Paraffin-Based Process for Producing Layered Composites with Cellular Microstructures

A paraffin-based process that results in high-strength bimaterial ceramic layered composites is reported. The process facilitates rolling, folding, and shape retention at room temperature and allows the transition from a laminar to a cellular microstructure during deformation. The strength of sintered alumina/zirconia/alumina composites reached 700 MPa, higher than that of conventional zirconia-toughened alumina composites containing dispersed particles

Myogenic Progenitor Cells Control Extracellular Matrix Production by Fibroblasts during Skeletal Muscle Hypertrophy

Satellite cells, the predominant stem cell population in adult skeletal muscle, are activated in response to hypertrophic stimuli and give rise to myogenic progenitor cells (MPCs) within the extracellular matrix (ECM) that surrounds myofibers. This ECM is composed largely of collagens secreted by interstitial fibrogenic cells, which influence satellite cell activity and muscle repair during hypertrophy and aging. Here we show that MPCs interact with interstitial fibrogenic cells to ensure proper ECM deposition and optimal muscle remodeling in response to hypertrophic stimuli. MPC-dependent ECM remodeling during the first week of a growth stimulus is sufficient to ensure long-term myofiber hypertrophy. MPCs secrete exosomes containing miR-206, which represses Rrbp1, a master regulator of collagen biosynthesis, in fibrogenic cells to prevent excessive ECM deposition. These findings provide insights into how skeletal stem and progenitor cells interact with other cell types to actively regulate their extracellular environments for tissue maintenance and adaptation

Potential Benefits of Combined Statin and Metformin Therapy on Resistance Training Response in Older Individuals

Metformin and statins are currently the focus of large clinical trials testing their ability to counter age-associated declines in health, but recent reports suggest that both may negatively affect skeletal muscle response to exercise. However, it has also been suggested that metformin may act as a possible protectant of statin-related muscle symptoms. The potential impact of combined drug use on the hypertrophic response to resistance exercise in healthy older adults has not been described. We present secondary statin analyses of data from the MASTERS trial where metformin blunted the hypertrophy response in healthy participants (\u3e65 years) following 14 weeks of progressive resistance training (PRT) when compared to identical placebo treatment (n = 94). Approximately one-third of MASTERS participants were taking prescribed statins. Combined metformin and statin resulted in rescue of the metformin-mediated impaired growth response to PRT but did not significantly affect strength. Improved muscle fiber growth may be associated with medication-induced increased abundance of CD11b+/CD206+ M2-like macrophages. Sarcopenia is a significant problem with aging and this study identifies a potential interaction between these commonly used drugs which may help prevent metformin-related blunting of the beneficial effects of PRT

Long-Lasting Impairments in Quadriceps Mitochondrial Health, Muscle Size, and Phenotypic Composition Are Present After Non-Invasive Anterior Cruciate Ligament Injury

Introduction: Despite rigorous rehabilitation aimed at restoring muscle health, anterior cruciate ligament (ACL) injury is often hallmarked by significant long-term quadriceps muscle weakness. Derangements in mitochondrial function are a common feature of various atrophying conditions, yet it is unclear to what extent mitochondria are involved in the detrimental sequela of quadriceps dysfunction after ACL injury. Using a preclinical, non-invasive ACL injury rodent model, our objective was to explore the direct effect of an isolated ACL injury on mitochondrial function, muscle atrophy, and muscle phenotypic transitions. Methods: A total of 40 male and female, Long Evans rats (16-week-old) were exposed to non-invasive ACL injury, while 8 additional rats served as controls. Rats were euthanized at 3, 7, 14, 28, and 56 days after ACL injury, and vastus lateralis muscles were extracted to measure the mitochondrial respiratory control ratio (RCR; state 3 respiration/state 4 respiration), mitochondrial reactive oxygen species (ROS) production, fiber cross sectional area (CSA), and fiber phenotyping. Alterations in mitochondrial function and ROS production were detected using two-way (sex:group) analyses of variance. To determine if mitochondrial characteristics were related to fiber atrophy, individual linear mixed effect models were run by sex. Results: Mitochondria-derived ROS increased from days 7 to 56 after ACL injury (30–100%, P \u3c 0.05), concomitant with a twofold reduction in RCR (P \u3c 0.05). Post-injury, male rats displayed decreases in fiber CSA (days 7, 14, 56; P \u3c 0.05), loss of IIa fibers (day 7; P \u3c 0.05), and an increase in IIb fibers (day 7; P \u3c 0.05), while females displayed no changes in CSA or phenotyping (P \u3e 0.05). Males displayed a positive relationship between state 3 respiration and CSA at days 14 and 56 (P \u3c 0.05), while females only displayed a similar trend at day 14 (P = 0.05). Conclusion: Long-lasting impairments in quadriceps mitochondrial health are present after ACL injury and play a key role in the dysregulation of quadriceps muscle size and composition. Our preclinical data indicate that using mitoprotective therapies may be a potential therapeutic strategy to mitigate alterations in muscle size and characteristic after ACL injury

