Unloaded Group Research Articles

Effects of excessive sodium chloride loading in the spontaneously diabetic torii (SDT) fatty rats, a preclinical model of type 2 diabetes mellitus

Type 2 diabetes mellitus represents an international health concern with its growing number of patients worldwide. At the same time, excessive salt consumption is also seen as a major cause of diseases such as hypertension and may expedite renal complications in diabetic patients. In this study, we investigated the effects of excessive sodium chloride supplementation on the kidney of the Spontaneously Diabetic Torii-Leprfa (SDT fatty) rat, an obese type 2 diabetes model. Male and female SDT fatty rats and normal Sprague-Dawley (SD) rats at 5 weeks of age were loaded with 0.3% sodium chloride (NaCl) in drinking water for 13 weeks. Blood serum and urinary parameters were observed throughout the experiment and kidney samples were examined in histopathological and genetical analyses. Significant changes on the body weight, blood pressure, urine volume, creatinine clearance, blood urea nitrogen (BUN), relative gene expressions of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), monocyte chemotactic protein-1 (MCP-1) and transforming growth factor-β (TGF-β) were observed in the salt-loaded male SDT fatty rats. Urinary L-type fatty acid-binding protein (L-FABP) and albumin levels were higher observed in the salt-loaded male SDT fatty rats throughout the period, but urinary albumin levels in the female SDT fatty rats remain unchanged. In the kidney, slight Armani-Ebstein changes, tubular degeneration, hyaline cast, and inflammatory cell infiltration were observed in female SDT fatty rats while the levels of some changes were higher in the salt-loaded group. The kidney of the salt-loaded male SDT fatty rats demonstrated a higher degree of lesions compared to the female group and the male unloaded group. Histopathological changes in salt-loaded SDT fatty rats show that excessive salt consumption may act as a diabetic pathology exacerbation factor, but the pathology may be influenced by gender difference. Urinary L-FABP levels may act as a useful biomarker to detect slight tubular damages in the kidney. Excessive salt loading was shown to exacerbate the renal injury in SDT fatty rats.

Effects of unloader bracing on clinical outcomes and articular cartilage regeneration following microfracture of isolated chondral defects: a randomized trial.

To determine whether the use of an unloading brace can increase the thickness of cartilage regenerate after microfracture surgery. This is a randomized (1:1) controlled clinical trial. Twenty-four patients who underwent microfracture between 2012 and 2015 were identified and were randomly assigned to an unloading brace group or a no-brace group. All patients were kept non-weight bearing for the first eight weeks after surgery and then patients in the intervention group began using an unloading brace for an average of 63.9 (SD = 41.6) days to protect clot stability by exerting a varus or valgus force on the knee to decrease the force on the knee's lateral or medial compartment, respectively. Quality of the cartilage repair was assessed with knee magnetic resonance imaging to determine repair tissue thickness (primary outcome), repair tissue volume, and T2 relaxation times at 12 and 24months after surgery. Clinical outcomes were evaluated with KOOS, Tegner, SF12, and Lysholm questionnaires at six, 12 and 24months after surgery. Three patients were lost to follow-up, resulting in 21 patients ultimately analyzed. The unloading brace repair tissue was greater than the no-brace group in volume (26.8 ± 23.7 mm3 vs - 8.4 ± 22.7 mm3, p = 0.005) and thickness (0.2 ± 0.2mm versus - 0.4 ± 0.3mm, p = 0.001) at 12months and in cartilage thickness in the unloading brace group at 24months (0.4 ± 0.4mm versus - 0.1 ± 0.3mm, p = 0.029). There was a positive correlation between wearing the brace longer and improved 6-month KOOS symptom scores (r = 0.82, p = 0.013), 6-month KOOS QOL scores (r = 0.80, p = 0.017), 6-month Tegner scores (r = 0.94, p = 0.002), and Tegner score changes from baseline to 6months (r = 0.80, p = 0.032). This study found a significant mid-term increase in cartilage repair tissue thickness following unloading bracing in patients recovering from microfracture for isolated chondral defects. II.

Effects of Unloaded Sprint and Heavy Sled Training on Sprint Performance in Physically Active Women.

