ABSTRACT
Mesenchymal stromal cells are promising candidates for regenerative applications upon treatment of bone defects. Bone marrow-derived stromal cells (BMSCs) are limited by yield and donor morbidity but show superior osteogenic capacity compared to adipose-derived stromal cells (ASCs), which are highly abundant and easy to harvest. The underlying reasons for this difference on a proteomic level have not been studied yet. Human ASCs and BMSCs were characterized by FACS analysis and tri-lineage differentiation, followed by an intraindividual comparative proteomic analysis upon osteogenic differentiation. Results of the proteomic analysis were followed by functional pathway analysis. 29 patients were included with a total of 58 specimen analysed. In these, out of 5148 identified proteins 2095 could be quantified in >80% of samples of both cell types, 427 in >80% of ASCs only and 102 in >80% of BMSCs only. 281 proteins were differentially regulated with a fold change of >1.5 of which 204 were higher abundant in BMSCs and 77 in ASCs. Integrin cell surface interactions were the most overrepresented pathway with 5 integrins being among the proteins with highest fold change. Integrin 11a, a known key protein for osteogenesis, could be identified as strongly up-regulated in BMSC confirmed by Western blotting. The integrin expression profile is one of the key distinctive features of osteogenic differentiated BMSCs and ASCs. Thus, they represent a promising target for modifications of ASCs aiming to improve their osteogenic capacity and approximate them to that of BMSCs.
Subject(s)
Adipose Tissue/cytology , Cell Differentiation , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Osteogenesis , Proteomics , Adult , Cancellous Bone/cytology , Female , Humans , Male , Middle Aged , Proteome/metabolism , Subcutaneous Fat/cytologyABSTRACT
A strategy to reduce implant-related infections is the inhibition of the initial bacterial implant colonization by biomaterials containing silver (Ag). The antimicrobial efficacy of such biomaterials can be increased by surface enhancement (nanosilver) or by creating a sacrificial anode system for Ag. Such a system will lead to an electrochemically driven enhanced Ag ion release due to the presence of a more noble metal. Here we combined the enlarged surface of nanoparticles (NP) with a possible sacrificial anode effect for Ag induced by the presence of the electrochemically more noble platinum (Pt) in physical mixtures of Ag NP and Pt NP dispersions. These Ag NP/Pt NP mixtures were compared to the same amounts of pure Ag NP in terms of cell biological responses, i.e. the antimicrobial activity against Staphylococcus aureus and Escherichia coli as well as the viability of human mesenchymal stem cells (hMSC). In addition, Ag NP was analyzed by ultraviolet-visible (UV-vis) spectroscopy, cyclic voltammetry, and atomic absorption spectroscopy. It was found that the dissolution rate of Ag NP was enhanced in the presence of Pt NP within the physical mixture compared to a dispersion of pure Ag NP. Dissolution experiments revealed a fourfold increased Ag ion release from physical mixtures due to enhanced electrochemical activity, which resulted in a significantly increased toxicity towards both bacteria and hMSC. Thus, our results provide evidence for an underlying sacrificial anode mechanism induced by the presence of Pt NP within physical mixtures with Ag NP. Such physical mixtures have a high potential for various applications, for example as antimicrobial implant coatings in the biomedicine or as bactericidal systems for water and surface purification in the technical area.
