Auf der Suche nach dem optimalen Material : Möglichkeiten und Grenzen der Knochenheilung

Operative Maßnahmen zur Behandlung von knöchernen Defekten müssen bei der Versorgung von Unfallverletzten, aber auch nach der Entfernung von Knochentumoren und -zysten tagtäglich in der Unfallchirurgie durchgeführt werden. Überschreitet das Ausmaß der Defekte eine kritische Größe, kann der Körper diesen Substanzverlust nicht mehr aus eigener Kraft regenerieren, was die statischen und dynamischen Skelettfunktionen beträchtlich einschränkt. Dies ist für die betroffenen Patienten mit weitreichenden Einbußen an Lebensqualität verknüpft. Daher ist es klinisch notwendig, knöcherne Substanzverluste ab einer bestimmten Größe mit einem Material zu ersetzen, das die körpereigenen Regenerationsmechanismen unterstützt, damit letztendlich die ursprüngliche Skelettarchitektur wiederhergestellt wird. Die Überprüfung der Gewebeverträglichkeit von neu entwickelten Knochenersatzmaterialien und die Charakterisierung der Mechanismen, die für ihren Abbau im Körper verantwortlich sind, bilden einen Forschungsschwerpunkt in der Experimentellen Unfallchirurgie der Universität Gießen

Auf der Suche nach dem optimalen Material : Möglichkeiten und Grenzen der Knochenheilung

Operative Maßnahmen zur Behandlung von knöchernen Defekten müssen bei der Versorgung von Unfallverletzten, aber auch nach der Entfernung von Knochentumoren und -zysten tagtäglich in der Unfallchirurgie durchgeführt werden. Überschreitet das Ausmaß der Defekte eine kritische Größe, kann der Körper diesen Substanzverlust nicht mehr aus eigener Kraft regenerieren, was die statischen und dynamischen Skelettfunktionen beträchtlich einschränkt. Dies ist für die betroffenen Patienten mit weitreichenden Einbußen an Lebensqualität verknüpft. Daher ist es klinisch notwendig, knöcherne Substanzverluste ab einer bestimmten Größe mit einem Material zu ersetzen, das die körpereigenen Regenerationsmechanismen unterstützt, damit letztendlich die ursprüngliche Skelettarchitektur wiederhergestellt wird. Die Überprüfung der Gewebeverträglichkeit von neu entwickelten Knochenersatzmaterialien und die Charakterisierung der Mechanismen, die für ihren Abbau im Körper verantwortlich sind, bilden einen Forschungsschwerpunkt in der Experimentellen Unfallchirurgie der Universität Gießen

PMMA vertebroplasty in patients with malignant vertebral destruction of the thoracic and lumbar spine

Object: Patients with osteolytic metastases frequently suffer from serious local and radicular pain. Pathophysiologically, local pain arises from skeletal instability, whereas radicular pain originates from compression of nerve roots by local tumor growth. Causal treatment of osteolytic metastases in disseminated malignant disease is very difficult. Resection of vertebrae, in combination with ventro-dorsal stabilization, is a complex treatment for patients with a limited life expectancy. Percutaneous polymethylmethacrylate (PMMA) vertebroplasty is a new and easy method of relieving patients' pain. In addition, it is both cost effective and safe. Pain is reduced immediately after treatment. Due to the regained vertebral stability, early mobilization of the patients is possible

A Case Report

The objective of this case report is to introduce a customized CAD/CAM freeze- dried bone allograft (FDBA) block for its use in Guided Bone Regeneration (GBR) procedures for severely deficient maxillary bones. Additionally, a special newly developed remote incision technique is presented to avoid wound dehiscence. The results show optimal integration behavior of the FDBA block after six months and the formation of new vital bone. Thus, the results of the present case report confirm the use of the customized CAD/CAM bone block for augmentation of complex defects in the maxillary aesthetic zone as a successful treatment concept. View Full-Tex

Effects of dendritic core-shell glycoarchitectures on primary mesenchymal stem cells and osteoblasts obtained from different human donors

