Multifunctional Lanthanide-Doped Core/Shell Nanoparticles: Integration of Upconversion Luminescence, Temperature Sensing, and Photothermal Conversion Properties.

Multifunctional integration on single upconversion nanoparticles (UCNPs), such as the simultaneous achievement of imaging, sensing, and therapy, will be extremely attractive in various application fields. Herein, we demonstrated that single core/shell NaGdF4:Yb/Er-based UCNPs (<10 nm) with a highly Yb3+ or Nd3+ doped shell simultaneously exhibited good upconversion luminescence (UCL), temperature sensing, and photothermal conversion properties under 980 or 808 nm excitation, respectively. The spatial separation between the emission/sensing core and the heating shell was able to tailor the competition between the light and heat generation processes, and hence higher UCL efficiency and enhanced heating capability were achieved by introducing the rational core/shell design. Especially, Nd3+-sensitized core/shell nanoparticles were excitable to the laser at a more biocompatible wavelength of 808 nm, and hence the heating effect of water was greatly minimized. The heating and sensing capabilities of Nd3+-sensitized core/shell UCNPs with smaller sizes (<10 nm) were confirmed in aqueous environment under single 808 nm laser excitation, implying their promising applications in imaging-guided and temperature-monitored photothermal treatments.

Loading-Induced Reduction in Sclerostin as a Mechanism of Subchondral Bone Plate Sclerosis in Mouse Knee Joints During Late-Stage Osteoarthritis.

OBJECTIVE: To establish an unbiased, 3-dimensional (3-D) approach that quantifies subchondral bone plate (SBP) changes in mouse joints, and to investigate the mechanism that mediates SBP sclerosis at a late stage of osteoarthritis (OA). METHODS: A new micro-computed tomography (micro-CT) protocol was developed to characterize the entire thickness of the SBP in the distal femur of a normal mouse knee. Four mouse models of severe joint OA were generated: cartilage-specific Egfr-knockout (Egfr-CKO) mice at 2 months after surgical destabilization of the medial meniscus (DMM), Egfr-CKO mice with aging-related spontaneous OA, wild-type (WT) mice at 10 months after DMM, and WT mice at 14 weeks after DMM plus hemisectomy of the meniscus (DMMH) surgery. As an additional model, mice with knockout of the sclerostin gene (Sost-KO) were subjected to DMMH surgery. Knee joints were examined by micro-CT, histology, and immunohistochemical analyses. RESULTS: Examination of the mouse distal femur by 3-D micro-CT revealed a positive correlation between SBP thickness and the loading status in normal knees. In all 4 mouse models of late-stage OA, SBP sclerosis was restricted to the areas under severely eroded articular cartilage. This was accompanied by elevated bone formation at the bone marrow side of the SBP and a drastic reduction in the levels of sclerostin in osteocytes within the SBP. Unlike in WT mice, no further increase in the thickness of the SBP was observed in response to DMMH in Sost-KO mice. CONCLUSION: Since focal stress on the SBP underlying sites of cartilage damage increases during late stages of OA, these findings establish mechanical loading-induced attenuation of sclerostin expression and elevation of bone formation along the SBP surface as the major mechanisms characterizing subchondral bone phenotypes associated with severe late-stage OA in mice.

Water deficit affected flavonoid accumulation by regulating hormone metabolism in Scutellaria baicalensis Georgi roots.

The content of flavonoids especially baicalin and baicalein determined the medical quality of Scutellaria baicalensis which is a Chinese traditional medicinal plant. Here, we investigated the mechanism responsible for the content and composition of flavonoids in S. baicalensis under water deficit condition. The transcription levels of several genes which are involved in flavonoid biosynthesis were stimulated by water deficit. Under water deficit condition, fifteen up-regulated proteins, three down-regulated proteins and other six proteins were detected by proteomic analysis. The identified proteins include three gibberellin (GA)- or indoleacetic acid (IAA)-related proteins. Decreased endogenous GAs level and increased IAA level were observed in leaves of S. baicalensis which was treated with water deficit. Exogenous application of GA or α-naphthalene acelic acid (NAA) to plants grown under water deficit conditions led to the increase of endogenous GAs and the decrease of IAA and flavonoids, respectively. When the synthesis pathway of GA or IAA in plants was inhibited by application with the inhibitors, flavonoid levels were recovered. These results indicate that water deficit affected flavonoid accumulation might through regulating hormone metabolism in S. baicalensis Georgi.

Flavonoids and antioxidative enzymes in temperature-challenged roots of Scutellaria baicalensis Georgi.

