Drug delivery systems (DDS) are essentially pharmaceutical products for human therapy, typically involving a mixture of active ingredients and excipients. Based upon quantitative characterization of structure, the thesis introduces the concept of classifying the architecture of DDS into four levels by their spatial scale and the life time period. The primary level is recognised as the static structure of the whole dosage form with a size from μm to cm with the final structure generated by formulation design. The secondary level categorises the structures of particles or sub-units to form a DDS with sizes from nm to mm as key units in processing such as mixing, grinding, granulation and packing; The tertiary level represents the dynamic structures of DDS during the drug release phase in vitro or in vivo incorporating the structure size range from nm to mm, which undergo changes during dissolution, swelling, erosion or diffusion. The spatial scale for the quaternary level is defined as the meso or micro scale architecture of active and non-active molecules within a DDS with sizes from Å to μm for the molecular structure of drug and excipients. Methods combining X-ray tomography, image processing, and 3D reconstructions have been devised and evaluated to study systematically pharmaceutical structures and correlate them with drug release kinetics of DDS. Based on the quantitative structural information of pharmaceutical intermediates and dosage forms, it is possible now to correlate structures with production processing, behaviour and function, and the static and dynamic structures of DDS with the release kinetics. Thus, a structure-guided methodology has been established for the research of DDS. / Chinese Academy of Sciences

No description available.

Injuries of the hands and wrist are prevalent in many occupations requiring repetitive tasks and may be further aggravated by advancing age; these injuries are termed work related musculoskeletal disorders (WMSDs). Prior studies using an innovative operant rat model of reaching and grasping as a model of WMSDs demonstrated exposure dependent changes in forelimb bones of young adult rats performing repetitive tasks ≤ 3 months. No one has yet to examine if aging enhances forelimb bone degradative changes occurring with WMSDs, or if forelimb bones adapt or degrade further in response to moderate versus high demand repetitive tasks performed for prolonged time periods (up to 24 months). Bone remodeling is a normal biological process that allows bones to adjust to strains. Unfortunately, both aging and inflammation can deregulate the balance between bone resorption and formation. Aging mammals display increased baseline inflammatory-cytokine levels, both systemically and at the tissue level. Several inflammatory cytokines have been shown to stimulate osteoclastogenesis leading to bone resorption and reduced bone formation. We have reported increased production of inflammatory cytokines in serum and musculotendinous tissues of aged animals performing a repetitive reaching and grasping tasks for up to 12 weeks, warranting further examination of whether aged rats performing these tasks have increased bone resorptive changes, compared to young adult rats. We hypothesized that aging would enhance bone degradative changes in our model as a consequence of increased bone inflammatory responses to a moderate demand repetitive task. Therefore, our first aim was to examine forearm grip strength, trabecular and cortical bone quality, and inflammatory cytokine levels in radii of mature (14-18 mo of age) and young adult (2.5-6.5 mo of age) female Sprague Dawley rats after performance of a high repetition low force (HRLF) task for 12 weeks, compared to each other and age-matched controls. We found that mature rats performing a moderate demand repetitive task for 12 weeks had decreased bone formation and quality, particularly cortical bone quality, compared to young adult rats performing the same task, with increased inflammatory and decreased anti-inflammatory responses, and perhaps lower grip strength, as likely contributors. An adaptive bone response was observed in young adult animals performing a moderate level task of high repetition low force for 12 weeks. In contrast, a previous study showed bone degradative changes in young adult rats performing a high demand task of high repetition high force task for 12 weeks. Osteocytes are the mechanosensing cells of bones, and disruption or changes to their environment can lead to apoptosis or molecular changes. In models of forced bone loading to bone fatigue, osteocyte apoptosis increases sclerostin levels and osteoclast recruitment. Increased sclerostin also leads to increased RANKL production. In contrast, low level loading for a short period reduces sclerostin levels and encourages bone formation. We hypothesized that long-term muscle loading at high repetition low force loads would induce further bone adaptation, but that long-term high repetition high force muscle loading would result in detrimental bone loss, as well as alterations in these two bone remodeling proteins, RANKL and sclerostin. Therefore, our second aim was to determine if prolonged performance of a moderate demand upper extremity reaching and grasping task by young adult rats would continue to enhance forelimb bone formation and quality. We hypothesized that continued performance of a high repetition low force (HRLF) task for 24 weeks would lead to increased bone formation. We also hypothesized that RANKL and sclerostin, two proteins that have not been investigated in our rat model of WMSDs, would be reduced in rats performing a HRLF task for 24 weeks, as the bones reach adaptation. We found that 24 week HRLF rats showed several indices of bone formation and adaptation to the task; as well as reduced sclerostin immunoexpression, compared to controls, a reduction that likely contributed to the enhanced bone formation. To expand on this investigation, in our third aim, we investigated the impact of performance of a high repetition high force (HRHF) task for 18 weeks on young adult rat forelimb bones, and on sclerostin and RANKL levels. We observed detrimental trabecular bone remodeling in the radius, including decreased trabeculae bone volume, number and thickness, increased trabecular separation and anisotropy, and a transition to rod-shaped trabeculae in 18-week HRHF task animals, compared to food restricted control rats. In the 18-week HRHF rats, osteoclast numbers increased and osteoblast numbers decreased, concomitant with increased osteocyte apoptosis and empty lacunae, compared to control rats. Also, mRNA and protein levels of RANKL increased and sclerostin decreased in the 18-week HRHF rats, compared to to control rats. Thus, prolonged performance of a high demand task of high repetition high force induced detrimental trabecular bone changes. The increased RANKL likely contributed to these changes, and although sclerostin level decreased, a change that should contribute to enhanced osteoblast activity, bone formation was not rescued. In conclusion, prolonged performance of a HRLF task by young adult rats leads to reduced sclerostin levels and increased bone formation and bone quality. Aged rats performing the same HRLF task showed increased bone degradative changes that might increase fracture risk. In contrast, prolonged performance of a HRHF task by young adult rats leads to increased bone resorption and degradation, changes associated with RANKL expression. Sclerostin levels were reduced by the HRHF task, but failed to rescue bone formation. / Cell Biology

