Abstract Nickel-titanium shape memory metal Nitinol (NiTi) is a new kind of implant material, which provides a possibility to prepare functional implants activated at body temperature and withstands kinking better than conventional metals. Applications utilizing these unique properties are a target of active research interest. Host reactions to NiTi and to the forces created by functional implants should also be studied. A functional NiTi intramedullary nail, which causes a bending force on the bone, was developed for correcting bone deformities. In the present studies, the action of the device was inverted to induce a bone deformity instead of correcting one, in order to test the hypothesis that bone modelling can be controlled using such functional nail. Implanting the nail into the medullary cavity of rat femur for twelve weeks caused bowing of the bone, retardation of its longitudinal growth, and thickening of the bone and the cortex. In another study the effects of functional and straight nails were compared. Bowing of the bone and significant overall thickening of the bone and the cortex were associated only with the functional nail, while the straight nail induced only minor thickening of the bone. Retardation of longitudinal growth was seen in both groups, and this may have been caused by perforating the distal epiphyseal plate during the nailing. Finite element model of the bone-nail combination was also created. Porous NiTi was studied as a bone graft substitute by filling a bone defect in the distal femoral metaphysis of a rat bone with porous NiTi implants of different porosities. After 30 weeks, porosity of 66.1% (mean pore size (MPS) 259μm) showed the best bone-implant contact (51%). However, porosity of 46.6% (MPS 505μm) with 39% bone-implant contact was not significantly inferior in this respect and showed a significantly lower incidence of fibrosis within the implant and thus seemed to be the best choice for a bone graft substitute, out of the porosities tested here. The porosity of 59.2% (MPS 272μm) showed lower contact values. NiTi tendon suture material was studied by implanting NiTi sutures into rabbit tendon and subcutaneous tissues for two, six, and twelve weeks. NiTi proved to be stronger than polyester, which served as control material. The encapsulating membrane was minimal with both materials, suggesting good biocompatibility in tendon tissue. The implantation did not affect the strength properties of either material. On the basis of the present studies, NiTi provides a possibility to develop new kinds of implants for correcting bone deformities, for filling bone defects in weight-bearing locations and a good candidate for a tendon suture material.

Abstract Reindeer antler was studied for its possible use as a bone implant material. A molecular biological study showed that antler contains a growth factor promoting bone formation. Ectopic bone formation assay showed that antler is not an equally effective inducer as allogenic material. Ectopic bone formation assay was optimised for biocompatibility studies of orthopaedic NiTi implants. Ti-6Al-4V and stainless steel were used as reference materials. The assay showed differences in bone mineral densities, with superior qualities in NiTi. The rate of endochondral ossification varied between the implants, NiTi ossicles had larger cartilage and bone areas than ossicles of the two other materials. The cytocompatibility of NiTi was studied with three different methods. Cell viability, cell adhesion and TGF-β1 concentration were assessed in ROS-17/2.8 cell cultures. Cells grown on NiTi had better viability than cells grown on pure nickel or stainless steel. Cell attachment on the materials was studied with paxillin staining of focal contacts. The number of focal contacts was clearly higher in cells grown on NiTi than in cells grown on pure titanium, pure nickel or stainless steel. TGF-β1 concentration was measured with ELISA. The results showed that there was only some minor variation between NiTi, pure titanium and stainless steel. Nickel showed a lower TGF-β1 concentration. Taken together, these results suggest that NiTi is well tolerated by ROS-17/2.8 cells. The cytocompatibility of stainless steel is not so good as that of NiTi. The same tests were used to study the effects of the surface roughness of the implant on cytocompatibility. Three different surface roughness grades were compared in cell cultures on NiTi and titanium alloy discs. Titanium alloy was subjected to two different heat treatments, to compare the effects of the treatments on cytocompatibility. The studies showed that NiTi had a lesser impact on cell viability and attachment than titanium alloy. Further, rough NiTi was found to be a better tolerated surface than the others. In this study, heat treatment of titanium alloy at +850° C did not interfere with cell viability or attachment, as did the +1050° C treatment of the alloy. On the contrary, TGF-β1 concentrations decreased on the +850° C treated alloy and were approximately same on the +1050° C treated alloy and on NiTi.