This study aimed to compare the effects of unresisted versus heavy sled sprint training (0% vs 40% body mass [BM]) on sprint performance in women. Moreover, the effects of the aforementioned loads on resisted sprint and jump performance were analyzed. Twenty-eight physically active women were randomly allocated into 2 groups: unloaded sprint training group (G0%, n = 14), and resisted sprint training with 40% BM group (G40%, n = 14). Pretraining and posttraining assessments included countermovement jump, unloaded 30-m sprint, and 20-m sprint with 20%, 40%, 60%, and 80% BM. Times to cover 0 to 10 (T10), 0 to 20 (T20), 0 to 30 (T30), 10 to 20 (T10-20), 20 to 30 (T20-30), and 10 to 30 m (T10-30) were recorded. Both groups were trained once a week for 8 weeks and completed the same training program, but with different loads (0% vs 40% BM). No significant time × group interactions were observed. For unloaded sprint performance, G0% showed significant (P = .027) decreases only in T10-20, while G40% attained significant decreases in T30 (P = .021), T10-30 (P = .015), and T20-30 (P = .003). Regarding resisted sprint performance, G0% showed significant (P = .010) improvements only for the 20% BM condition. The G40% group attained significant improvements in all loading conditions (20%, 40%, 60%, and 80% BM). Both groups showed significant improvements (P < .001) in countermovement jump height. In physically active women, no significant differences in sprint and countermovement jump performance were detected after 8 weeks of resisted and unresisted sprint training programs. Future studies should, therefore, be devoted to how sprint training should be individualized to maximize performance.

Effects of reloading after chronic neuromuscular inactivity on the three-dimensional capillary architecture in rat soleus muscle

The purpose of the study was to investigate the effects of ambulatory reloading following hindlimb unloading on the three-dimensional (3D) capillary architecture of rat soleus muscle. In this study, 15 male Sprague–Dawley rats were used. The rats were randomly assigned to the following 3 groups: a normal weight bearing control group (CON), 14 days of hindlimb unloading group (HU), and 14 days of hindlimb unloading followed by 7 days of ambulatory reloading group (HU-RL). The capillary diameter and volume were measured using confocal laser microscopy, and capillary number was determined by two-dimensional (2D) capillary staining in the soleus muscle of each group. The capillary diameter and volume as well as the capillary number were significantly lower in the HU group than in the CON group and significantly higher in the HU-RL group than in the HU group. These results provided novel information about the effectiveness of reloading following unloading on not only the 2D increase in capillary number but also the 3D capillary remodeling in the diameter and volume within the unloaded soleus muscle.

Polycaprolactone nanofiber coated with chitosan and Gamma oryzanol functionalized as a novel wound dressing for healing infected wounds

This study was conducted to design and evaluate a wound dressing based on a polycaprolactone (PCL) nanofiber coated with gamma oryzanol (GO) and chitosan (CS) in mice model. All the dressings were prepared by electrospinning method, and their morphology and physical properties were investigated. The mice were divided into five groups and treated with I) PCL-sole (PCL), III) PCL-mupirocin (PCL-M), IV) PCL-GO, IV) PCL-CS, and V) PCL-CS-GO. Wound area, total bacterial count, histopathological parameters, and expressions of IL-1β, TNF-α, IL-10, MMP-9, EGF, and VEGF were assessed. The fibers were randomly distributed in PCL group, but loading CS and GO increased the complexity and placing on the dressings. PCLs loaded with GO and CS showed lower viscosity, surface tension, and fiber diameter and higher conductivity and water contact angle compared to unloaded PCLs (P < 0.05). The treatment with PCLs loaded with mupirocin, CS, and GO significantly reduced wound area and total bacterial count (P < 0.05). Loading PCLs with mupirocin, CS, and GO decreased the expressions of IL-1β, TNF-α, MMP-9, but increased the expressions of IL-10 and VEGF compared to the unloaded PCL group (P < 0.05). The most optimal responses to wound healing and physical parameters belonged to the PCL-CS-GO group.

The Effects of Cyclic Loading and Motion on the Implant-Cement Interface and Cement Mantle of PEEK and Cobalt-Chromium Femoral Total Knee Arthroplasty Implants: A Preliminary Study.

This study investigated the fixation of a cemented PEEK femoral TKA component. PEEK and CoCr implants were subjected to a walking gait cycle for 10 million cycles (MC), 100,000 cycles or 0 cycles (unloaded control). A method was developed to assess the fixation at the cement–implant interface, which exposed the implants to a fluorescent penetrant dye solution. The lateral condyles of the implants were then sectioned and viewed under fluorescence to investigate bonding at the cement–implant interface and cracking of the cement mantle. When tested for 100,000 cycles, debonding of the cement–implant interface occurred in both PEEK (61%) and CoCr (13%) implants. When the duration of testing was extended (10 MC), the percentage debonding was further increased for both materials to 88% and 61% for PEEK and CoCr, respectively. The unloaded PEEK specimens were 79% debonded, which suggests that, when PEEK femoral components are cemented, complete bonding may never occur. Analysis of cracks in the cement mantle showed an absence of full-thickness cracks in the unloaded control group. For the 100,000-cycle samples, on average, 1.3 and 0.7 cracks were observed for PEEK and CoCr specimens, respectively. After 10 MC, these increased to 24 for PEEK and 19 for CoCr. This was a preliminary study with a limited number of samples investigated, but shows that, after 10 MC under a walking gait, substantial debonding was visible for both PEEK and CoCr implants at the cement–implant interface and no significant difference in the number of cement cracks was found between the two materials.

