A osteorradionecrose é uma séria e debilitante consequência da radioterapia da cabeça e pescoço, definida como uma área óssea que não sofre reparação após a irradiação. O objetivo deste trabalho foi analisar o efeito in vivo da presença de células mesenquimais da medula óssea carreadas em gel em defeitos provocados na calvária de ratos submetidos a um modelo experimental de osteorradionecrose. Foi realizada a obtenção de células osteoblásticas da medula óssea de ratos cultivadas in vitro e devidamente caracterizadas quanto ao seu fenótipo por meio de ensaios bioquímicos como proliferação celular, atividade de fosfatase alcalina e sua detecção in situ, além da detecção e quantificação de nódulos mineralizados. Posteriormente, ratos wistar foram submetidos ao protocolo de osteorradionecrose (irradiação com 20 Gy em dose única) e pareados com controles para em seguida serem realizados defeitos nas calvárias para inserção de células osteoblásticas da medula óssea previamente cultivadas in vitro juntamente com gel carreador e realizada a sutura. Os animais foram divididos em 4 grupos: controle (C), controle + células osteoblásticas (CC); osteorradionecrose (IR) osteorradionecrose + células osteoblásticas (IRc). Ao final de 4 semanas os animais foram sacrificados e realizados os seguintes ensaios: (1) análise histológica com base em cortes histológicos descalcificados (2) análise tomográfica por meio de micro-CT dos defeitos previamente criados. Os dados obtidos foram submetidos ao teste de normalidade e posteriormente à análise estatística para p<0,05. As células mesenquimais derivadas da medula óssea apresentaram fenótipo característico de células osteoblásticas após serem cultivadas em meio osteogênico, com aumento da proliferação, atividade de fosfatase alcalina e formação de nódulos mineralizados quando comparado com as células cultivadas em meio basal. A análise qualitativa dos cortes histológicos corados em hematoxilina-eosina e tricrômico de Masson demonstrou maior neoformação óssea no grupo IRc quando comparado ao grupo IR e similar ao grupos controles. A análise tomográfica revelou um aumento na espessura trabecular, densidade de conectividade, número trabecular e superfície óssea no grupo IRc em relação ao grupo IR. Os resultados sugerem que a inserção de células mesenquimais diferenciadas em osteoblastos favorece a neoformação de defeitos ósseos na presença de osteorradionecrose. / Osteoradionecrosis is a serious and debilitating consequence of head and neck radiotherapy, defined as a bone area that does not repair after irradiation. The purpose of this investigation was to analyze in vivo the presence of mesenchymal cells from bone marrow in calvariae defects of rats submitted to osteoradionecrosis. Cells were collected from rat femur bone marrow to perform characterization of osteoblastic phenotype by means of biochemical assays such as cell proliferation, alkaline phosphatase activity and its in situ detection and mineralization. Afterwards, male wistar rats were submitted to osteoradionecrosis protocol (20 Gy in a single dose) and paired with control animals. After 30 days, there were performed calvariae defects and placement of osteoblastic cells previously cultured with a gel vehicle inside the defects, which were sutured properly. The animals were divided in 4 groups: control (C), control + osteoblastic cells (CC), osteoradionecrosis (IR) osteoradionecrosis + osteoblastic cells (Irc). After 30 days, the animals were euthanized to perform the following analysis: (1) Histological evaluation by means of decalcified slide sections stained with hematoxilin-eosin and Masson trichrome; (2) Tomographic evaluation by means of adequate parameters. Data obtained were submitted to normality test and statistical analysis for p<0,05. The mesenchymal cells from bone marrow presented osteoblastic phenotype after being cultured in osteogenic medium, with higher ALP detection and activity, as well as an increase of mineralized nodules when compared to cells cultured in basal medium. Histological analysis showed that irradiation impaired bone neoformation and affected bone marrow composition, as well as the presence of osteoblasts and osteocytes. On the other hand, cell therapy in group IRc improved bone neoformation when compared to group IR, showing similarity to control groups. Tomographic analysis revealed an increase in trabecular thickness, density of connectivity, trabecular number and bone surface when compared to group IR. The results suggest that the placement of mesenchymal cells differentiated in osteoblasts may improve bone neoformation of defects created after the onset of osteoradionecrosis.