Subject(s)
Anti-Bacterial Agents/chemistry , Metal Nanoparticles/chemistry , Platinum/chemistry , Silver/chemistry , Anti-Bacterial Agents/pharmacology , Bacteria/drug effects , Cell Survival/drug effects , Electrochemistry , Electrodes , Humans , Mesenchymal Stem Cells , Silver/pharmacology , SolubilityABSTRACT
The development of antibacterial implant surfaces is a challenging task in biomaterial research. We fabricated a highly antibacterial bimetallic platinum (Pt)/silver(Ag) nanopatch surface by short time sputtering of Pt and Ag on titanium. The sputter process led to a patch-like distribution with crystalline areas in the nanometer-size range (1.3-3.9 nm thickness, 3-60 nm extension). Structural analyses of Pt/Ag samples showed Ag- and Pt-rich areas containing nanoparticle-like Pt deposits of 1-2 nm. The adhesion and proliferation properties of S. aureus on the nanopatch samples were analyzed. Consecutively sputtered Ag/Pt nanopatches (Pt followed by Ag) induced enhanced antimicrobial activity compared to co-sputtered Pt/Ag samples or pure Ag patches of similar Ag amounts. The underlying sacrificial anode mechanism was proved by linear sweep voltammetry. The advantages of this nanopatch coating are the enhanced antimicrobial activity despite a reduced total amount of Ag/Pt and a self-limited effect due the rapid Ag dissolution.
Subject(s)
Anti-Bacterial Agents , Membranes, Artificial , Nanostructures/chemistry , Platinum , Silver , Staphylococcus aureus/growth & development , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Electrodes , Platinum/chemistry , Platinum/pharmacology , Silver/chemistry , Silver/pharmacologyABSTRACT
Schizophrenia is a devastating disease that arises on the background of genetic predisposition and environmental risk factors, such as early life stress (ELS). In this study, we show that ELS-induced schizophrenia-like phenotypes in mice correlate with a widespread increase of histone-deacetylase 1 (Hdac1) expression that is linked to altered DNA methylation. Hdac1 overexpression in neurons of the medial prefrontal cortex, but not in the dorsal or ventral hippocampus, mimics schizophrenia-like phenotypes induced by ELS. Systemic administration of an HDAC inhibitor rescues the detrimental effects of ELS when applied after the manifestation of disease phenotypes. In addition to the hippocampus and prefrontal cortex, mice subjected to ELS exhibit increased Hdac1 expression in blood. Moreover, Hdac1 levels are increased in blood samples from patients with schizophrenia who had encountered ELS, compared with patients without ELS experience. Our data suggest that HDAC1 inhibition should be considered as a therapeutic approach to treat schizophrenia.
Subject(s)
Histone Deacetylase 1/metabolism , Schizophrenia/enzymology , Stress, Psychological/enzymology , Adult , Aged , Animals , DNA Methylation , Female , Hippocampus/enzymology , Histone Deacetylase 1/blood , Histone Deacetylase 1/genetics , Histone Deacetylase Inhibitors/pharmacology , Humans , Male , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Mice, Transgenic , Middle Aged , Phenotype , Prefrontal Cortex/enzymology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Schizophrenia/etiology , Schizophrenia/genetics , Stress, Psychological/complications , Stress, Psychological/genetics , Young AdultABSTRACT
Bimetallic alloyed silver-platinum nanoparticles (AgPt NP) with different metal composition from Ag10Pt90 to Ag90Pt10 in steps of 20 mol% were synthesized. The biological effects of AgPt NP, including cellular uptake, cell viability, osteogenic differentiation and osteoclastogenesis as well as the antimicrobial activity towards Staphylococcus aureus and Escherichia coli were analyzed in comparison to pure Ag NP and pure Pt NP. The uptake of NP into human mesenchymal stem cells was confirmed by cross-sectional focused-ion beam preparation and observation by scanning and transmission electron microscopy in combination with energy-dispersive x-ray analysis. Lower cytotoxicity and antimicrobial activity were observed for AgPt NP compared to pure Ag NP. Thus, an enhanced Ag ion release due to a possible sacrificial anode effect was not achieved. Nevertheless, a Ag content of at least 50 mol% was sufficient to induce bactericidal effects against both Staphylococcus aureus and Escherichia coli. In addition, a Pt-related (≥50 mol% Pt) osteo-promotive activity on human mesenchymal stem cells was observed by enhanced cell calcification and alkaline phosphatase activity. In contrast, the osteoclastogenesis of rat primary precursor osteoclasts was inhibited. In summary, these results demonstrate a combinatory osteo-promotive and antimicrobial activity of bimetallic Ag50Pt50 NP.