Periodontal ligament proliferation and expressions of bone biomolecules upon orthodontic preloading: Clinical implications for tooth autotransplantation.

ObjectivePreservation of the periodontal ligament (PDL) is vital to the success of tooth autotransplantation (TAT). Increased PDL volumes and facilitated tooth extraction have been observed upon orthodontic preloading. However, it is unclear whether any changes occur in the expressions of bone biomolecules in the increased PDL volumes. This study aimed to determine the expressions of runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), receptor activator of nuclear factor kappa-B ligand (RANKL), and osteoprotegerin (OPG) in PDL upon preloading.MethodsSeventy-two premolars from 18 patients were randomly assigned to experimental groups that received a leveling force for 1, 2, or 4 weeks or to a control unloaded group. Following extraction, PDL volumes from 32 premolars of eight patients (21.0 ± 3.8 years) were evaluated using toluidine blue staining. The expressions of the biomolecules in the PDL from 40 premolars of ten patients (21.4 ± 4.0 years) were analyzed via immunoblotting.ResultsThe median percentage of stained PDL was significantly higher at 2 and 4 weeks after preloading than in the unloaded condition (p < 0.05). The median RUNX2 and ALP expression levels were significantly higher at 2 and 4 weeks after preloading than in the unloaded condition (p < 0.05), whereas the median RANKL/OPG ratios were significantly higher at 1 and 4 weeks after preloading (p < 0.05).ConclusionsOrthodontic preloading for 4 weeks enhances PDL volumes as well as the expressions of RUNX2, ALP and the RANKL/OPG ratio in the PDL, suggesting this loading period is suitable for successful TAT.

In Vitro and In Vivo Toxicity and Biodistribution of Paclitaxel-Loaded Cubosomes as a Drug Delivery Nanocarrier: A Case Study Using an A431 Skin Cancer Xenograft Model

Cubosomes with an internal three-dimensional (3D) periodic and porous particulate nanostructure have emerged as a promising drug delivery system for hydrophobic small molecules as well as large biomolecules over the past several decades. Limited understanding of their safety profiles and biodistribution, however, hinders clinical translation. This study used monoolein-based cubosomes stabilized by Pluronic F127 and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene glycol)] polymers to encapsulate paclitaxel (PTX) as a model drug and investigated the in vitro cytotoxicity, in vivo acute response, and whole body biodistribution of the developed nanoparticles. Comparison of the PTX and nanoparticle cytotoxicity in two-dimensional and 3D spheroid cell models revealed distinct differences, with the cells in the 3D model found to be more tolerable to unloaded PTX as well as the PTX-loaded nanoparticle form. One-time intraperitoneal (i.p.) injection of unloaded cubosomes were generally well tolerated up to 400 mg/kg. Using the A431 skin cancer xenograft model, in vivo imaging studies showed the preferential accumulation of PTX-loaded cubosomes at the tumor sites following i.p. injection. Lastly, average tumor size was reduced by approximately 50% in the nanoparticle-based treatment group compared to the unloaded PTX drug group. The study provides significant information on the biological response of cubosomes and highlights their potential as a versatile drug delivery platform for safe and effective delivery of chemotherapeutic drugs.

Effects of orthodontic forces on bone turnover biomarkers in peri-miniscrew crevicular fluid: A systematic review

Peri-miniscrew crevicular fluid (PMCF) analysis of biomarkers representing bone formation or resorption could provide a non-invasive way to monitor bone turnover around miniscrews and the response to force loading. Our objective was to systematically investigate the relevant available evidence. Search without restrictions in eight databases and hand searching until March 2020 took place. We searched for prospective human studies measuring the levels of markers of bone formation and resorption in PMCF under the effect of orthodontic forces. Following study retrieval and selection, relevant data was extracted and the risk of bias was assessed following the Cochrane Collaboration guidelines. Four studies, two randomized and two non-randomized, were finally identified, following miniscrews for a period up to 90 days. Loading of miniscrews led to a transient increase in C-telopeptide of type I collagen amounts. Temporary increases were also observed for the enzymes: alkaline phosphatase and aspartate aminotransferase. Under the effect of orthodontic loading the total amount of Receptor Activator of Nuclear Factor-κB Ligand (RANKL) in the PMCF consistently increased compared to the unloaded group, at all sampling points. These changes led to a stable decrease in the osteoprotegerin (OPG)/RANKL ratio under force application throughout the study period, as OPG in this group, together with OPG and RANKL in the unloaded group, remained mostly unchanged. No differences were detected for the total OPG quantity between the two loading groups. The levels of bone specific alkaline phosphatase and chondroitin sulfate did not change significantly during observation. All studies presented some issues of concern regarding the risk of bias. Biomarkers of bone turnover in PMCF showed variable responses following orthodontic loading. Overall, the findings were suggestive of adaptive bone alterations to physiologic force stimuli.

