Resumo: Nesta pesquisa foram investigados aspectos anatômicos e biométricos da face flexora dos dígitos de 20 novilhas mestiças da raça Nelore, com idades entre 24 e 36 meses, criadas em sistema extensivo. O material foi proveniente de matadouro sob Fiscalização do Serviço de Inspeção Oficial e mantido sob congelamento a -18°C até o momento da dissecação. Após tricotomia e limpeza das regiões distais ao metacarpo e metatarso, as faces palmar e plantar dos dígitos foram dissecadas in natura e as estruturas anatômicas identificadas e medidas com auxílio de paquímetro universal. As mensurações foram feitas em milímetros (mm) e incluíram o comprimento, a largura e a espessura das estruturas anatômicas digitais. A técnica de dissecação in natura foi considerada útil para a execução deste estudo e as estruturas identificadas e mensuradas incluíram os ligamentos acessórios distais dos paradígitos, ligamentos anulares palmares e plantares, bainha digital tendínea, ligamentos anulares proximais e distais, ligamentos interdigitais distais, tendões flexores digitais superficiais e profundos e a manica flexoria. Os dados biométricos obtidos indicam diferenças (p≤0,05) entre membros, antímeros e dígitos, especialmente nas estruturas anatômicas localizadas distalmente. Devido à escassez de estudos biométricos sobre o assunto, os achados deste trabalho podem servir como valores de referência para a faixa etária de novilhas avaliadas e contribuir em pesquisas morfológicas futuras / Abstract: In the present work anatomic and biometric studies of the distal aspect in the flexor aspect of digits were carried out for 20 cross-breed heifer of Nelore breed aging around 24-36 months raised in an extensive regiment. The materials came from shamble under the Official Fiscalization of Inspect Service and were kept under freezing at -18°C until dissection. After the clipper and cleaning the distal areas, from metacarpus to metatarsus, the structures were dissected and measured by using a universal caliper rule (mm) on length (proximal-distal), widht (abaxial-axial) and thickness (dorso-palmar or dorso-plantar). The dissected structures included the paradigits distal accessories ligaments, palmar and plantar annular ligaments and tendon digital sheath, proximal and distal annular ligaments, interdigital distal ligament, superficial digital flexor tendon and deep digital flexor tendon and manica flexoria. The measured data indicate differences (p≤0,05) among the limb, right, left and respective digits mainly in the more distal analyzed structures. Due to scarcity of biometric studies about this topic, the data found in this work might be useful as a reference for the analyzed group, as well in the anatomic detailed structures which are essential to understanding many of the pathological process of locomotor apparatus in cattle / Orientador: José Wanderley Cattelan / Coorientador: Silvana Martinez Baraldi Artoni / Banca: Julio Carlos Canola / Banca: Claudia Acosta Duarte / Mestre

Flexible fillers has recently been implemented as corrosion protection for post-tensioning tendons used in bridge structures in Florida. There are two different explanations why corrosion could take place: 1. water is able to reach the steel 2. Microbiologica l ly Influenced Corrosion. The aim of this research is to evaluate corrosion protection effectiveness of five differe nt microcrystalline waxes under different environmental conditions. Specimens tested ranged from 7-wire steel strands to single wires (12-16 cm). Another aim is the appraisal of wax degradation by fungi species. Single wires coated with each of the investigated protection materials, were sprayed with suspensions of three different fungi species and a mix of them. For single wires, independent of the environmental condition the specimen with more corrosion was Nontribos, as well as the filler coated wires contaminated with Fungi. Fungi species investigated were able to utilize the waxes as carbon source and caused differe nt extents of MIC. / Includes bibliography. / Thesis (M.S.)--Florida Atlantic University, 2017. / FAU Electronic Theses and Dissertations Collection

O presente trabalho aborda a utilização da protensão não aderente em edifícios residenciais e comerciais de concreto, focando os aspectos referentes às soluções com protensão aderente e não aderente e em concreto armado, para o consumo de materiais, notadamente: concreto, fôrmas e armaduras. São considerados três diferentes sistemas estruturais, sendo estes: laje plana maciça apoiada sobre pilares; laje plana nervurada apoiada sobre pilares e laje nervurada apoiada em vigas faixa protendidas sobre pilares. São apresentados estudos destes casos com a utilização da ferramenta computacional de análise estrutural TQS, a qual é baseada na análise por grelha. São comparados os índices de consumo de materiais para os dois referidos sistemas de protensão, discutindo os limites de sua utilização. Como resultados, o estudo fornece conclusões satisfatórias para utilização da protensão, tanto aderente quanto não aderente, em relação ao concreto armado. Ainda, o sistema de protensão aderente mostrou-se ligeiramente mais econômico, do ponto de vista de consumo de materiais, porém, é um sistema