The effect of albumin and γ-globulin on synovial fluid lubrication: Implication for knee joint replacements.

Total knee arthroplasty has become a routine procedure for patients suffering from joint diseases. Although the number of operations continuously increases, a limited service-life of implants represents a persisting challenge for scientists. Understanding of lubrication may help to suitably explain tribological processes on the way to replacements that become durable well into the third decade of service. The aim of the present study is to assess the formation of protein lubricating film in the knee implant. A developed knee simulator was used to observe the contact of real femoral and transparent polymer tibial component using fluorescent microscopy. The contact was lubricated by various protein solutions with attention to the behaviour of albumin and γ-globulin. In order to suitably mimic a human synovial fluid, hyaluronic acid and phospholipids were subsequently added to the solutions. Further, the change in shape and the migration of the contact zone were studied. The results showed considerable appearance differences of the contact over the swing phase of the simplified gait cycle. Regarding film formation, a strong interaction of the various molecules of synovial fluid was observed. It was found that the thickness of the lubricating layer stabilizes within around 50 s. Throughout the contact zone, protein agglomerations were present and could be clearly visualised using the applied optical technique.

[Strong Antibacterial Efficacy of Titanium Surfaces Modified by Nanotubes and Silver Nanoparticles]. / Silné antibakteriální vlastnosti kovového povrchu modifikovaného nanotrubicemi titanu a nanostríbrem.

PURPOSE OF THE STUDY Nano-structuring and nano-silver have been extensively studied for improving the antibacterial ability of implants due to their powerful antibacterial activity; however, there is no clinical application as yet. The aim of the study was to determine the antibacterial, antiadhesive and cytotoxic features of Ti6Al4V modified with nano-texturing and silver nano-particles. MATERIAL AND METHODS The nanoparticles were applied on polished and nano-textured Ti6Al4V using sonoreduction. The surface topography, roughness, friction coefficients, hardness and elastic modulus values for prepared top layers were established. The materials were tested for antibacterial and antiadhesion activity using reference bacterial strains (Staphylococcus epidermidis CCM 7221, Staphylococcus aureus MRSA 4591, Enterococcus faecalis CCM 4224, Escherichia coli CCM 3954) and their cytocompatibility. RESULTS A strong antibacterial activity of samples treated with nano-texture and/or silver nanoparticles compared to all the tested bacterial strains at 24 hours was proven. This antibacterial activity was diminishing in relation to Staphylococcus aureusand Enterococcus faecalisat 48 and 72 hours but remained very effective against Staphylococcus epidermidisand Escherichia coli. We also demonstrated antibiofilm activity for samples treated with silver nanoparticles and nano-tubes in experiments lasting 24 and 72 hours. DISCUSSION Our main findings are in agreement with those reported in recent literature. The implant surfaces treated with nano-texture in combination with silver nanoparticles exhibit strong antibacterial and antibiofilm characteristics. Despite there is conclusive evidence of strong antibacterial functioning, why these implant modifications have not been widely applied in clinical practice remains a question. While many obstacles including legislative procedures required for clinical implementation are more or less known, it should be clearly demonstrated that this surface modification does neither harm the patient nor interfere with the long-term survivorship of the implants before their wide-range clinical application. CONCLUSIONS Surface modification of Ti6Al4V with nano-texturing and silver nanoparticles resulted in strong antibacterial and modest antibiofilm effects. Thus, our results confirmed the technological potential of nano-texturing and silver nanoparticles for the improvement of antibacterial properties of implants. Key words:prosthetic joint infection, anti-infective biomaterials, titanium alloy, silver nanoparticles, nanotubes, prevention of infection.

On the observation of lubrication mechanisms within hip joint replacements. Part I: Hard-on-soft bearing pairs.

The present study describes the lubrication mechanisms within artificial hip joints considering real conformity of rubbing surfaces. Part I is focused on hard-on-soft material combination, introducing the fundamentals of lubrication performance. These pairs have not been explored in terms of in situ observation before. The contact of metal femoral component articulating with transparent polymer acetabular cup was studied using a hip joint simulator. The film formation was evaluated by fluorescent microscopy method. Various model synovial fluids were employed while the key constituents, i.e. albumin, γ-globulin, and hyaluronic acid were fluorescently stained to determine its role in film formation process. Two types of the tests were performed. The first dynamic test aimed on the development of film thickness under constant load during motor driven swinging motion mimicking flexion-extension. Subsequently, a combined test was designed consisting of the three phases; static part with loading/unloading phase (1), pendulum swinging till spontaneous damping of the motion due to friction (2), and static observation under the constant load (3). The results clearly confirmed that the interaction of constituents of synovial fluid plays a dominant role and substantially influences the lubrication conditions. In particular, the main finding coming from the present study is that γ-globulin together with hyaluronic acid form relatively thin stable boundary layer enabling the enhanced adsorption of albumin, thus increasing the lubricant film. Part II of the present study is focused on hard-on-hard pairs while the main differences in film formation process are highlighted among others.

