Application of Human Epineural Conduit Supported with Human Mesenchymal Stem Cells as a Novel Therapy for Enhancement of Nerve Gap Regeneration

Various therapeutic methods have been suggested to enhance nerve regeneration. In this study, we propose a novel approach for enhancement of nerve gap regeneration by applying human epineural conduit (hEC) supported with human mesenchymal stem cells (hMSC), as an alternative to autograft repair. Restoration of 20 mm sciatic nerve defect with hEC created from human sciatic nerve supported with hMSC was tested in 4 experimental groups (n = 6 each) in the athymic nude rat model (Crl:NIH-Foxn1rnu): 1 - No repair control, 2 - Autograft control, 3 - Matched diameter hEC filled with 1 mL saline, 4 - Matched diameter hEC supported with 3 × 106 hMSC. Assessments included: functional tests: toe-spread and pinprick, regeneration assessment by immunofluorescence staining: HLA-1, HLA-DR, NGF, GFAP, Laminin B, S-100, VEGF, vWF and PKH26 labeling; histomorphometric analysis of myelin thickness, axonal density, fiber diameter and myelinated nerve fibers percentage; Gastrocnemius Muscle Index (GMI) and muscle fiber area ratio. Best sensory and motor function recovery, as well as GMI and muscle fiber area ratio, were observed in the autograft group, and were comparable to the hEC with hMSC group (p = 0.038). Significant improvements of myelin thickness (p = 0.003), fiber diameter (p = 0.0296), and percentage of myelinated fibers (p < 0.0001) were detected in hEC group supported with hMSC compared to hEC with saline controls. At 12-weeks after nerve gap repair, hEC combined with hMSC revealed increased expression of neurotrophic and proangiogenic factors, which corresponded with improvement of function comparable with the autograft control. Application of our novel hEC supported with hMSC provides a potential alternative to the autograft nerve repair. Graphical Abstract

Aquatic Training after Joint Immobilization in Rats Promotes Adaptations in Myotendinous Junctions

The myotendinous junction (MTJ) is the muscle-tendon interface and constitutes an integrated mechanical unit to force transmission. Joint immobilization promotes muscle atrophy via disuse, while physical exercise can be used as an adaptative stimulus. In this study, we aimed to investigate the components of the MTJ and their adaptations and the associated elements triggered with aquatic training after joint immobilization. Forty-four male Wistar rats were divided into sedentary (SD), aquatic training (AT), immobilization (IM), and immobilization/aquatic training (IMAT) groups. The samples were processed to measure fiber area, nuclear fractal dimension, MTJ nuclear density, identification of telocytes, sarcomeres, and MTJ perimeter length. In the AT group, the maintenance of ultrastructure and elements in the MTJ region were observed; the IM group presented muscle atrophy effects with reduced MTJ perimeter; the IMAT group demonstrated that aquatic training after joint immobilization promotes benefits in the muscle fiber area and fractal dimension, in the MTJ region shows longer sarcomeres and MTJ perimeter. We identified the presence of telocytes in the MTJ region in all experimental groups. We concluded that aquatic training is an effective rehabilitation method after joint immobilization due to reduced muscle atrophy and regeneration effects on MTJ in rats.

The effect of regular resistance exercise, vitamin D, and calcium supplements on the gastrocnemius muscle in rats in the post-menopausal period: An experimental study

Background: Menopause is the natural termination of menstruation which affects the quality and important aspects of women’s life. Objective: To evaluate the effect of regular resistance training (Ex) with vitamin D (Vit. D) and calcium (Ca) supplements in the postmenopausal period on muscle tissue in rats. Materials and Methods: In this experimental study, 72 female Wistar rats (8-10-wk old) were randomly divided into control, placebo, Vit. D, Ca, Ex, Ca + Vit. D, Ex + Ca, Ex + Vit. D, and Ex + Ca + Vit. D groups. Control and placebo groups were fed with a standard diet and sesame oil, respectively. Two months after the ovariectomy, Ex, Ca (35 mg/kg), and Vit. D (10000 IU) were administered in all groups except the control. The number of muscle and inflammatory cells, fiber diameter, endomysium thickness, and degenerative collagen fiber area were assessed through hematoxylin-eosin staining. Results: Muscle cell number was increased in the Ex + Vit. D + Ca, Vit. D + Ex, and Vit. D groups compared to the control group; also, inflammatory cell number showed significant increase in the Ex + Vit. D + Ca (12 ± 5.46), Vit. D + Ex (14 ± 3.25), Ex (13 ± 4.08), Vit. D (11 ± 3.26), Ca + Vit. D (10 ± 1.01), and Ca + Ex (9 ± 2.87) groups. Muscle fiber diameter in the Ex + Vit. D + Ca and Vit. D + Ex groups was higher than the other groups. Endomysium thickness was significantly decreased in the Ex + Vit. D + Ca and Vit. D + Ex groups compared to the control and placebo groups (p < 0.001). Degenerative collagen fiber area showed a significant increase in the Ex + Vit. D + Ca and Vit. D + Ex groups (p ≤ 0.001) comparison with the control group. Conclusion: Regular resistance exercise, Vit. D, and Ca supplements can improve muscle morphological features in the postmenopausal period. Key words: Menopause, Muscle, Vitamin D, Calcium, Exercise.

