Local Mechanical Stimuli Regulate Bone Formation and Resorption in Mice at the Tissue Level

Bone is able to react to changing mechanical demands by adapting its internal microstructure through bone forming and resorbing cells. This process is called bone modeling and remodeling. It is evident that changes in mechanical demands at the organ level must be interpreted at the tissue level where bone (re)modeling takes place. Although assumed for a long time, the relationship between the locations of bone formation and resorption and the local mechanical environment is still under debate. The lack of suitable imaging modalities for measuring bone formation and resorption in vivo has made it difficult to assess the mechanoregulation of bone three-dimensionally by experiment. Using in vivo micro-computed tomography and high resolution finite element analysis in living mice, we show that bone formation most likely occurs at sites of high local mechanical strain (p<0.0001) and resorption at sites of low local mechanical strain (p<0.0001). Furthermore, the probability of bone resorption decreases exponentially with increasing mechanical stimulus (R² = 0.99) whereas the probability of bone formation follows an exponential growth function to a maximum value (R² = 0.99). Moreover, resorption is more strictly controlled than formation in loaded animals, and ovariectomy increases the amount of non-targeted resorption. Our experimental assessment of mechanoregulation at the tissue level does not show any evidence of a lazy zone and suggests that around 80% of all (re)modeling can be linked to the mechanical micro-environment. These findings disclose how mechanical stimuli at the tissue level contribute to the regulation of bone adaptation at the organ level.     

Mechanical response and conformational changes of alpha-actinin domains during unfolding: a molecular dynamics study

Alpha-actinin is a cytoskeleton-binding protein involved in the assembly and regulation of the actin filaments. In this work molecular dynamics method was applied to investigate the mechanical behaviour of the human skeletal muscle α-actinin. Five configurations were unfolded at an elongation speed of 0.1 nm/ps in order to investigate the conformational changes occurring during the extension process. Moreover, a sensitivity analysis at different velocities was performed for one of the R2–R3 spectrin-like repeat configuration extracted in order to evaluate the effect of the pulling speed on the mechanical behaviour of the molecule. Two different behaviours were recognized with respect to the pulling speed. In particular, at speed higher than 0.025 nm/ps a continuous rearrangement without evident force peaks was obtained, on the contrary at lower speed evident peaks in the range 500–750 pN were detected. R3 repeat resulted more stable than R2 during mechanical unfolding, due to the lower hydrophobic surface available to the solvent. The characterization of the R2–R3 units can be useful for the development of cytoskeleton network models based on stiffness values obtained by analyses performed at the molecular level.     

Effect of thermal shock in the development of androgenic plants of <Emphasis Type="Italic">Anemone coronaria</Emphasis> L.: influence of genotype and flower parameters

Anemone is an allogamous species marketed as a cut flower. Pure lines formation in this species is expensive and inefficient because progeny suffers from inbreeding depression. Pure lines obtained from plant anthers are a widely used material to produce new hybrids in short time. The aim of this study is the optimization of production of A. coronaria androgenic plants. Flower buds of eight cultivars were measured, disinfected for 20′ in 1% sodium hypochlorite and rinsed with distilled sterile water. Development stage of microspores and their size were evaluated. Anthers were distributed in Petri plates containing a double layer constituted of Nitsch and Nitsch agarized substrate enriched with activated charcoal (1%) and with or without sucrose (3%) and the same substrate in liquid phase without activated charcoal. Anthers were shocked with a preculture at 5 or 33 °C for 5 days and then incubated in the dark at 23 °C. After 21 weeks, the number of regenerants was counted. Plants were transferred into the flask with modified MS substrate and placed at 18 °C. After 3 months of in vitro culture, the survival rate of plants was evaluated. Among the varieties, significant differences in anther number, microspore size, and presence of abnormal microspores were observed. Heat shock pretreatment at 33 °C increases the number and survival of embryos and plants, but the varieties respond to the thermal shock in different ways. The influence of genotype and the maturation phase of flower buds, anthers and microspores are discussed related to the androgenic plant development.     