Subject(s)
Anti-Bacterial Agents/pharmacology , Metal Nanoparticles , Osteogenesis/drug effects , Platinum/pharmacology , Silver/pharmacology , Anti-Bacterial Agents/chemistry , Cell Differentiation/drug effects , Cell Line , Escherichia coli/drug effects , Escherichia coli Infections/drug therapy , Humans , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/drug effects , Metal Nanoparticles/chemistry , Platinum/chemistry , Silver/chemistry , Staphylococcal Infections/drug therapy , Staphylococcus aureus/drug effectsABSTRACT
In current diagnostic systems, schizophrenia and bipolar disorder are still conceptualized as distinct categorical entities. Recently, both clinical and genomic evidence have challenged this Kraepelinian dichotomy. There are only few longitudinal studies addressing potential overlaps between these conditions. Here, we present design and first results of the PsyCourse study (N = 891 individuals at baseline), an ongoing transdiagnostic study of the affective-to-psychotic continuum that combines longitudinal deep phenotyping and dimensional assessment of psychopathology with an extensive collection of biomaterial. To provide an initial characterization of the PsyCourse study sample, we compare two broad diagnostic groups defined by the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) classification system, that is, predominantly affective (n = 367 individuals) versus predominantly psychotic disorders (n = 524 individuals). Depressive, manic, and psychotic symptoms as well as global functioning over time were contrasted using linear mixed models. Furthermore, we explored the effects of polygenic risk scores for schizophrenia on diagnostic group membership and addressed their effects on nonparticipation in follow-up visits. While phenotypic results confirmed expected differences in current psychotic symptoms and global functioning, both manic and depressive symptoms did not vary between both groups after correction for multiple testing. Polygenic risk scores for schizophrenia significantly explained part of the variability of diagnostic group. The PsyCourse study presents a unique resource to research the complex relationships of psychopathology and biology in severe mental disorders not confined to traditional diagnostic boundaries and is open for collaborations.
Subject(s)
Mental Disorders/diagnosis , Mental Disorders/psychology , Psychotic Disorders/diagnosis , Adult , Aged , Bipolar Disorder/diagnosis , Bipolar Disorder/psychology , Diagnostic and Statistical Manual of Mental Disorders , Female , Humans , Longitudinal Studies , Male , Middle Aged , Phenotype , Psychopathology/methods , Psychotic Disorders/psychology , Research Design , Schizophrenia/diagnosis , Schizophrenic PsychologyABSTRACT
The cellular uptake and dissolution of trigonal silver nanoprisms (edge length 42 ± 15 nm, thickness 8 ± 1 nm) and mostly spherical silver nanoparticles (diameter 70 ± 25 nm) in human mesenchymal stem cells (hMSC's) and human keratinocytes (HaCaT cells) were investigated. Both particles are stabilized by polyvinylpyrrolidone (PVP), with the prisms additionally stabilized by citrate. The nanoprisms dissolved slightly in pure water but strongly in isotonic saline or at pH 4, corresponding to the lowest limit for the pH during cellular uptake. The tips of the prisms became rounded within minutes due to their high surface energy. Afterward, the dissolution process slowed down due to the presence of both PVP stabilizing Ag{100} sites and citrate blocking Ag{111} sites. On the contrary, nanospheres, solely stabilized by PVP, dissolved within 24 h. These results correlate with the finding that particles in both cell types have lost >90% of their volume within 24 h. hMSC's took up significantly more Ag from nanoprisms than from nanospheres, whereas HaCaT cells showed no preference for one particle shape. This can be rationalized by the large cellular interaction area of the plateletlike nanoprisms and the bending stiffness of the cell membranes. hMSC's have a highly flexible cell membrane, resulting in an increased uptake of plateletlike particles. HaCaT cells have a membrane with a 3 orders of magnitude higher Young's modulus than for hMSC. Hence, the energy gain due to the larger interaction area of the nanoprisms is compensated for by the higher energy needed for cell membrane deformation compared to that for spheres, leading to no shape preference.