com produtividade inferior às soluções com protensão não aderente, tornando esta última solução, a mais adotada no cotidiano dos escritórios de cálculo de engenharia civil dentre as citadas / The present work deals the use of the prestressed unbonded tendons in residential and commercial concrete buildings, pointing out aspects to the application with bonded and unbonded prestressing and reinforced concrete, regarding the consumption such as concrete, molds and reinforcing steel. Three different structural systems are considered: flat plate and columns; waffle slab and columns and waffle slab (non prestressed) in prestressed strip beams supported by columns. Case studies are presented with the use of the software of structural analysis TQS, which is based on the grillage analogy method. The consumption of materials is compared for the two referred prestressing systems, discussing the limits of their application. Based on the developed analyses, the study supplies satisfactory conclusions for use of the prestressing systems (bonded and unbonded), compared to reinforced concrete systems. With relation to the consumption of materials, the use of bonded tendons is a bit more cost effective, however, with smaller productivity than tendons with prestressed unbonded tendons, being the last one the more usual prestressing systems. Still, the system of prestressed bonded tendons was shown more economical, of the point of view of materials consumption, however, it is a system with inferior productivity to the solutions with prestressed unbonded tendons adopted in civil engineering offices nowadays

Un des problèmes majeurs contribuant à la réduction de la mobilité chez la personne âgée est la hausse de l’occurrence des chutes. La capacité à maintenir l’équilibre ou la stabilité posturale a été précédemment associée à la structure et aux propriétés mécaniques des tendons du membre inférieur. Cette étude fut menée afin d’évaluer les effets de l’intensité d’entrainement et de l’âge sur les changements de l’architecture tendineuse et ses propriétés mécaniques ainsi que sur les adaptations musculaires du membre inférieur. Ce projet avait ainsi pour objectif de comparer les effets de deux conditions d’entrainement pour un volume équivalent (intensité modérée (55% d’une répétition maximale (1RM) vs élevée (80% de 1RM)) sur deux groupes musculaires différents (quadriceps vs triceps sural), sur les adaptations des tendons d’Achille et patellaire associés aux adaptations de ces groupes musculaires respectifs. Enfin, le dernier objectif de cette étude était de montrer si des changements de la balance posturale et de la capacité de mouvement pouvaient s’expliquer par les évolutions de l’architecturale et de propriétés mécaniques des structures musculaires et tendineuses avec l’âge. Dix hommes jeunes (Age : 24.8 ± 3.6) et 27 séniors (Age : 69.9 ± 4.5) sédentaires ont été recrutés et ont participé à un programme d’entrainement en résistance de 12 semaines (3 fois/semaine) sur les muscles du triceps sural et du quadriceps. Le groupe de jeunes (n=10) ainsi qu’un groupe de séniors (n=13) ont participé à un programme d’entrainement modéré correspondant à 55% de 1RM, tandis qu’un deuxième groupe de seniors s’est vu imposer une intensité d’entrainement de 80% de 1RM (n=14). Chaque groupe a reçu exactement le même volume d'entraînement sur les muscles quadriceps et triceps sural en utilisant des machines de musculation guidées : la presse à jambes, l'extension des jambes et la machine à mollets assis. Afin de pouvoir obtenir les paramètres nécessaires à cette étude, l’utilisation d’ergomètres, d’images échographiques et IRM et d’un système de capture de mouvement ont été nécessaires. En comparant deux populations de jeunes et de séniors, cette étude a ainsi permis de quantifier une diminution de la force, couplée ou non suivant le tendon considéré à une diminution des propriétés intrinsèques du matériau tendineux. L’obtention de l’architecture musculaire a permis de construire les courbes d’évolutions de la section de chacun des muscles du quadriceps et du triceps sural pour les populations jeunes et séniors. Les deux conditions d’entrainement nous ont permis de mettre en évidence une amélioration des propriétés mécaniques des tendons d’Achille et patellaire, et plus sensiblement le tendon d’Achille, sur les deux populations jeunes et séniors sans toutefois observer de gain supplémentaire pour une intensité élevée. Des gains similaires suite à la période d’entrainement ont pu être observés chez les séniors sur les volumes des muscles du triceps sural et du quadriceps sans distinction de l’intensité considérée. L’analyse du mouvement nous a permis de mettre en évidence l’amélioration de la stabilité posturale et une évolution de la stratégie de flexion du tronc lors d’un lever de chaise suite à l’entrainement chez les séniors sans bénéfice supplémentaire entre une intensité modérée et élevée. De plus, les effets de l’âge sur les propriétés mécaniques des tendons ont pu être corrélés avec les performances liées aux exercices de stabilité posturale, de saut et de lever de chaise. Ce travail a donc permis de quantifier les effets de l’âge sur les capacités musculaires, tendineuses et de mouvement. Cette