No description available.

Alterações na articulação emporomandibular (ATM) comumente geram desequilíbrios musculares que estão associados à alterações no tecido ósseo. Esta articulação pode sofrer a influência de traumas, fatores congênitos ou desordens de crescimento. Estudos sobre alterações de crescimento do complexo maxilomandibular decorrentes de problemas da ATM são escassos. O objetivo deste trabalho foi avaliar por meio da microtomografia os efeitos da remoção do disco articular e a remoção conjugada do disco e cartilagem articular no crescimento e na microarquitetura óssea da mandíbula de ratos. Trinta ratos da raça Wistar com um mês de idade foram divididos em três grupos: CTR (controle operado); RD (remoção de disco articular) e RDC (remoção conjugada do disco e cartilagem articular). Apenas o lado direito foi operado; o lado esquerdo permaneceu intacto. Após dois meses de acompanhamento, os ratos foram sacrificados e as hemimandíbulas escaneadas em microtomógrafo A remoção do disco articular e a remoção conjugada do disco e cartilagem articular alteram o volume e microestrutura do osso trabecular da mandíbula de ratos jovens. Estas duas intervenções provocaram uma queda na qualidade de parâmetros da microestrutura do trabeculado do processo angular e diminuição do crescimento da hemimandíbula do lado operado. / Changes in the temporomandibular joint (TMJ) lead to muscle dysfunctions that are associated with bone changes. This joint region can be influenced by trauma, congenital factors or growth disorders. Studies linking TMJ problems and growth alterations are scarce. The aim of this study was to evaluate the effects of the articular disc removal or articular disc and cartilage removal on the bone microarchitecture and mandibular growth of young rats. Thirty Wistar rats (one month old) were divided into three groups: CTR (sham operated); RD (disc removal) and RDC (disc and cartilage removal). Only the right side was operated, keeping the left side intact. After two months, the rats were sacrificed and the mandibles scanned on micro-CT for quantitative analysis. Some microstructural parameters were altered by the disc removal or disc and cartilage removal. The right side presented lower growth than the left side.