ABSTRACT
BACKGROUND: In contrast to other tissues, bone has the remarkable ability to heal without scarring. After union of the fracture, the remodeled bone ideally does not differ from the original bone, especially in terms of biomechanical properties. The healing of a fracture resembles the embryonic development of bone. Depending on the biomechanical properties of the fracture, bone heals directly or indirectly, which refers to the formation of cartilage as a step before new bone appears. Currently, treatment of the patient is often limited to anatomical reduction and optimization of the fracture environment with respect to biomechanics. PROSPECTS: Future treatment strategies, however, could include systemic medication that could be especially beneficial for patients at risk of complications in fracture healing. The aim of this review is to provide an overview on the process of fracture healing and to depict possibilities for current and future treatment strategies.
Subject(s)
Fracture Healing/physiology , Biomechanical Phenomena , Bone Morphogenetic Proteins/physiology , Bone Regeneration/physiology , Bone and Bones/physiopathology , Bony Callus/physiopathology , Humans , OsteogenesisABSTRACT
The aim of this study was to reproduce the physico-mechanical antibacterial effect of the nanocolumnar cicada wing surface for metallic biomaterials by fabrication of titanium (Ti) nanocolumnar surfaces using glancing angle sputter deposition (GLAD). Nanocolumnar Ti thin films were fabricated by GLAD on silicon substrates. S. aureus as well as E. coli were incubated with nanostructured or reference dense Ti thin film test samples for one or three hours at 37 °C. Bacterial adherence, morphology, and viability were analyzed by fluorescence staining and scanning electron microscopy and compared to human mesenchymal stem cells (hMSCs).Bacterial adherence was not significantly different after short (1 h) incubation on the dense or the nanostructured Ti surface. In contrast to S. aureus the viability of E. coli was significantly decreased after 3 h on the nanostructured film compared to the dense film and was accompanied by an irregular morphology and a cell wall deformation. Cell adherence, spreading and viability of hMSCs were not altered on the nanostructured surface. The results show that the selective antibacterial effect of the cicada wing could be transferred to a nanostructured metallic biomaterial by mimicking the natural nanocolumnar topography.
Subject(s)
Anti-Bacterial Agents/pharmacology , Nanostructures , Titanium/pharmacology , Animals , Bacterial Adhesion/drug effects , Escherichia coli/cytology , Escherichia coli/drug effects , Hemiptera , Humans , Microbial Viability/drug effects , Nanostructures/ultrastructure , Staphylococcus aureus/cytology , Staphylococcus aureus/drug effects , Surface PropertiesABSTRACT
The German Association for Psychiatry and Psychotherapy (DGPPN) has committed itself to establish a prospective national cohort of patients with major psychiatric disorders, the so-called DGPPN-Cohort. This project will enable the scientific exploitation of high-quality data and biomaterial from psychiatric patients for research. It will be set up using harmonised data sets and procedures for sample generation and guided by transparent rules for data access and data sharing regarding the central research database. While the main focus lies on biological research, it will be open to all kinds of scientific investigations, including epidemiological, clinical or health-service research.
