This in vivo study presents the preliminary results of the use of a novel piezoelectric actuator for orthopedic application. The innovative use of the converse piezoelectric effect to mechanically stimulate bone was achieved with polyvinylidene fluoride actuators implanted in osteotomy cuts in sheep femur and tibia. The biological response around the osteotomies was assessed through histology and histomorphometry in nondecalcified sections and histochemistry and immunohistochemistry in decalcified sections, namely, through Masson's trichrome, and labeling of osteopontin, proliferating cell nuclear antigen, and tartrate-resistant acid phosphatase. After one-month implantation, total bone area and new bone area were significantly higher around actuators when compared to static controls. Bone deposition rate was also significantly higher in the mechanically stimulated areas. In these areas, osteopontin increased expression was observed. The present in vivo study suggests that piezoelectric materials and the converse piezoelectric effect may be used to effectively stimulate bone growth.

a b s t r a c tBone mass distribution and structure are dependent on mechanical stress and adaptive response at cellular and tissue levels. Mechanical stimulation of bone induces new bone formation in vivo and increases the metabolic activity and gene expression of osteoblasts in culture. A wide variety of devices have been tested for mechanical stimulation of cells and tissues in vitro. The aim of this work was to experimentally validate the possibility to use piezoelectric materials as a mean of mechanical stimulation of bone cells, by converse piezoelectric effect. To estimate the magnitude and the distribution of strain, finite numerical models were applied and the results were complemented with the optical tests (Electronic Speckle Pattern Interferometric Process). In this work, osteoblasts were grown on the surface of a piezoelectric material, both in static and dynamic conditions at low frequencies, and total protein, cell viability and nitric oxide measurement comparisons are presented.

The impacts of sprinkler irrigation on infiltration, runoff and sediment loss of ten representative soils of Southern Portugal were assessed by laboratory sprinkler irrigation simulation tests. All soils showed very low permeability to applied water. The mechanical impact of water droplets enhanced soil dispersion and further lowered their infiltration capacity, particularly for high clay plus silt content soils that showed the poorest results. As a consequence, high runoff and sediment losses were also measured, primarily with the first irrigation. More moderate losses were observed thereafter. Soils with higher sand particle size fractions better absorbed the energy impact of droplets and showed higher infiltration rates and lower runoff and sediment losses. Polyacrylamide (PAM) applied to the soils through the irrigation water acted as a binding and settling agent to increase soils aggregate stability and infiltration and reduce runoff and sediment losses. Slope increase, from 2Á5 to 5%, decreased overall soils infiltration by 7% and increased runoff and sediment losses by 10 and 27%, respectively. Exposed to the same change in slope, PAM application boosted overall infiltration of treated soils to a 24% difference and increased runoff by only 10%. It had a less positive effect on sediment loss, the 5% slope being responsible for a 52% increase. In agreement with this the tests showed that, compared to the control, exposure of PAM-treated soil on 2Á5 and 5% slopes enhanced overall infiltration to 457 and 642% respectively, reduced runoff by 25% on both cases and lessened sediment loss by 39 and 27%. The demonstrated ability of PAM to influence surface soil conditions of specific soils can be used to reduce the environmental risks associated with the intensive use of sprinkler irrigation in Southern Portugal. It offers a safe, practical and non-intrusive management alternative to current costly, labour-and energy-intensive practices of increasing the number of machine turns and building storage basins to control runoff and soil erosion.

Carbon nanotubes are highly versatile materials; new applications using them are continuously being developed. Special attention is being dedicated to the possible use of multiwalled carbon nanotubes in biomaterials contacting with bone. However, carbon nanotubes are also controversial in regards to effects exerted on living organisms. Carbon nanotubes can be used to improve the tribological properties of polymer/composite materials. Ultrahigh molecular weight polyethylene (UHMWPE) is a polymer widely used in orthopedic applications that imply wear and particle generation. We describe here the response of human osteoblast-like MG63 cells after 6 days of culture in contact with artificially generated particles from both UHMWPE polymer and multiwalled carbon nanotubes (MWCNT)/UHMWPE nanocomposites. This novel composite has superior wear behavior, having thus the potential to reduce the number of revision hip arthroplasty surgeries required by wear failure of acetabular cups and diminish particle-induced osteolysis. The results of an in vitro study of viability and proliferation and interleukin-6 (IL-6) production suggest good cytocompatibility, similar to that of conventional UHMWPE (WST-1 assay results are reported as percentage of control ± SD: UHMWPE = 96.19 ± 7.92, MWCNT/UHMWPE = 97.92 ± 8.29%; total protein: control = 139.73 ± 10.78, UHMWPE = 137.07 ± 6.17, MWCNT/UHMWPE = 163.29 ± 11.81 µg/mL; IL-6: control = 90.93 ± 10.30, UHMWPE = 92.52 ± 11.02, MWCNT/UHMWPE = 108.99 ± 9.90 pg/mL). Standard cell culture conditions were considered as control. These results, especially the absence of significant elevation in the osteolysis inductor IL-6 values, reinforce the potential of this superior wearresistant composite for future orthopedic applications, when compared to traditional UHMWPE.