Shear bond strength between veneering ceramics and ceria-stabilized zirconia/alumina

STATEMENT OF PROBLEM: Ceria-stabilized tetragonal ZrO(2)/Al(2)O(3) nanocomposite (Ce-TZP/A) offers superior properties compared to yttria-stabilized zirconia (Y-TZP). However, the bond quality to veneering ceramics has not been investigated. PURPOSE: The purpose of this study was to evaluate the bond strength of different veneering ceramics to Ce-TZP/A. MATERIAL AND METHODS: Cubes of Ce-TZP/A (NANOZR) (edge length, 10 mm) were layered with veneering ceramics (5 mm in thickness) with or without application of a liner and sheared at the interface. The effect of different surface treatments (polished with 3-mum diamond paste or airborne-particle abraded) was evaluated with 1 veneering ceramic (Cerabien ZR). Shear bond strength of 5 additional veneering ceramics (IPS e.max, Initial ZR, Triceram, Vintage ZR, or VITA VM 9) to polished Ce-TZP/A was measured (n=10). Polished Y-TZP (Hint-ELs ZrO(2) HIP) veneered with 2 ceramics (Cerabien ZR, Vintage ZR) served as the control. Mean shear bond strength values (MPa) were calculated. The means were statistically analyzed with 2-way ANOVA for the effect of surface treatment and liner, 2-way ANOVA for the effect of different veneering ceramic brands and liner, and 3-way ANOVA for the effect of substrate, veneering ceramic brands, and liner, as well as 1-way ANOVA for the differences between the veneering ceramics. A post hoc Scheffé test was used (alpha=.05). RESULTS: The effects of surface treatment (P=.007) or application of liner (P<.001) were significant. Shear bond strength with different veneering ceramics showed bond strength values with means ranging between 14.2 +/-1.7 MPa (IPS e.max with liner) and 27.5 +/-4.2 MPa (VITA VM 9). A significant difference was found between the results of shear bond tests with Y-TZP and Ce-TZP/A (P=.022). The application of a liner on Y-TZP had no significant effect. CONCLUSIONS: Airborne-particle abrasion is not necessary to enhance the shear bond strength of the evaluated veneering ceramics to Ce-TZP/A. Liners impair the shear bond strength of veneering ceramics to Ce-TZP/A

Characterisation of the immune repertoire of a humanised transgenic mouse through immunophenotyping and high-throughput sequencing

Immunoglobulin loci-transgenic animals are widely used in antibody discovery and increasingly in vaccine response modelling. In this study, we phenotypically characterised B-cell populations from the Intelliselect® Transgenic mouse (Kymouse) demonstrating full B-cell development competence. Comparison of the naïve B-cell receptor (BCR) repertoires of Kymice BCRs, naïve human, and murine BCR repertoires revealed key differences in germline gene usage and junctional diversification. These differences result in Kymice having CDRH3 length and diversity intermediate between mice and humans. To compare the structural space explored by CDRH3s in each species' repertoire, we used computational structure prediction to show that Kymouse naïve BCR repertoires are more human-like than mouse-like in their predicted distribution of CDRH3 shape. Our combined sequence and structural analysis indicates that the naïve Kymouse BCR repertoire is diverse with key similarities to human repertoires, while immunophenotyping confirms that selected naïve B-cells are able to go through complete development