Optimal Mechanical Unloading in Left Ventricular Assist Device Recipients Relates to Progressive Up-Titration in Pump Speed

Left ventricular (LV) assist devices (LVADs) are known to elicit reverse remodeling by mechanically unloading the left ventricle. Current guidelines target a reduction in LV end-diastolic diameter (LVEDD) of15% compared with pre-LVAD dimensions; however, there is significant heterogeneity in the degree of unloading achieved. We sought to investigate factors associated with mechanical unloading at 6months of LVAD support. Data were retrospectively collected for 75 LVAD recipients at five time points: pre-LVAD, within 14days post-LVAD, and at 1, 3, and 6months post-LVAD. The percentage change in LVEDD between the pre-LVAD and 6months post-LVAD time points was termed ΔLVEDD. Optimal LV unloading was defined as ΔLVEDD of ≥15% at 6months. Patients who achieved optimal unloading (group A, n=30) were compared with patients who did not (group B, n=45). At 6months, optimally unloaded patients (group A) demonstrated higher fractional shortening (15%±10% vs 10%±7%, P=.007), lower rates of moderate or severe mitral regurgitation (10% vs 33%, P=.02), and lower pulmonary capillary wedge pressure (9±4 vs 16±7mm Hg, P=.02). Right ventricular dysfunction was more prevalent at 6months in poorly unloaded (group B) patients (73% vs 43%, P=.008). Between hospital discharge and 6months, the percentage increase in pump speed (Δ revolutions per minute) washigher in group A patients (4.4%±3.7% vs 0.1%±2.6%, P<.001). In a multivariate analysis, Δ revolutions per minute and tricuspid annular systolic velocity (S') at 6months were independently associated with 6-month ΔLVEDD. Recipients of LVADs who undergo progressive pump speed up-titration during outpatient follow-up are more likely to sustain optimal LV unloading. Progressive LVAD-related right ventricular failure is prevalent in suboptimally unloaded patients.

Effects of Warming Up With Lower-Body Wearable Resistance on Physical Performance Measures in Soccer Players Over an 8-Week Training Cycle.

Bustos, A, Metral, G, Cronin, J, Uthoff, A, and Dolcetti, J. Effects of warming up with lower-body wearable resistance on physical performance measures in soccer players over an 8-week training cycle. J Strength Cond Res 34(5): 1220-1226, 2020-Warm-ups provide an opportune time to integrate specific movements to improve performance. This study aimed to examine the effects of adding wearable resistance (WR) lower-limb loading to a warm-up on physical performance measures in soccer athletes. Thirty-one national-level soccer players (aged 16-18 years) were matched for speed and allocated to either a WR training (WRT = 15) or an unloaded (CON = 16) group. Both groups performed the same warm-up 2-3x·wk for 8 weeks with the WRT group wearing 200- to 600-g loads on their calves. Pre-training, mid-training, and post-training data were collected for 10- and 20-m sprint times, repeated sprint ability, and vertical countermovement jump (CMJ) and horizontal countermovement jump (standing long jump [SLJ]) performance. Wearable resistance training improved pre-training to post-training 10- and 20-m sprint times more than the unloaded training (effect size [ES] = -1.06 to -0.96, respectively; 60.0-66.7 vs. 18.8-37.5% > smallest worthwhile change [SWC]). Both groups decreased CMJ over the first 4 weeks (ES ≥ 0.45) and increased CMJ performance over the second 4 weeks of training (ES ≥ 0.27). Both the WRT and CON groups improved SLJ performance after the 8-week training block (ES = 0.85 and 0.93, respectively; 86.7 and 62.5% > SWC, respectively), yet no differences were identified between groups. These findings indicate that 8 weeks (23 sessions) of WR training appears to elicit practically meaningful improvements in accelerated sprinting and horizontal jumping performance. Strength and conditioning practitioners should consider including WR in sports where sprinting and horizontal force production are critical performance indicators.

The Influence of Progressive Static Load on the Ability of Dental Implants to Withstand Overloading Forces: An Experimental Study in Dogs.