étude nous a également permis de mettre en évidence un seuil d’intensité d’entrainement (55% de 1RM) à partir duquel les personnes âgées ne semblent pas montrer de gain additionnel pour les systèmes musculaires et tendineux. Ce travail permet donc de proposer une optimisation de l’activité physique prescrite à la personne âgée ou vieillissante. / The ability to maintain balance has previously been associated with the structure and mechanical properties of the tendons of the lower limb. In order to evaluate the effects of training intensity (moderate vs. high intensity) and age on changes in tendon architecture (Achilles and patellar) and its mechanical properties, 10 young men and 27 sedentary seniors participated in a 12-week resistance training program (3 times/week) on the muscles of the triceps surae (TS) an quadriceps (QF). The young group and a senior group participated in a moderate training program corresponding to 55% of IRM (maximum repetition), while a second group of seniors received a training intensity of 80% of IRM. Each group received the same volume of training on the TS and QF muscles using guided weight machines. The use of ergometers, ultrasound and MRI images and a motion capture system were required. A decrease in strength, coupled or not depending on the tendon under consideration, and a decrease in the properties of the tendon material have been quantified with age. Evolution curves of each of the QF and TS muscles for both populations were constructed. The two training conditions showed an improvement in the mechanical properties of the Achilles and patellar tendons, and more significantly the Achilles, on both populations without any additional gain for a high intensity. Similar gains after training between the two intensities could be observed in seniors on muscle volumes. The improvement of postural stability and an evolution of the strategy during a chair lift were observed in seniors without any additional benefit between the two training intensities. This made it possible to quantify the effects of age on muscle, tendon and movement abilities by highlighting a threshold of training intensity (55% of IRM) from which seniors do not seem to show any additional gain. This work therefore makes it possible to propose an optimization of the physical activity prescribed to the elderly person

肌腱損傷發生率高，並且癒合結果很不理想，因為少量的肌腱細胞缺乏有效的修復能力，僅僅通過瘢痕形成來癒合, 肌腱瘢痕癒合難以恢復原本的肌腱組織結構及力學特性。目前，國內外臨床上治療肌腱損傷的方法很多，包括藥物、物理治療、手術等，這些並不能獲得滿意的療效。因此，如何採用肌腱組織工程技術迅速、安全、有效的修復肌腱損傷已成為運動醫學領域急需解決的重要問題。 / 有研究表明，骨髓間充質幹細胞、表皮成纖維細胞、肌腱細胞和胚胎幹細胞通過肌腱組織工程技術用於肌腱修復及再生取得了不錯的療效。但是，這些來源的細胞存在分化效率低，形成畸胎瘤和異位骨化等風險。近來，有研究報導可從人、小鼠、大鼠和兔的肌腱組織中分離培養出幹細胞，可作為肌腱組織工程種子細胞的一種新選擇，用於肌腱修復和再生。對於間充質幹細胞的成肌腱分化，有研究報導結締組織生長因子（CTGF）和抗壞血酸（維生素C的一種形式）在膠原及細胞外基質合成、調節細胞成肌腱分化方面扮演者重要的角色。 / 本研究的旨在：（1）在大鼠髕腱損傷模型中，證實肌腱幹細胞可作為一種新的幹細胞來源用於肌腱修復；（2）檢驗結締組織生長因子和抗壞血酸能在體外促進肌腱幹細胞的成肌腱分化；（3）嘗試通過肌腱幹細胞的成肌腱分化過程在體外構建不含外源性支架的肌腱樣組織；（4）探索該肌腱樣組織在大鼠髕腱損傷模型中是否可以促進肌腱癒合。 / 在大鼠急性髕腱損傷動物模型中，與對照組相比，肌腱幹細胞組具有更好的膠原排列，顯著增高的最大張力和楊氏模量，表明肌腱幹細胞可作為一種新的幹細胞來源用於肌腱損傷的修復。結締組織生長因子和抗壞血酸體外誘導肌腱幹細胞2周後，可顯著增加Tenomodulin, Scleraxis, Thbs4, I型膠原等肌腱相關基因的表達以及膠原蛋白的合成，說明結締組織生長因子和抗壞血酸可促進肌腱幹細胞的成肌腱分化。被結締組織生長因子和抗壞血酸誘導兩周後，肌腱幹細胞可形成了細胞膜樣結構，將這種細胞膜纏繞在迴紋針上，構建成肌腱樣組織，其具有相對疏鬆的細胞外基質和雜亂排列其中的肌腱幹細胞，以及表達Tenomodulin，I型膠原和III型膠原。將該肌腱樣組織移植到裸鼠體內8周和12周可形成新生肌腱組織，梭形細胞縱行分佈在平行的膠原纖維之間，並表達Tenomodulin，I型膠原和III型膠原蛋白。在大鼠髕腱損傷動物模型中，與對照組相比較，該肌腱樣組織可通過恢復肌腱組織結構及生物力學特性來促進肌腱癒合。 / 總的來說，本研究證實肌腱幹細胞可作為一種新的幹細胞來源用於肌腱組織工程促進肌腱再生。結締組織生長因子和抗壞血酸可調控肌腱幹細胞的成肌腱分化，並形成細胞膜結構。該細胞膜結構可在體外構建出不含外源性支架的肌腱樣組織，進而在裸鼠體內形成新生肌腱，並且在大鼠髕腱損傷模型中可有效的促進損傷肌腱的癒合。這種不含外源性支架的肌腱樣組織有希望成為肌腱組織工程技術的新手段，在肌腱再生和肌腱修復的臨床應用及基礎研究方面有廣泛的前景。 / Tendon injuries are common and tendon healing outcome is poor, because tendon contains few cells with limited capacities for self-repair/regeneration. The current treatments on tendon injuries including drugs, physiotherapy, and surgery are not ideal and there is a need for the development of novel tissue-engineering strategies for tendon repair. / Previous studies have shown positive effects of bone marrow-derived mesenchymal stem cells (BMSCs), dermal fibroblast, tenocytes, and embryonic stem cells-derived MSCs for tendon repair/regeneration. However, these cells have limitations including insufficient differentiation; risk of teratoma and ectopic bone formation etc. Recently, stem cells have been isolated from tendons of human, mouse, rat and rabbit and considered as a new alternative cell source for tendon tissue engineering (TDSCs). For tenogenic differention of MSCs, connective tissue growth factor (CTGF) and ascorbic acid (one form of vitamin C) are reported to play important roles in promoting collagen and other extracellular matrixes (ECM) production, and regulating the MSCs differentiation towards tenogenic pathway. / The aims of the current study are: (1) To investigate the use of TDSCs in tendon repair in a rat acute patellar tendon injury model; (2) To test the effects of CTGF and ascorbic acid on tenogenic differentiation of TDSCs in vitro; (3) To construct scaffold-free tendon-like tissues in vitro using tenogenically differentiated TDSCs; (4) To promote tendon healing by engineered tendon-like tissues in a rat acute patellar tendon injury model. / In the rat acute patellar tendon injury model, in contract to control