The beneficial effect of cold atmospheric plasma on parameters of molecules and cell function involved in wound healing in human osteoblast-like cells in vitro.

The aim of this study was to analyse the effect of cold atmospheric plasma (CAP) on human osteoblast-like cells in vitro. Additionally, underlying intracellular mechanisms were to be studied. Human osteoblast-like (MG63) cells were exposed to CAP for 60 s. The effects of CAP on key molecules essential for the wound healing response were studied using real-time PCR, ELISA and immunocytochemistry. For studying intracellular signalling pathways, MAP kinase MEK 1/2 was blocked. Cell viability was analysed by an XTT assay and with an EVE automated cell counter. Cell migration was examined by an in vitro wound healing assay.CAP exposition on osteoblast-like cells caused a significant upregulation of interleukin (IL)-1ß, IL-6, IL-8, tumor necrosis factor (TNF)α, cyclooxygenase (COX)2, collagen (COL) 1α, matrix metalloproteinase (MMP)1, Ki67, proliferating-cell-nuclear-antigen (PCNA) and chemokine ligand (CCL)2 mRNA expression at 1 day. Interestingly, after blocking of MAP kinase, CAP-induced upregulation of Ki67 was inhibited by 57%. Moreover, CAP treatment improved significantly osteoblast-like cell viability as compared to untreated cells at 1 day. Beneficial effect of CAP treatment was shown by an in vitro wound healing assay, displaying a significant faster wound closure. Our findings provide evidence that CAP exposure effects gene and protein regulation in human osteoblast-like cells. Furthermore, CAP treatment has a positive impact on wound closure in an in vitro setting and might improve existing concepts of hard tissue regeneration in the future.

Implication of low level inflammation in the insulin resistance of adipose tissue at late pregnancy.

Insulin resistance is a characteristic of late pregnancy, and adipose tissue is one of the tissues that most actively contributes to the reduced maternal insulin sensitivity. There is evidence that pregnancy is a condition of moderate inflammation, although the physiological role of this low-grade inflammation remains unclear. The present study was designed to validate whether low-grade inflammation plays a role in the development of insulin resistance in adipose tissue during late pregnancy. To this end, we analyzed proinflammatory adipokines and kinases in lumbar adipose tissue of nonpregnant and late pregnant rats at d 18 and 20 of gestation. We found that circulating and tissue levels of adipokines, such as IL-1ß, plasminogen activator inhibitor-1, and TNF-α, were increased at late pregnancy, which correlated with insulin resistance. The observed increase in adipokines coincided with an enhanced activation of p38 MAPK in adipose tissue. Treatment of pregnant rats with the p38 MAPK inhibitor SB 202190 increased insulin-stimulated tyrosine phosphorylation of the insulin receptor (IR) and IR substrate-1 in adipose tissue, which was paralleled by a reduction of IR substrate-1 serine phosphorylation and an enhancement of the metabolic actions of insulin. These results indicate that activation of p38 MAPK in adipose tissue contributes to adipose tissue insulin resistance at late pregnancy. Furthermore, the results of the present study support the hypothesis that physiological low-grade inflammation in the maternal organism is relevant to the development of pregnancy-associated insulin resistance.

Efficient nutrient removal from swine manure in a tubular biofilm photo-bioreactor using algae-bacteria consortia.

Concentrated animals feeding operations (CAFOs) often pose a negative environmental impact due to the uncontrolled spreading of manure into soils that ends up in the release of organic matter and nutrients into water bodies. Conventional aerobic methods treating CAFOs wastewater require intensive oxygenation, which significantly increases the operational costs. The alternative proposed in this research is the application of micro-algae based systems by taking advantage of the cost-effective in situ oxygenation via photosynthesis. A 4.9 L enclosed tubular biofilm photo-bioreactor was inoculated with an algal-bacterial consortium formed by the micro-algae Chlorella sorokiniana and a mixed bacterial culture from an activated sludge process. C. sorokiniana delivers the O(2) necessary to accomplish both organic matter and ammonium oxidation. The reactor was fed with diluted swine wastewater containing 180, 15 and 2,000 mg/L of NH(4) (+)-N, soluble P and total COD, respectively. The photo-bioreactor exhibited good and sustained nutrient removal efficiencies (up to 99% and 86% for NH(4) (+) and PO(4) (3-), respectively) while total COD was removed up to 75% when the biofilm was properly established. Liquid superficial velocities up to 0.4 m/s (achieved by culture broth recirculation) hindered the formation of a stable biofilm, while operation at velocities lower than 0.1 m/s supported stable process performance. The high shear stress imposed by the centrifugal recirculation pump disintegrated the large aggregates detached from the biofilm, which resulted in a poor settling performance and therefore poor COD removal efficiencies. Enclosed biofilm photo-bioreactors therefore offer a potentially more economical alternative to conventional tertiary treatments process.

The activity of exoglycosidases in the synovial membrane and knee fluid of patients with rheumatoid arthritis and juvenile idiopathic arthritis.