To examine the effect of controlled progressive orthodontic loading on bone around implants subjected to overloading forces. Bilateral edentulous alveolar ridges were created in the posterior maxilla of five beagle dogs and left to heal for an 8-week period, after which 40 implants were placed. In the overloading group (OL), 16 implants were inserted and left to osseointegrate for 16 weeks; impressions were made, and metal crowns were mounted on with supraocclusal contacts with the antagonist teeth. Implants were exposed to dynamic overloading for 16 weeks. In the progressive loading + overloading group (PL+OL), 16 implants were left to osseointegrate for 8 weeks, and custom abutment cores were fabricated and coupled by pairs with Ni-Ti orthodontic springs. Ascending static forces of 100g, 200g, and 300g were each applied for a 3-week period, for a total 9-week progressive loading period. Thereafter, metal crowns with supraocclusal contacts were adapted, and a 16-week overloading protocol for implants was followed as for the overloading group. In the unloaded control group (UL), eight implants were inserted and left uncovered and unloaded for 32 weeks, that is, until the end of the experimental period, at which point all 40 implants were removed with the surrounding bone. Histologic, histomorphometric, and statistical analysis followed. Higher bone-to-implant contact percentage was reported for the OL group (P = .006) and PL+OL group (P < .001) compared with the UL group. Between the OL and PL+OL groups, the addition of progressive loading did not increase the bone-to-implant contact percentage (P = .225). Bone density 1 mm and 2 mm distant to the threads did not differ significantly between the three groups. Significantly lower crestal bone resorption was detected around OL group implants (P = .006) and PL+OL group implants (P = .004) compared with the UL group implants. The implant success rate was 87.5% for the UL group, 67.5% for the OL group, and 87.5% for the PL+OL group. The application of controlled progressive orthodontic loading on osseointegrated implants preceding overloading forces did not increase bone-to-implant contact. When applied, overloading significantly increased bone-to-implant contact compared with the unloaded implants. A significantly higher implant success rate was reported in the PL+OL group compared with the OL group.

Dynamic Compressive Loading Improves Cartilage Repair in an In Vitro Model of Microfracture: Comparison of 2 Mechanical Loading Regimens on Simulated Microfracture Based on Fibrin Gel Scaffolds Encapsulating Connective Tissue Progenitor Cells

Background: Microfracture of focal chondral defects often produces fibrocartilage, which inconsistently integrates with the surrounding native tissue and possesses inferior mechanical properties compared with hyaline cartilage. Mechanical loading modulates cartilage during development, but it remains unclear how loads produced in the course of postoperative rehabilitation affect the formation of the new fibrocartilaginous tissue. Purpose: To assess the influence of different mechanical loading regimens, including dynamic compressive stress or rotational shear stress, on an in vitro model of microfracture repair based on fibrin gel scaffolds encapsulating connective tissue progenitor cells. Study Design: Controlled laboratory study. Methods: Cylindrical cores were made in bovine hyaline cartilage explants and filled with either (1) cartilage plug returned to original location (positive control), (2) fibrin gel (negative control), or (3) fibrin gel with encapsulated connective tissue progenitor cells (microfracture mimic). Constructs were then subjected to 1 of 3 loading regimens: (1) no loading (ie, unloaded), (2) dynamic compressive loading, or (3) rotational shear loading. On days 0, 7, 14, and 21, the integration strength between the outer chondral ring and the central insert was measured with an electroforce mechanical tester. The central core component, mimicking microfracture neotissue, was also analyzed for gene expression by real-time reverse-transcription polymerase chain reaction, glycosaminoglycan, and double-stranded DNA contents, and tissue morphology was analyzed histologically. Results: Integration strengths between the outer chondral ring and central neotissue of the cartilage plug and fibrin + cells groups significantly increased upon exposure to compressive loading compared with day 0 controls (P = .007). Compressive loading upregulated expression of chondrogenesis-associated genes (SRY-related HGMG box-containing gene 9 [SOX9], collagen type II α1 [COL2A1], and increased ratio of COL2A1 to collagen type I α1 [COL1A1], an indicator of more hyaline phenotype) in the neotissue of the fibrin + cells group compared with the unloaded group at day 21 (SOX9, P = .0032; COL2A1, P < .0001; COL2A1:COL1A1, P = .0308). Fibrin + cells constructs exposed to shear loading expressed higher levels of chondrogenic genes compared with the unloaded condition, but the levels were not as high as those for the compressive loading condition. Furthermore, catabolic markers (MMP3 and ADAMTS 5) were significantly upregulated by shear loading (P = .0234 and P < .0001, respectively) at day 21 compared with day 0. Conclusion: Dynamic compressive loading enhanced neotissue chondrogenesis and maturation in a simulated in vitro model of microfracture, with generation of more hyaline-like cartilage and improved integration with the surrounding tissue. Clinical Relevance: Controlled loading after microfracture may be beneficial in promoting the formation of more hyaline-like cartilage repair tissue; however, the loading regimens applied in this in vitro model do not yet fully reproduce the complex loading patterns created during clinical rehabilitation. Further optimization of in vitro models of cartilage repair may ultimately inform rehabilitation protocols.

