the occurrence of remineralization under these conditions. Collagen did not lose its ability to chelate Ca2+, and these interactions allowed collagen to rearrange into a conformation similar to that of sound dentin. This process appeared slower in HBSS than DPBS, as also shown by the lower degree of maturation of the inorganic phase enucleated in the former medium (amorphous calcium phosphate versus B-type carbonated apatite). Collagen appeared to act as a spatial constraint to crystal deposition, affecting crystallinity and carbonate content of the enucleated phase. Remineralization was found to strongly depend on the calcium releasing ability of the cements. The fast formation of a rough apatite biocoating may represent a favorable clinical condition in the context of mineralized tissue regeneration.

Analysis of the soybean metallothionein system under free radical stress: protein modification connected to lipid membrane damage.

Metallothioneins are small Cys-rich peptides capable of coordinating metal ions, and proposed to be involved in radical stress. The four Zn(ii)-GmMT complexes of soybean (Glycine max) were recombinantly synthesised and exposed to oxidative (HO˙) and reductive (H˙ atoms and eaq-) stress conditions. Gamma-irradiation was used to simulate the endogenous formation of the reactive species in both aqueous solutions and unsaturated lipid vesicle suspensions, a biomimetic model that showed that tandem protein/lipid damage occurs, in particular under reductive radical stress. This is due to the formation of diffusible sulphur-centred radicals, which migrate from the aqueous phase to the lipid bilayer and are thus able to transform the cis double bond of the oleate moiety into the trans isomer. Among the amino acid residues present in GmMTs, Cys is one of the most sensitive residues towards the attack of free radicals, thus suggesting metal-clusters to be good interceptors of free radicals. Also Met, Tyr and Phe residues are sensitive amino acid sites of attack under both oxidative and reductive conditions. The modification of the Zn(ii)-GmMT complexes, in particular isoform 2, was monitored by Raman spectroscopy and mass spectrometry. Free radical stress on the Zn(ii)-GmMT complexes is able to induce significant structural changes such as partial deconstruction and/or rearrangement of the metal clusters, but not the complete demetallation of the proteins nor breaking of the backbone, thus confirming their capability to act as protectors under free radical stress conditions.

Silk fibres grafted with 2-hydroxyethyl methacrylate (HEMA) and 4-hydroxybutyl acrylate (HBA) for biomedical applications.

Silk fibroin may be chemically modified by grafting, with the purpose of improving its properties according to the desired function. In this study, silk fabrics from Bombyx mori silk fibres were grafted with 2-hydroxyethyl methacrylate (HEMA), as well as a binary mixture of HEMA and 4-hydroxybutyl acrylate (HBA). The samples were then electrospun from trifluoroacetic acid and treated with aqueous methanol. The% weight gains ascribable to HEMA and HBA were successfully determined through Raman spectroscopy. PolyHEMA made the fibres more hydrophilic and hindered crystallization into ß-sheet only upon electrospinning and treatment with aqueous methanol; the presence of the HBA component in the grafting mixture did not further decrease the ability of silk fibroin to rearrange into ß-sheet, due to its low contents (below 5%) under the used experimental conditions. Fibrillation partially occurred in the grafted fabrics; the electrospun samples maintained their nanostructured morphology. The surface of the substrates under investigation was compatible with cell attachment and growth, which were higher for the nanofibres. Cell adhesion and proliferation may be modulated by varying the surface chemistry and topography of the fabrics; grafting improved the surface properties of silk fibroin for enhanced functional performance in view of biomedical applications.