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Biomedical subjects

Sabine Wenisch

Publications and source records attributed to Sabine Wenisch.

9 recordsLinked to original sources

The effects of combined gentamicin-hydroxyapatite coating for cementless joint prostheses on the reduction of infection rates in a rabbit infection prophylaxis model.

Infections remain a critical issue in total joint arthroplasty. Addition of antibiotics to bone cement was shown to significantly improve antimicrobial prophylaxis in cemented joint arthroplasty. In cementless joint arthroplasty a comparable prophylaxis by local antibiotics has not been possible yet. The aim of the current study was to investigate the antimicrobial effect of two different gentamicin-hydroxyapatite (HA) coatings for cementless prostheses in a rabbit infection model. Staphylococcus aureus with a dose of 10(7) CFUs was inoculated into the intramedullary canal of the tibia of 30 rabbits followed by the implantation of standard steel HA K-wires (n=10), steel K-wires coated with a gentamicin-HA combination (n=10), and steel K-wires coated with a gentamicin-RGD-HA combination (n=10), respectively. The animals were sacrificed after 28 days and clinical, histological and microbiological assessment on the bone and on the removed K-wire itself by agar plating and DNA-pulsed field gel electrophoresis were carried out to detect infection. There was a statistically significant reduction of infection rates by both gentamicin-coating types (0 infections in both groups) compared to standard HA coating (7 infections in 8 animals; 2 animals were lost due to acute diarrhea) (p<0.001). An excellent correlation between agar plating testing results of the K-wires and of the bone samples was found. Detailed histology showed cortical lysis, abscess and sequester formation in the infected animals. Both gentamicin-coating types showed significant improvement of infection prophylaxis compared to standard HA coating and, therefore, this coating technology could help to improve infection prophylaxis in cementless total joint arthroplasty. In further studies biocompatibility of the coatings has to be assessed.

Animals↗

Immunochemical, ultrastructural and electrophysiological investigations of bone-derived stem cells in the course of neuronal differentiation.

Numerous reports have highlighted the use of mesenchymal stem cells (MSC) for tissue engineering because of the capacity of the cells to differentiate along the osteogenic, chondrogenic or adipogenic pathway. As MSC also display neuronal morphologies under appropriate culture conditions, the differentiation capacity of stem cells seems to be more complex than initially thought, but it requires careful characterization of the cells. This is especially the case because recently it has been suggested that neuronal differentiation of stem cells is only an artifact. Here, we investigate the sequence of ultrastructural changes of bone-derived stem cells during neuronal induction and compare these data with immunocytochemical and electrophysiological properties of the cells. For further comparative analyses, stem cells were incubated with non-neurologically inducing stressors. The stem cells were harvested from human osseous debris and were characterized morphologically, immunocytochemically and by using FACS. After 6 h of neuronal induction, the cells had assumed neuronal morphologies and expressed neuron-specific enolase, beta-III-tubulin, neurofilament-H and HNK-1, while only a subpopulation expressed CD15 and synaptophysin. However, electrical signaling could not be detected, neither spontaneously nor after electrical stimulation. Nevertheless, transmission electron microscopy revealed cellular features of neuritogenesis and synaptogenesis in the course of neuronal induction and suggested that the cells have features similar to those observed in immature neurons. Based upon the results, it can be concluded that neuronal induction had initiated the early steps of neuronal differentiation, while exposure of the cells to non-neurological stressors had caused necrotic alterations.

Bone and Bones↗

Glycerol-l-lactide coating polymer leads to delay in bone ingrowth in hydroxyapatite implants.

Glycerol-l-lactide as coating polymer for the delivery of basic fibroblast growth factor (bFGF) from hydroxyapatite (HA) ceramic implants was shown to lead to significant delay in bone ingrowth into the implants compared to implants without the coating polymer. The purpose of this work was to study bone ingrowth in HA ceramic implants with and without the coating polymer but without growth factors to enable differentiation between a locking effect of the pores by the polymer and the fact of inactivation of the growth factors by the polymer, which could both be possible for the delay. A defect was created in the subchondral region of both femurs in 24 miniature-pigs and was either filled by the HA implants with or without the coating polymer. Histomorphometry showed a significant delay in bone ingrowth in the polymer coated implants both after 6 and 12 weeks. Detailed histology revealed that the HA pores were completely "locked" by the polymer leading to complete loss of the osteoconductive properties of the HA. Also electron microscopy showed filling of the HA pores by the polymer. Therefore, it can be concluded that glycerol-l-lactide should not be used to coat HA ceramic implants due to significant delay in bone ingrowth.

