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BACKGROUND CONTEXT: Lumbar interbody fusion is an effective treatment for unstable spinal segments. However, the time needed to establish a solid bony interbody fusion between the two vertebrae may be longer than twelve months after surgery. During this time window, the instrumented spinal segment is assumed to be at increased risk for instability related complications such as cage migration or subsidence. It is hypothesized that the design of new interbody cages that enable direct osseointegration of the cage at the vertebral endplates, without requiring full bony fusion between the two vertebral endplates, might shorten the time window that the instrumented spinal segment is susceptible to failure.PURPOSE: To quantify the bone ingrowth and resulting segmental stability during consolidation of lumbar interbody fusion using two different cage types.STUDY DESIGN: Preclinical ovine model.METHODS: Seven skeletally mature sheep underwent bi-segmental lumbar interbody fusion surgery with one conventional polyether ether ketone (PEEK) cage, and one newly developed trussed titanium (TT) cage. After a postoperative time period of 13 weeks, non-destructive range of motion testing and histological analysis were performed. Additionally, sample specific finite element (FE) analysis was performed to predict the stability of the interbody fusion region alone.RESULTS: Physiological movement of complete spinal motion segments did not reveal significant differences between the segments operated with PEEK and TT cages. The onset of creeping substitution within the cage seemed to be sooner for PEEK cages, which led to significantly higher bone volume over total volume (BV/TV) compared to the TT cages. TT cages showed significantly more direct bone to implant contact (BIC). Although the mean stability of the interbody fusion region alone was not statistically different between the PEEK and TT cages, the variation within the cage types illustrated an all-or-nothing response for the PEEK cages while a more gradual increase in stability was found for the TT cages.CONCLUSIONS: Spinal segments operated with conventional PEEK cages were not different from those operated with newly developed TT cages in terms of segmental stability but did show a different mechanism of bone ingrowth and attachment. Based on the differences in development of bony fusion, we hypothesize that TT cages might facilitate increased early segmental stability by direct osseointegration of the cage at the vertebral endplates without requiring complete bony bridging through the cage.CLINICAL SIGNIFICANCE: Interbody cage type affects the consolidation process of spinal interbody fusion. Whether different consolidation processes of spinal interbody fusion result in clinically significant differences requires further investigation.

Calcium phosphate (CaP) coatings can enhance the performance of bone implants in compromised conditions, such as osteoporosis. Therefore, this study compared non-coated vs. CaP-coated (n = 8) titanium implants in osteoporotic ovariectomized (OVX) rats. Bone volume (BV) was assessed using micro-computer tomography (micro-CT) and three-dimensional (3D) histology, in three zones from the implant surface. Bone remodeling was assessed using fluorochrome labels and osteoclast staining. Micro-CT and 3D histology showed a BV reduction in OVX animals, of respectively 22.4 and 10.5%. BV was significantly increased inside all zones around CaP coatings, especially in the inner zone of the OVX animals. Fluorochrome labels were predominantly seen when the coating was applied. Osteoclasts were mainly found in the area remote from the surface of non-coated implants in control animals. For the coated implants, osteoclasts were distributed evenly, and present in direct vicinity of the surface. In conclusion, 3D histology is a suitable technique to obtain data and insight into bone architecture around implants at relatively high resolution. Bone formation was significantly reduced in osteoporotic animals. CaP coatings resulted in a higher BV directly around implants installed in osteoporotic animals, enhanced turnover, and a shift of remodeling activity toward the implant surface. (c) 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 3463-3473, 2015.

Thermosensitive injectable hydrogels for bone regeneration consisting of chitosan, sodium beta-glycerophosphate (Na-β-GP) and alkaline phosphatase (ALP) were enriched with oxygenated perfluorodecalin (PFD), a liquid hydrophobic perfluorochemical with high oxygen affinity, in order to improve cell growth on the hydrogels. Furthermore, influence of PFD concentration on hydrogel physicochemical properties relevant for bone regeneration, namely gelation speed, radiopacity and homogenicity, was investigated. Addtionally, ALP-mediated and non-ALP-mediated mineralization were evaluated by incubation in 0.1 M calcium glycerophosphate and simulated body fluid. 2% (w/v) chitosan hydrogels containing 2.5 mg/ml ALP were enriched with PFD at five concentrations, namely 0 (control), 0.069, 0.138, 0.207 and 0.276 ml/ml hydrogel, denoted A, B, C, D and E, respectively. Rheometrical investigations revealed that gelation speed increased with increasing PFD concentration. Micro-CT analysis revealed homogenicity of all sample groups except E and that radiopacity increased in the order B>C>A>D>E. ALP-mediated and non-ALP-mediated mineralization were not affected adversely by PFD. Growth of human adipose tissue-derived mesenchymal stem cells (ADSC) encapsulated in hydrogels was markedly higher in sample groups containing PFD, i.e. B–E. Hence, incorporation of oxygenated PFD can improve the suitability of hydrogels as bone regeneration materials.