Lubrication within hip replacements - Implication for ceramic-on-hard bearing couples.

The objective of the present study is to clarify the lubrication processes within artificial joints considering the ceramic femoral heads focusing on the role of particular proteins. Two optical methods were employed; colorimetric interferometry and fluorescent microscopy. The experiments were conducted in ball-on-disc configuration, where the ball is made from ceramic (Sulox(TM), BIOLOX(®)delta) and the disc from optical glass. The measurements were realized under pure rolling, partial negative and partial positive sliding, to get a complex information about the protein film behaviour under various conditions. Moreover, two different speeds were investigated; 5.7 and 22 mm/s, respectively. The contact was lubricated by saline solutions containing albumin and γ-globulin in a ratio 2:1, while the total protein concentration was 10.5 mg/ml. Under pure rolling conditions, the film thickness gradually increases with time/rolling distance independently of material and rolling speed, while the dominant fluid constituent is albumin. In the case of negative sliding, the film formation is time/distance/speed dependent. At lower speed, both proteins contribute to film thickness; at higher speed, the effect of γ-globulin is not substantial. When the disc is faster, the character of film formation is similar to the metal component in the case of Sulox ceramic. Biolox ceramic shows a different behaviour, while for both materials, the contribution of γ-globulin increases with increasing speed. As most of the results can be well explained in terms of specific proteins, it can be concluded that the experimental approach is suitable for the investigation of protein film formation considering the ceramic materials.

Effects of silver nanoparticles on human dermal fibroblasts and epidermal keratinocytes.

Biomedical application of silver nanoparticles (AgNPs) has been rapidly increasing. Owing to their strong antimicrobial activity, AgNPs are used in dermatology in the treatment of wounds and burns. However, recent evidence for their cytotoxicity gives rise to safety concerns. This study was undertaken as a part of an ongoing programme in our laboratory to develop a topical agent for wound healing. Here, we investigated the potential toxicity of AgNPs using normal human dermal fibroblasts (NHDF) and normal human epidermal keratinocytes (NHEK) with the aim of comparing the effects of AgNPs and ionic silver (Ag-I). Besides the effect of AgNPs and Ag-I on cell viability, the inflammatory response and DNA damage in AgNPs and Ag-I-treated cells were examined. The results showed that Ag-I were significantly more toxic than AgNPs both on NHDF and NHEK. Non-cytotoxic concentrations of AgNPs and Ag-I did not induce DNA strand breaks and did not affect inflammatory markers, except for a transient increase in interleukin 6 levels in Ag-I-treated NHDF. The results showed that AgNPs are more suitable for the intended application as a topical agent for wound healing up to the concentration 25 µg/mL.

[Determination of a Friction Coefficient for THA Bearing Couples]. / Stanovení soucinitele trení hlavic a jamek TEP kycle.