Subject(s)
Cooperative Behavior , Mental Disorders , Psychiatry , Psychotherapy/methods , Psychotherapy/standards , Cohort Studies , Female , Germany , Humans , Male , Mental Disorders/diagnosis , Mental Disorders/psychology , Mental Disorders/therapy , Societies, MedicalABSTRACT
PURPOSE: Freshly prepared autologous plasma clots may serve as a carrier matrix for expanded multipotent mesenchymal stromal cells (MSCs) or bone marrow cells. By varying the calcium concentration, plasma clots with different properties can be produced. The purpose of this in vitro study was to determine the optimal calcium concentrations for the clotting process, intra-clot cell viability, and clot lysis. METHODS: Different plasma clots were prepared by adding an equal volume of RPMI1640 (with or without MSCs) to citrate plasma (either containing platelets or platelet-free). Clotting was initiated by the addition of CaCl(2) (10 g/100 ml H(2)O, 10 % solution). The final concentration of CaCl(2) ranged from 1 to 10 % by volume of plasma. Viability and distribution of the MSCs were analysed by calcein-AM/propidium iodide staining. MSC-embedded plasma clots were dissolved with trypsin (0.25 %), and recovered cells were further incubated for 1 week under cell culture conditions. RESULTS: The viability of MSCs embedded in clots formed by the addition of 1-8 % by volume CaCl2 was not affected by incubation of up to 1 week. In contrast, clots produced by higher volumes of CaCl(2) solutions (9-10 % by volume of plasma) showed decreased numbers of viable cells. Intra-clot cell proliferation was highest in clots produced by addition of 5 % CaCl(2) by plasma volume. Osteocalcin release was not influenced in platelet-free plasma but decreased in platelet-containing plasma. Morphological analysis of stained recovered MSCs revealed that lysis of the plasma clot did not affect cell morphology or subsequent spontaneous proliferation. CONCLUSIONS: Clot formation and clot stability can be controlled by changing the concentration of CaCl(2) added to plasma. The addition of 5 % CaCl(2) produced a plasma clot with optimal results for stem cell delivery.
Subject(s)
Blood Coagulation/drug effects , Bone Marrow Transplantation/methods , Calcium Chloride/pharmacology , Cell Survival/drug effects , Mesenchymal Stem Cells/drug effects , Plasma , Cell Culture Techniques/methods , Cell Proliferation , Dose-Response Relationship, Drug , Humans , Osteocalcin/biosynthesisABSTRACT
Six types of titanium dioxide particles with defined size, shape, and crystal structure (polymorphic form) were prepared: nanorods (70 × 25 nm2), rutile sub-microrods (190 × 40 nm2), rutile microspheres (620 nm), anatase nanospheres (100 nm), anatase microspheres (510 nm), and amorphous titania microspheres (620 nm). All particles were characterized by scanning electron microscopy, X-ray powder diffraction, dynamic light scattering, infrared spectroscopy, and UV spectroscopy. The sub-toxic cell-biological response to these particles by NR8383 macrophages was assessed. All particle types were taken up well by the cells. The cytotoxicity and the induction of reactive oxygen species (ROS) were negligible for all particles up to a dose of 100 µg mL-1, except for rutile microspheres which had a very rough surface in contrast to anatase and amorphous titania microspheres. The particle-induced cell migration assay (PICMA; based on chemotaxis) of all titanium dioxide particles was comparable to the effect of control silica nanoparticles (50 nm, uncoated, agglomerated) but did not show a trend with respect to particle size, shape, or crystal structure. The coating with carboxymethylcellulose (CMC) had no significant biological effect. However, the rough surface of rutile microspheres clearly induced pro-inflammatory cell reactions that were not predictable by the primary particle size alone.
