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Biomedical subjects

U Nöth

Publications and source records attributed to U Nöth.

At least 19 recordsLinked to original sources
PubMed2006

[A novel questionnaire to assess activity in patients after hip arthroplasties].

BACKGROUND: Until now, the activity of the lower limb could only be exactly measured with expensive electronic pedometers. The aim of this study was to develop a feasible questionnaire to measure the activity of patients after arthroplasty. METHODS: The "Daily Activity Questionnaire" (DAQ) was developed in several steps and the evaluation was carried out in three groups of patients with osteoarthritis of the hip (160 patients and 855 investigated days). The psychometric characteristics were verified. RESULTS: The retest reliability (ICC) of the DAQ is in a range comparable to the electronic pedometer StepWatch. The testing of the criterion validity showed high correlations between the StepWatch and the DAQ (r=0.743). CONCLUSION: Because of its high correlations to the load changes actually performed, the DAQ is especially suited to answer arthroplasty-related questions. In all patient groups, projected load changes between 1.8 and 2.4 million per year were found, which by far exceed the generally accepted test standards for endoprostheses.

Activities of Daily Living↗
PubMed2005

Anterior cruciate ligament constructs fabricated from human mesenchymal stem cells in a collagen type I hydrogel.

BACKGROUND: Disruptions of the anterior cruciate ligament (ACL) of the knee joint are common and are currently treated using ligament or tendon grafts. In this study, we tested the hypothesis that it is possible to fabricate an ACL construct in vitro using mesenchymal stem cells (MSC) in combination with an optimized collagen type I hydrogel, which is in clinical use for autologous chondrocyte transplantation (ACT). METHODS: ACL constructs were molded using a collagen type I hydrogel containing 5 x 10(5) MSC/mL and non-demineralized bone cylinders at each end of the constructs. The constructs were kept in a horizontal position for 10 days to allow the cells and the gel to remodel and attach to the bone cylinders. Thereafter, cyclic stretching with 1 Hz was performed for 14 days (continuously for 8 h/day) in a specially designed bioreactor. RESULTS: Histochemical analysis for H and E, Masson-Goldner and Azan and immunohistochemical analysis for collagen types I and III, fibronectin and elastin showed elongated fibroblast-like cells embedded in a wavy orientated collagenous tissue, together with a ligament-like extracellular matrix in the cyclic stretched constructs. No orientation of collagen fibers and cells, and no formation of a ligament-like matrix, could be seen in the non-stretched control group cultured in a horizontal position without tension. RT-PCR analysis revealed an increased gene expression of collagen types I and III, fibronectin and elastin in the stretched constructs compared with the non-stretched controls. DISCUSSION: In conclusion, ACL-like constructs from a collagen type I hydrogel, optimized for the reconstruction of ligaments, and MSC have been fabricated. As shown by other investigators, who analyzed the influence of cyclic stretching on the differentiation of MSC, our results indicate a ligament-specific increased protein and gene expression and the formation of a ligament-like extracellular matrix. The fabricated constructs are still too weak for animal experiments or clinical application and current investigations are focusing on the development of a construct with an internal augmentation using biodegradable fibers.

Animals↗
PubMed2004

[In vitro fabrication of tendon substitutes using human mesenchymal stem cells and a collagen type I gel].

BACKGROUND AND PURPOSE: Reconstruction of tendon tissue is problematic in many cases. Since direct tendon suture is often impossible, major reconstruction with the use of free tendon transplants or tendon transposition is necessary. Important motor units often have to be sacrificed for reconstructive purposes. In this study we investigated whether long tendon-like substitutes can be fabricated in vitro from human mesenchymal stem cells (MSCs) and a collagen type I gel when cultured under cyclic stretching conditions. MATERIAL AND METHODS: MSCs were obtained from bone marrow aspirates of the iliac crest. Cells were suspended in a collagen type I gel and polymerized in a glass-cylinder with defined size. The fabricated tendon substitutes underwent static stretching for 14 days followed by cyclic stretching for 21 days in a special manufactured bioreactor. Non-stretched substitutes served as a control. RESULTS: Macroscopically the stretched tendon substitutes showed an increased opacity and a smoother surface structure compared to the non-stretched control. The stretched substitutes displayed more spindle-shaped, longitudinal orientated cells, a tendon-like organization of the collagen matrix, and a parallel organization of the collagen fibers when stained with Hematoxylin/Eosin and Elastica. CONCLUSION: Long tendon substitutes could be fabricated from MSCs and a collagen type I gel by cyclic stretching and showed tendon-like parallel collagen fibers and spindle-shaped cells. The use of MSCs in combination with adequate scaffold materials has great therapeutic potential for the development of autologous transplantable tendon substitutes.