group, TDSCs treated group showed better alignment of collagen fibers and the significantly higher ultimate stress and Young’s modulus, indicating TDSCs may be an alternative cell source for tendon repair. The effects of CTGF and ascorbic acid on tenogenic differentiation of TDSCs were also confirmed with higher expression of tendon related markers such as Tenomodulin, Scleraxis, Thbs4, Type I Collagen, etc; with higher production of collagenous proteins. After treatment with CTGF and ascorbic acid for 2 weeks, TDSCs can form cell sheets, which can be harvested, rolled up on a U-shaped spring to form tendon-like tissues in culture, which had loose extracellular matrices and randomly distributed TDSCs and also expressed Tenomodulin, Type I & III collagen. Following transplantation of the engineered tendon-like tissue in nude mice for 8 and 12 weeks, neo-tendon tissues were formed, with thin and parallel collagen fibrils and extracellular matrices of Tenomodulin, Type I & III collagen. Finally in the rat patellar tendon window injury model, data suggested that the engineered tendon-like tissue could promote tendon healing with significantly improved histological features and biomechanical properties comparing to the control group. / In conclusion, our study has indicated that TDSCs can be an alternative cell source in tendon tissue engineering for tendon regeneration. The tenogenic differentiation of TDSCs, induced by CTGF and ascorbic acid in vitro, produces cell sheets, which can be constructed tendon-like tissues in vitro; to form neo-tendon and repair tendon injuries in vivo. The use of engineered scaffold-free tendon tissue for tendon tissue engineering has potentials in clinical application for tendon repair/regeneration. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Ni, Ming. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 107-126). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / DEDICATION --- p.I / ACKNOWLEDGEMENT --- p.II-III / TABLE OF CONTENTS --- p.IV-IX / PUBLICATIONS --- p.X-XII / ABBREVIATION --- p.XIII-XV / ABSTRACT (ENGLISH) --- p.XVI-XVIII / ABSTRACT (CHINESE) --- p.XIX-XX / Chapter CHAPTER 1 --- Introduction --- p.1 / Chapter 1.1 --- Epidemiology of tendon injury --- p.1 / Chapter 1.2 --- Healing process of tendon injury --- p.1 / Chapter 1.3 --- Tendon tissue engineering for tendon repair --- p.2 / Chapter 1.4 --- Stem cells in tendon repair --- p.2 / Chapter 1.5 --- Tenogenic differentiation of tendon derived stem cells --- p.7 / Chapter 1.6 --- Growth factors for tenogenic differentiation --- p.8 / Chapter 1.7 --- Vitamin C for tenogenic differentiation --- p.9 / Chapter 1.8 --- Summary --- p.10 / Chapter CHAPTER 2 --- Hypothesis, Objectives and Study Design --- p.11 / Chapter 2.1 --- Hypothesis --- p.11 / Chapter 2.1.1 --- Overall hypothesis --- p.11 / Chapter 2.1.2 --- Specific hypothesis --- p.11 / Chapter 2.2 --- Objectives --- p.12 / Chapter 2.3 --- Study design --- p.12 / Chapter 2.3.1 --- Study I --- p.12 / Chapter 2.3.2 --- Study II --- p.14 / Chapter 2.3.3 --- Study III --- p.14 / Chapter 2.3.4 --- Study IV --- p.17 / Chapter CHAPTER 3 --- Tendon-derived Stem Cells (TDSCs): A New Cell Source for Tendon Repair (Study I) --- p.19 / Chapter 3.1 --- Materials and Methods --- p.19 / Chapter 3.1.1 --- Isolation and characterization of rat GFP-TDSCs --- p.19 / Chapter 3.1.2 --- Animal surgery --- p.20 / Chapter 3.1.3 --- Ultrasound imaging --- p.25 / Chapter 3.1.4 --- Histology --- p.27 / Chapter 3.1.5 --- Biomechanical test --- p.27 / Chapter 3.1.6 --- Ex vivo fluorescence imaging --- p.28 / Chapter 3.1.7 --- Data analysis --- p.29 / Chapter 3.2 --- Results --- p.29 / Chapter 3.2.1 --- Gross observation of the injured knee and patellar tendon --- p.29 / Chapter 3.2.2 --- Histology of regenerated tendon tissue --- p.30 / Chapter 3.2.3 --- Biomechanical test of regenerated tendon tissue --- p.32 / Chapter 3.2.4 --- Ex vivo fluorescence imaging of GFP-TDSCs --- p.33 / Chapter 3.2.5 --- Ultrasound imaging of wound gap volume --- p.34 / Chapter 3.3 --- Discussion --- p.35 / Chapter 3.4 --- Conclusion --- p.50 / Chapter CHAPTER 4 --- Tenogenic Differentiation of Tendon-derived Stem Cells (TDSCs) (Study II) --- p.51 / Chapter 4.1 --- Materials and Methods --- p.51 / Chapter 4.1.1 --- Tenogenic differentiation of tendon-derived stem cells (TDSCs) --- p.51 / Chapter 4.1.2 --- Quantification of collagenous proteins --- p.51 / Chapter 4.1.3 --- Quantitative Real Time PCR (qRT-PCR) --- p.52 / Chapter 4.1.4 --- Data analysis --- p.54 / Chapter 4.2 --- Results --- p.55 / Chapter 4.2.1 --- Quantification of collagenous proteins --- p.55 / Chapter 4.2.2 --- Tenogenic, osteogenic and chondrogenic markers mRNA expression --- p.57 / Chapter 4.2.3 --- Tendon extracellular matrix markers mRNA expression --- p.57 / Chapter 4.3 --- Discussion --- p.59 / Chapter 4.4 --- Conclusion --- p.66 / Chapter CHAPTER 5 --- Engineered Scaffold-free Tendon Tissue Produced by Tendon-derived Stem Cells (TDSCs) Cell Sheet (Study III) --- p.67 / Chapter 5.1 --- Materials and Methods --- p.67 / Chapter 5.1.1 --- In vitro engineered scaffold-free tendon tissue by TDSCs cell sheet --- p.67 / Chapter 5.1.2 --- In vivo neo-tendon formation using engineered scaffold-free