Intermolecular interactions between B. mori silk fibroin and poly(l-lactic acid) in electrospun composite nanofibrous scaffolds.

In this study, composite nanofibrous scaffolds were obtained by electrospinning a trifluoroacetic acid solution containing B. mori silk fibroin (SF) and poly(l-lactic acid) (PLLA) in a 1:1 weight ratio. SF, PLLA and SF/PLLA nanofibres were prepared with average diameter sizes of 360±90nm, 470±240nm and 580±220nm, respectively, as assessed by SEM analysis. Vibrational and thermal analyses showed that upon blending in the SF/PLLA nanofibres, the crystallisation of PLLA was hindered by the presence of SF, which crystallized preferentially and underwent conformational changes that did not significantly change its prevailing ß-sheet structure. The two components were thermodynamically compatible and the intermolecular interactions between them were revealed for the first time. Human keratinocytes were cultured on nanofibres and their viability and proliferation were determined. Preliminary in vitro tests showed that the incorporation of SF into the PLLA component enhanced cell adhesion and proliferation with respect to the unfunctionalised material. SF has been successfully used to modify the biomaterial properties and confirmed to be an efficient bioactive protein to mediate cell-biomaterial interaction.

Structural study on methacrylamide-grafted Tussah silk fibroin fibres.

Tussah silk fibroin fibres were modified by grafting with methacrylamide (MAA), with weight gains ranging between 2.6% and 71.4%. Raman and IR spectroscopic analyses showed that upon grafting the fibres underwent slight conformational changes towards a more unordered state, due to the covalent and hydrogen bonds interactions occurring between the polymer (polyMAA) and the amorphous domains of silk fibres. To test the stability towards alkaline hydrolysis, the untreated and MAA-grafted silk fibres (weight gain of 71.4%) were immersed in NaOH 5% at 50°C for different times; the IR and Raman spectroscopic techniques were utilized to elucidate the degradation mechanism as well as the rearrangements of the fibres induced by the treatment. Upon hydrolysis, both the untreated and grafted fibres underwent an enrichment in ß-sheet conformation, due to the preferential removal of the unordered domains. As a result of the covalent interactions with silk fibroin, the polymer increased its stability towards alkaline hydrolysis, since its complete solubilization was avoided and the transformation of its CONH2 groups into COO(-) and COOH was delayed. Vibrational spectroscopy proved to be a valid technique to investigate the mechanism and the effects of the hydrolytic attack, which are both fundamental to design new-generation silk-based materials.

May the surface roughness of the retrieved femoral head influence the wear behavior of the polyethylene liner?

This study was aimed at determining the surface degradation occurred on retrieved ceramic and metallic heads, as well as the influence of the head surface quality on the wear of the polyethylene counterface. To this purpose, 14 ceramic and 14 metallic femoral heads retrieved at revision surgery were examined. Scanning electron microscopic analysis provided visual evidence that some metallic heads presented crescent wear more often than the ceramic ones; the former showed a higher volumetric loss (as determined by Coordinate Measuring Machine) than the latter, but less negative Rsk values. This apparent lack of correlation between volumetric loss (i.e., wear factor) and roughness data may be explained by considering that they are two temporally variant parameters. No significant differences were observed between the Ra values of the two sets of femoral heads. The cups articulating against metal heads were characterized by higher mean wear volumes than those articulating against alumina although this difference was not statistically significant; metal heads displayed significantly higher mean wear volumes than alumina heads. The micro-Raman analysis of the cup articulated against the most worn alumina femoral head showed an orthorhombic into monoclinic phase transformation that was not observed in the cups coupled to metal heads. The obtained results showed that the surface finishing of the femoral head (in terms of Rsk values) determined the morphological changes experienced by the ultra-high-molecular-weight polyethylene crystalline phase at the molecular level. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1374-1385, 2016.

Ring closure of azo compounds to 1,2-annulated benzimidazole derivatives and further evidence of reversibility of the azo-coupling reaction.

The reaction between 1,3,5-tris(N,N-dialkylamino)benzene derivatives and 2 equiv of p-substituted benzenediazonium salts gives dicationic species which collapse to new benzimidazole derivatives with expulsion of p-substituted anilines. The presence of electron-withdrawing groups on the benzenediazo moiety of the dicationic species plays a key role in this unexpected ring closure reaction. The observed chemical behavior has been rationalized in terms of the already reported reversibility of azo coupling reactions and provided further evidence for it.

In vitro effects on mobile polyethylene insert under highly demanding daily activities: stair climbing.