ABSTRACT
OBJECTIVES: Autologous transplants are still the means of choice for bypass surgery. In addition to good tolerability, there is a reduced thrombogenicity and fewer neointima hyperplasia compared to artificial materials. However, since viable transplants are limited, attempts are being made to improve existing artificial vascular prosthesis material. Next to the reduction of thrombogenicity, a rapid endothelialization of the vascular graft should reduce intimal hyperplasia and thus prevent stenoses. The effect of newly developed silicon oxide coatings on the growth of endothelial cells was therefore the goal of this work in a cell culture study. METHODS: A woven, uncoated polyethylene terephthalate (PET) vessel prosthesis was used. The coating process was carried out in a low-pressure plasma reactor in a multi-step process. After preparation of the vacuum chamber hexamethyldisiloxane (HDMSO) with oxygen was evaporated using argon plasma. By this an approx. 1 nm thin adhesion promoter layer was separated from plasma and HMDSO. The silicone oxide barrier layer was applied to the PET vessel samples. The carbon content of the layer could be selectively altered by changing the HMDSO oxygen flow ratio, resulting in coatings of 100 nm, 500 nm, and 1,000 nm. In addition, two different oxygen-to-HMDSO ratios were used. To achieve a carbon coating as low as possible, the ratio was set to 200:1. A carbon-rich layer was obtained with the 1:1 setting. The various coatings were then examined for their surface texture by scanning electron microscopy (SEM) as well as by cell culture experiments for cell viability and growth using EA.hy 926 cells. RESULTS: SEM showed no changes in the surface morphology; however a layer thickness of 1,000 nm showed peeled off coating areas. Alamar blue assays showed a significantly higher metabolic activity (p=0.026) for the coating 500 nm, ratio 200:1 compared to untreated control samples and a significantly lower metabolic activity (p=0.037) of the coating 500 nm, ratio 1:1 compared to the coating 500 nm, ratio 200:1. This underlines the apparent tendency of the 1:1 coating to inhibit the metabolic activity of the cells, while the 200:1 coating increases the activity. Fluorescence microscopy after calcein acetoxymethyl ester (AM) staining showed no significant difference between the different coatings and the uncoated PET material. However, a tendency of the increased surface growth on the coating 500 nm, ratio 200:1, is shown. The coatings with the ratio 1:1 tend to be less densely covered. CONCLUSIONS: The results of this work indicate a great potential in the silicon coating of vascular prosthesis material. The plasma coating can be carried out easy and gently. Cell culture experiments demonstrated a tendency towards better growth of the cells on the 200:1 ratio coating and a poorer growth on the carbon-rich coating 1:1 compared to the uncoated material. The coating with silicon oxide with a thickness of 500 nm and an oxygen-HMDSO ratio of 200:1, a particularly low-carbon layer, appears to be a coating, which should therefore be further investigated for its effects on thrombogenicity and intimal hyperplasia.
ABSTRACT
PURPOSE: The induced membrane technique (IMT) is a two-stage surgical procedure for reconstruction of bone defects. Bone grafting (second stage of IMT) is recommend after 4-8 weeks assuming the highest bioactivity of IMs. However, larger studies concerning the biology and maturation of IMs and a potential time dependency of the bioactivity are missing. Therefore, aim of this study was the time-dependent structural and cellular characterization of cement spacer IMs concomitantly to an analysis of membrane bioactivity. METHODS: IMs from 60 patients (35-82 years) were obtained at different maturation stages (1-16 weeks). IMs were studied by histology and co-culture with mesenchymal stem cells (MSC). IM lysates were analyzed by ELISA and protein microarray. RESULTS: Increasing vascularization and fibrosis were found in membranes older than 4 and 7 weeks, respectively. MSC grew out from all membranes and all membranes enhanced proliferation of cultured MSC. Osteocalcin and osteopontin (in membrane lysates or induced in MSC by membrane tissue) were found over all time points without significant differences. In contrast to alkaline phosphatase activity, increasing levels of osteoprotegerin were found in membranes. CONCLUSION: The histological structure of IMs changes during growth and maturation, however, biologically active MSC and factors related to osteogenesis are found over all time points with minor changes. Thus, membranes older than 8 weeks exert regenerative capacities comparable to the younger ones. The postulated narrow time frame of 4-8 weeks until bone grafting can be questioned and surgeons may choose timing for the second operation more independently and based on other clinical factors.
Subject(s)
Mesenchymal Stem Cells , Osteogenesis , Coculture Techniques , Humans , Membranes , Mesenchymal Stem Cells/metabolism , OsteocalcinABSTRACT
The current Coronavirus Disease 19 (COVID-19) pandemic has exemplified the need for simple and efficient prevention strategies that can be rapidly implemented to mitigate infection risks. Various surfaces have a long history of antimicrobial properties and are well described for the prevention of bacterial infections. However, their effect on many viruses has not been studied in depth. In the context of COVID-19, several surfaces, including copper (Cu) and silver (Ag) coatings have been described as efficient antiviral measures that can easily be implemented to slow viral transmission. In this study, we detected antiviral properties against Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) on surfaces, which were coated with Cu by magnetron sputtering as thin Cu films or as Cu/Ag ultrathin bimetallic nanopatches. However, no effect of Ag on viral titers was observed, in clear contrast to its well-known antibacterial properties. Further enhancement of Ag ion release kinetics based on an electrochemical sacrificial anode mechanism did not increase antiviral activity. These results clearly demonstrate that Cu and Ag thin film systems display significant differences in antiviral and antibacterial properties which need to be considered upon implementation.