Abstract The improvement of implant biocompatibility is constantly under investigation. Titanium is a standard biomaterial that performs well in dental and orthopedic implantations. However, detrimental adverse effects resulting from e.g. biomaterial properties, inflammatory responses and surgical procedures occasionally occur. Coating the biomaterials aims at increasing the proportion of successful operations. Pectins, large plant cell wall polysaccharides, are innovative, modifiable, and potentially anti-inflammatory candidates for biomaterial nanocoatings. In this thesis, covalently-grafted pectin fragments (modified hairy regions, MHRs) modified either in vitro (from apple) or in vivo (from potato) were tested. Cell culture vessels and titanium substratum coated with the apple-MHRs, MHR-A and a further-tailored fragment type, MHR-B, were compared with controls for their ability to support proliferation and differentiation of osteoclasts and osteoblasts. Cells grew and differentiated on MHR-B and on the control surfaces; MHR-A did not perform well in these assays. Genetically-engineered potato MHRs did not support bone cell growth to the same extent as apple MHR-B, but nonetheless the possibility to manipulate cellular proliferation with specific in vivo – modifications of pectins was introduced. When implanted into rat soft tissues, neither of the apple MHRs provoked severe acute inflammatory reactions, which indicates good in vivo - tolerance of these botanical macromolecules. These studies illustrate the biocompatibility of MHRs, and the directions towards which they could be further tailored. In terms of clinical use, their tolerability in vivo is especially significant.

This diploma thesis is focused on biocompatible polymers degradation study. Copolymers were studied based on poly(lactic-co-glycolic) acid and poly(ethylene glycol) PLGA-PEG-PLGA and further these copolymers modified with itaconic acid ITA-PLGA-PEG-PLGA-ITA. This paper investigated the influence of pH phosphate solution on the degradation of polymers. Degradation of polymers occurred at 37 °C in phosphate solution with pH 4.2, 7.4 and 9.2. High performance liquid chromatography with UV-VIS detection of diode-array type was used for quantitative determination of lactic acid and glycolic acid as the final degradation products. For qualitative identification of additional degradation products were used tandem connection liquid chromatography and mass spectrometry. Gel permeation chromatography with refractive index detector was used to determine the molecular weight decrease polymer chain after the degradation.

Polylactide (PLA) is a biodegradable and biocompatible polymer which is attracting much attention for environmental issues imposed by the petroleum-based polymers. PLA can be used as medical polymer in surgical sutures, implants tissue and many other areas. However, one of the main shortcomings of PLA is its brittleness in nature and relatively poor mechanical properties, which often limits its further application. It is generally accepted for polymeric materials that some mechanical properties of oriented structures can be improved as the molecular weight of PLA increases. The outcome of this thesis will provide the knowhow to achieve ultrahigh molecular weight of polylactides, and further to improve the mechanical properties and extend its range of applications. In this work, different catalytic systems for the synthesis of ultra-high molecular weight (UHMW) polylactide are considered. For the catalytic systems considered, the reaction conditions and initiators are investigated. The resulting molecular characteristics and mechanical properties of the synthesized polymers will be evaluated. On the contrary to the brittle nature of PLA, Poly(ε-carprolactone) (PCL) is elastic and flexible with a relatively low melting point (60 oC) and low glass transition temperature (-60 oC). Hence, ultra-high molecular weight PCL will be also synthesized by using the same catalytic systems employed for achieving UHMWPLAs. PCL is also used in different biomedical applications, such as in scaffolds for tissue engineering. It is well documented that the complementary physical properties of PLA and PCL have the potential to enhance toughness of PLA. To enhance the toughness and mechanical properties of the block copolymers attempt is made to synthesize ultra-high molar mass of the two polymers in the block copolymer. But their molar masses (and consequently their mechanical properties) are always on the low side. For this reason, the synthesis of high molecular mass PLA and PCL multiblocks will be attempted. Furthermore, it is interesting to study the synthesis of high molar masses PLLA and PDLA stereoblocks especially their ability to crystallize during the polymerization and test the possibility to prepare stereocomplex only during synthesis. The resulting molecular characteristics and mechanical properties of the synthesized multiblock-polymers will be also evaluated.