O sistema vascular de monocotiledôneas, incluindo Commelinaceae, tem sido estudado desde o sec. 17, sendo uma das maiores dificuldades em estudar esse sistema a grande quantidade de feixes vasculares e a complexidade de suas conexões, especialmente nas espécies que possuem plexo vascular nodal. As variações no sistema vascular não se restringem à relação caule/folha, uma vez que as raízes também podem apresentar certo grau de complexidade. Neste trabalho buscamos entender e apresentar uma arquitetura vascular em Commelinaceae baseado na reconstrução direta de análises tridimensionais dos sistemas caulinar e radicular. Para isto, foram analisadas amostras de caule (20 espécies) e raiz tuberosa (7 espécies) da família Commelinaceae, a partir de representantes das duas tribos de Commelinoideae (Commelineae e Tradescantieae). O rastreamento do sistema vascular no ápice caulinar mostrou que os feixes vasculares caulinares (em fase procambial) começam a ser distinguidos no 4º fitômero, a partir do ápice caulinar. A atividade meristemática do periciclo e o incremento foliar foram os principais fatores de espessamento primário do caule. A partir de análises tridimensionais complementares, como microscopia confocal, microtomografia computadorizada,vetorização gráfica e diafanização whole mount foram construídos modelos tridimensionais da arquitetura vascular em Commelinaceae. Nestes modelos foi possível verificar uma categoria de feixes ainda não relatada para Commelinaceae e que os feixes periféricos não são interrompidos ou terminam cegamente na periferia da medula. Três padrões de plexo vascular nodal são propostos, nos quais todos os feixes vasculares se conectam por meio de traqueídes. Para raiz tuberosa, dados morfométricos foram obtidos com auxílio do software ImageJ e vetorização gráfica foi utilizada para rastreamento do sistema vascular. Os resultados mostram quatro variações anatômicas do cilindro vascular e a quantificação morfométrica das características responsáveis por estas variações / The vascular system of monocotyledons, including Commelinaceae, has been studied since 17 th century, and one of the greatest difficulties in studying these systems is the great number of vascular bundles and the complexity of their connections, especially the species that have vascular nodal plexus. The variations in the vascular system are not restricted to the stem-leaf relation, since the roots can also present some degree of complexity. In this work we seek to understand and present a vascular architecture for Commelinaceae based on the direct reconstruction of 3D analyzes of shoot and root systems. Samples of stem (20 species) and tuberous root (7 species) of the Commelinaceae were analyzed including two tribes of Commelinoideae (Commelineae e Tradescantieae). The tracing of the vascular system in the shoot apex showed that the vascular bundles (in the procambial phase) begin to be distinguished in the 4th phytomer, from the apex. The meristematic activity of the pericycle (in the procambial phase) and the leaf increment were the main factors of primary stem thickening. From complementary 3D analyzes, such as confocal microscopy, computer microtomography, graphic vectorization and whole mount diaphanization, three-dimensional models of the vascular architecture were constructed from Commelinaceae. In these models it was possible to verify a category of bundles not yet reported for Commelinaceae and that the peripheral bundles are not interrupted or end blindly in the periphery of the vascular cylinder. Three patterns of nodal vascular plexus are proposed, in which all vascular bundles are connected by means of tracheids. For tuberous root, morphometric data were obtained by the aid of ImageJ software and graphic vectorization was used to trace the vascular system. The results show four anatomical variations of the vascular cylinder and the morphometric quantification of the characteristics responsible for these variations