Maternal Undernutrition Alters The Skeletal Muscle Development and Methylation of Myogenic Factors in Progeny: A Randomized Controlled Trial

Maternal malnutrition alters protein synthesis and muscle development in offspring. Epigenetic modification is one of the mechanisms involved in this developmental programming. However, the role of the methylation of myogenic factors remains unclear. Twenty-four pregnant goats were assigned to a control (100% of the nutrients requirement, n = 12) or a restricted group (60% of the nutrients requirement, n = 12) between 0.3 and 0.7 gestation (G). Descendants were harvested at 0.7 G and at 91 days after birth to analyze the fiber area and fiber type of femoris muscle and to examine the effects on the mTOR signal pathway and methylation of the promoter regions of myogenic factors. Maternal undernutrition increased the fiber area of the vastus muscle in the fetuses (P < 0.05). Undernutrition enhanced (P < 0.01) the proportions of MyHCI and MyHCIIA fibers in the vastus muscle of fetuses and semitendinosus (ST) muscle of kids, while the proportion of MyHCIIX fibers were decreased (P < 0.01) in the restricted fetuses and kids. The mRNA expression levels of EYA1 and MYOZ2 in the restricted fetuses were downregulated (P < 0.05), but no difference was observed in the restricted kids (P > 0.05). The percentage of DNA methylation at the + 530 CpG site of the MYF5 gene in the restricted fetuses was increased (P < 0.05), but the methylation percentages of the MYF5 gene at the +274,280 CpG site and of the MYOD gene at the +252 CpG site in the restricted kids were reduced (P < 0.05). The mTOR protein signals were downregulated (P < 0.05) in the restricted fetuses and kids. These results indicated that maternal undernutrition altered the fiber type of femoris muscle in offspring, but its relationship with the methylation in the promoter regions of myogenic genes needs to be further studied.

Anatomical and ontogenetic influences on muscle density

Physiological cross-sectional area (PCSA), an important biomechanical variable, is an estimate of a muscle’s contractile force potential and is derived from dividing muscle mass by the product of a muscle’s average fascicle length and a theoretical constant representing the density of mammalian skeletal muscle. This density constant is usually taken from experimental studies of small samples of several model taxa using tissues collected predominantly from the lower limbs of adult animals. The generalized application of this constant to broader analyses of mammalian myology assumes that muscle density (1) is consistent across anatomical regions and (2) is unaffected by the aging process. To investigate the validity of these assumptions, we studied muscles of rabbits (Oryctolagus cuniculus) in the largest sample heretofore investigated explicitly for these variables, and we did so from numerous anatomical regions and from three different age-cohorts. Differences in muscle density and histology as a consequence of age and anatomical region were evaluated using Tukey’s HSD tests. Overall, we observed that older individuals tend to have denser muscles than younger individuals. Our findings also demonstrated significant differences in muscle density between anatomic regions within the older cohorts, though none in the youngest cohort. Approximately 50% of the variation in muscle density can be explained histologically by the average muscle fiber area and the average percent fiber area. That is, muscles with larger average fiber areas and a higher proportion of fiber area tend to be denser. Importantly, using the age and region dependent measurements of muscle density that we provide may increase the accuracy of PCSA estimations. Although we found statistically significant differences related to ontogeny and anatomical region, if density cannot be measured directly, the specific values presented herein should be used to improve accuracy. If a single muscle density constant that has been better validated than the ones presented in the previous literature is preferred, then 1.0558 and 1.0502 g/cm3 would be reasonable constants to use across all adult and juvenile muscles respectively.

Mechanical Properties of Carbon-Fabric-Reinforced High-Strength Matrices

Fabric-reinforced cementitious matrices (FRCM) are promising technologies that respond to today’s architectural approaches. However, due to their high strength and ductility, they are starting to be implemented in buildings as strengthening systems. In this experimental study, the amount of fiber along the load direction in high-strength cementitious matrices and the effects of the fiber orientation on FRCM mechanical properties were studied. A total of four different composites were produced with two fabrics and two matrices. Tensile and flexural tests were carried out on composites. Within the scope of microstructure studies, scanning electron microscope micrographs were obtained and analyzed, along with microtopography sections. The main result obtained from the study indicates that as the fiber area in the direction of the load increases, the load order carried in this direction increases. However, this increase does not have to be proportional to the fiber area used in the direction of the load. The fiber coating and coating matrix interface play important roles in a composite’s performance. The carbon fibers can be used more efficiently by using them along the load direction and the loads in the matrix can be transferred to the carbon fibers by creating a larger fiber–matrix interface area.