Alterações na articulação emporomandibular (ATM) comumente geram desequilíbrios musculares que estão associados à alterações no tecido ósseo. Esta articulação pode sofrer a influência de traumas, fatores congênitos ou desordens de crescimento. Estudos sobre alterações de crescimento do complexo maxilomandibular decorrentes de problemas da ATM são escassos. O objetivo deste trabalho foi avaliar por meio da microtomografia os efeitos da remoção do disco articular e a remoção conjugada do disco e cartilagem articular no crescimento e na microarquitetura óssea da mandíbula de ratos. Trinta ratos da raça Wistar com um mês de idade foram divididos em três grupos: CTR (controle operado); RD (remoção de disco articular) e RDC (remoção conjugada do disco e cartilagem articular). Apenas o lado direito foi operado; o lado esquerdo permaneceu intacto. Após dois meses de acompanhamento, os ratos foram sacrificados e as hemimandíbulas escaneadas em microtomógrafo A remoção do disco articular e a remoção conjugada do disco e cartilagem articular alteram o volume e microestrutura do osso trabecular da mandíbula de ratos jovens. Estas duas intervenções provocaram uma queda na qualidade de parâmetros da microestrutura do trabeculado do processo angular e diminuição do crescimento da hemimandíbula do lado operado. / Changes in the temporomandibular joint (TMJ) lead to muscle dysfunctions that are associated with bone changes. This joint region can be influenced by trauma, congenital factors or growth disorders. Studies linking TMJ problems and growth alterations are scarce. The aim of this study was to evaluate the effects of the articular disc removal or articular disc and cartilage removal on the bone microarchitecture and mandibular growth of young rats. Thirty Wistar rats (one month old) were divided into three groups: CTR (sham operated); RD (disc removal) and RDC (disc and cartilage removal). Only the right side was operated, keeping the left side intact. After two months, the rats were sacrificed and the mandibles scanned on micro-CT for quantitative analysis. Some microstructural parameters were altered by the disc removal or disc and cartilage removal. The right side presented lower growth than the left side.

The TRUShape 3D Conforming File (TRUShape), a novel, S-shaped nickel-titanium (NiTi) rotary file, was developed to facilitate cleaning and shaping of irregular-shaped root canals. The purpose of this study was to evaluate the shaping ability of TRUShape compared to Vortex Blue (VB) when used in non-round, oval-shaped root canals by micro-computed tomography (MCT). Thirty single-rooted human mandibular premolar teeth with radiographically similar root canal size and curvature were randomly allocated to two groups (N=15), and mechanically prepared with TRUShape or VB. Each tooth was submitted to MCT at 20 μm resolution at three time intervals: before shaping, and after shaping to an intermediate apical size 30 and a final apical size 40. Three-dimensional data sets were superimposed and evaluated for root canal volume, surface area, and treated surface. Matched axial slices in the apical, middle, and coronal root thirds were evaluated for area, roundness, and canal transportation expressed as center of mass shift (CMS). Data were statistically analyzed using parametric and non-parametric tests. Root canal volumes increased similarly and significantly overall (p< 0.001; from an initial volume of 7.3±3.5 mm3 to an intermediate volume of 8.7±3.1 mm3 and a final volume of 9.9±3.0 mm3). Treated canal surface was significantly larger in the TRUShape group at both apical sizes 30 and 40 with 72±15% vs. 55±23% and 85±12% vs. 71±20% non-static voxels for TRUShape and VB, respectively (p< 0.05). Canal transportation was less than 100 μm in all but 8 out of 90 cross sections and was not significantly different between groups. This MCT study demonstrated the TRUShape 3D Conforming File to be effective in the mechanical preparation, specifically, the surface treatment, of single-rooted premolars with non-round, oval-shaped root canals. Funding was provided by the American Association of Endodontists (AAE) Foundation and the Department of Endodontics at the University of Iowa, College of Dentistry.

The understanding of multiphase flow properties is essential for the exploitation of hydrocarbon reserves in a reservoir; these properties in turn are dependent on the geometric properties and connectivity of the pore space. The determination of the pore size distribution in carbonate reservoirs remains challenging; carbonates exhibit complex pore structures comprising length scales from nanometers to several centimeters. A major challenge to the accurate evaluation of these reservoirs is accounting for pore scale heterogeneity on multiple scales. This is the topic of this thesis. Conventionally, this micron scale information is achieved either by building stochastic models using 2D images or by combining log and laboratory data to classify pore types and their behaviour. None of these capture the true 3D connectivity vital for flow characterisation. We present here an approach to build realistic 3D network models across a range of scales to improve property estimation through employment of X-ray micro-Computed Tomography (μCT) and Focussed Ion Beam Tomography (FIBT). The submicron, or microporous, regions are delineated through a differential imaging technique undertaken on x-ray CT providing a qualitative description of microporosity. Various 3-Phase segmentation methods are then applied for quantitative characterisation of those regions utilising the attenuation coefficient values from the 3D tomographic images. X-ray micro-CT is resolution limited and can not resolve the detailed geometrical features of the submicron pores. FIB tomography is used to image the 3D pore structure of submicron pores down to a scale of tens of nanometers. We describe the experimental development and subsequent image processing including issues and difficulties resolved at various stages. The developed methodology is implemented on cores from producing wackstone and grainstone reservoirs. Pore network models are generated to characterise the 3D interconnectivity of pores. We perform the simulations of petrophysical properties (permeability and formation resistivity) directly on the submicron scale image data. Simulated drainage capillary pressure curves are matched with the experimental data. We also present some preliminary results for the integration of multiscale pore information to build dual-scale network models. The integration of multiscale data allows one to select appropriate effective medium theories to incorporate sub-micron structure into property calculations at macro scale giving a more realistic estimation of properties.