Item does not contain fulltext Hydrogels of biocompatible calcium-crosslinkable polysaccharide gellan gum (GG) were enriched with bioglass particles to enhance (i) mineralization with calcium phosphate (CaP); (ii) antibacterial properties and (iii) growth of bone-forming cells for future bone regeneration applications. Three bioglasses were compared, namely one calcium-rich and one calcium-poor preparation both produced by a sol-gel technique (hereafter referred to as A2 and S2, respectively) and one preparation of composition close to that of the commonly used 45S5 type (hereafter referred to as NBG). Incubation in SBF for 7 d, 14 d and 21 d caused apatite formation in bioglass-containing but not in bioglass-free samples, as confirmed by FTIR, XRD, SEM, ICP-OES, and measurements of dry mass, i.e. mass attributable to polymer and mineral and not water. Mechanical testing revealed an increase in compressive modulus in samples containing S2 and NBG but not A2. Antibacterial testing using biofilm-forming meticillin-resistant staphylococcus aureus (MRSA) showed markedly higher antibacterial activity of samples containing A2 and S2 than samples containing NBG and bioglass-free samples. Cell biological characterization using rat mesenchymal stem cells (rMSCs) revealed a stimulatory effect of NBG on rMSC differentiation. The addition of bioglass thus promotes GG mineralizability and, depending on bioglass type, antibacterial properties and rMSC differentiation.

Item does not contain fulltext Calcium phosphate cements (CPCs) use the simultaneous presence of several calcium phosphates phases. This is done to generate specific bulk and in vivo properties. This work has processed and evaluated novel multiphasic CPCs containing dual tricalcium phosphate (TCPs) phases. Dual TCPs containing alpha- and beta-TCP phases were obtained by thermal treatment. Standard CPC (S-CPC) was composed of alpha-TCP, anhydrous dicalcium phosphate and precipitated hydroxyapatite, while modified CPC (DT-CPC) included both alpha- and beta-TCP. Physicochemical characterization of these CPCs was based on scanning electron microscopy, X-ray diffraction, specific surface area (SSA) and particle size (PS) analysis and mechanical properties. This characterization allowed the selection of one DT-CPC for setting time, cohesion and biological assessment compared with S-CPC. Biological assessment was carried out using a tibial intramedullary cavity model and subcutaneous pouches in guinea pigs. Differences in the surface morphology and crystalline phases of the treated TCPs were detected, although PS analysis of the milled CPC powders produced similar results. SSA analysis was significantly higher for DT-CPC with alpha-TCP treated at 1100 degrees C for 5h. Poorer mechanical properties were found for DT-CPC with alpha-TCP treated at 1000 degrees C. Setting time and cohesion, as well as the in vivo performance, were similar in the selected DT-CPC and the S-CPC. Both CPCs created the desired host reactions in vivo. 01 september 2012

Item does not contain fulltext Quantification of the amount of newly formed bone is an essential part of bone regeneration studies. Histomorphometry, based on histological sections of plastic-embedded specimens, is the most frequently applied technique in this assessment. Before performing image analysis, a specific region of interest (ROI) has to be determined. Based on the histological procedure, different areas within the ROI can be discriminated and assigned to relevant tissue structures. However, in literature not much attention is paid to the effect of the histological procedures on the final outcome of the histomorphometrical measurements on bone regeneration. In this study, the histomorphometrical bone formation of the intramedullary cavity of the guinea pig tibia, filled with calcium phosphate cement, was quantified in plastic-embedded and paraffin-embedded specimens and in specimens analyzed with scanning electron microscopy in the backscattering mode (SEM-BS). The data showed that the histological procedure significantly affected the measured bone amount. Therefore, it is recommended that scaffold characteristics are carefully considered in selecting a proper technique for the analysis of bone formation in bone tissue engineering studies. The results of this study identified high-resolution SEM-BS and elastic van Gieson staining of decalcified histological sections as recommendable techniques for evaluating bone formation. 01 mei 2012

Contains fulltext : 79581.pdf (Publisher’s version ) (Open Access) The aim of this work was to introduce high-resolution computed tomography (micro-CT) for scaffolds made from soft natural biomaterials, and to compare these data with the conventional techniques scanning electron microscopy and light microscopy. Collagen-based scaffolds were used as examples. Unlike mineralized tissues, collagen scaffolds do not provide enough X-ray attenuation for micro-CT imaging. Therefore, various metal-based contrast agents were applied and evaluated using two structurally distinct scaffolds, one with round pores and one with unidirectional lamellae. The optimal contrast techniques for obtaining high-resolution three-dimensional images were either a combination of osmium tetroxide and uranyl acetate, or a combination of uranyl acetate and lead citrate. The data obtained by micro-CT analysis were in line with data obtained by light and electron microscopy. However, small structures (less than a few mum) could not be visualized due to limitation of the spot size of the micro-CT apparatus. In conclusion, reliable three-dimensional images of scaffolds prepared from soft natural biomaterials can be obtained using appropriate contrast protocols. This extends the use of micro-CT analysis to soft materials, such as protein-based biomaterials.