tendon tissue in nude mouse model --- p.67 / Chapter 5.1.3 --- Histology and immunohistochemistry staining --- p.68 / Chapter 5.1.4 --- In vivo fluorescence imaging --- p.69 / Chapter 5.1.5 --- Data analysis --- p.70 / Chapter 5.2 --- Results --- p.70 / Chapter 5.2.1 --- Gross observation of TDSCs cell sheet and engineered scaffold-free tendon tissue --- p.70 / Chapter 5.2.2 --- Histological and immunohistochemical characteristics in engineered scaffold-free tendon tissue --- p.71 / Chapter 5.2.3 --- Gross observation and in vivo fluorescence imaging of neo-tendon tissue --- p.74 / Chapter 5.2.4 --- Histology of neo-tendon tissue --- p.75 / Chapter 5.2.5 --- Immunohistochemistry staining in neo-tendon tissue --- p.76 / Chapter 5.3 --- Discussion --- p.78 / Chapter 5.4 --- Conclusion --- p.82 / Chapter CHAPTER 6 --- Use of Engineered Scaffold-free Tendon Tissue for Tendon Repair (Study IV) --- p.83 / Chapter 6.1 --- Materials and methods --- p.83 / Chapter 6.1.1 --- Animal surgery --- p.83 / Chapter 6.1.2 --- Ex vivo fluorescence imaging --- p.84 / Chapter 6.1.3 --- Histology and immunohistochemistry staining --- p.85 / Chapter 6.1.4 --- Biomechanical test --- p.86 / Chapter 6.1.5 --- Ultrasound imaging --- p.87 / Chapter 6.1.6 --- Data Analysis --- p.87 / Chapter 6.2 --- Results --- p.88 / Chapter 6.2.1 --- Gross observation of the injured knee and patellar tendon --- p.88 / Chapter 6.2.2 --- Histology of regenerated tendon tissue --- p.89 / Chapter 6.2.3 --- Tendon specific and ECM markers expression in regenerated tendon tissue --- p.91 / Chapter 6.2.4 --- Osteogenic and chondrogenic specific markers expression in neo-tendon tissue --- p.93 / Chapter 6.2.5 --- The fate of the transplanted engineered scaffold-free tendon tissue --- p.93 / Chapter 6.2.6 --- Biomechanical test of regenerated tendon tissues --- p.94 / Chapter 6.3 --- Discussion --- p.96 / Chapter 6.4 --- Conclusion --- p.102 / Chapter CHAPTER 7 --- General Conclusions --- p.103 / Chapter 7.1 --- General discussion --- p.103 / Chapter 7.2 --- General conclusions --- p.105 / FUNDING --- p.106 / REFERENCES --- p.107 / APPENDIX --- p.127

Functional integration of bone with soft tissues such as tendon is essential for joint motion and musculoskeletal function. This is evident in the rotator cuff of the shoulder, which consists of four muscles and their associated tendons that connect the humerus and scapula. The cuff functions to stabilize the shoulder joint, and actively controls shoulder kinematics. Rotator cuff injuries often occur as a result of tendon avulsion at the tendon-bone interface, with more than 250,000 cuff repair surgeries performed annually in the United States. However, these procedures are associated with a high failure rate, as re-tears often occur due to the lack of biological fixation of the tendon to bone post-surgery. Instead of regenerating the tendon-bone interface, current repair techniques and augmentation grafts focus on improving the load bearing capability of the repaired rotator cuff. Biologically, the supraspinatus tendon inserts into bone via a biphasic fibrocartilaginous transition, exhibiting region-dependent changes in its compositional, structural and mechanical properties, which enables efficient load transfer from tendon to bone as well as multi-tissue homeostasis. Inspired by the native tendon-bone interface, we have designed and evaluated a biomimetic bilayer scaffold, comprised of electrospun poly (lactide-co-glycolide) (PLGA) nanofibers seamlessly integrated with PLGA-hydroxyapatite (HA) fibers, in order to engineer tendon-bone integration. The objective of this thesis is to explore the key design parameters that are critical for integrative tendon-bone repair using this biphasic scaffold as a model. Specifically, intrinsic to the scaffold, effects of fiber alignment, fiber diameter, mineral distribution, and polymer composition on integrative rotator cuff tendon-bone healing were evaluated in vivo using a rat model. Results indicated that an aligned, nanofiber-based scaffold with a distinct order of non-mineralized and mineralized regions will lead to insertion regeneration and integrative tendon-bone repair. Additional tissue engineering design parameters such as healing time and animal model were also tested. It was observed that the biphasic scaffold exhibited a stable long term response, as the mechanical properties of rat shoulders repaired by this scaffold remained comparable to that of the control at 20 weeks post-surgery. This scaffold was also evaluated in a large animal model (sheep), in which a clinically-relevant rotator cuff repair procedure was implemented with the biphasic scaffold. Results demonstrated the scaffold lead to integrative rotator cuff repair through the regeneration of the enthesis in both small and large animal models. In summary, through a series of in vivo studies, the work of this thesis has identified the critical tissue engineering parameters for integrative and functional rotator cuff tendon repair. More importantly, the design principles elucidated here are anticipated to have a broader impact in the field of tissue engineering, as they can be readily applied towards the regeneration of other soft-hard tissue interfaces.