Subject(s)
COVID-19 , Silver , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Antiviral Agents/pharmacology , Copper/chemistry , Copper/pharmacology , Humans , SARS-CoV-2 , Silver/chemistry , Silver/pharmacologyABSTRACT
BACKGROUND: In recent years, plasma treatment of medical devices and implant materials has gained more and more acceptance. Inactivation of microorganisms by exposure to ultraviolet (UV) radiation produced by plasma discharges and sterilization of medical implants and instruments is one possible application of this technique. The aim of this study was to evaluate the effectiveness of this sterilization technique on silicone implant material. METHODS: Bacillus atrophaeus spores (10(6) colony-forming units [CFUs]) were sprayed on the surfaces of 12 silicone implant material samples. Four plasma sets with different gas mixtures (argon [Ar], argon-oxygen [Ar:O(2)], argon-hydrogen [Ar:H(2)] and argon-nitrogen [Ar:N(2)]) were tested for their antimicrobial properties. Post-sterilization mechanical testing of the implant material was performed in order to evaluate possible plasma-induced structural damage. RESULTS: The inductively coupled low-pressure plasma technique can achieve fast and efficient sterilization of silicone implant material without adverse materials effects. All four gas mixtures led to a significant spore reduction, and no structural damage to the implant material could be observed.
Subject(s)
Plasma Gases , Prostheses and Implants/microbiology , Sterilization/methods , Argon , Bacillus/drug effects , Colony Count, Microbial , Hydrogen , Nitrogen , Oxygen , Pressure , Silicones , Spores, Bacterial/drug effects , Spores, Bacterial/growth & developmentABSTRACT
Silica fibers with a dimension of 0.3 µm â 3.2 µm2 nm were prepared by a modified Stöber synthesis as model particles. The particles were characterized by scanning electron microscopy, elemental analysis, thermogravimetry and X-ray powder diffraction. Their uptake by macrophages (THP-1 cells and NR8383 cells) was studied by confocal laser scanning microscopy and scanning electron microscopy. The uptake by cells was very high, but the silica fibers were not harmful to NR8383 cells in concentrations up to 100 µg mL-1. Only above 100 µg mL-1, significant cell toxic effects were observed, probably induced by a high dose of particles that had sedimented on the cells and led to the adverse effects. The chemotactic response as assessed by the particle-induced migration assay (PICMA) was weak in comparison to a control of agglomerated silica particles. The as-prepared fibers were fully X-ray amorphous but crystallized to ß-cristobalite after heating to 1000 °C and converted to α-cristobalite upon cooling to ambient temperature. The fibers had sintered to larger aggregates but retained their elongated primary shape. The particle cytotoxicity towards THP-1 cells was not significantly enhanced by the crystallization.