In this study, the biocompatibility of a calcium phosphate (CaP) cement incorporating poly (D,L-lactic-co-glycolic acid) (PLGA) microparticles was evaluated in a subcutaneous implantation model in rats. Short-term biocompatibility was assessed using pure CaP discs and CaP discs incorporating PLGA microparticles (20% w/w) with and without preincubation in water. Long-term biocompatibility was assessed using CaP discs incorporating varying amounts (5, 10, or 20% w/w) and diameter sizes (small, 0-50 mum; medium, 51-100 mum, or large, 101-200 mum) of PLGA microparticles. The short-term biocompatibility results showed a mild tissue response for all implant formulations, irrespective of disc preincubation, during the early implantation periods up to 12 days. Quantitative histological evaluation revealed that the different implant formulations induced the formation of similar fibrous tissue capsules and interfaces. The results concerning long-term biocompatibility showed that all implants were surrounded by a thin connective tissue capsule (

Item does not contain fulltext The importance of polyamines in prostatic growth and differentiation has prompted studies to evaluate the clinical relevance of the ornithine decarboxylase/polyamine system in prostatic cancer. These studies show that differences in biological behaviour of prostatic (cancer) cells are associated with changes in polyamine levels and/or the activity of their metabolic enzymes. Faulty antizyme regulation of polyamine homoeostasis may play an important role in the growth and progression of prostatic carcinoma. Treatment of human prostate carcinoma cells with inhibitors of polyamine metabolic enzymes or polyamine analogues induces cell growth arrest or (apoptotic) cell death. Our recent in vitro studies using conformationally restricted polyamine analogues show that these compounds inhibit cell growth, probably by inducing antizyme-mediated degradation of ornithine decarboxylase. Sensitivity of human prostate cancer cells for these compounds was increased in the absence of androgens. These results suggest that these analogues might have chemotherapeutic potential in case prostatic cancer has become androgen-independent. Pilot data in an in vivo model show that these analogues have effects on tumour cell proliferation, vascularity, blood perfusion and tissue hypoxia. Overall, these studies show that polyamines may serve as important biomarkers of prostatic malignancy and provide a promising target for chemotherapy of prostatic cancer.

Calcium phosphate cements (CPCs) including poly(D,L-lactic-co-glycolic) acid (PLGA) microparticles are promising candidates for bone regenerative applications. Previous studies with CPC/PLGA demonstrated that the material is non-toxic, biocompatible and osteoconductive. However, the outcome of these studies was based on healthy individuals and consequently does not provide information on bone substitute material performance in a compromised situation, such as osteoporosis. Therefore, this study comparatively evaluated the performance of injectable CPC/PLGA in healthy (SHAM) and osteoporotic rats (OVX) using a rat femoral condyle defect with implantation periods of 4 and 12 weeks. It was hypothesized that in OVX rats the degradation of CPC/PLGA would increase due to a higher osteoclastic activity present in osteoporotic animals and that the obtained space would be rapidly filled with newly formed bone. The results revealed an accelerated degradation of the used CPC/PLGA in osteoporotic animals, but bone formation was less compared to that in healthy animals at 4 and 12 weeks after implantation. In addition, after 4 weeks, the amount of newly formed bone under osteoporotic conditions was less in the femoral condyle defect compared to that present in a non-defect, osteoporotic control femoral condyle, but equal after 12 weeks. On the other hand, in healthy animals, the amount of newly formed bone in the femoral condyle defect was equal to that present in a non-defect control femoral condyle at 4 weeks, while higher after 12 weeks. This indicates that bone regeneration at a defect site under osteoporotic conditions is slower, but can reach native amounts after longer time periods. Consequently, bone regenerative treatments under osteoporotic conditions seem to require additional empowerment of bone substitute materials.

Contains fulltext : 51473.pdf (Publisher’s version ) (Closed access) High-resolution three-dimensional data about the bone response to oral implants can be obtained by using microfocus computer tomography. However, a disadvantage is that metallic implants cause streaking artifacts due to scattering of X-rays, which prevents an accurate evaluation of the interfacial bone-to-implant contact. It has been suggested that the use of thin titanium coatings deposited on polymeric implants can offer an alternative option for analyzing bone contact using micro-CT imaging. Consequently, the aim of the current study was to investigate bone behavior to titanium-coated polymethylmethacrylate (PMMA) implants by micro-CT and histological evaluation. For the experiment titanium-coated PMMA implants were used. The implants had a machined threaded appearance and were provided with a 400-500 nm thick titanium coating. The implants were inserted in the right or left tibia of 10 goats. After an implantation period of 12 weeks the implants were retrieved and prepared for micro-computer tomography (microCT), light microscopy, and X-ray microanalysis. The micro-CT showed that the screw-threads and typical implant configuration were well maintained through the installation procedure. Overall, histological responses showed that the titanium-coated implants were well tolerated and caused no atypical tissue response. In addition, the bone was seen in direct contact with the titanium-coated layer. The X-ray microanalysis results confirmed the light microscopical data. In conclusion, the obtained results proof the final use of titanium-coated PMMA implants for evaluation of the bone-implant response using microCT. However, this study also confirms that for a proper analysis of the bone-implant interface the additional use of microscopical techniques is still required.