Na Endodontia, a microtomografia (CT) tem sido amplamente utilizada como método de análise tridimensional. Atualmente, três movimentos mecanizados para o preparo do canal radicular tem sido introduzidos na rotina endodôntica, oscilatório, rotatório e recíproco. Existem diferentes sistemas de limas acionadas por motores capazes de realizar esses movimentos. Sendo assim, os objetivos deste trabalho são utilizar a CT para avaliar, em canais mesiais achatados de molares inferiores, a qualidade e modelagem final de preparo do canal radicular, utilizando-se sistemas oscilatório, recíproco e rotatório e avaliar a obturação do canal radicular com cone único ou ondas contínuas de condensação, no que diz respeito ao volume de material obturador e de espaços vazios. Os resultados não mostraram diferença estatísitica significativa no volume de desgaste entre os grupos testados. Para a porcentagem de superfície não tratada os grupos oscilatório e rotatório tocaram mais paredes nos terços cervical e médio do que o grupo recíproco. A técnica de ondas contínuas de condensação e cone único apresentaram preenchimento de obturação semelhantes nos terços médio e apical, sendo que no terço cervical a técnica de ondas contínuas deixou menor porcentagem de espaços vazios. / Endodontics in the microtomography (CT) has been widely used as a method for three-dimensional analysis. Currently, three mechanized movements for root canal preparation has been introduced in endodontic, oscillatory, rotational and reciprocal routine. There are different files systems driven by motors capable of these movements. Thus, the objectives of this work are using CT to evaluate, in flattened mesial canals of mandibular molars, the quality and final modeling of root canal preparation, using oscillatory, reciprocal and rotational systems and evaluate the root canal filling with single cone or continuous waves of condensation, with respect to the volume of filling material and voids. The results showed no significant difference in estatísitica wear volume between the tested groups. For the percentage of untreated surface and the oscillating rotating groups played more walls in the cervical and middle thirds of the reciprocal group. The technique of continuous wave of condensation and single cone showed similar fillings fill in the middle and apical thirds, and in the cervical third of the technique of continuous wave left lower percentage of voids.

A osteorradionecrose é uma séria e debilitante consequência da radioterapia da cabeça e pescoço, definida como uma área óssea que não sofre reparação após a irradiação. O objetivo deste trabalho foi analisar o efeito in vivo da presença de células mesenquimais da medula óssea carreadas em gel em defeitos provocados na calvária de ratos submetidos a um modelo experimental de osteorradionecrose. Foi realizada a obtenção de células osteoblásticas da medula óssea de ratos cultivadas in vitro e devidamente caracterizadas quanto ao seu fenótipo por meio de ensaios bioquímicos como proliferação celular, atividade de fosfatase alcalina e sua detecção in situ, além da detecção e quantificação de nódulos mineralizados. Posteriormente, ratos wistar foram submetidos ao protocolo de osteorradionecrose (irradiação com 20 Gy em dose única) e pareados com controles para em seguida serem realizados defeitos nas calvárias para inserção de células osteoblásticas da medula óssea previamente cultivadas in vitro juntamente com gel carreador e realizada a sutura. Os animais foram divididos em 4 grupos: controle (C), controle + células osteoblásticas (CC); osteorradionecrose (IR) osteorradionecrose + células osteoblásticas (IRc). Ao final de 4 semanas os animais foram sacrificados e realizados os seguintes ensaios: (1) análise histológica com base em cortes histológicos descalcificados (2) análise tomográfica por meio de micro-CT dos defeitos previamente criados. Os dados obtidos foram submetidos ao teste de normalidade e posteriormente à análise estatística para p<0,05. As células mesenquimais derivadas da medula óssea apresentaram fenótipo característico de células osteoblásticas após serem cultivadas em meio osteogênico, com aumento da proliferação, atividade de fosfatase alcalina e formação de nódulos mineralizados quando comparado com as células cultivadas em meio basal. A análise qualitativa dos cortes histológicos corados em hematoxilina-eosina e tricrômico de Masson demonstrou maior neoformação óssea no grupo IRc quando comparado ao grupo IR e similar ao grupos controles. A análise tomográfica revelou um aumento na espessura trabecular, densidade de conectividade, número trabecular e superfície óssea no grupo IRc em relação ao grupo IR. Os resultados sugerem que a inserção de células mesenquimais diferenciadas em osteoblastos favorece a neoformação de defeitos ósseos na presença de osteorradionecrose. / Osteoradionecrosis is a serious and debilitating consequence of head and neck radiotherapy, defined as a bone area that does not repair after irradiation. The purpose of this investigation was to analyze in vivo the presence of