Investigation on the Effect of Low Temperature on Impact Properties of Ti-6Al-4V Titanium Alloy

Cryogenic cutting is becoming an attractive machining method for difficult-to-cut materials. However, it’s very difficult to analyze directly their cutting mechanism at low temperature. In order to better understand the various physical phenomena in the cryogenic cutting of titanium alloy, the Charpy impact test of Ti-6Al-4V titanium alloy at low temperatures (as low as -196 °C) was undertaken in this work. The Charpy absorbed energy of Ti-6Al-4V titanium alloy at low temperatures was investigated firstly. Then, by observing the microscopic and macroscopic morphology of the fracture, the impact properties and fracture modes of Ti-6Al-4V titanium alloy at low temperatures were analyzed. It was found that the impact toughness of Ti-6Al-4V titanium alloy reduces when the temperature decreases from 20 °C to -196 °C, and the fracture appears a tendency to become brittle. Meanwhile, three kinds of areas, i.e. shear lip area, fiber area, and radiation area, were found on the fracture morphology at each temperature. Those areas correspond to the shear fracture zone, crack initiation zone, and crack extension zone, respectively. With the decrease in temperature, the proportion of fiber area decreases, and the radiation area appears and increases gradually. However, fiber areas were still observed on the macroscopic morphology of the fracture under − 196 °C, which suggests that Ti-6Al-4V titanium alloy still has the ability to deform plastically at such low temperatures. The research result in this work provide a fundamental support for analyzing the cutting mechanism of Ti-6Al-4V titanium alloy at low temperatures.

THE EFFECT OF POLYSILAZANE ON THE DENSIFICATION AND MECHANICAL PROPERTIES OF SICF/SIC COMPOSITES

The effect of polysilazane on the densification and mechanical properties of SiCf/SiC composites was examined thoroughly. The polysilazane was infiltrated into the fiber area, followed by pyrolysis at 1450°C for 4 h in an argon atmosphere. Furthermore, the SiC fabrics contain the pyrolyzed polysilazane was infiltrated by β-SiC slurry and sintered at 1750°C for 2 h in an argon atmosphere. The density of SiCf/SiC composites with and without the addition of polysilazane was 2.65 and 3.05 g/cm3, respectively. The pores in the fiber area were observed for the SiCf/SiC composites with the addition of polysilazane. However, the fine-grained size in the matrix area was obtained by the addition of polysilazane. The average grain size of the SiCf/SiC composite with and without the addition of polysilazane was 253 and 740 nm, respectively. Indeed, the hardness of the matrix area for SiCf/SiC composites with the addition of polysilazane (31.6 ± 1.8 GPa) was higher compared to SiCf/SiC composites without polysilazane (27.4 ± 1.6 GPa).

Beef Tenderness Prediction by a Combination of Statistical Methods: Chemometrics and Supervised Learning to Manage Integrative Farm-To-Meat Continuum Data

This trial aimed to integrate metadata that spread over farm-to-fork continuum of 110 Protected Designation of Origin (PDO)Maine-Anjou cows and combine two statistical approaches that are chemometrics and supervised learning; to identify the potential predictors of beef tenderness analyzed using the instrumental Warner-Bratzler Shear force (WBSF). Accordingly, 60 variables including WBSF and belonging to 4 levels of the continuum that are farm-slaughterhouse-muscle-meat were analyzed by Partial Least Squares (PLS) and three decision tree methods (C&RT: classification and regression tree; QUEST: quick, unbiased, efficient regression tree and CHAID: Chi-squared Automatic Interaction Detection) to select the driving factors of beef tenderness and propose predictive decision tools. The former method retained 24 variables from 59 to explain 75% of WBSF. Among the 24 variables, six were from farm level, four from slaughterhouse level, 11 were from muscle level which are mostly protein biomarkers, and three were from meat level. The decision trees applied on the variables retained by the PLS model, allowed identifying three WBSF classes (Tender (WBSF ≤ 40 N/cm2), Medium (40 N/cm2 < WBSF < 45 N/cm2), and Tough (WBSF ≥ 45 N/cm2)) using CHAID as the best decision tree method. The resultant model yielded an overall predictive accuracy of 69.4% by five splitting variables (total collagen, µ-calpain, fiber area, age of weaning and ultimate pH). Therefore, two decision model rules allow achieving tender meat on PDO Maine-Anjou cows: (i) IF (total collagen < 3.6 μg OH-proline/mg) AND (µ-calpain ≥ 169 arbitrary units (AU)) AND (ultimate pH < 5.55) THEN meat was very tender (mean WBSF values = 36.2 N/cm2, n = 12); or (ii) IF (total collagen < 3.6 μg OH-proline/mg) AND (µ-calpain < 169 AU) AND (age of weaning < 7.75 months) AND (fiber area < 3100 µm2) THEN meat was tender (mean WBSF values = 39.4 N/cm2, n = 30).