肌腱修復一直是一個難題，因為依靠現在的治療很難將肌腱功能恢復到正常水平，近年來肌腱幹細胞的分離和發現為肌腱修復提供了新的策略。但是在利用肌腱幹細胞修復肌腱之前，我們應該瞭解肌腱幹細胞的哪些方面呢？ / 不同來源的成體幹細胞雖然具備相似的幹細胞特性，但是他們仍然具有組織特異性和功能的差異。這就意味選擇合適的細胞來源對於肌腱再生和肌腱組織工程有特殊意義。所以我們認為與骨髓間充質幹細胞相比，肌腱幹細胞具備特殊的幹細胞特性。迄今為止，還沒有研究比較肌腱幹細胞和骨髓間充質幹細胞的幹細胞特性。臨床應用要求幹細胞在體外增殖培養，體外的微環境也會影響幹細胞的幹性和治療潛能，所以我們還並不清楚肌腱幹細胞的幹性在體外培養中維持多久。成功的幹細胞治療需要深入理解組織特異性幹細胞的體內特徵和他們在組織修復中的作用。肌腱幹細胞的体内特徵还有没详细研究过，而且也不知道這些內源性幹細胞是否參與肌腱修復。 / 所以為了更好地利用肌腱幹細胞進行肌腱修復，本研究的總體目標是比較肌腱幹細胞和骨髓間充質幹細胞的幹細胞特性，同時從臨床角度考慮研究肌腱幹細胞體外幹性的維持。進一步研究鑒定肌腱幹細胞的體內特徵，並且探索他們在肌腱癒合中的作用。本研究將會探討我們應該瞭解關於肌腱幹細胞的體內和體外特性。 / 在第一部分研究中， 我們從同一隻GFP大鼠中分離出肌腱幹細胞和骨髓間充質幹細胞。經過比較，我們發現肌腱幹細胞与骨髓間充質幹細胞相比具备更高的克隆形成能力，增殖速度，更強的多向分化能力和更高的肌腱相关的基因表达。所以肌腱幹細胞表現出更好的幹性，可能是比骨髓间充质干细胞更好的用于肌腱再生的细胞来源。 / 在第二部分研究中，我們發現肌腱幹細胞伴隨體外傳代培養細胞衰老β-半乳糖苷酶活性增高，而同時間充質幹細胞標誌物和多向分化能力降低，所以研究人員和臨床醫生在利用肌腱幹細胞進行組織工程時需要考慮在體外傳代培養中他們的幹性的變化。 / 在第三部分研究中，IdU標記滯留細胞方法用於在體內標記幹細胞。我們發現休眠的幹細胞以IdU標記滯留細胞的形式存在於肌腱中，相比肌腱本體更多標記滯留細胞位於和肌腱腱鞘和肌腱骨結合部位。其中我們發現在肌腱腱鞘中的標記滯留細胞位於血管周圍的微環境血管，所以血管周圍的微環境可能是肌腱幹細胞來源之一。肌腱損傷后，位於損傷區域的標記滯留細胞的數量，增殖標誌物，肌腱相關標誌物， 多能性標誌物，和微血管相關標誌物都有明顯增加，意味著標記滯留細胞可能通過遷移，增殖和分化參與肌腱修復。 / 綜上所述，我們的結果為理解肌腱幹細胞的體外幹性特徵和在體外培養中的幹性變化以及体内肌腱幹細胞的鑒定提供了新的解釋，這有利于未來促進肌腱幹細胞的組織工程應用於肌腱修復。 / Tendon repair remains a great challenge due to current therapies cannot restore normal tendon function. Tendon-derived stem cells (TDSCs) have been isolated from tendon tissues and characterized in vitro in recent studies and provide new strategies for tendon repair. But what should we know about tendon stem cells before we use them to repair injured tendon? / Although stem cells that originate from different tissues share some common stem cell characteristics, they might also exhibit some tissue unique properties and hence functional differences. Therefore, we hypothesized that TDSCs have unique stemness properties compared with bone marrow-derived stem cells (BMSCs). There has been no study to compare the stemness properties of TDSCs and BMSCs. Clinical applications often require the in vitro expansion of stem cells. In vitro microenvironment also affects the stemness properties and therapeutic potential of stem cells. It is not clear if the stemness properties of TDSCs can be maintained and how long that they can be preserved during in vitro expansion. Moreover, successful stem cell-based repair therapies will require an understanding of tissue specific stem cells in vivo and their roles in the tissue repair. Tendon stem cells have not been described in details in vivo and it is unknown whether these endogenous stem cells participate in the tendon healing. / Therefore, in order to better make use of TDSCs for tendon repair, the objective of this study is to characterize the stemness properties of TDSCs compared with BMSCs and also to investigate the stemness limitation of TDSCs during culture in vitro for clinical use purpose. Furthermore, this study aims to identify the putative tendon stem cells in vivo and their role in tendon healing. This study would tell how much we should know about tendon stem cells in vitro and in vivo. / In the first part of the study, TDSCs and BMSCs were isolated from the same GFP Sprague-Dawley rat. TDSCs showed higher mensenchymal and pluripotent stem cell makers; clonogenicity; proliferative capacity; and tenogenic, osteogenic, chondrogenic, and adipogenic differentiation markers and multi-lineage differentiation potential than BMSCs. Compared with BMSCs, TDSCs shows great stemness properties and might be an alternative cell source for tendon regeneration. / In the second part of this study, the senescence-associated β-galactosidase activity of TDSCs increased while their stem cell-related marker expression and the multi-lineage differentiation potential decreased during in vitro passaging. It suggests that researchers and clinicians need to consider the changes of stemness properties of TDSCs when multiplying them in vitro for tissue engineering. / In the third part of the study, IdU label-retaining method was used for the labeling of stem cells in vivo. We have identified quiescent stem cells as IdU label retaining cells (LRCs) at the peritenon, tendon mid-substance and tendon-bone junction. More LRCs were found at the peri-tenon and tendon-bone junction compared to the mid-substance. Some LRCs could be identified in the peri-vascular niche in the peri-tenon, suggesting that peri-vascular niche is one source of tendon stem cells. After injury, The LRC number and the expression of proliferative, tendon-related, pluripotency and pericyte-related markers in LRCs in the window wound increased, indicating that LRCs might be involved in tendon repair via cell migration, proliferation and differentiation. / In conclusion, our results have provided new findings about the understanding of tendon-derived stem cells including their stemness properties and their changes during the in vitro culture, as well as in vivo identity of tendon stem cells, which might facilitate the application of TDSCs in tissue engineering for tendon repair in the future. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Tan, Qi. / Thesis (Ph.D.) Chinese University of Hong Kong, 2014. / Includes bibliographical references (leaves 130-162). / Abstracts also in Chinese.