Early Weight-Bearing Improves Cartilage Repair in an in vitro Model of Microfracture: Comparison of Two Mechanical Loading Regimens on Simulated Microfracture Based onFibrin Gel Scaffolds Encapsulating Bone Marrow Mesenchymal Stem Cells

Objectives: Microfracture of focal chondral defects produces fibrocartilage, which inconsistently integrates with the surrounding native tissue and possesses inferior mechanical properties compared to hyaline cartilage. Mechanical loading modulates cartilage during development, but it remains unclear how loads produced in the course of postoperative rehabilitation affect the formation of the new fibrocartilaginous tissue. The purpose of this study was to assess the influence of different mechanical loading regimens simulating weight-bearing or passive motion exercises on an in vitro model of microfracture repair based on fibrin gel scaffolds encapsulating mesenchymal stem cells (MSCs). Methods: Cylindrical cores were made in bovine hyaline cartilage explants and filled with either: (1) cartilage plug returned to original location (positive control), (2) fibrin gel (negative control), or (3) fibrin gel with encapsulated bone marrow-derived MSCs (BM-MSCs) (microfracture mimic). Constructs were then subjected to one of three loading regimens, including (1) no loading (i.e., unloaded) (2) dynamic compressive loading, or (3) rotational shear loading. On days 0, 7, 14, and 21, the integration strength between the outer chondral ring and the central insert was measured with an electroforce mechanical tester. The central core component, mimicking microfracture neotissue, was also analyzed for gene expression by real-time RT-PCR, glycosaminoglycan and dsDNA contents, and tissue morphology by histology. Results: Integration strengths between the outer chondral ring and central neotissue of the cartilage plug and fibrin + BM-MSC groups significantly increased upon exposure to compressive loading, compared to day 0 controls (p= 0.007). Compressive loading upregulated expression of chondrogenesis-associated genes (SOX9, collagen type II, and collagen type II: I, an indicator of more hyaline phenotype) in the neotissue of the fibrin + BM-MSC group, as compared to the unloaded group at day 21 (SOX9, p =0.0032; COL2A1, p &lt;0.0001; COL2A1/COL1A1, p = 0.0308,). Fibrin + BM-MSC constructs exposed to shear loading expressed higher levels of chondrogenic genes as compared to the unloaded condition, but not as high as the compressive loading condition. Furthermore, catabolic markers (MMP3 and ADAMTS 5) were significantly upregulated by shear loading (p = 0.0234 and p&lt; 0.0001, respectively) at day 21, as compared to day 0. Conclusion: Dynamic compressive loading enhanced neotissue chondrogenesis and maturation in a simulated in vitro model of microfracture, with generation of more hyaline-like cartilage and improved integration with the surrounding tissue. Early weight-bearing after microfracture may be beneficial in promoting the formation of more hyaline-like cartilage repair tissue, whereas range of motion exercise by continuous passive motion without weight-bearing might not be as effective, or even negatively affect the formation of the repair tissue during post-surgery rehabilitation.

Механизмы поддержания экспрессии медленного миозина в волокнах постуральной мышцы при стимуляции опорных афферентов на фоне гравитационной разгрузки

The study was aimed to investigate the role of support afferentation in support of functioning the signaling mechanisms responsible for slow-type oxidative genes expression during postural soleus muscle unloading. For the investigation of the support afferentation the plantar mechanical stimulation was used. Male Wistar rats were divided into six groups (8 animals in each): two cage control groups (C), two groups hindlimb unloaded for 1 day or 3 days (1 HS, 3HS) and two hindlimb-unloaded groups with plantar mechanical stimulation 1HS+МСО; 3HS+МСО. Animals from stimulated groups received plantar stimulation during 4 hours a day, mimicking the pattern of normal animal walking. Plantar mechanical stimulation led to prevention of slow myosin I(β) and myh7b mRNAs expression decreases after 1 day of unloading. After three days of hindlimb unloading the expression of slow myosin I(β) mRNA and myh7b mRNA significantly increased in plantar stimulated group, and the expression of SOX6 mRNA in this group did not differ from control, although it was significantly increased in pure unloaded group. So, plantar mechanical stimulation prevents unloading-induced MyHC I/myh7b/SOX6 inactivation, inhibiting slow-to-fast fiber-type shift.