Animals↗

The tissue response to an alkylene bis(dilactoyl)-methacrylate bone adhesive.

Gluing is an attractive technique to fix small bone fragments. However, to date no bone adhesive could be established successfully for all day clinical use. The purpose of this experimental study was to investigate the biocompatibility of a new bone glue based on alkylene bis(dilactoyl)-methacrylate in 36 rabbits. Monocondylar osteotomy of the distal femur was performed and bone glue was applied into the osteotomy gap in 24 rabbits. The remaining 12 animals served as controls. In all rabbits the osteotomy was subsequently stabilized by K-wire osteosynthesis. Six animals of the glue group and 3 controls were euthanized after 7, 21, 42, and 84 days, respectively. Fracture healing and degradation pattern of the glue was studied using histological, histomorphometrical, scanning electron microscopical, and radiological methods. Good resorption of the glue by mononuclear and multinucleated giant cells without prolonged inflammatory processes was observed in the glue group. Histomorphometrical analysis did not reveal any significant differences in fracture healing between the glue and control group at any time. Complete remodelling of the former osteotomy gap was found in all rabbits after 84 days. This bioresorbable bone adhesive exhibited good biocompatibility and its degradation did not interfere with physiological fracture healing.

Animals↗

New blood vessel formation and expression of VEGF receptors after implantation of platelet growth factor-enriched biodegradable nanocrystalline hydroxyapatite.

Vascular endothelial growth factor (VEGF) plays a key role for the interaction of osteoblasts and endothelial cells and, therefore, is an important factor for the osteointegration of bone substitutes. The aim of the current work was to study the effects of platelet growth factors (PLF) on new blood vessel formation and VEGF-receptors expression pattern in bone defects filled with nanocrystalline hydroxyapatite (HA) paste in miniature-pigs. Conventional histology, RT-PCR for VEGF and receptors mRNA, Western blot analysis, immunohistochemical staining and quantitative assessment of newly formed vessels was performed. HA enriched with platelet growth factor (HA/PLF+) led to an up-regulation of VEGF-R1 synthesis, a slightly enhanced number of newly formed vessels with higher sprouting activity compared with HA without PLF (HA/PLF-) filling defects. These observation are most likely attributable to a stimulating effect of TGF-ss from the platelet factor on VEGF expression in osteoblasts.

Animals↗

Human reaming debris: a source of multipotent stem cells.

The biological characteristics of human reaming debris (HRD) generated in the course of surgical treatment of long bone diaphyseal fractures and nonunions are still a matter of dispute. Therefore, the objective of the present investigation has been to characterize the intrinsic properties of human reaming debris in vitro. Samples of reaming debris harvested from 12 patients with closed diaphyseal fractures were examined ultrastucturally and were cultured under standard conditions. After a lag phase of 4-7 days, cells started to grow out from small bone fragments and established a confluent monolayer within 20-22 days. The cells were characterized according to morphology, proliferation capacity, cell surface antigen profile, and differentiation repertoire. The results reveal that human reaming debris is a source of multipotent stem cells which are able to grow and proliferate in vitro. The cells differentiate along the osteogenic pathway after induction and can be directed toward a neuronal phenotype, as has been shown morphologically and by the expression of neuronal markers after DMSO induction. These findings have prompted interest in the use of reaming debris-derived stem cells in cell and bone replacement therapies.

Adult↗

Effect of glycerol-L-lactide coating polymer on bone ingrowth of bFGF-coated hydroxyapatite implants.