by Wang Wen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 113-124). / Abstracts in English and Chinese. / ABSTRACT --- p.i / FLOWCHART --- p.vi / ACKNOWLEDGEMENT --- p.x / ABBREVIATIONS --- p.xi / INDEX FOR FIGURES --- p.xii / INDEX FOR TABLES --- p.xv / TABLE OF CONTENTS --- p.xvi / Chapter 1. --- INTRODUCTION --- p.1 / Chapter 1.1 --- PATELLAR TENDINOSIS --- p.1 / Chapter 1.1.1 --- Introduction --- p.1 / Chapter 1.1.2 --- Epidemiology of Patellar Tendinosis --- p.3 / Chapter 1.1.3 --- Etiology of Patellar Tendinosis --- p.3 / Chapter 1.1.4 --- Manifestations of Patellar Tendinosis --- p.4 / Chapter 1.1.5 --- Imaging Examination on Patellar Tendinosis --- p.4 / Chapter 1.1.6 --- Clinical Diagnosis of Patellar Tendinosis --- p.6 / Chapter 1.1.7 --- Management of Patellar Tendinosis … --- p.6 / Chapter 1.2 --- ANATOMY AND HISTOLOGY OF PATELLAR TCNDON --- p.7 / Chapter 1.3 --- STRUCTURE AND METABOLISM OF TENDON --- p.9 / Chapter 1.3.1 --- Tenocytes --- p.9 / Chapter 1.3.2 --- Extra-cellular Matrix --- p.11 / Chapter 1.3.2.1 --- Collagen --- p.11 / Chapter 1.3.2.2 --- Proteoglycans --- p.12 / Chapter 1.4 --- ROLES OF GROWTH FACTORS TENDON HEALING AND REPAIR --- p.14 / Chapter 1.4.1 --- Platelet-Derived Growth Factor --- p.14 / Chapter 1.4.2 --- Transforming Growth Factor-beta --- p.15 / Chapter 1.5 --- HISTOPATHOLOGY OF PATELLAR TENDINOSIS --- p.16 / Chapter 1.6 --- STUDY PLAN --- p.17 / Chapter 1.6.1 --- Characterization on Hypercellularity --- p.18 / Chapter 1.6.2 --- Characterization on Disorganization and Loosening of Collagen --- p.18 / Chapter 1.6.3 --- Characterization on Inflammatory Trace --- p.20 / Chapter 1.6.4 --- Characterization on Growth Factors in Tendinosis --- p.21 / Chapter 1.7 --- OBJECTIVES --- p.22 / Chapter 2. --- MATERIALS AND METHODS --- p.27 / Chapter 2.1 --- HUMAN TISSUES --- p.27 / Chapter 2.1.1 --- Patellar Tendinosis Tissues --- p.27 / Chapter 2.1.1.1 --- Diagnosis of patellar tendinosis --- p.27 / Chapter 2.1.1.2 --- Recruitment of patients --- p.27 / Chapter 2.1.4 --- Healthy Patellar Tendon tissues --- p.28 / Chapter 2.2 --- TISSUES COLLECTION AND PREPARATION --- p.28 / Chapter 2.3 --- HISTOLOGICAL STUDY ON HUMAN SPECIMENS --- p.28 / Chapter 2.3.1 --- Haematoxyline and Eosin Staining --- p.29 / Chapter 2.3.2 --- Safranin O Staining --- p.29 / Chapter 2.3.2.1 --- Reagents preparation --- p.29 / Chapter 2.3.2.2 --- Experimental procedure --- p.30 / Chapter 2.3.5 --- Polarization Microscopy --- p.30 / Chapter 2.4 --- IMMUNOHISTOCHEMICAL STAINING --- p.30 / Chapter 2.4.1 --- Reagents Preparation --- p.31 / Chapter 2.4.2 --- Experimental Procedure --- p.33 / Chapter 2.5 --- IMAGE ANALYSIS --- p.35 / Chapter 2.5.1 --- Equipment --- p.35 / Chapter 2.5.2 --- Procedures --- p.35 / Chapter 2.6 --- IN SITU ZYMOGRAPHY --- p.37 / Chapter 2.6.1 --- Reagents Preparation --- p.37 / Chapter 2.6.2 --- Experimental Procedure --- p.38 / Chapter 2.7 --- STATISTIC ANALYSIS.… --- p.39 / Chapter 3. --- RESULTS --- p.42 / Chapter 3.1 --- HUMAN SAMPLES --- p.42 / Chapter 3.1.1 --- Patellar tendinosis patients --- p.42 / Chapter 3.1.2 --- Healthy control group --- p.43 / Chapter 3.2 --- HISTOLOGICAL STUDY ON HUMAN SPECIMENS --- p.43 / Chapter 3.2.1 --- Gross Morphology --- p.43 / Chapter 3.2.2 --- Haematoxyline and Eosin Staining --- p.44 / Chapter 3.2.3 --- Safranin O Staining --- p.44 / Chapter 3.2.4 --- Polarization Microscopy --- p.44 / Chapter 3.3 --- IMAGE ANALYSIS --- p.45 / Chapter 3.3.1 --- Immunohistochemistry of PCNA --- p.45 / Chapter 3.3.2 --- Immunohistochemistry of hsp47 --- p.46 / Chapter 3.3.3 --- Immunohistochemistry of Procollogen Type I --- p.47 / Chapter 3.3.4 --- Immunohistochemistry of MMP1 --- p.47 / Chapter 3.3.5 --- Immunohistochemistry of TIMP1 --- p.48 / Chapter 3.3.6 --- Immunohistochemistry of COX-2 --- p.49 / Chapter 3.3.7 --- Immunohistochemistry of TGFP --- p.49 / Chapter 3.3.8 --- Immunohistochemistry of PDGFbb --- p.50 / Chapter 3.3.9 --- Immunohistochemistry of PDGFRβ --- p.51 / Chapter 3.3.10 --- Summary of Image Analysis of Immunohistochemical staining --- p.51 / Chapter 3.4 --- IN SITU ZYMOGRAPHY --- p.52 / Chapter 4. --- DISCUSSION --- p.93 / Chapter 4.1 --- DIAGNOSIS OF PATELLAR TENDINOSIS --- p.93 / Chapter 4.2 --- HYPERCELLULARITY IN PATELLAR TENDINOSIS --- p.95 / Chapter 4.3 --- COLLAGEN DISTURBANCE IN PATELLAR --- p.97 / Chapter 4.4 --- INFLAMMATORY RESPONSE IN PATELLAR TENDINOSIS --- p.100 / Chapter 4.5 --- THE EXPRESSION OF GROWTH FACTORS IN PATELLAR TENDINOSIS --- p.102 / Chapter 4.6 --- PROPOSED PATHOGENESIS FOR PATELLAR TENDINOSIS --- p.105 / Chapter 4.7 --- LIMITATION OF THIS STUDY --- p.108 / Chapter 4.8 --- FUTURE STUDY --- p.109 / Chapter 5. --- CONCLUSION --- p.111 / BIBLIOGRAPHY --- p.113