Bioreactors↗
PubMed2002

Multilineage differentiation of adult human bone marrow progenitor cells transduced with human papilloma virus type 16 E6/E7 genes.

We have established a new adult human bone marrow-derived cell line hMPC 32F, stably transduced with human papilloma virus type 16 E6/E7 genes, that displays mesenchymal multilineage differentiation ability in vitro. The hMPC 32F cells exhibited a population doubling time of 22 h and have been maintained in culture for about 20 passages. When cultured in conditions promoting osteogenic, adipogenic, or chondrogenic differentiation, hMPC 32F cells expressed mature differentiated phenotypes. These include (1) osteoblastic phenotype characterized by upregulated alkaline phosphatase (ALP) expression and extracellular matrix mineralization, (2) adipocytic phenotype with the presence of intracellular lipid droplets, and (3) chondrocytic phenotype of round cells surrounded by a sulfated proteoglycan-rich matrix. In addition, the hMPC 32F cells expressed differentiation lineage-specific genes, as detected by RT-PCR. Furthermore, osteogenic and adipogenic cultures responded to regulatory factors such as transforming growth factor-beta1 (TGF-beta1) and 1alpha, 25-dihydroxyvitamin D3 (1,25(OH)2D3). Thus, continuous treatment of osteogenic cultures for 2 weeks with TGF-beta1 decreased ALP activity and mRNA expression and inhibited osteocalcin mRNA expression and matrix mineralization, whereas l,25(OH)2D3 had an additive, stimulatory effect. In adipogenic cultures, treatment with TGF-beta1 for 2 weeks markedly inhibited adipogenesis whereas 1,25(OH)2D3 had no obvious effect. Finally, clonal analysis of hMPC 32F cells revealed a high percentage of multipotent clones, although clones of more restricted differentiation potential were also present. These characteristics of the hMPC 32F cell line suggest their pluripotent, progenitor, and nontransformed nature and indicate their potential application for studying the mechanisms governing developmental potential of adult human bone marrow mesenchymal progenitor cells.

Adipocytes↗
PubMed2001

A novel class of amitogenic alginate microcapsules for long-term immunoisolated transplantation.

In the light of results of clinical trials with immunoisolated human parathyroid tissue Ba2+-alginate capsules were developed that meet the requirements for long-term immunoisolated transplantation of (allogeneic and xenogeneic) cells and tissue fragments. Biocompatibility of the capsules was achieved by subjecting high-M alginate extracted from freshly collected brown algae to a simple purification protocol that removes quantitatively mitogenic and cytotoxic impurities without degradation of the alginate polymers. The final ultra-high-viscosity, clinical-grade (UHV/CG) product did not evoke any (significant) foreign body reaction in BB rats or in baboons. Similarly, the very sensitive pERK assay did not reveal any mitogenic impurities. Encapsulated cells also exhibited excellent secretory properties under in vitro conditions. Despite biocompatible material, pericapsular fibrosis is also induced by imperfect capsule surfaces that can favor cell attachment and migration under the release of material traces. This material can interact with free end monomers of the alginate polymers under formation of mitogenic advanced glycation products. Smooth surfaces, and thus topographical biocompatibility of the capsules (visualized by atomic force microscopy), can be generated by appropriate crosslinking of the UHV/CG-alginate with Ba2+ and simultaneous suppression of capsule swelling by incorporation of proteins and/or perfluorocarbons (i.e., medically approved compounds with high oxygen capacity). Perfluorocarbon-loaded alginate capsules allow long-term non-invasive monitoring of the location and the oxygen supply of the transplants by using 19F-MRI. Transplantation studies in rats demonstrated that these capsules were functional over a period of more than two years.

Alginates↗
PubMed2000

[Cytotoxicity study of high gold content Degutan surfaces of various degrees of roughness with fibroblasts (BALB 3T3) and osteoblasts (hFOB 1.19)].

The cytotoxicity of Degutan surfaces with different degrees of roughness, and the effect of surface structures on osteoblast proliferation and differentiation, was investigated with standardised cell culture systems. Fibroblast cell lines (BALB/3T3) and osteoblast cell lines (hFOB 1.19) were used. The number and variability of the cells were determined for assessment of proliferation and alkaline phosphatase activity, collagen I and osteocalcin production were used as parameters for differentiation. In the early phase, the largest numbers of cells and greatest proliferation were measured on polished Degutan surfaces. In the late phase, however, larger numbers of cells and a greater degree of proliferation were to be seen on sandblasted and sandblasted/heat-treated Degutan surfaces. No differences were found for collagen I, osteocalcin production or alkaline phosphatase activity. Neither the osteoblasts nor the fibroblasts revealed a toxic effect of Degutan. The results for osteoblast differentiation correlate with recent studies on identical structured titanium surfaces. In view of the immeasurable amount of ion release, Degutan may be considered an ideal model for an inert material surface.