Influence of cement thickness on resin-zirconia microtensile bond strength

PURPOSE: The aim of this study was to evaluate the influence of resin cement thickness on the microtensile bond strength between zirconium-oxide ceramic and resin cement. MATERIALS AND METHODS: Thirty-two freshly extracted molars were transversely sectioned at the deep dentin level and bonded to air-abraded zirconium oxide ceramic disks. The specimens were divided into 8 groups based on the experimental conditions (cement type: Rely X UniCem or Panavia F 2.0, cement thickness: 40 or 160 µm, storage: thermocycled or not). They were cut into microbeams and stored in 37℃ distilled water for 24 h. Microbeams of non-thermocycled specimens were submitted to a microtensile test, whereas those of thermocycled groups were thermally cycled for 18,000 times immediately before the microtensile test. Three-way ANOVA and Sheffe's post hoc tests were used for statistical analysis (α=95%). RESULTS: All failures occurred at the resin-zirconia interface. Thermocycled groups showed lower microtensile bond strength than non-thermocycled groups (P.05). The number of adhesive failures increased after thermocycling in all experimental conditions. No cohesive failure was observed in any experimental group. CONCLUSION: When resin cements of adhesive monomers are applied over air-abraded zirconia restorations, the degree of fit does not influence the resin-zirconia microtensile bond strength.ope

Immunohistochemical Identification of Human Skeletal Muscle Macrophages

Macrophages have well-characterized roles in skeletal muscle repair and regeneration. Relatively little is known regarding the role of resident macrophages in skeletal muscle homeostasis, extracellular matrix remodeling, growth, metabolism and adaptation to various stimuli including exercise and training. Despite speculation into macrophage contributions during these processes, studies characterizing macrophages in non-injured muscle are limited and methods used to identify macrophages vary. A standardized method for the identification of human resident skeletal muscle macrophages will aide in the characterization of these immune cells and allow for the comparison of results across studies. Here, we present an immunohistochemistry (IHC) protocol, validated by flow cytometry, to distinctly identify resident human skeletal muscle macrophage populations. We show that CD11b and CD206 double IHC effectively identifies macrophages in human skeletal muscle. Furthermore, the majority of macrophages in non-injured human skeletal muscle show a ‘mixed’ M1/M2 phenotype, expressing CD11b, CD14, CD68, CD86 and CD206. A relatively small population of CD11b+/CD206- macrophages are present in resting skeletal muscle. Changes in the relative abundance of this population may reflect important changes in the skeletal muscle environment. CD11b and CD206 IHC in muscle also reveals distinct morphological features of macrophages that may be related to the functional status of these cells

A Muscle Cell-Macrophage Axis Involving Matrix Metalloproteinase 14 Facilitates Extracellular Matrix Remodeling with Mechanical Loading

The extracellular matrix (ECM) in skeletal muscle plays an integral role in tissue development, structural support, and force transmission. For successful adaptation to mechanical loading, remodeling processes must occur. In a large cohort of older adults, transcriptomics revealed that genes involved in ECM remodeling, including matrix metalloproteinase 14 (MMP14), were the most upregulated following 14 weeks of progressive resistance exercise training (PRT). Using single-cell RNA-seq, we identified macrophages as a source of Mmp14 in muscle following a hypertrophic exercise stimulus in mice. In vitro contractile activity in myotubes revealed that the gene encoding cytokine leukemia inhibitory factor (LIF) is robustly upregulated and can stimulate Mmp14 expression in macrophages. Functional experiments confirmed that modulation of this muscle cell-macrophage axis facilitated Type I collagen turnover. Finally, changes in LIF expression were significantly correlated with MMP14 expression in humans following 14 weeks of PRT. Our experiments reveal a mechanism whereby muscle fibers influence macrophage behavior to promote ECM remodeling in response to mechanical loading