Characteristics of torque production of the lower limb are significantly altered after 2 hours of treadmill load carriage

Load carriage is seldom completed in isolation, meaning load bearers need to be physically capable of physical activity after the load carriage task. This study aims to examine changes in lower limb muscle strength, as measured by torque production across a range of joint angles as a result of prolonged load carriage. Thirty-four healthy participants underwent two hours of loaded or unloaded treadmill load carriage, with lower limb muscle function variables assessed pre and post activity. The loaded group had a mass of (Mean(range)) 76.45 (27.12)kg, stature: 178.56 (17.63)cm, age: 23(6)yrs, and comprised of 13 males and 3 females. While the unloaded group had a body mass of 73.69(24.19)kg, stature: 178.89(18.49)cm, age: 22(5)yrs and comprised of 14 males and 4 females. Significant reductions across a range of parameters were observed. Characterised by reductions at the optimum muscle length for torque output, with all aspects demonstrating large (knee extension at 180˚·s-1: 0.51 Standardised SD, knee extnsion at 60˚·s-1: 0.98 standardised SD) or extremely large individual differences (knee flexion at 180˚·s-1: 2.17 standardised SD). These findings suggest after the completion of the load carriage task participants are in a significantly reduced physical state, which may have implications for secondary tasks.

PO-095 Effects of dynamic foot stimulation on GSK-3 beta signaling pathway in rat soleus muscle under hindlimb unloading

Objective It is known that under simulated microgravity GSK-3 beta phosphorylation (Ser 9) is decreased [Mirzoev et al., 2016] which is associated with the activation of its kinase activity. GSK-3 beta activation may lead to NFATc1 export from myonuclei resulting in a slow-to-fast myosin shift in soleus muscle and may cause a decrease in muscle oxidative capacity and protein synthesis [Glass, 2003, Theeuwes et al, 2017].&#x0D; It was demonstrated that GSK-3 beta Ser 9 phosphorylation is decreased in response to nitric oxide synthase inhibition [Martins et al., 2011]. Under unloading conditions, the content of NO as well as nitric oxide synthase is attenuated in rat soleus muscle [Tidball et al., 1998; Lomonosova et al, 2011]. Dynamic foot stimulation (DFS) of the soles of the feet results in an increase of neuromuscular activation [Muller et al., 2005] of the lower limb muscles and prevents nitric synthase content decrease during 7-day exposure to dry immersion [Moukhina et al, 2004; Shenkman et al., 2004]. The aim of our study was to analyze the effect of rat dynamic foot stimulation during early unloading on GSK-3 beta phosphorylation and some of its downstream targets.&#x0D; Methods Male Wistar rats were randomly assigned to vivarium control, 1-day unloading and 1-day&#x0D; unloading with DFS. The pressure stimulation protocol mimicked the normal animal walking (104 mm Hg pressure, 4 Hz frequency and 250 ms signal duration) for a total of 20 min followed by a 10-min rest interval for 4 hours.&#x0D; Results We found that 1-day unloading caused a significant decrease (p˂ 0.025) in NFATc1 nuclear content as well as slow myosin (MHC I (beta) isoform) mRNA expression. In the 1-day unloading group, the level of glycogen synthase 1 phosphorylation (Ser 641), which is a direct GSK-3 beta target, was significantly higher than that in the control group, although a decrease in GSK-3 beta Ser 9 phosphorylation in the 1-day unloaded group was not statistically significant compared to the control group. However, in the DFS-treated unloaded rats the level of GSK-3 beta phosphorylation was significantly higher than in the untreated unloaded rats. NFATc1 nuclear content, slow myosin mRNA expression and glycogen synthase 1 phosphorylation (Ser 641) did not differ from the control group. The glycogen content in soleus muscles of both unloaded groups was higher than in the control group, which is in accordance with previous studies [Henriksen and Tischler, 1988], but in DFS subjected unloaded group the glycogen content was higher compared to the untreated unloaded group.&#x0D; Conclusions Thus, we found that dynamic foot stimulation during 1-day hindlimb unloading leads to GSK-3 beta inactivation and prevention of both NFATc1 myonuclei export and the decrease in MyHC I beta expression.&#x0D; The study was supported by the Russian Foundation for Basic Research grant #17-29-1029 and Basic Research Program of the IBMP, RAS. The authors express their deep gratitude to Prof. Inessa B. Kozlovskaya for useful recommendations concerning the design of the study.