PURPOSE: Wear and survival of total joint replacements do not depend on the duration of the implant in situ, but rather on the amount of its use, i.e. the patient's activity level. With this in mind, the present study was driven by two questions: (1) How does total knee replacement (TKR) respond to the simulation of daily highly demanding activities? (2) Are certain activities to be advised against or, on the contrary, useful to implanted patients, in order to reduce wear of TKR and its related problems? METHODS: One set of the same total knee prosthesis (TKP), equal in design and size, was tested on a three-plus-one knee joint simulator for two million cycles using a highly demanding daily load waveform, replicating a stair-climbing movement. The results were compared with a set of TKP previously tested with the ISO level walking task. A digital microscope was used to characterise the superficial structure of all the TKPs. Gravimetric and micro-Raman spectroscopic analyses were carried out on the polyethylene inserts. Visual comparison with in vivo explants was carried out. RESULTS: The average volumetric mass loss after two million cycles was 44 ± 6 mm(3). Microscope examinations showed some deep scratches along the flexion/extension movements for all the components. Also, the metallic backside surface showed intense non-linear scratches and the polyethylene counterface was characterised by some craters. A decrease in crystallinity, induced by mechanical stress was observed on all polyethylene components and was quantitatively confirmed by the orthorhombic fraction αo value. CONCLUSIONS: The results of this study demonstrated that the forces and motion sustained by the knee are highly activity-dependent. Moreover, this test confirmed that under more severe conditions, the material properties change according to a different wear mechanism and a decrease in crystallinity occurs. Loading characteristics for specific activities should be considered for the design of functional and robust TKRs.

Combined effect of the body mass index and implant size on the wear of retrieved total knee prostheses.

Total Knee Arthroplasty is a well-established surgical procedure performed to relieve pain and to restore function in knee osteoarthritis. A proper choice of the implant size is mandatory in order to guarantee the success of the implant and to respect the bone stock of tibial plateau and femoral condyles. The aim of this study was to investigate the combined effect of Body Mass Index and implant size on the wear damage of retrieved knee implants, a still debated subject. To this purpose, twelve total knee prostheses of the same design (NexGen, Zimmer, Warsaw, Indiana) but with different sizes were investigated. These prostheses were all cemented fixed bearing inserts and posterior stabilised. Roughness analyses were performed on femoral components to assess surface modifications. Micro-Raman spectroscopic characterization was performed on polyethylene inserts to evaluate crystallinity changes. A worsening of the surface and a nonhomogeneous aspect was observed in the prostheses characterized by a high ratio between preoperative Body Mass Index and implant size, which we called Ф factor. Delamination, a good indicator for polyethylene osteolysis, was more pronounced in the inserts having a high Ф than in those with a low Ф. Micro-Raman analyses showed more significant crystallinity changes in the prostheses of lower size, in agreement with the results previously obtained in vitro. The findings of the present study suggested that the Ф factor can be considered a predictor factor of the Total Knee Arthroplasty overall performance. Potentially, it could be used as a discriminating factor to redact Total Knee Arthroplasty outcome and prevent complications.

Toward the interpretation of the combined effect of size and body weight on the tribological performance of total knee prostheses.

PURPOSE: The research questions of the present study were: (1) Is total knee prosthesis wear behaviour influenced by implant size, body weight and their combined effect? (2) Are these findings significant and helpful from a clinical point of view? METHODS: Two very different sizes of the same total knee prosthesis (TKP), previously tested with ISO 14243 parameters, were tested on a knee simulator for a further two million cycles using a modified ISO 14243 load waveform. Roughness examination was performed on the metallic components. Gravimetric and micro-Raman spectroscopic analyses were carried out on the polyethylene inserts. RESULTS: The average volumetric mass loss was 69 ± 3 mm(3) and 88 ± 4 mm(3) for smaller and bigger size, respectively. Bigger TKPs are little influenced by an increased load, while the wear trend of the smaller TKP showed a redoubled slope, and more significant morphology changes were observed. However, the two sizes seem to behave similarly when subjected to a load increase of 15 %; the slope of the volumetric mass loss trend was comparable for the two sets of inserts, which did not appear significantly different also at the molecular level. Roughness average parameters of the lateral femoral condyle support this evidence. CONCLUSIONS: It can be asserted that the body weight and implant size are relevant to the understanding of TKP wear behaviour. A post-implantation body weight increase in a patient with smaller knee dimensions could results in more critical effects on prosthesis long-term performance.

Study on the interaction between gliadins and a coumarin as molecular model system of the gliadins-anthocyanidins complexes.

To clarify the conformational changes of gliadins (Glia) upon complexation with anthocyanidins (in particular cyanidin, Cya), the interaction of Glia with a coumarin derivative (3-ethoxycarbonylcoumarin, 3-EcC), having a benzocondensed structure similar to that of Cya, has been investigated by NMR, IR, and Raman spectroscopy under acidic and neutral conditions. Raman spectra showed that both molecules produce a similar effect on the Glia structure, i.e. an increase in the α-helix conformation and a decrease in ß-sheet and ß-turns content. In the presence of both molecules, this effect is more marked; the spectroscopic results showed that both Cya and 3-EcC interact with Glia and 3-EcC favors the complex formation with Glia. The results obtained in this study provide new insights into anthocyanidins-Glia interactions and may have relevance to human health, in the field of the attempts to modify gluten proteins to decrease allergen immunoreactivity.