The active compounds in the roots of Scutellaria baicalensis Georgi, a traditional Chinese medicinal plant, are mainly flavonoids which have anti-inflammatory, antitumour, and anti-HIV activity, respectively. The increasing annual average temperature has rendered the S. baicalensis plants grown in some ancient producing regions no longer suitable for their medicinal usage. Hydrogen peroxide plays an important role in root responses to abnormal temperature in S. baicalensis. Baicalin and baicalein and antioxidative enzymes were anticipated to detoxify H2O2 in S. baicalensis. Here, we show that abnormal temperatures (10 and 40 degrees C) decreased the content of flavonoids as compared with the normal temperature (30 degrees C), and the transcripts of UDP-glucuronate:baicalein 7-O-glucuronosyltransferase and beta-glucuronidase involved in the interconversion between baicalin and baicalein were affected by the 40-degrees C treatment. High temperature also increased the activities of catalase and peroxidase. Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that the transcript levels of peroxidase 2, peroxidase 3, monodehydroascorbate reductase 2, and dehydroascorbate reductase were significantly increased under high-temperature conditions. The respective genes would be candidates for improvement of the adaptation of S. baicalensis plants to abnormal temperatures and for regulation of the contents of the active compounds.

[Effects of PEG stress on flavonoids accumulation and related gene expression in suspension of Scutellaria baicalensis].

OBJECTIVE: To study the effects of PEG stress on baicalin, baicalein accumulation induced by an increased concentration of PEG solution and the related genes' expression in suspension of Scutellaria baicalensis. METHOD: The content of baicalin, baicalein in suspension of S. baicalensis was determined by HPLC. The related genes' expression was analyzed by semi-quantitative PCR. RESULT: The content of proline in suspension of S. baicalensis was promoted by PEG treatment. Ten percent PEG treatment promoted the expression of PAL and the content of baicalein in experimental material via a drought stress. 20% PEG solution treatment promoted the expression of UBGAT. At the same time, the increased activity of APX inhibited the progress of eliminating reactive oxygen by baicalein, which induced the transformation from baicalein to baicalin. CONCLUSION: Active ingredient in suspension of S. baicalensis was promoted significantly via a stress of light concentration of PEG solution.

Validation of the Boussinesq equation for use in traction field determination.

Cell traction force plays an important role in many biological processes. Several traction force microscopy methods have been developed to determine cell traction forces based on the Boussinesq solution. This approach, however, is rooted in a half-space assumption. The purpose of this study was to determine the error induced in the half-space assumption using a finite element method (FEM). It demonstrates that displacement error between the FEM and the Boussinesq equation can be used to measure the accuracy of the Boussinesq equation, although singularity exists in the loading point. For one concentrated force, significant difference between the FEM and the Boussinesq equation occurs in the whole field; this difference decreases with an increase in the plate thickness. However, in the case of the balanced forces, the offset of the balanced forces decreases the errors in the middle area. Overall, this study demonstrates that increasing the thickness of the polyacrylamide gel is important for reducing the error of the Boussinesq equation when determining the displacement field of the gel under loads.

Development of a subject-specific model to predict the forces in the knee ligaments at high flexion angles.

Recent clinical evidence has suggested that tasks performed in kneeling or squatting postures place the knee at a higher risk for injury because loads across the knee might overload the ligaments. The objective of this study was to develop a subject-specific model of the knee that is kinematically driven to predict the forces in the major ligaments at high flexion angles. The geometry of the femur, tibia, and fibula and the load-elongation curves representing the structural properties of the ACL, PCL, LCL, and MCL served as inputs to the model, which represented each ligament as a nonlinear elastic spring. To drive the model, kinematic data was obtained while loads were applied to the same cadaveric knee at four flexion angles. The force in each ligament during the recorded kinematic data allowed an optimization procedure to determine the location of the ligament attachment sites on each bone and their reference lengths. The optimization procedure could successfully minimize the differences between the experimental and predicted forces only when the kinematics at 90°, 120°, and 140° of flexion were utilized. This finding suggests that the ligaments at the knee function differently at high-flexion angles compared to low flexion angles and separate models must be used to examine each range of motion. In the future, the novel experimental and computational methodology will be used to construct additional models and additional knee kinematics will be input to help elucidate mechanisms of injury during tasks performed in kneeling or squatting postures.

Genetic stability, active constituent, and pharmacoactivity of Salvia miltiorrhiza hairy roots and wild plant.