3T3 Cells↗
PubMed2000

Non-invasive evaluation of the location, the functional integrity and the oxygen supply of implants: 19F nuclear magnetic resonance imaging of perfluorocarbon-loaded Ba2+-alginate beads.

19F nuclear magnetic resonance imaging (MRI) can be used as a non-invasive tool to simultaneously determine the location, the integrity and the oxygen supply of Ba2+-alginate implants. This requires that the beads (implants) are pre-loaded with the perfluorocarbon compound F-44E. Implantation of solid 19F-labelled beads into the peritoneum, below the kidney capsule or into the muscle of Wistar WU rats demonstrated that these beads could be detected by 19F-MRI for up to 18 months after implantation. This indicated that F-44E is not considerably released from the beads during implantation. The signal to noise ratio of liquid-core beads was higher by a factor of 4 than the signal to noise ratio of solid beads, but liquid-core beads were more fragile and also too large for implantation under the kidney capsule and into the intramuscular tissue. Quantitative 2-dimensional 19F-T1 maps (resolution 0.5 x 0.5 mm) could be deduced from 19F-MRI measurements. These T1-maps correlated to the local pO2-values. The partial oxygen pressure estimated in F-44E-loaded Ba2+-alginate beads showed that the oxygen supply inside the beads was very poor when they were implanted below the kidney capsule or into the peritoneal cavity. These low pO2-values obtained for the renal subcapsular site and the peritoneum may explain the failure of previous immunoisolated islet transplantation studies using these locations.

Alginates↗
PubMed1999

19F-MRI in vivo determination of the partial oxygen pressure in perfluorocarbon-loaded alginate capsules implanted into the peritoneal cavity and different tissues.

Semipermeable hydrogels formed with a biocompatible alginate solution and Ba(2+) ions protect encapsulated cells and tissues from a foreign immune system. For the viability and metabolic activity of the encapsulated materials, a sufficient oxygen supply inside the capsules is necessary. Quantitative (19)F-MRI was performed on perfluorocarbon-loaded alginate capsules implanted into the peritoneal cavity, the musculus quadriceps femoris, and beneath the kidney capsule of rats, in order to determine in vivo the partial oxygen pressure (pO(2)) inside the capsules at these implantation sites. The temporal behavior of the pO(2) values was observed for at least 3 months. The most stable values over time were observed in the kidney, where inter-rat pO(2) differences were considerable. In the muscle, the values were very high directly after implantation and decreased to nearly zero after 2 weeks. In the peritoneal cavity, values changed randomly over a wide range between different rats and over time. Magn Reson Med 42:1039-1047, 1999.

Alginates↗
PubMed1998

[Standardized tests of bone implant surfaces with an osteoblast cell culture system. I. Orthopedic standard materials].

The effect of standard orthopaedic materials on proliferation and differentiation of osteoblasts was examined using a standardised cell culture system. Osteoblasts hFOB 1.19 were cultured on stainless steel (SS), a chromium-cobalt-molybdenum alloy (CrCoMb) and commercially pure titanium (cpTi) for 12 days. Cell culture polystyrene (PS) was used as a reference. Cell numbers and cell viability were used as parameters of proliferation. Cell differentiation was assessed using alkaline phosphatase activity, collagen I and osteocalcin production. The parameters of proliferation showed earlier maximum values on PS and cpTi, while proliferation was delayed on SS and CrCoMb. The highest values of differentiation were found on cpTi. The development of alkaline phosphatase activity showed two peaks reflecting apoptosis and redifferentiation. The cell culture system hFOB 1.19 is thus suitable for revealing differences in proliferation and differentiation of osteoblasts on standard orthopaedic materials. The results correlate with previous in vivo findings. Using this system, the dynamic effect of the material surface on the differentiation process of osteoblasts can be demonstrated.

Cell Differentiation↗
PubMed1997

Quantitative magnetic resonance imaging of capillary water permeability and regional blood volume with an intravascular MR contrast agent.