On the antibacterial activity of roots of Capparis spinosa L.

A decoction of Capparis spinosa L. roots, widely used in the traditional folk medicine of southern Italy, was prepared and submitted to antibacterial activity tests, which showed an interesting bacteriostatic activity on the growth of Deinococcus radiophilus. Heterocyclic compounds were also recovered from the chloroformic extract of the roots.

Interaction between gliadins and anthocyan derivatives.

The interaction of gliadins with some anthocyanins (e.g. myrtillin, malvin, keracyanin, callistephin) and anthocyanidins (e.g. delphinidin, pelargonidin, cyanidin) has been analysed in aqueous solution at pH condition of the stomach, in which these compounds are initially metabolized. NMR, FT-IR and UV-Vis spectroscopic methods have been employed to determine the anthocyanin binding mode. The spectroscopic data seem to indicate that anthocyans are located along the polypeptide chains of gliadins in a generical molecular interaction between the two moieties. Our data do not exclude that hydrogen bonding interaction too is operating. Anthocyan-gliadins complexes are very soluble in acidic conditions. The results provide new insights into anthocyan-protein interaction and may have relevance to human health.

Meisenheimer-Wheland complexes between 1,3,5-tris(N,N-dialkylamino)benzenes and 4,6-dinitrotetrazolo[1,5-a]pyridine. Evidence of reversible C-C coupling in the S(E)Ar/S(N)Ar reaction.

Reactions between a superelectrophilic carbon reagent, 4,6-dinitrotetrazolopyridine, and 1,3,5-tris(N,N-dialkylamino)benzenes, a supernucleophilic carbon reagent series, afford C-C coupling products which are "double sigma-complexes" (W-M), Wheland-like on the 1,3,5-tris(N,N-dialkylamino)benzene moiety and Meisenheimer-like on the 4,6-dinitrotetrazolopyridine moiety. These complexes were moderately stable at low temperature, and they were characterized by NMR spectroscopy methods. (1)H NMR experiments at variable temperature strongly indicate that the formation of these complexes by a nucleophile/electrophile attack is a reversible process.

Evidence of reversibility in azo-coupling reactions between 1,3,5-tris(N,N-dialkylamino)benzenes and arenediazonium salts.

The reactions between strongly electron-rich aromatic substrates (1,3,5-tris(N,N-dialkylamino)benzenes, neutral carbon super nucleophiles) and diazonium salts produce moderately stable sigma complexes (Wheland complexes). The reactivity of Wheland complexes with electrophiles (other diazonium salts, or 4,7-dinitrobenzofuroxan) produces exchange reactions in the electrophilic part: the better electrophile replaces the less powerful electrophile. In the same way, in Wheland complexes with the 1,3,5-tris(morpholinyl)benzene, the 1,3,5-tris(piperidinyl)benzene replaces the less powerful nucleophile 1,3,5-tris(morpholinyl)benzene. Evidence is reported here indicating that for the title system the reaction of the attack of the electrophilic reagent producing Wheland complexes is a reversible process. The final products of the diazo-coupling reactions undergo a further attack of some diazonium salts. From the final products of the double diazo-coupling reactions (diazo compounds), we collected evidence that is a clear instance of complete reversibility of the diazo-coupling reaction.

Evidence for the intermediacy of Wheland-Meisenheimer complexes in SEAr reactions of aminothiazoles with 4,6-dinitrobenzofuroxan.

Reactions of DNBF with a series of 2-aminothiazoles (1 a-f) to afford thermodynamically stable C-bonded sigma-adducts have been investigated in acetonitrile. A most significant finding emerged on recording NMR spectra immediately after mixing of equimolar amounts of DNBF and the unsubstituted 2-aminothiazole (1 a) in Me2SO: namely, that the formation of 9 a is preceded by that of a short-lived intermediate species X. From the 1H NMR parameters characterizing this intermediate, as well as the dependence of its lifetime on the experimental conditions-the presence of excess DNBF over 1 a increases the lifetime of X while an excess of base (1 a) accelerates its conversion into 9 a--it is convincingly demonstrated that the structure of X combines the presence of a positively charged Wheland complex moiety (with regard to the thiazole ring) with that of a negatively charged Meisenheimer complex moiety (with regard to the benzofuroxan system). So far, only one intermediate of this type (noted WM) has been successfully characterized, in the reactions of DNBF with 1,3,5-tris(N,N-dialkylamino)benzenes. Among the key features supporting the intermediacy of X along the reaction coordinate leading to 9 a is the fact that the reactions of DNBF with 1 a in the presence of an alcohol (MeOH, EtOH, nPrOH) produce new adducts arising from the addition of an alcohol molecule to the thiazole moiety of WM-1 a. Reflecting the presence of three chiral centres, these species are formed as mixtures of several diastereomers that could all be characterized in their racemic forms in ethanol. These findings generalize the previous report on the formation of Wheland-Meisenheimer carbon-carbon complexes in homocyclic series.