京都大学 / 新制・課程博士 / 博士(工学) / 甲第24625号 / 工博第5131号 / 新制||工||1981(附属図書館) / 京都大学大学院工学研究科材料化学専攻 / (主査)教授 松原 誠二郎, 教授 中尾 佳亮, 教授 杉野目 道紀 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DGAM

Current articular cartilage tissue engineering endeavors, using synthetic polymers as scaffolds, have been somewhat successful. However, the use of these materials has not yielded a satisfactory, functional replacement for articular cartilage. Therefore, this project focuses on an alternative to these materials, chitosan, which is a naturally occurring biopolymer. The first project objective was to fabricate and analyze bulk, porous chitosan scaffolds, based on total porosity, average pore diameter, mechanical integrity, and degradation susceptibility. Secondly, scaffolds were evaluated in terms of their ability to support neochondrogenesis, including assessments of cell attachment and viability, cell morphology, and the biosynthesis of proteoglycan and type-II collagen-rich extracellular matrix. Results indicated that chitosan scaffolds possessing an interconnecting, porous structure could be easily created through a simple freezing and lyophilization process, and these scaffolds did support neochondrogenesis. Results suggest chitosan may be a useful alternative to synthetic polymers for use in cartilage tissue engineering applications.

No description available.

Yes / Monolayers of neurons and glia have been employed for decades as tools for the study of cellular physiology and as the basis for a variety of standard toxicological assays. A variety of three dimensional (3D) culture techniques have been developed with the aim to produce cultures that recapitulate desirable features of intact. In this study, we investigated the effect of preparing primary mouse mixed neuron and glial cultures in the inert 3D scaffold, Alvetex. Using planar multielectrode arrays, we compared the spontaneous bioelectrical activity exhibited by neuroglial networks grown in the scaffold with that seen in the same cells prepared as conventional monolayer cultures. Two dimensional (monolayer; 2D) cultures exhibited a significantly higher spike firing rate than that seen in 3D cultures although no difference was seen in total signal power (<50 Hz) while pharmacological responsiveness of each culture type to antagonism of GABAAR, NMDAR and AMPAR was highly comparable. Interestingly, correlation of burst events, spike firing and total signal power (<50 Hz) revealed that local field potential events were associated with action potential driven bursts as was the case for 2D cultures. Moreover, glial morphology was more physiologically normal in 3D cultures. These results show that 3D culture in inert scaffolds represents a more physiologically normal preparation which has advantages for physiological, pharmacological, toxicological and drug development studies, particularly given the extensive use of such preparations in high throughput and high content systems.

Yes / Healthcare applications are known to have a considerable environmental impact and the use of bio-based polymers has emerged as a powerful approach to reduce the carbon footprint in the sector. This research aims to explore the suitability of using a new sustainable polyester blend (Floreon™) as a scaffold directed to aid in musculoskeletal applications. Musculoskeletal problems arise from a wide range of diseases and injuries related to bones and joints. Specifically, bone injuries may result from trauma, cancer, or long-term infections and they are currently considered a major global problem in both developed and developing countries. In this work we have manufactured a series of 3D-printed constructs from a novel biopolymer blend using fused deposition modelling (FDM), and we have modified these materials using a bioceramic (wollastonite, 15% w/w). We have evaluated their performance in vitro using human dermal fibroblasts and rat mesenchymal stromal cells. The new sustainable blend is biocompatible, showing no differences in cell metabolic activity when compared to PLA controls for periods 1-18 days. FloreonTM blend has proven to be a promising material to be used in bone tissue regeneration as it shows an impact strength in the same range of that shown by native bone (just under 10 kJ/m2) and supports an improvement in osteogenic activity when modified with wollastonite. / We would like to acknowledge the Medical Research Council in the UK (MRC) for funding this research throughout a MRC Proximity to Discovery award (P2D) with grant number MC_PC_16084. We would also like to acknowledge CONACYT for funding DH RamosRodriguez’s work.