Los estímulos mecánicos son uno de los factores que afectan a la diferenciación celular en el proceso de regeneración del tejido óseo, por lo tanto, en el desarrollo de andamios para ingeniería de tejidos, se pueden aplicar las cargas mecánicas con el fin de inducir la actividad de las células. Cuando se aplican cargas mecánicas, los estímulos mecánicos específicos transmitidos a las células a nivel microscópico pueden estudiarse mediante técnicas numéricas. El objetivo de esta tesis fue estudiar la mecanoregulación de la diferenciación de tejido en andamios de fosfato de calcio utilizando modelos de elementos finitos basados en micro tomografía axial computarizada.Dos muestras de materiales porosos basados en fosfato de calcio fueron utilizadas. Se desarrollaron mallas de elementos finitos congruentes, discretizando la fase sólida y los macro poros interconectados, con el fin de tener en cuenta la morfología irregular de los andamios.En primer lugar, se estudió la distribución de los estímulos mecánicos. La fase sólida y el fluido intersticial se simularon como material elástico lineal y como fluido Newtoniano, respectivamente. Se simuló una compresión del 0.5% en el sólido y un fluido con velocidades de entrada de 1, 10 y 100 µm/s en los poros. Se encontraron distribuciones de deformación similares en las paredes ambos materiales, con valores máximos de 1.6% en compresión y de 0.6% en tracción. En algunos poros, la velocidad del fluido aumentó a 100 y 1000 veces la velocidad de entrada. Este estudio mostró como estímulos mecánicos macroscópicos pueden causar distintos niveles de estímulos mecánicos microscópicos dentro los andamios, debido a la morfología.A continuación se realizó un estudio en el tiempo de la diferenciación de tejido en un andamio sometido a condiciones in vitro. La compresión y la perfusión se modelaron como en el estudio anterior. Se simularon una compresión del 0.5% y una velocidad de entrada de fluido constante de 10 µm/s o una presión de entrada de fluido constante de 3 Pa. La deformación cortante octaédrica y el esfuerzo cortante del fluido se utilizaron como estímulos mecano-regulatorios basándose en la teoría de Prendergast et al. (1997). Al aplicar velocidad constante, se predijeron fluctuaciones entre los estímulos equivalentes a la formación de tejido y a la muerte celular, debido al aumento en el esfuerzo cortante del fluido cuando el tejido comienza a llenar los poros. Sin embargo, al aplicar presión constante, se predijo estímulo equivalente a la diferenciación de tejido óseo en la mitad del volumen de los poros. Estos resultados sugieren que para permitir la diferenciación de tejido, la velocidad del fluido debe disminuirse cuando el tejido empieza a mineralizarse.Finalmente, se llevó acabo un estudio en el tiempo de la angiogénesis y de la diferenciación de tejido en un andamio bajo condiciones in vivo. La deformación cortante octaédrica y la velocidad relativa del fluido se utilizaron como estímulos mecano-regulatorios. Las fases sólida y porosa fueron tratadas como materiales poroelásticos. Se simuló la actividad individual de las células. Compresiones de 0.5 y 1% fueron simuladas. La mayoría de los vasos crecieron en los poros de la periferia del andamio y se bloquearon por las paredes. Se formaron redes capilares similares independientemente de la magnitud de deformación utilizada. Al aplicar 0.5% de compresión, estímulos correspondientes a la formación de hueso se predijeron en el 70% del volumen de los poros, sin embargo, sólo el 40% del volumen se llenó de osteoblastos debido a la falta de oxigeno. Este estudio mostró el efecto de la falta de vascularización en el centro del andamio en la diferenciación de tejido.Ese tipo de estudios, combinados con estudios in vitro, deberían contribuir a la comprensión del proceso de diferenciación de los tejidos dentro de los andamios y por lo tanto a la mejora de los métodos de diseño de andamios. / Mechanical stimuli are one of the factors that affect cell differentiation in the process of bone tissue regeneration; therefore, in the development of scaffolds for tissue engineering, mechanical loads can be applied in order to induce cell activity. The specific mechanical stimuli transmitted to cells at a microscopic level when mechanical loads are applied can be studied using numerical techniques. The objective of this thesis was to study the mechanoregulation of tissue differentiation within calcium phosphate scaffolds using micro computed tomographed based finite element models.Two samples of porous calcium phosphate based materials were used. Congruent finite element meshes, with the solid phase and the interconnected pores discretized, were developed in order to account for the scaffold irregular morphology.First, a study of the distribution of mechanical stimuli was performed. The solid phase and the fluid flow within the pores were modeled as linear elastic solid material and Newtonian fluid respectively. Compressive strains of 0.5% of total deformation applied to the solid and interstitial fluid flows with inlet velocities of 1, 10 and 100 µm/s applied to the pores were simulated. Similar strain distributions for both materials were found, with compressive and tensile strain maximal values of 1.6% and 0.6% respectively. For the fluid flow models, the fluid velocity in some of the scaffold pores increased to 100 and 1000 times the inlet velocity. This study showed how mechanical loads and fluid flow applied to the scaffolds caused different levels of mechanical stimuli within the samples according to the morphology of the materials.Next, a study of the mechanoregulation of tissue differentiation over time in a scaffold subjected to in vitro loads was performed. The solid phase and the fluid flow were modeled as in the study described above. Compressive strain of 0.5% and fluid flow with constant inlet velocity of 10 µm/s or constant inlet pressure of 3 Pa were applied. Octahedral shear strain and fluid shear stress were used as mechano-regulatory stimuli based on the theory of Prendergast et al. (1997). When a constant velocity was simulated, fluctuations between stimuli equivalent to tissue formation and cell death were predicted due to the increase in the fluid shear stress when tissue started to fill the pores. However, when constant pressure was applied, stimuli equivalent to bone formation were predicted in about half of the pore volume. These results suggest that in order to allow tissue differentiation within a scaffold, the fluid velocity should be decreased when tissue starts mineralizing.Finally, a study of the angiogenesis and the mechanoregulation of tissue differentiation over time in a scaffold subjected to in vivo conditions was performed. Octahedral shear strain and relative fluid velocity were used as mechano-regulatory stimuli. The solid and pore phases were treated as poroelastic materials. Individual cell activity was simulated within the pore domain. Compressive strains of 0.5 and 1% of total deformation were simulated. Most vessels grew in the pores at the periphery of the scaffolds and were blocked by the scaffold walls. Similar capillary networks were formed independently of the magnitude of the mechanical strain applied. When 0.5% of strain was applied, 70% of the pore volume was affected by mechano-regulatory stimuli corresponding to bone formation; however, because of the lack of oxygen, only 40% of the volume was filled with osteoblasts. This study showed the effect of the lack of vascularization in the center of the scaffold on the tissue differentiation.Such kind of studies, combined with in vitro studies, should contribute to the understanding of the process of tissue differentiation within the constructs and therefore to the improvement of scaffold design methods.