mesenchymal cells from bone marrow in calvariae defects of rats submitted to osteoradionecrosis. Cells were collected from rat femur bone marrow to perform characterization of osteoblastic phenotype by means of biochemical assays such as cell proliferation, alkaline phosphatase activity and its in situ detection and mineralization. Afterwards, male wistar rats were submitted to osteoradionecrosis protocol (20 Gy in a single dose) and paired with control animals. After 30 days, there were performed calvariae defects and placement of osteoblastic cells previously cultured with a gel vehicle inside the defects, which were sutured properly. The animals were divided in 4 groups: control (C), control + osteoblastic cells (CC), osteoradionecrosis (IR) osteoradionecrosis + osteoblastic cells (Irc). After 30 days, the animals were euthanized to perform the following analysis: (1) Histological evaluation by means of decalcified slide sections stained with hematoxilin-eosin and Masson trichrome; (2) Tomographic evaluation by means of adequate parameters. Data obtained were submitted to normality test and statistical analysis for p<0,05. The mesenchymal cells from bone marrow presented osteoblastic phenotype after being cultured in osteogenic medium, with higher ALP detection and activity, as well as an increase of mineralized nodules when compared to cells cultured in basal medium. Histological analysis showed that irradiation impaired bone neoformation and affected bone marrow composition, as well as the presence of osteoblasts and osteocytes. On the other hand, cell therapy in group IRc improved bone neoformation when compared to group IR, showing similarity to control groups. Tomographic analysis revealed an increase in trabecular thickness, density of connectivity, trabecular number and bone surface when compared to group IR. The results suggest that the placement of mesenchymal cells differentiated in osteoblasts may improve bone neoformation of defects created after the onset of osteoradionecrosis.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The removal of pre-existing root canal filling is basic requirement for endodontic retreatment in order to reveal necrotic tissue debris and bacteria which may be responsible for periapical inflammation and by the treatment failure. The aim of this study was to evaluate the efficacy of passive ultrasonic activation (PUA) with use of organic solvent as an assistant method in removal of filling material during endodontic retreatment and determine a protocol for its utilization, using high-resolution micro-Computed Tomography (micro-CT). Thirty curved mesial roots of mandibular molars with isthmus area were instrumented and filled with gutta-percha and EndoFill (Dentsply-Maillefer, Ballaigues, Switzerland) through Tagger s hybrid technique. After 30 days, the filling material was removed with the ProTaper Retreatment rotary system (Dentsply-Maillefer, Ballaigues, Switzerland) and the root canals were prepared with the F1-F4 ProTaper instruments (Dentsply-Maillefer, Ballaigues, Switzerland). The specimens will be randomized into two groups according to the final irrigation protocol. In the Manual Group, the root canals were irrigated and agitated manually with Orange Oil solvent (Citrol, Biodinâmica, Ibiporã, Brazil), 2,5% Sodium Hypochlorite (NaOCl) (Biodinâmica, Ibiporã, Brazil) and 17% Ethylenediaminetetraacetic Acid (EDTA) (Biodinâmica, Ibiporã, Brazil). In the PUA Group, the specimens were submitted to PUA with Orange Oil solvent, 2,5% NaOCl and 17% EDTA. High-resolution micro-CT scans was used to select the sample, analyze the root filling and measure the volume of the remaining root filling material after each stage of the retreatment. The volume of remaining filling material was established and transformed into percentage in relation to total volume of filling material. The dates were statistically analyzed with a significance level set at α = 0.05. The Mann Whitney test was used to compare the percentage of remaining filling material between groups, for canal thirds in each retreatment step. The Friedman test was used to verify differences among root filling material and the successive retreatment stages at each canal segment in same group. The differences amongst the coronal, middle and apical third in the each group were analyzed with Friedman test. The Dunn test was performed as the post-hoc multiple comparison method. No significant differences between groups when rotary instrumentation (Stage 1) was considered (P > 0.05). None of the irrigation protocols removed completely the remains of the root canal filling. The use of Manual e PUA protocols decreases statistically the amount of remaining filling material compared to Stage 1 (P = 0.000). In the PUA Group, the root canals showed significantly less percentage remaining filling material compared with Manual Group, for all thirds (P < 0.05). The PUA protocol left similar amount of filling material among the apical, middle and cervical thirds (P > 0.05). None of the irrigation protocols tested was able to completely remove all gutta-percha/sealer from the curved canals and with isthmus area. However, the results of this study suggest that in root canals with complex anatomy that require endodontic retreatment, the PUA with orange oil, NaOCl and EDTA can be a great aid in the removal of the remaining root filling material left after removal of the bulk of filling material. / A remoção do material obturador pré-existente é um requisito fundamental para a realização do retratamento endodôntico a fim de revelar restos de tecido necrótico e bactérias que podem ser responsáveis pela inflamação periapical e pela falha do tratamento. O objetivo deste estudo foi avaliar a eficácia da Ativação Ultrassônica Passiva (AUP) com uso de solvente orgânico, como um método auxiliar na remoção do material obturador durante o retratamento endodôntico e determinar um protocolo para sua utilização, através da Microtomografia Computadorizada (micro-CT). Trinta raízes mesiais curvas e com istmo de dentes molares inferiores foram instrumentadas e obturadas com guta percha e cimento EndoFill (Dentsply-Maillefer, Ballaigues, Suíça) através da técnica híbrida de Tagger. Após 30 dias, o material obturador foi removido com o sistema rotatório de desobturação ProTaper Retratamento (Dentsply-Maillefer, Ballaigues, Suíça) e o canal repreparado com os instrumentos F1-F4 do sistema rotatório ProTaper Universal (Dentsply-Maillefer, Ballaigues, Suíça). Os espécimes foram aleatoriamente divididos em dois grupos (n=15) de acordo com o protocolo de irrigação final. No Grupo Manual, os canais radiculares foram irrigados e agitados manualmente com solvente a base de Óleo de Laranja (Citrol, Biodinâmica, Ibiporã, Brasil), Hipoclorito de Sódio (NaOCl) 2,5% (Citrol, Biodinâmica, Ibiporã, Brasil) e Ácido Etilenodiaminotetracético (EDTA) 17% (Citrol, Biodinâmica, Ibiporã, Brasil) . No Grupo AUP, os espécimes foram submetidos à AUP com Óleo de Laranja, NaOCl 2,5% e EDTA 17%. A seleção da amostra, a análise da obturação e a mensuração do volume de material obturador remanescente após cada estágio do retratamento foram realizadas através da micro-CT. O volume de material obturador remanescente foi estabelecido e transformado em porcentagem em relação ao volume total de material obturador. Os dados foram submetidos à análise estatística com nível de significância fixado em α = 0.05. O teste de Mann Whitney foi usado para comparar a porcentagem de material obturador remanescente entre os grupos, em cada fase do retratamento. O teste de Friedman foi usado para analisar diferenças entre o volume da obturação e os estágios do retratamento dentro de um mesmo grupo. Diferenças entre os terços cervical, médio e apical dentro do mesmo grupo foram analisadas com o teste de Friedman. O teste de Dunn foi usado como post hoc para múltiplas comparações. Não houve diferença estatisticamente significante entre os grupos quando a instrumentação rotatória (Estágio 1) foi considerada (P > 0.05). Nenhum dos protocolos de irrigação removeu completamente os remanescentes de material obturador em todos os espécimes. Tanto o protocolo da AUP quanto o Manual diminuíram a quantidade de remanescente de guta percha e cimento em relação ao Estágio 1 (P = 0.001). No grupo AUP, os canais radiculares mostraram significantemente menos porcentagem de remanescentes de material obturador que o grupo Manual, para todos os terços (P < 0.05). O protocolo da AUP deixou quantidade similar de material remanescente em todos os terços do canal radicular (P > 0.05). Nenhum dos protocolos de irrigação testados foi capaz de remover completamente a guta percha/cimento de canais radiculares curvos e com área de istmo. Contudo, os resultados obtidos neste estudo sugerem que em canais radiculares com anatomia complexa que necessitam de retratamento endodôntico, a AUP com óleo de laranja, NaOCl e EDTA pode ser um grande auxiliar na remoção dos remanescentes de material obturador deixados após a desobturação do canal radicular.