Basic fibroblast growth factor (bFGF)-coated hydroxyapatite (HA) cylinders showed good bony incorporation in a previously conducted animal study. However, some cylinders exhibited focal inhomogeneous bone ingrowth. The purpose of the current study was to test whether glycerol-L-lactide polymer coating could improve release properties and bone incorporation of bFGF-coated HA implants. bFGF-coated HA cylinders with or without coating polymer were investigated for in vitro release of bFGF by an immuno-ligand-assay and also for bone ingrowth in miniature pigs after 42 and 84 days. Release from bFGF polymer composites was lower for the first 3 days compared to the other group but was more homogenous and detectable amounts were still found after 20 days. There was significant delay in bone ingrowth of the polymer implants in which even after 84 days bone ingrowth was not completed, whereas in the other group incorporation after 42 days occurred. Detailed histology revealed filling of the HA pores with the polymer, making ingrowth of the surrounding host bone impossible. Only after 84 days starting resorption of the polymer accompanied by bone ingrowth was found. The current study showed that glycerol-L-lactide is not suitable for coating of HA implants due to polymer induced "locking" of HA pores.

Animals↗

Bone ingrowth in bFGF-coated hydroxyapatite ceramic implants.

This experimental study was performed to evaluate angiogenesis, bone formation, and bone ingrowth in response to osteoinductive implants of bovine-derived hydroxyapatite (HA) ceramics either uncoated or coated with basic fibroblast growth factor (bFGF) in miniature pigs. A cylindrical bone defect was created in both femur condyles of 24 miniature pigs using a saline coated trephine. Sixteen of the 48 defects were filled with HA cylinders coated with 50 microg rhbFG, uncoated HA cylinders, and with autogenous transplants, respectively. Fluorochrome labelled histological analysis, histomorphometry, and scanning electron microscopy were performed to study angiogenesis, bone formation and bone ingrowth. Complete bone ingrowth into bFGF-coated HA implants and autografts was seen after 34 days compared to 80 days in the uncoated HA group. Active ring-shaped areas of fluorochrome labelled bone deposition with dynamic bone remodelling were found in all cylinders. New vessels could be found in all cylinders. Histomorphometric analysis showed no difference in bone ingrowth over time between autogenous transplants and bFGF-coated HA implants. The current experimental study revealed comparable results of bFGF-coated HA implants and autogenous grafts regarding angiogenesis, bone synthesis and bone ingrowth.

Animals↗

[Brain emboli in the lungs of cattle].

There is no information whether the BSE agent is introduced into the human food chain through contamination of the lungs of cattle with central nervous system tissue (CNS). Studies in the United Kingdom and in the USA showed that CNS tissue could contaminate the lungs after using pneumatic powered air injection stunners (e.g. "The Knocker") or after pithing. Thus, pithing was forbidden in the European Union since January 2001. In German abattoirs conventional cartridge-fired stunners (e.g. model by Schermer) are usually applied. Pithing was used up to December 2000 in approx. 75% of the German abattoirs. In the present study 323 lungs of cattle were analysed for CNS. The lungs were derived from cattle exclusive stunned by use of the knocker from Schermer. 60% of the lungs contained emboli which were tested with immuno chemistry as well as immuno histochemistry to detect CNS. Two of 108 pooled samples showed a faint immuno reaction in the anti-NSE and anti-GFAP immunoblot. Further two particles showed a faint reaction for NSE and GFAP in immuno histochemistry, thus suggesting the presence of CNS. Even though CNS tissue could not be shown in the histological investigation, we used our findings to estimate the worst case scenario for human BSE exposure risk (HER) by lung contaminated by CNS emboli. The content of CNS in the samples was estimated to be about 0.11% when the respective immuno reactions were calibrated against standards containing known brain concentrations. Under the assumption that only one lung in the pooled samples was contaminated with BSE-infected central nervous tissue, the HER was calculated to reach a maximum of 2.2 x 10(-5) CoID50/consumer after consumption of a sausage with a portion of 10% lung. The results of our study suggest that the contamination of the lung with CNS after using a conventional cartridge-fired stunner cannot be excluded, however, the incidence appears to be very low. In addition, presumed CNS emboli, if at all, are microscopically small. Furthermore the incidence of BSE in Germany is very low and lungs of cattle are usually not consumed. Thus we can judge the potential for human oral exposure after consumption of lungs of cattle which were stunned in Germany to be extremely low. A final assessment, however, is impossible as there is no knowledge about the minimum infectious dose for humans.

Abattoirs↗