Growth factor release and enhanced encapsulated periodontal stem cells viability by freeze-dried platelet concentrate loaded thermo-sensitive hydrogel for periodontal regeneration

Periodontium regeneration is a highly challenging process as it requires the regeneration of three different tissues simultaneously. The aim of this study was to develop a composite material that can be easily applied and can sufficiently deliver essential growth factors and progenitor cells for periodontal tissue regeneration.Freeze-dried platelet concentrate (FDPC) was prepared and incorporated in a thermo-sensitive chitosan/β-glycerol phosphate (β-GP) hydrogel at concentrations of 5, 10, or 15 mg/ml. The viscosity of the hydrogels was investigated as the temperature rises from 25 °C to 37 °C and the release kinetics of transforming growth factor (TGF-β1), platelet-derived growth factor (PDGF-BB) and insulin-like growth factor (IGF-1) were investigated at four time points (1 h, 1 day, 1 week, 2 weeks). Periodontal ligament stem cells (PDLSCs) were isolated from human third molars and encapsulated in the different hydrogel groups. Their viability was investigated after 7 days in culture in comparison to standard culture conditions and non FDPC-loaded hydrogel.Results showed that loading FDPC in the hydrogel lowered the initial viscosity in comparison to the unloaded control group and did not affect the sol-gel transition in any group. All FDPC-loaded hydrogel groups exhibited sustained release of TGF-β1 and PDGF-BB for two weeks with significant difference between the different concentrations. The loading of 10 and 15 mg/ml of FDPC in the hydrogel increased the PDLSCs viability significantly compared to the unloaded hydrogel and was comparable to the standard culture conditions.Accordingly, it may be concluded that loading FDPC in a chitosan/β-GP hydrogel can offer enhanced injectability, a sustained release of growth factors and increased viability of encapsulated stem cells which can be beneficial in periodontium tissue regeneration.

Iron overload involved in the enhancement of unloading-induced bone loss by hypomagnetic field.

During deep-space exploration missions, astronauts will be exposed to abnormal space environments including microgravity and hypomagnetic field (HyMF) that is 10,000 times weaker than geomagnetic field (GMF). It is well known that microgravity in space can induce bone loss; however, it is ill-defined whether HyMF involved in this process. Herein, we aimed to investigate the combined effects of HyMF and microgravity on bone loss. A mouse model of hindlimb suspension (HLU) was adopted to simulate microgravity-induced bone loss, that was exposed to a hypomagnetic field of <300 nanotesla (nT) generated by a geomagnetic field-shielding chamber. Besides, a recent study showed that HLU induced bone loss was orchestrated by iron overload. Therefore, the changes of iron content in unloading-induced bone loss under HyMF condition were detected simultaneously. The results showed HyMF exacerbated the loss of bone mineral content (BMC), induced more detrimental effects on microstructure of cancellous bone but not cortical bone and yielded greater negative effects on biomechanical characteristics in mice femur under unloading status. Concomitantly, there was more iron accumulation in serum, liver, spleen and bone in the combined treatment group than in the separate unloading group or HyMF exposure group. These results showed that HyMF promoted additional bone loss in mice femur during mechanical unloading, and the potential mechanism may be involved in inducing iron overload of mice.

Therapeutic Drug Monitoring of Teicoplanin in Haematological Malignancy Patients with Febrile Neutropenia and Optimizing Dosage Regimens.

This study used high-performance liquid chromatography to measure 202 teicoplanin plasma trough concentrations (Cmin ) in 114 haematological malignancy patients with febrile neutropenia. Patients were divided into two groups according to the mean initial dose (MID) over the first 3 days of treatment: (i) MID = 533.33 mg/day (loading dose group, 400 mg q12h for three doses followed by 400 mg qd, n = 62) and (ii) MID < 533.33 mg/day (unloaded or underloaded group, n = 52). During the first 3 days after treatment, the overall Cmin was higher in group 1 than in group 2 (10.96 ± 5.44 mg/L versus 6.31 ± 3.73 mg/L, mean ± S.D.; p = 0.002), as was the qualifying rate of Cmin > 10 mg/L (54.5% versus 11.1%, p = 0.001), and the probability of Cmin < 5 mg/L was lower in group 1 than in group 2 (13.6% versus 40.7%, p = 0.037). After 3 days, the average Cmin and qualifying rates did not differ significantly between the two groups, and the average Cmin was <10 mg/L in both groups. Binary logistic regression analysis revealed that creatinine clearance (p = 0.004) and MID (p = 0.010) could affect Cmin during the first 3 days of treatment and that age (p = 0.022) only could affect Cmin after 3 days. In conclusion, it is necessary to apply loading dose to achieve teicoplanin Cmin > 10 mg/L rapidly and, from a pharmacokinetic/pharmacodynamic perspective, 600 mg is recommended as loading and maintenance dose for these patients when AUC24 /minimum inhibitory concentration > 345.