Na Endodontia, a microtomografia (CT) tem sido amplamente utilizada como método de análise tridimensional. Atualmente, três movimentos mecanizados para o preparo do canal radicular tem sido introduzidos na rotina endodôntica, oscilatório, rotatório e recíproco. Existem diferentes sistemas de limas acionadas por motores capazes de realizar esses movimentos. Sendo assim, os objetivos deste trabalho são utilizar a CT para avaliar, em canais mesiais achatados de molares inferiores, a qualidade e modelagem final de preparo do canal radicular, utilizando-se sistemas oscilatório, recíproco e rotatório e avaliar a obturação do canal radicular com cone único ou ondas contínuas de condensação, no que diz respeito ao volume de material obturador e de espaços vazios. Os resultados não mostraram diferença estatísitica significativa no volume de desgaste entre os grupos testados. Para a porcentagem de superfície não tratada os grupos oscilatório e rotatório tocaram mais paredes nos terços cervical e médio do que o grupo recíproco. A técnica de ondas contínuas de condensação e cone único apresentaram preenchimento de obturação semelhantes nos terços médio e apical, sendo que no terço cervical a técnica de ondas contínuas deixou menor porcentagem de espaços vazios. / Endodontics in the microtomography (CT) has been widely used as a method for three-dimensional analysis. Currently, three mechanized movements for root canal preparation has been introduced in endodontic, oscillatory, rotational and reciprocal routine. There are different files systems driven by motors capable of these movements. Thus, the objectives of this work are using CT to evaluate, in flattened mesial canals of mandibular molars, the quality and final modeling of root canal preparation, using oscillatory, reciprocal and rotational systems and evaluate the root canal filling with single cone or continuous waves of condensation, with respect to the volume of filling material and voids. The results showed no significant difference in estatísitica wear volume between the tested groups. For the percentage of untreated surface and the oscillating rotating groups played more walls in the cervical and middle thirds of the reciprocal group. The technique of continuous wave of condensation and single cone showed similar fillings fill in the middle and apical thirds, and in the cervical third of the technique of continuous wave left lower percentage of voids.

Osteocytes, the most abundant cell within bone, have been linked to the processes of mechanosensation and transduction. Based upon relatively limited empirical evidence, variations in their abundance and morphology have been linked to sex, age, biomechanics and disease. In order to better elucidate lacunar variation within a healthy cohort, samples from 30 women aged 20-86 and 36 men aged 18-92 were studied utilizing synchrotron radiation micro-CT. Initial studies of normal variation within the femoral proximal shaft cross-section found high variation in lacunar density (up to ~54%) and associated morphological differences linked to biomechanical regions. In women, a non-significant trend in lacunar density reduction was apparent with age; however, a significant reduction in lacunar volume with age (~30%) was observed. Also noted were differences in lacunar morphology, with the lacunae of younger women characterized as flatter and less equant than their older counterparts. The males, who demonstrated lacunar density decline with age and a tendency towards more equant and less elongate lacunae, did not share these characteristics. Intriguingly, the previously noted reductions in lacunar volume were not observed in males. The results of this research indicate that normal variation in osteocyte lacunar parameters is high. To our knowledge the observation that lacunar volume differs in women with age is novel, potentially resulting from preferential surface infilling within the extracellular space. The functional impact of this infilling is unclear but such a change in scale likely impacts the mechanosensing function of the osteocyte network. This hypothesis warrants further investigation as, if confirmed, it would represent a profound negative impact on the osteocyte network and may provide new insights into age-related bone loss.

Preclinical micro-computed tomography (microCT) images are of utility for 3D morphological bone evaluation, which is of great interest in cancer detection and treatment development. This work introduces a compression strategy for microCTs that allocates specific substances in different Volumes of Interest (Vols). The allocation procedure is conducted by the Hounsfield scale. The Vols are coded independently and then grouped in a single DICOM-compliant file. The proposed method permits the use of different codecs, identifies and transmit data corresponding to a particular substance in the compressed domain without decoding the volume(s), and allows the computation of the 3D morphometry without needing to store or transmit the whole image. The proposed approach reduces the transmitted data in more than 90% when the 3D morphometry evaluation is performed in high density and low density bone. This work can be easily extended to other imaging modalities and applications that work with the Hounsfield scale. (C) 2015 Elsevier Ltd. All rights reserved.