Subject(s)
Macrophages , Silicon Dioxide , Crystallization , Microscopy, Electron, Scanning , Particle Size , Silicon Dioxide/toxicity , X-Ray DiffractionABSTRACT
PURPOSE: Mesenchymal stem cells (multipotent human mesenchymal stromal cells, MSC) are currently the most promising cell type for regenerative medicine. For a clinical approach, it is necessary to develop and establish methods for expansion, differentiation, and delivery. METHODS: A completely autologous plasma clot containing peripheral blood mononuclear cells (PBMC) was tested for the osteopromotive activity towards expanded human mesenchymal stem cells in vitro. The plasma clot was prepared from anticoagulated blood plasma after addition of isolated leukocytes and calcium chloride. Plasma clots after the gelation were added to subconfluently growing MSC or used in a transwell system. Cell proliferation, the activity of alkaline phosphatase, the release of osteoprotegerin, C-terminal procollagen peptide, as well as osteocalcin, the analysis of matrix mineralization as well as bone nodule formation were analyzed up to 3 weeks. RESULTS: In contrast to plasma clots with no exogenously added leukocytes, the presence of PBMC within the plasma clot significantly promoted osteogenic differentiation of MSC correlated to the time period of incubation. Proliferation of MSC was decreased at maximal mineralization time points. In addition, the osteopromotive activity was identified as soluble factor/factors by transwell assay system. There was a decrease in osteoprotegerin when the cells were cultured in the presence of plasma clots compared to control cell cultures without plasma clots. The osteocalcin expression was continuously higher after culture in the presence of plasma clots and significantly higher after 2- and 3-week after culture in the presence of leukocyte-containing plasma clots compared to 1-week cell culture. Differences in the concentration of the C-terminal procollagen peptide were not measured. CONCLUSIONS: The direct inoculation of an autologous mononuclear cell fraction (which contains leukocytes and MSC), e.g., isolated from a bone marrow aspirate or a different source into an autologous plasma gel, may be a further new strategy for bone fracture therapy.
Subject(s)
Extracellular Matrix/physiology , Leukocytes/physiology , Mesenchymal Stem Cells/physiology , Stromal Cells/physiology , Blood Coagulation/physiology , Cell Differentiation , Cell Proliferation , Cells, Cultured , Fibrin/physiology , Humans , Plasma/physiologyABSTRACT
OBJECTIVES: The purpose was to evaluate inflammatory and microcirculatory reactions after implantation of various calcium phosphate bone substitutes in an in vivo model. METHODS: Calcium phosphate-based bone substitutes were implanted in dorsal skinfold chambers of mice. Intravital fluorescence microscopy was performed to measure inflammatory and microcirculatory reactions based on functional vessel density (FVD), capillary leakage, and relative white blood cell velocity (rWBCV). RESULTS: An increase of FVD was observed in all groups and the capillary leakage grew with a level of significance (p < 0.001). The fraction of rolling and sticking leukocytes (rWBCV) was highest at the beginning of the trial and decreased during the course. CONCLUSIONS: There are differences in microvascular soft tissue reactions between various calcium phosphate bone substitutes, but inflammatory reactions were moderate, and the results revealed no reasons which explain the sporadic failure of the tested substances under clinical conditions.
Subject(s)
Bone Substitutes , Calcium Phosphates , Capillary Leak Syndrome/pathology , Foreign-Body Reaction/pathology , Microcirculation/physiology , Microvessels/pathology , Animals , Blood Flow Velocity , Cell Adhesion/physiology , Female , Leukocyte Count , Leukocyte Rolling , Mice , Mice, Inbred BALB C , Neovascularization, Physiologic/physiology , Skin/blood supply , Skin Window TechniqueABSTRACT
Biocidal compounds that quickly kill bacterial cells and are then deactivated in the surrounding without causing environmental problems are of great current interest. Here, we present new biodegradable antibacterial polymers based on polyionenes with inserted ester functions (PBI esters). The polymers are prepared by polycondensation reaction of 1,4-dibromobutene and different tertiary diaminodiesters. The resulting PBI esters are antibacterially active against a wide range of bacterial strains and were found to quickly kill these cells within 1 to 10 min. Because of hydrolysis of the ester groups, the PBI esters are degraded and deactivated in aqueous media. The degradation rate depends on the backbone structure and the pH. The structure of the polymers also controls the deactivation mechanism. While the more hydrophilic polymers require hydrolyses of only 19 to 30% of the ester groups to become practically inactive, the more hydrophobic PBI esters require up to 85% hydrolysis to achieve the same result. Thus, depending on the environmental conditions and the chemical nature, the PBI esters can be active for only 20 min or for at least one week.