A novel method is presented to simultaneously measure the permeability surface area product of water (PS), also known as capillary diffusion capacity, and the regional blood volume (RBV). It is based on magnetic resonance imaging of the longitudinal relaxation times of tissue and blood at different concentrations of an intravascular MR contrast agent. PS and RBV were measured in vivo in different regions of the brain and the skeletal muscle of the rat. The average PS values (n = 5) obtained in cerebral cortex, corpus callosum, hippocampus, thalamus, jaw muscle, and tongue muscle were 3.31 +/- 0.20, 1.81 +/- 0.25, 3.37 +/- 0.36, 3.68 +/- 0.44, 10.6 +/- 1.1, and 14.1 +/- 2.51 ml x min(-1) x g(-1), respectively. The corresponding average RBV values were 1.63 +/- 0.18, 1.22 +/- 0.25, 3.30 +/- 0.37, 3.03 +/- 0.36, 1.66 +/- 0.30, and 1.38 +/- 0.33 ml x 100 g(-1). These results are in good agreement with previously reported literature values obtained by means of autoradiography.

Animals↗
PubMed1997

Perfluoro-15-crown-5-ether labelled macrophages in adoptive transfer experimental allergic encephalomyelitis.

In this serial in vivo study, macrophages labelled with perfluoro-15-crown-5-ether (15C5) were monitored in rats after inducing adoptive transfer experimental allergic encephalomyelitis (AT-EAE). AT-EAE is an animal model of multiple sclerosis and is characterized by inflammatory infiltrates in the central nervous system (CNS) and breakdown of the blood-brain-barrier. A particular feature of AT-EAE are macrophage infiltrates. Purpose of this study was to monitor the invasive and evasive phase of the macrophages in AT-EAE by using 3-dimensional 19F magnetic resonance imaging (3D 19F-MRI). In the early stage of the disease, a much stronger 19F-signal intensity was observed in AT-EAE-rats than in healthy control rats in the tissue adjacent to CNS regions severely affected by inflammatory infiltrates, and thereafter the 19F-signal intensity was decreasing over the time. However, no 19F-signal could be observed in the CNS itself neither in AT-EAE-rats nor in control rats. According to these findings it is assumed that we monitored the evasion of the macrophages from the region of inflammation.

Animals↗
PubMed1995

Compensation of diffusion effects in T2 measurements.

Measurements of the transverse relaxation time T2 are usually conducted with the Carr Purcell Meiboom Gill (CPMG) pulse sequence, which causes T2-weighted magnetization. Diffusion effects are a common source of error in measurements of this kind, because the incoherent motion of spins in external magnetic field gradients distorts T2 weighting of the transverse magnetization. As a result, inaccurate T2-values are obtained. In this work, we present a method which completely compensates for the effect of diffusion.

Diffusion↗
PubMed1995

Calculation of signal intensities in hybrid sequences for fast NMR imaging.

A number of techniques that recently have been used for fast NMR-imaging are based on a hybrid sequence of echo planar imaging (EPI) and FLASH imaging: after each NMR excitation several k-space lines are measured. The complete k-space is covered by performance of several excitations. It has been observed that there is usually an optimal hybrid sequence that maximizes the signal-to-noise ratio. In this work, a method is presented that allows a determination of the optimal sequence as a function of the relaxation times T1 and T2*.

Echo-Planar Imaging↗
PubMed1995

In vivo measurement of partial oxygen pressure in large vessels and in the reticuloendothelial system using fast 19F-MRI.

Quantitative in vivo 19F-MRI was performed in a rat model to monitor partial oxygen pressure (pO2) using a perfluorocarbon (PFC) emulsion as contrast agent. On Days 1, 4, and 8 postinjection of the PFC emulsion, transaxial T1 and pO2 maps were acquired of the abdomen of rats that were consecutively ventilated with pure oxygen, air, and a mixture of 10% oxygen and 90% nitrogen. The images had a resolution of 0.75 mm x 0.75 mm x 2 mm and a total acquisition time of 24 min. In these images it was possible to distinguish between different vessels and hepatic and splenic tissue in the selected imaging plane. Serial 19F-MRI measurements on the different days postinjection of the PFC allowed to determine separately the pO2 of arterial and venous blood and the intracellular pO2 in macrophages of the liver and spleen.

Animals↗
PubMed1995

Fast T2-mapping with snapshot flash imaging.

It has been shown recently that quantitative T2-maps may be measured by acquisition of a series of at least eight T2-weighted SNAPSHOT FLASH images. These measurements require a relaxation delay of 10-15 s after each T2-weighted image for the complete relaxation of the spin system. This results in long measuring times. The method presented in this paper allows a considerable reduction of the measuring time by combining the T2-sequence with a fast T1-measurement. Quantitative T1- and T2-maps may be acquired simultaneously in less than 30 s. The method was tested in a phantom experiment. In an in vivo application, relaxation times of different tissues in the abdomen of a rat were measured.

Abdomen↗