Cystic fibrosis (CF) is an inherited disease leading to disrupted function of the cystic fibrosis transmembrane conductance regulator (CFTR) anion channel. CF affects many organ systems including the pancreas, liver, intestine, sweat glands, and gallbladder. The leading cause of morbidity and mortality, however, is lung disease. A porcine model of CF was developed, and over time it develops lung disease that recapitulates many of the characteristics observed in humans with CF including airway remodeling, mucus accumulation, infection, and inflammation. At birth, and despite the absence of inflammation and infection, the CF pig airways exhibit a host of abnormalities including tracheal cartilage ring defects, abnormal appearing smooth muscle bundles, reduced trachea diameter, and reduced mainstem bronchi diameter. The primary objectives of this study were to construct an experimental method that allowed for the attainment of airway size information at multiple inflation pressures, to assess the extent of airway narrowing in the newborn CF porcine lung at 20 cmH2O, to determine the tracheal lobe volume for CF and non-CF, and to perform morphologic assessment of the parenchymal airspaces for CF and non-CF newborn pigs. Micro-computed tomography (micro-CT) was selected as the primary analysis tool. The volumetric, high resolution data sets of micro-CT provided a means to virtually track airways through the three dimensional space of the lung, and to image airways as small as 250 microns in diameter. Due to experimental constraints, only one lobe was analyzed: the tracheal lobe; it is the porcine equivalent of the human right upper lobe. Each excised tracheal lobe was cannulated and micro-CT scanned five times. Each lobe was scanned at multiple inflation pressures ranging from 0 to 20 cmH2O. The airways were segmented with a custom designed, substantially-automated computer algorithm. Quantitative analysis of airway size was done with the Pulmonary Workstation 2 software package. At a pressure of 20 cmH2O, the CF airway narrowing was most pronounced in the large airways of the tracheal lobe, and the percent difference in airway cross sectional area between CF and non-CF lessened for airways of smaller size. The volume of the newborn CF pig tracheal lobe was approximately twenty percent smaller than non-CF, but no differences were observed in tracheal lobe airspace histology between the groups. Airway size deviations at birth imply developmental abnormalities in utero that are dependent upon CFTR function. Additionally, the observation that reduced airway caliber exists only in relatively large airways suggests a time-dependent role of CFTR on airway development, as the large airways develop before the small ones in utero. These findings may provide insight to the early pathogenesis of CF lung disease.

It is well established that bone is capable of adapting to changes in its environment; however, little is known regarding how environmental stimuli, specifically loading, are associated with the internal 3D microarchitecture of cortical bone. The aim of this thesis was to develop a small animal model that can be used to experimentally test hypotheses regarding bone adaptation. High resolution micro-CT was validated and employed as a novel method for the visualization and quantification of rat cortical bone microarchitecture in 3D. The use of this imaging method allowed for the measurement of primary vascular canal orientation in 3D, which had never been achieved before. Using this measure along with an immobilization model for unloading allowed me to test how loading is associated with the orientation of these vascular canals. Normally ambulating rat bones (from 10 female rats) had a canal structure that was 9.9° more longitudinal than their immobilized counterparts. This finding that loading has an effect on primary canal orientation brought to light the need to induce remodeling and therefore, secondary vascular canals, in the rat to increase its novelty as a model for looking at bone adaptation. Remodeling was induced by increasing the calcium demands of female rats, either through a calcium restricted diet (n=2) or pregnancy and lactation coupled with a calcium restricted diet (n=2). Mean cortical thickness for the calcium restricted rats and the pregnant and lactating rats that were on a calcium restricted diet were 622 µm and 419 µm, respectively. The mean BMU count for calcium restricted rats seemed to be higher than that of the pregnant and lactating rats; however, the calcium restricted rats seemed to have a lower BMU density. Once this full-scale study is executed the rat will provide a more representative model for studying human bone adaptation.