As interest in using carbon nanotubes for developing biologically compatible systems continues to grow, biological inspiration is stimulating new directions for in vivo approaches. The ability to integrate nanotechnology-based systems in the body will provide greater successes if the implanted material is made to mimic elements of the biological milieu especially through tuning physical and chemical characteristics. Here, we demonstrate the highly successful capacity for in vivo implantation of a new carbon nanotube-based composite that is, itself, integrated with a hydroxyapatite-polymethyl methacrylate to create a nanocomposite. The success of this approach is grounded in finely tailoring the physical and chemical properties of this composite for the critical demands of biological integration. This is accomplished through controlling the surface modification scheme, which affects the interactions between carbon nanotubes and the hydroxyapatite-polymethyl methacrylate. Furthermore, we carefully examine cellular response with respect to adhesion and proliferation to examine in vitro compatibility capacity. Our results indicate that this new composite accelerates cell maturation through providing a mechanically competent bone matrix; this likely facilitates osteointegration in vivo. We believe that these results will have applications in a diversity of areas including carbon nanotube, regeneration, chemistry, and engineering research.

Experimental results obtained in Southern Portugal from a dry-farmed mature olive tree orchard recently converted to drip irrigation are described. Water use and response to two irrigation management practices by olive trees was monitored with sap flow compensation heat pulse sensors, 'Watermark' granular matrix block sensors and a capacitance probe.The 80-plus-year-old mature olive tree orchard planted on a 12 m by 12 m spacing layout was converted in 2005 from dry-farming to drip irrigation and subjected to two water treatments: trees irrigated daily to supply for crop water demand and trees irrigated beforeflowering, during pit-hardening and before crop-harvesting. Sap flow sensors were implanted in sample trees at three different positions around the trunk and measurements were taken at 30 min intervals during 4 months, from April to mid-August of 2005. Tree transpiration rates were estimated as average of sap flow rates. When trees were fully irrigated, the observed differences in daily sap flow rate amplitude were explained by the natural trees difference in canopy cover, plant height and conductance of water vapour sites. However, when deficit irrigation was prescribed and, when the trees stopped being irrigated, they gradually lost their ability to adequately respond to the evaporative demands of the day, showing smaller variations in amplitudes sap flow. After irrigation ceased in May 15, transpiration rate gradually decreased from its maximum of 7 l h À1 , when trees were fully irrigated and soil water content was near to field capacity, to values of less than 3 l h À1 by July 3 as the soil water content gradually acted as the transpiration limiting factor.Transpiration rates recovered after irrigation was re-introduced on July 4. Although low in the non-irrigation period, transpiration rates never dropped to zero and stayed between 37 and 50 l d À1 from May 27 to June 9, as trees were able to extract soil water in the absence of irrigation. Olive trees maintained transpiration to levels as high as 50 l d À1 suggesting that long after irrigation is suppressed, a considerable amount of water held in the soil is made available to the trees. Differences in evapotranspiration and transpiration rates during the same period also indicated that olive trees, making use of the extensive root system developed in the 12 m by 12 m tree spacing, were able to extract soil water and maintain transpiration levels as high as 50 l d À1 , while soil water balance indicated tree evapotranspiration rates close to zero. This particular ability of dry-farmed olive trees to remove water held in the soil under adverse conditions of very low soil moisture and uncertainties associated with the real volume of soil effectively explored by the root system, make profile probe sensors, regardless of their accuracy, unsuitable for control of water uptake and management of dry-farmed olive orchards recently converted to irrigation. Likewise, ARTICLE IN PRESS

Bone is a composite with piezoelectric properties. Bone mass and structure are dependent on mechanical stress and adaptive response at cellular and tissue levels, but the role piezoelectricity plays in bone physiology is yet to be understood. Physical activity enhances bone density, through mechanical stimulation. Osteocytes and osteoblasts are essential for mechanosensing and mechanotransduction. Strategies have been tested for mechanical stimulation of cells and tissues in vitro. The aim of this work was to experimentally validate the use of piezoelectric materials as a mean of directly straining bone cells by converse piezoelectric effect. To estimate the magnitude of stress/strain, finite numerical models were applied and theoretical data was complemented by optic experimental data. Osteoblasts were then grown on the surface of the piezoelectric material and cell response studied.

Oxidative stress plays a central role in physiological and pathological bone conditions. Its role in signalment and control of bone cell population differentiation, activity, and fate is increasingly recognized. The possibilities of its use and manipulation with therapeutic goals are virtually unending. However, how redox balance interplays with the response to mechanical stimuli is yet to be fully understood. The present work summarizes current knowledge on these aspects, in an integrative and broad introductory perspective.