Salvia miltiorrhiza is an annual plant growing in China, Mongolia, Korea and some other Asian countries. The extract from S. miltiorrhiza roots has been used for supporting healthy cardiovascular and circulatory systems during the last decade. The active constituents of S. miltiorrhiza from different areas vary significantly, and the wild resources are overexploited. To adapt the demand for active constituents of S. miltiorrhiza against cardiovascular-related diseases, alternative materials need to be developed. The aim of the present work was to investigate the possibility of S. miltiorrhiza hairy roots as the alternative materials. The results showed that S. miltiorrhiza hairy roots are genetically stable. The contents of salvianolic acid B and tanshinone IIA, two main active constituents in hairy roots, determined by the assessment of combining flow cytometry and phytochemical analysis, are comparable to or significantly lower than in wild plant roots. The extract from S. miltiorrhiza hairy roots also had similar protection activity for hypoxia and reoxygenation injury in rat cardiac myocytes like that from wild plant roots. S. miltiorrhiza hairy roots may be alternative materials to obtain the drug or healthy food for cardiovascular-related diseases.

A new image processing technique for determination of cell-generated deformations on substrata.

This paper introduces a new image processing technique that determines the displacement field of a given substrate from "null-force" and "force-loaded" images. In this method, fluorescent elements used to track motion, which will be referred to as beads, can be seen in these images by locating the gray value that is normally distributed around their central point. Next comes a two-step process of matching the beads with displacements. The first step matches the beads with a small displacement using the correlation function of the characteristic pixels. Based on results from this initial step, another correlation function determines a pair of beads with a relatively large displacement. The entire matching process is done in this way, gradually working from the small displacement to the large one. Finally, using the cubic spline weight function, the whole displacement field is interpolated and filtered out of those displacements, which were initially found with the matched beads. Applying this new method on the cell migration yields satisfying results. Based on the particle tracking, the displacement field obtained by this new image processing technique has clear physical meaning. More importantly, this new method completes the matching of the displacement using the features of the displacement field, thus avoiding the direct matching with the image gray values for the relatively large strain of the substrate around the cell. Accordingly, it greatly decreases mismatching, making data checking unnecessary.

Development of micropost force sensor array with culture experiments for determination of cell traction forces.

Cell traction forces (CTFs) are critical for cell motility and cell shape maintenance. As such, they play a fundamental role in many biological processes such as angiogenesis, embryogenesis, inflammation, and wound healing. To determine CTFs at the sub-cellular level with high sensitivity, we have developed high density micropost force sensor array (MFSA), which consists of an array of vertically standing poly(dimethylsiloxane) (PDMS) microposts, 2 microm in diameter and 6 microm in height, with a center-to-center distance of 4 microm. In combination with new image analysis algorithms, the MFSA can achieve a spatial resolution of 40 nm and a force sensitivity of 0.5 nN. Culture experiments with various types of cells showed that this MFSA technology can effectively determine CTFs of cells with different sizes and traction force magnitudes.

Dynamic finite element modeling of poroviscoelastic soft tissue.

Clinical evidences relative to biomechanical factors have demonstrated their important contribution to the behaviour of soft tissues. Finite element (FE) analysis is used to study the mechanical behaviour of soft tissue because it can provide numerical solutions to problems that are intractable to analytic solutions. This study focuses on the development of a FE model of a poroelastic biological tissue, which incorporates the viscoelastic material behaviour, finite deformation and inertial effect. The FE formulation is based on the weak form derived from the governing equation, and Newmark-beta method as well as Newton's method is incorporated into the implicit non-linear solutions. One-dimensional analytical solutions were used to verify the theoretical formulation and the numerical implementation of the proposed model. This study was further extended to analyze two-dimensional biomechanical models and the results clearly demonstrate the importance of including finite deformation, viscoelasticity and inertial effects.

Determining substrate displacement and cell traction fields--a new approach.

This paper presents a new approach for the traction force microscopy (TFM) method which determines traction forces exerted by adherent cells on a thin, elastic polyacrylamide gel embedded with fluorescent microbeads. In this enhanced TFM method, a pattern recognition technique is first applied to match the pair of microbead embedded images before and after deformation, which subsequently provides the displacement field of the elastic substrate. Once the displacement field is obtained, the 3-D finite element method (FEM) is used to compute cell traction forces. The new TFM has been applied to determine traction forces of human tendon fibroblasts. Compared to existing TFM methods, the present method has the following advantages: (1) its displacement field obtained is associated with microbead movements; (2) it considers the finite thickness of the thin polyacrylamide gel and is therefore free from the infinite half-space approximation adopted by existing TFM methods; and (3) its computation procedure for determining cell traction forces is fast.