Estruturas com protensão não aderente estão sendo utilizadas como uma alternativa na tecnologia de projeto e execução de edifícios. Este trabalho apresenta a análise numérica de estruturas com protensão não aderente. Para este fim, foi desenvolvido um programa computacional onde implementou-se um modelo não linear físico e geométrico através do método dos elementos finitos. O comportamento dos materiais é descrito por um modelo elasto-viscoplástico. No concreto, são utilizados elementos finitos isoparamétricos tridimensionais. Para representar o seu comportamento após a fissuração é utilizado o modelo de fissuras distribuídas. As armaduras são incluídas através do modelo incorporado, utilizando-se de elementos unidimensionais isoparamétricos.As armaduras passivas são consideradas como uma linha de material mais rígido no interior do elemento de concreto, existindo uma aderência perfeita entre o concreto e o aço. Nas armaduras não aderentes, é considerada a compatibilidade de deslocamentos entre os materiais apenas nas ancoragens, sendo que a armadura pode movimentar-se livremente no interior do concreto. O modelo não linear geométrico, utilizado para o concreto e para a armadura, foi desenvolvido com base na formulação Lagrangeana Total, considerando grandes deslocamentos e pequenas deformações. Para verificar a precisão do modelo computacional, compararam-se resultados numéricos com valores experimentais disponíveis na literatura. / Unbonded prestressed concrete structures have been increasingly used as an alternative in the technology of design and construction of buildings. This work presents a numerical analysis of unbonded prestressed concrete structures. To accomplish this, a computational program has been developed in which a physical and geometrical nonlinear model was implemented through the finite element method. Materials behavior has been described through an elasto-viscoplastic model. In the concrete, a threedimensional isoparametric finite element has been used. To represent its behavior after cracking, the smeared cracking model has been used to. The prestressing tendons and reinforcement have been included according with the embedded model approach by the use of one-dimensional isoparametric elements. The reinforcement has been considered in the model as a line of a stiffer material inside the concrete element, with a perfect bonding between concrete and steel. As for the unbonded tendons, displacement compatibility between materials has been considered only at the anchorages, but they are allowed to move freely along their length inside the concrete. The geometric nonlinear model that has been used for the concrete, reinforcement and tendons has been developed according to the Total Lagrangean formulation, considering large displacements and small strains. In order to evaluate the accuracy of the computational model, numerical results have been compared with experimental values available in the literature.

As the incidence of diagnosed carpal tunnel syndrome continues to increase, an understanding of the mechanism(s) of insult to the median nerve which leads to its development becomes ever more imperative. Knowledge of the exact cause of CTS could lead to improved diagnostic and treatment methods, or more importantly, to better preventative measures. The goal of this study was to investigate movements and interactions of structures within the carpal tunnel during wrist flexion and hand loading in order to obtain information about a specific mechanism of insult to the median nerve. Symptomatic and normal subjects were compared to observe differences in the interactions of the median nerve and its surrounding structures. A new methodology was developed to facilitate the evaluation of these populations.