Essential Oil Composition and Volatile Profile of Seven Helichrysum Species Grown in Italy

Helichrysum genus consists of about 600 species widespread throughout the world, especially in South Africa and in the Mediterranean area. In this study the aroma profile (HS‐SPME) and the EO compositions of seven Helichrysum species (H. cymosum, H. odoratissimum, H. petiolare, H. fontanesii, H. saxatile, H. sanguineum, and H. tenax) were evaluated. All the plants were grown in Italy under the same growth conditions. The volatile constituents, particularly monoterpenes, depended by the plant's genotype and ecological adaptation. This study represents the first headspace evaluation on the selected plants and the results evidenced that monoterpenes represented the main class of constituents in five of the seven species analysed (from 59.2% to 95.0%). The higher content in sesquiterpene hydrocarbons was observed in the Mediterranean species of H. sanguineum (68.0%). Only H. saxatile showed relative similar abundance of monoterpenes and sesquiterpene hydrocarbons. The essential oil composition of the majority of examined species are characterised by high percentage of sesquiterpenes (especially β‐caryophyllene and δ‐cadinene) ranging from 51.3% to 92.0%, except for H. cymosum, H. tenax, and H. sanguineum leaves where monoterpenes predominated (from 51.7% to 74.7%).     

Mechanisms of reduced implant stability in osteoporotic bone

The determining factors for the fixation of uncemented screws in bone are the bone-implant interface and the peri-implant bone. The goal of this work was to explore the role of the peri-implant bone architecture on the mechanics of the bone-implant system. In particular, the specific aims of the study were to investigate: (i) the impact of the different architectural parameters, (ii) the effects of disorder, and (iii) the deformations in the peri-implant region. A three-dimensional beam lattice model to describe trabecular bone was developed. Various microstructural features of the lattice were varied in a systematic way. Implant pull-out tests were simulated, and the stiffness and strength of the bone-implant system were computed. The results indicated that the strongest decrease in pull-out strength was obtained by trabecular thinning, whereas pull-out stiffness was mostly affected by trabecular removal. These findings could be explained by investigating the peri-implant deformation field. For small implant displacements, a large amount of trabeculae in the peri-implant region were involved in the load transfer from implant to bone. Therefore, trabecular removal in this region had a strong negative effect on pull-out stiffness. Conversely, at higher displacements, deformations mainly localized in the trabeculae in contact with the implant; hence, thinning those trabeculae produced the strongest decrease in the strength of the system. Although idealized, the current approach is helpful for a mechanical understanding of the role played by peri-implant bone.     

Quantitative,structural, and image-based mechanical analysis of nonunion fracture repaired by genetically engineered mesenchymal stem cells

Stem cell-mediated gene therapy for fracture repair, utilizes genetically engineered mesenchymal stem cells (MSCs) for the induction of bone growth and is considered a promising approach in skeletal tissue regeneration. Previous studies have shown that murine nonunion fractures can be repaired by implanting MSCs over-expressing recombinant human bone morphogenetic protein-2 (rhBMP-2). Nanoindentation studies of bone tissue induced by MSCs in a radius fracture site indicated similar elastic modulus compared to intact murine bone, eight weeks post-treatment. In the present study we sought to investigate temporal changes in microarchitecture and biomechanical properties of repaired murine radius bones, following the implantation of MSCs. High-resolution micro-computed tomography (micro-CT) was performed 10 and 35 weeks post MSC implantation, followed by micro-finite element (micro-FE) analysis. The results have shown that the regenerated bone tissue remodels over time, as indicated by a significant decrease in bone volume, total volume, and connectivity density combined with an increase in mineral density. In addition, the axial stiffness of limbs repaired with MSCs was 2–1.5 times higher compared to the contralateral intact limbs, at 10 and 35 weeks post-treatment. These results could be attributed to the fusion that occurred in between the ulna and radius bones. In conclusion, although MSCs induce bone formation, which exceeds the fracture site, significant remodeling of the repair callus occurs over time. In addition, limbs treated with an MSC graft demonstrated superior biomechanical properties, which could indicate the clinical benefit of future MSC application in nonunion fracture repair.     