The biological impact of novel nano-scaled drug delivery vehicles in highly topical therapies of bone diseases have to be investigated in vitro before starting in vivo trials. Highly desired features for these materials are a good cellular uptake, large transport capacity for drugs and a good bio-compatibility. Essentially the latter has to be addressed as first point on the agenda. We present a study on the biological interaction of maltose-modified poly(ethyleneimine) (PEI-Mal) on primary human mesenchymal stem cell, harvested from reaming debris (rdMSC) and osteoblasts obtained from four different male donors. PEI-Mal-nanoparticles with two different molecular weights of the PEI core (5000 g/mol for PEI-5k-Mal-B and 25,000 g/mol for PEI-25k-Mal-B) have been administered to both cell lines. As well dose as incubation-time dependent effects and interactions have been researched for concentrations between 1 μg/ml to 1 mg/ml and periods of 24 h up to 28 days. Studies conducted by different methods of microscopy as light microscopy, fluorescence microscopy, transmission-electron-microscopy and quantitative assays (LDH and DC-protein) indicate as well a good cellular uptake of the nanoparticles as a particle- and concentration-dependent impact on the cellular macro- and micro-structure of the rdMSC samples. In all experiments PEI-5k-Mal-B exhibits a superior biocompatibility compared to PEI-25k-Mal-B. At higher concentrations PEI-25k-Mal-B is toxic and induces a directly observable mitochondrial damage. The alkaline phosphatase assay (ALP), has been conducted to check on the possible influence of nanoparticles on the differentiation capabilities of rdMSC to osteoblasts. In addition the production of mineralized matrix has been shown by von-Kossa stained samples. No influence of the nanoparticles on the ALP per cell has been detected. Additionally, for all experiments, results are strongly influenced by a large donor-to-donor variability of the four different rdMSC samples. To summarize, while featuring a good cellular uptake, PEI-5k-Mal-B induces only minimal adverse effects and features clearly superior biocompatibility compared to the larger PEI-25k-Mal-B

Effects of dendritic core-shell glycoarchitectures on primary mesenchymal stem cells and osteoblasts obtained from different human donors

The biological impact of novel nano-scaled drug delivery vehicles in highly topical therapies of bone diseases have to be investigated in vitro before starting in vivo trials. Highly desired features for these materials are a good cellular uptake, large transport capacity for drugs and a good bio-compatibility. Essentially the latter has to be addressed as first point on the agenda. We present a study on the biological interaction of maltose-modified poly(ethyleneimine) (PEI-Mal) on primary human mesenchymal stem cell, harvested from reaming debris (rdMSC) and osteoblasts obtained from four different male donors. PEI-Mal-nanoparticles with two different molecular weights of the PEI core (5000 g/mol for PEI-5k-Mal-B and 25,000 g/mol for PEI-25k-Mal-B) have been administered to both cell lines. As well dose as incubation-time dependent effects and interactions have been researched for concentrations between 1 μg/ml to 1 mg/ml and periods of 24 h up to 28 days. Studies conducted by different methods of microscopy as light microscopy, fluorescence microscopy, transmission-electron-microscopy and quantitative assays (LDH and DC-protein) indicate as well a good cellular uptake of the nanoparticles as a particle- and concentration-dependent impact on the cellular macro- and micro-structure of the rdMSC samples. In all experiments PEI-5k-Mal-B exhibits a superior biocompatibility compared to PEI-25k-Mal-B. At higher concentrations PEI-25k-Mal-B is toxic and induces a directly observable mitochondrial damage. The alkaline phosphatase assay (ALP), has been conducted to check on the possible influence of nanoparticles on the differentiation capabilities of rdMSC to osteoblasts. In addition the production of mineralized matrix has been shown by von-Kossa stained samples. No influence of the nanoparticles on the ALP per cell has been detected. Additionally, for all experiments, results are strongly influenced by a large donor-to-donor variability of the four different rdMSC samples. To summarize, while featuring a good cellular uptake, PEI-5k-Mal-B induces only minimal adverse effects and features clearly superior biocompatibility compared to the larger PEI-25k-Mal-B

Podoplanin immunopositive lymphatic vessels at the implant interface in a rat model of osteoporotic fractures