Fracture healing requires a series of events including inflammatory response and callus formation, callus remodeling and bone healing. Fracture healing is a complex process, there are several overlapping phases , including inflammation , cartilage formation and bone remodeling, there are many internal or external factors could impact on fracture healing, leading to delayed bone healing or non healing. The global androgen receptor knockout (GARKO) mice has been know to reduce bone mass in endochondral bone and osteoblast mineralization, but the impact for callus formation in fracture healing is still unclear. The goals of study is to investigate the role of androgen and androgen receptor in wild-type (WT) mice and GARKO mice after fracture healing during callus formation and bone mineralization and bone remodeling. Therefore, long-term animal experiments observed by micro-computed tomography to study the roles of androgen and androgen receptor on the process and mechanisms of fracture healing is necessary. We applied in vivo micro-computer tomography (Micro-CT) to build up the three-dimensional model images at different time points for wild-type mice and GARKO mice after fracture healing and observe the bone healing process of micro-structure of the development of callus during fracture healing. The callus tissue morphology observed by histological staining to study the proportion and position of collagen, fibrous tissue and bone. The results show that the healing of WT mice is better than GARKO mice. GARKO mice develop smaller callus size and less bone volume and show delayed healing. In general, orchiectomy (ORX) decreases callus size in WT mice but not in GARKO mice. However, the healing rate of elderly GARKO mice is not obvious in comparison with young GARKO mice. Together, our study demonstrated that the androgen and androgen receptor regulate fracture healing and play an important role in bone repair and healing. Our mouse model may be used for the therapeutic drug screening of bone fractures caused by osteoporosis.