PURPOSE OF THE STUDY: The wear of articular surfaces is considered one of the most important factors limiting the life of total hip arthroplasty (THA). It is assumed that the particles released from the surface of a softer material induce a complex inflammatory response, which will eventually result in osteolysis and aseptic loosening. Implant wear is related to a friction coefficient which depends on combination of the materials used, roughness of the articulating surfaces, internal clearance, and dimensions of the prosthesis. MATERIAL AND METHODS: The selected parameters of the bearing couples tested were studied using an experimental device based on the principle of a pendulum. Bovine serum was used as a lubricant and the load corresponded to a human body mass of 75 kg. The friction coefficient was derived from a curve of slowdown of pendulum oscillations. Roughness was measured with a device working on the principle of interferometry. Clearance was assessed by measuring diameters of the acetabular and femoral heads with a 3D optical scanner. The specimens tested included unused metal-on-highly cross-linked polyethylene, ceramic-on-highly cross-linked polyethylene and ceramic-on-ceramic bearing couples with the diameters of 28 mm and 36 mm. For each measured parameter, an arithmetic mean was calculated from 10 measurements. RESULTS: 1) The roughness of polyethylene surfaces was higher by about one order of magnitude than the roughness of metal and ceramic components. The Protasul metal head had the least rough surface (0.003 µm). 2) The ceramic-on-ceramic couples had the lowest clearance. Bearing couples with polyethylene acetabular liners had markedly higher clearances ranging from 150 µm to 545 µm. A clearance increased with large femoral heads (up to 4-fold in one of the couple tested). 3) The friction coefficient was related to the combination of materials; it was lowest in ceramic-on-ceramic surfaces (0.11 to 0.12) and then in ceramic-on-polyethylene implants (0.13 to 0.14). The friction coefficient is supposed to increase with a decreasing femoral head diameter. However, in the bearing couples with polyethylene liners manufactured by one company, paradoxically, the friction coefficient slightly increased with an increase in femoral head size from 28 mm to 36 mm. 4) The lowest friction moment (< 3.5 Nm) was found for ceramic-on-ceramic implants 28 mm in diameter; the highest values were recorded in metal-on-polyethylene bearing couples 36 mm in diameter (> 7 Nm). DISCUSSION: Although our study confirmed that the bearing couples produced by different manufacturers varied to some extent in the parameters studied, in our opinion, this variability was not significant because it was not within an order of magnitude in any of the tests. CONCLUSIONS: The study showed that both the friction coefficient and the friction moment are affected more by the combination of materials than by the diameter of a femoral head. The best results were achieved in ceramic-on-ceramic implants.

Antibacterial, cytotoxicity and physical properties of laser--silver doped hydroxyapatite layers.

Hydroxyapatite layers with silver doping from 0.06 at.% to 14 at.% were prepared by laser deposition. The films' physical properties such as morphology, composition, crystallinity, Young's modulus and microhardness were measured. Films were amorphous or polycrystalline in dependence on deposition temperature (from RT to 600 °C). Antibacterial properties were tested using Escherichia coli and Bacillus subtilis cells. The antibacterial efficacy changed with silver doping from 4% to 100%. Cytotoxicity was studied by a direct contact test. Depending on doping and crystallinity the films were either non-toxic or mildly toxic.

[Oxidative stress in patients after kidney transplantation]. / Oxidacní stres u pacientu po transplantaci ledviny.

Successful kidney transplantation (KT) leads not only to normalization of renal excretory function, but also to modification of the metabolic and endocrine kidney function. The most common cause of morbidity and mortality in patients after KT are cardiovascular diseases which development is also associated with oxidative stress (OS). KT and the posttransplantation period are associated with increased OS that could gradually decrease. Some immunosuppressive drugs also contribute to increase of OS, especially compounds from a group of calcineurin inhibitors and thus indirectly contribute to increased risk of cardiovascular complications.

[Oxidative stress in patients on regular hemodialysis and peritoneal dialysis]. / Oxidacní stres u pacientu s náhradou funkce ledvin hemodialýzou a peritoneální dialýzou.

Hemodialysis and peritoneal dialysis are methods of blood purification, which partially replaced excretory renal function in patients with chronic renal failure, which was depleted regime, dietary and pharmaco-therapeutic remedy, and who are not eligible for kidney transplantation. Both two methods are accompanied by increased oxidative stress. In peritoneal dialysis particularly the composition of dialysis solution contributes to oxidative stress. In extracorporeal hemodialysis the oxidative stress is associated with the character of hemodialysis membranes, non-specific loss of low molecular weight antioxidants, activation of leukocytes (oxidative burst), feroteraphy, supplementation with low molecular weight antioxidants and other factors. To improve and maintain the quality of life of dialysis patients, the continuous monitoring of oxidative stress-related parameters as non-traditional risk factors for cardiovascular complications development is suitable.

[Oxidative stress in kidney disease patients]. / Oxidacní stres u pacientu s onemocnením ledvin.

Patients with chronic kidney disease are exposed to oxidative stress (OS) that contributes to deterioration of health. Decrease in renal excretory capacity contributes to the accumulation of pro-oxidative substances that are detrimental not only to kidney but to the whole organism including the cardiovascular system. Components of antioxidant system play an important role in the elimination of the OS. The monitoring of antioxidant levels and products of oxidative damage in these patients and the correct interpretation of relationship between these markers and the function of kidney and other organs may contribute to the more effective treatment and health improvement of the patients.