Insertion of bone substitution materials accelerates healing of osteoporotic fractures. Biodegradable materials are preferred for application in osteoporotic patients to avoid a second surgery for implant replacement. Degraded implant fragments are often absorbed by macrophages that are removed from the fracture side via passage through veins or lymphatic vessels. We investigated if lymphatic vessels occur in osteoporotic bone defects and whether they are regulated by the use of different materials. To address this issue osteoporosis was induced in rats using the classical method of bilateral ovariectomy and additional calcium and vitamin deficient diet. In addition, wedge-shaped defects of 3, 4, or 5 mm were generated in the distal metaphyseal area of femur via osteotomy. The 4 mm defects were subsequently used for implantation studies where bone substitution materials of calcium phosphate cement, composites of collagen and silica, and iron foams with interconnecting pores were inserted. Different materials were partly additionally functionalized by strontium or bisphosphonate whose positive effects in osteoporosis treatment are well known. The lymphatic vessels were identified by immunohistochemistry using an antibody against podoplanin. Podoplanin immunopositive lymphatic vessels were detected in the granulation tissue filling the fracture gap, surrounding the implant and growing into the iron foam through its interconnected pores. Significant more lymphatic capillaries were counted at the implant interface of composite, strontium and bisphosphonate functionalized iron foam. A significant increase was also observed in the number of lymphatics situated in the pores of strontium coated iron foam. In conclusion, our results indicate the occurrence of lymphatic vessels in osteoporotic bone. Our results show that lymphatic vessels are localized at the implant interface and in the fracture gap where they might be involved in the removal of lymphocytes, macrophages, debris and the implants degradation products. Therefore the lymphatic vessels are involved in implant integration and fracture healing

Small changes in bone structure of female a7 nicotinic acetylcholine receptor knockout mice

BACKGROUND: Recently, analysis of bone from knockout mice identified muscarinic acetylcholine receptor subtype M3 (mAChR M3) and nicotinic acetylcholine receptor (nAChR) subunit a2 as positive regulator of bone mass accrual whereas of male mice deficient for a7-nAChR (a7KO) did not reveal impact in regulation of bone remodeling. Since female sex hormones are involved in fair coordination of osteoblast bone formation and osteoclast bone degradation we assigned the current study to analyze bone strength, composition and microarchitecture of female a7KO compared to their corresponding wild-type mice (a7WT). METHODS: Vertebrae and long bones of female 16-week-old a7KO (n = 10) and a7WT (n = 8) were extracted and analyzed by means of histological, radiological, biomechanical, cell- and molecular methods as well as time of flight secondary ion mass spectrometry (ToF-SIMS) and transmission electron microscopy (TEM). RESULTS: Bone of female a7KO revealed a significant increase in bending stiffness (p<0.05) and cortical thickness (p<0.05) compared to a7WT, whereas gene expression of osteoclast marker cathepsin K was declined. ToF-SIMS analysis detected a decrease in trabecular calcium content and an increase in C4H6N+ (p<0.05) and C4H8N+ (p<0.001) collagen fragments whereas a loss of osteoid was found by means of TEM. CONCLUSIONS: Our results on female a7KO bone identified differences in bone strength and composition. In addition, we could demonstrate that a7-nAChRs are involved in regulation of bone remodelling. In contrast to mAChR M3 and nAChR subunit a2 the a7-nAChR favours reduction of bone strength thereby showing similar effects as a7ß2-nAChR in male mice. nAChR are able to form heteropentameric receptors containing a- and ß-subunits as well as the subunits a7 can be arranged as homopentameric cation channel. The different effects of homopentameric and heteropentameric a7-nAChR on bone need to be analysed in future studies as well as gender effects of cholinergic receptors on bone homeostasis

Improved in vitro test procedure for full assessment of the cytocompatibility of degradable magnesium based on ISO 10993-5/-12

Magnesium (Mg)-based biomaterials are promising candidates for bone and tissue regeneration. Alloying and surface modifications provide effective strategies for optimizing and tailoring their degradation kinetics. Nevertheless, biocompatibility analyses of Mg-based materials are challenging due to its special degradation mechanism with continuous hydrogen release. In this context, the hydrogen release and the related (micro-) milieu conditions pretend to strictly follow in vitro standards based on ISO 10993-5/-12. Thus, special adaptions for the testing of Mg materials are necessary, which have been described in a previous study from our group. Based on these adaptions, further developments of a test procedure allowing rapid and effective in vitro cytocompatibility analyses of Mg-based materials based on ISO 10993-5/-12 are necessary. The following study introduces a new two-step test scheme for rapid and effective testing of Mg. Specimens with different surface characteristics were produced by means of plasma electrolytic oxidation (PEO) using silicate-based and phosphate-based electrolytes. The test samples were evaluated for corrosion behavior, cytocompatibility and their mechanical and osteogenic properties. Thereby, two PEO ceramics could be identified for further in vivo evaluations