OBJECTIVE: To determine the activities of the five exoglycosidases that catabolize glycoconjugates (proteoglycans, glycoproteins, and glycolipids) in the synovial membrane and knee joint fluid of patients with rheumatoid arthritis (RA) and juvenile idiopathic arthritis (JIA). METHODS: The following exoglycosidases were analysed with the p-nitrophenyl derivatives of appropriate sugars as substrates: hexosaminidase (HEX) and its isoenzymes A and B, beta-glucuronidase, beta-galactosidase, alpha-mannosidase, and alpha-fucosidase. RESULTS: Our results show that the activity of all exoglycosidases tested in the synovial membrane of patients with RA and JIA was significantly higher than in synovial fluid. We demonstrated that only the enzymatic activity of HEX was significantly higher in the tissue of patients with inflammatory diseases in comparison to the activity in the control group. CONCLUSION: These data support the concept that the synovial cells of patients with RA and JIA are the main source of exoglycosidases, which catabolize glycoconjugates of cartilage.

Activity of N-acetyl-beta-hexosaminidase and its isoenzymes in serum and synovial fluid from patients with different arthropathies.

OBJECTIVE: To evaluate the activity of N-acetyl-Beta-hexosaminidase (HEX) and its isoenzymes in the serum and synovial fluid of healthy volunteers and patients with an injury to the anterior cruciate ligament and/or meniscus (ACL) osteoarthritis (OA), juvenile idiopathic arthritis (JIA) and rheumatoid arthritis (RA). METHODS: The activity of HEX and its isoenzymes was determined according to Zwierz et al. method. Protein content was determined by the biuret method. RESULTS: The specific activity of HEX and its isoenzymes in the serum of patients with JIA showed a tendency to increase in comparison to the reference group. The specific activity of total HEX in the serum of RA patients was significantly increased in comparison to control. Our results show, that specific activity of HEX in synovial fluid, in the reference group 4.2 +/- 0.21 microkat/kg protein (0.25 unit/mg protein), is similar to activity in normal temporomandibular joint fluid (0.3 unit/mg protein). Therefore, we included this group in our research. In patients with OA and ACL injuries, HEX and its isoenzymes showed a tendency to increase in the specific activity in synovial fluid. The specific activity of HEX and its isoenzymes in the synovial fluid of patients with RA and JIA was significantly elevated in comparison to the control and the remaining groups. CONCLUSION: In the synovial fluid of patients with JIA and RA, the specific activity of HEX and its isoenzymes significantly increased in comparison to control and patients with diseases of a non-inflammatory etiology (OA and ACL). In the synovial fluid of control and diseased groups, HEX constituted a higher percent of total proteins than in serum.

The cleavage of vitamin E galactoside in the rat tissue homogenates.

The stability of alpha-tocopheryl beta-galactoside in the presence of endogenous galactosidases in selected tissue homogenates (liver, kidney, ileum and brain) was estimated. High degree release of alpha-tocopherol from alpha-tocopheryl beta-galactoside in tissues of ileum, kidney and brain was observed (82%, 75% and 72%, increase above endogenous alpha-tocopherol, respectively). A possible enzymatic mechanism of the galactoside decomposition was proposed.

Austenitic steel: a basic implant material in traumatic and orthopedic surgery.

In this article the author analyzed the quality of implants made of chromium-nickel-molybdenum austenitic steels used in traumatic and orthopedic surgery. The steel quality is discussed in reference to the recommendations in current standards. The problem of chemical composition and phase structure of the steel has been taken into consideration, along with its physical and chemical properties, with particular attention to the problem of implant corrosion. The complications observed in clinical practice are largely dependent on material factors and should be taken into account when reporting clinical outcomes.

The influence of electrostimulation with the use of direct and alternating current on the corrosion of Cr-Ni-Mo steel implants.

This paper presents results of researches of influence of different methods of electrostimulation of a bone union on the corrosion of implants made of AISI 316 L steel with the passive and passive-carbon layer deposited on their surface. The electrostimulation was carrying out for 28 days in Tyrode's physiological solution of temperature 36,6 +/- 10C. The pH was varying in the 7,6 - 8,6. A direct, pulsating and sinusoidal current was used for the electrostimulation. Mass decrements and corrosion damages of implants were estimated for individual methods of electrostimulation. Furthermore the paper presents results of infiltration of corrosion products. The corrosion products come into being as the result of stimulation of union bone with the use direct current and invasive methods. A ratio of the corrosion product infiltration was estimated with use of X-ray microanalysis.

[Stresses in self-compressing plates used for joining bone fragments]. / Naprezenia w plytkach samodociskowych stosowanych do zespalania odlamów kostnych.

The analysis of the state of stresses in the auto-compression plates after different stages of joining has been carried out. Distinct differences in the values of stresses in the specific zones of plates were detected. The highest level of stress was located near the holes, in the areas where prebending takes place. The main reason of the loss of the ability to carry introduced compressing forces in the joint and possibility of cracks initiation in plates during the operation has been clarified.