Three years of experience in biomanipulating a small eutrophic lake: Lago di Candia (Northern Italy)

Results obtained from a step by step approach to the biomanipulation of a natural lacustrine environment (Lago di Candia, Northern Italy) are presented. Since the diversion of the municipal sewage of the small town of Candia, runoff and precipitation have been the sole contributors of nutrient to the lake. Fish population is mainly characterized by rudd (Scardinius erythrophthalmus) overstocking and by a low density of large-mouth-bass (Micropterus salmoides) and pike (Esox lucius). During 1986 about 12t of rudd (1–2 year old) were removed from the lake. Considering 1986 as ‘control year’, average Secchi disc transparency improved from 2.3 m in 1986 to 3.3 m in 1988; phytoplankton biovolume decreased from 114 to 58 mm³ l⁻¹ but zooplankton biovolume increased from 8 to 11.5 mm³ l⁻¹. The results achieved show that a grodual biomanipulation treatment can have a satisfactory outcome, and has the advantage of not producing catastrophic situations either in the biotic or in the abiotic compartments of the lake.     

Ecophysiological and phytochemical responses of <Emphasis Type="Italic">Salvia sinaloensis</Emphasis> Fern. to drought stress

Salvia sinaloensis Fern. (sage) is a medicinal plant containing plant secondary metabolites (PSMs) with antioxidant properties. The current study investigated the effects of drought stress on S. sinaloensis morphological and ecophysiological traits, and active constituent production. Sage plants were cultivated in controlled conditions for 34 days and exposed to full irrigation as control, half irrigation, or no irrigation. Changes in growth index (G.I.), dry biomass, leaf water potential (LWP), physiological parameters, active compounds, volatilome (BVOCs) and essential oils (EOs) were determined. Not irrigated plants showed a decrease in total chlorophyll content (~???14.7%) and growth (G.I., ~???59.4%) from day 18, and dry biomass at day 21 (??56%), when the complete leaf withering occurred (LWP, ??1.10 MPa). Moderate drought stressed plants showed similar trends for chlorophyll content and growth but kept a constant LWP (??0.35 MPa) and dry biomass throughout the experiment, as control plants. Carotenoids were not affected by water regimes. The photosynthetic apparatus tolerated mild to severe water deficits, without a complete stomatal closure. Plants under both stress conditions increased the percentage of phenols and flavonoids and showed altered BVOC and EO chemical profiles. Interestingly Camphor, the main EO oxygenated monoterpene, increased in moderate stressed plants while the sesquiterpene hydrocarbon Germacrene D decreased. The same trend was seen in the headspace under stress severity. The data evidenced a possible role of the active molecules in the response of S. sinaloensis plants to drought stress. Taking together, these findings point at S. sinaloensis as a potential drought adaptive species, which could be used in breeding strategies to obtain sages with high quality PSMs, saving irrigation water.     

Spatial Genetic Structure of Campanula sabatia, a Threatened Narrow Endemic Species of the Mediterranean Basin

Campanula sabatia is endemic to NW Italy (Liguria) and it was included in the European Red list of endangered species due to the heavy human pressure on its habitat. AFLP markers were used to detect the genetic diversity within and among ten populations (totalling 83 individuals) representing the range of the species. In spite of the limited distribution of this endemic taxon, high levels in percentage of polymorphic bands (PPB), gene diversity (H _S and H _T) and Shannon??s information index (I) were detected both at population (PPB?=?60?%, H _S?=?0.1853, I?=?0.2836) and at species level (PPB?=?100?%, H _T?=?0.2415, I?=?0.3871). The coefficient of genetic differentiation among population (G??_ST) was 0.1935, while the level of gene flow (N?? _m) was estimated to be 2.0832. AMOVA analysis identified a genetic variation within populations of 83?% of the total. Bayesian clustering methods assigned individuals to two geographical groups partly found within the same population, probably due to a high rate of genetic exchange among its populations. Conservation measures are suggested on the basis of the genetic diversity detected to ensure an effective protection for this endemic species.