Academic literature on the topic 'Bones – Cytology'

Giant cell tumor of bone also known as osteoclastoma is a distinct clinical, roentgenographic and pathologic entity with specific characteristics. It is a benign but locally aggressive neoplasm of the epiphyseal or metaphyseal-epiphyseal region of long tubular bones extending to the articular surface. It usually occurs after skeletal maturation and is one of the rare bone tumors that more frequently affects women. We report here a case of giant cell tumor originating from the right patella diagnosed on ne needle aspiration cytology. DOI: http://dx.doi.org/10.3126/joim.v34i3.8920 Journal of Institute of Medicine, December, 2012; 34:58-60

Osteoblastoma is a rare bone tumor mostly affecting the young adults and commonly involving the spinal cord and long bones. Talus is the uncommon site of presentation, and if involved, then the neck is more commonly involved than the body of talus. The cytological diagnosis of osteoblastoma is limited, and to the best of our knowledge, its fine-needle aspiration (FNA) in the talus has still not been reported in the literature. The present case of osteoblastoma is, therefore, being reported due to the unusual presentation in elderly male in the body of talus and showing extensive involvement on X-ray. The case was initially diagnosed on FNA cytology excluding the possibility of giant-cell tumor and osteosarcoma. The case also highlights the importance of vigilant observation of subtle cytological features of this rare tumor which may be helpful in avoiding diagnostic pitfalls, especially at an uncommon site and with unusual presentation. An early precise diagnosis by cytology may be followed by appropriate treatment and thus avoiding any further complications.

Aneurysmal Bone Cyst (ABCs) is a cystic expansile lesion which is usually found in solitary locations in the metaphysis of long bones. The most common locations of ABC are long bones of lower limbs. There is little knowledge about the presence of this tumor in Clavicle. Here we present a case of a six year old male child who presented with complaint of swelling in left shoulder for four months. There was a dilemma in the diagnosis of the swelling due to its unusual presentation and appearance. The Fine Needle Aspiration Cytology was also inconclusive. The diagnosis of ABC was made only by excision biopsy. The Clavicle regenerated to nearly normal anatomy at 15 weeks follow up period. The patient is asymptomatic now and is able to perform all daily activities without any restrictions. The uniqueness of this case lies in the fact that this is the first case where ABC has been reported in Clavicular region in young Asian male of less than 10 years age. Keywords: ABC, Bone tumor, Expansile lesion.

A 16 years old girl presented with low back pain, fever and weakness in both lower limbs for 3 months. Plain radiograph shows lytic lesion in first sacral vertebra and multiple lung nodules. In suspicion of tuberculosis, antitubercular drugs were advised. But magnetic resonance imaging revealed right paraspinal soft tissue mass with multiple lesions in several vertebrae causing spinal canal compression alongwith lesions in iliac bones, sacrum, lung parenchyma and scalp tissue. Fine needle aspiration cytology and biopsy revealed Ewing’s sarcoma. Patient was referred to higher centre for chemotherapy/radiotherapy. Herein we report this rare case of extraosseous Ewing’s sarcoma with disseminated metastases masquerading as Pott’s spine. Bangladesh Journal of Medical Science Vol. 12 No. 03 July ’13 Page 334-338 DOI: http://dx.doi.org/10.3329/bjms.v12i3.15447

Adenoid cystic carcinoma (ACC) of the lacrimal gland is a rare disease, but it is the most common malignant epithelial tumor of the lacrimal gland, with poor prognosis. ACC of the lacrimal gland is notorious for its slow growth and tendency to recur despite surgical therapy and following radiotherapy and/or chemotherapy. This tumor type occasionally metastasize via haematogenous spread to lungs, brain and bone in decreasing order of frequency. When it happens, metastases are always multiple and wide. We report a case of a 28-year young man followed in our Hospital with ACC of right lacrimal gland resected on April 2014 and treated with surgery followed by post-operative radiotherapy (50Gy total dose). After one years of follow-up, Positron Emission Tomography (PET) scan reported multiple lytic lesions in distant bones specially to multiple sites of vertebral column. Fine Needle Aspiration Cytology (FNAC) confirmed metastasis of ACC. No evidence of other loco-regional or distant metastasis were described. Patient was treated with chemotherapy. After treatment, patient is actually in close follow up.Faridpur Med. Coll. J. Jul 2015;10(2): 84-86

Endometrial cancer is the most common gynecologic malignancy in the United States. However, bony metastasis is infrequent and exceptionally rare as the initial presentation. We report a case of a 77-year-old female with a clinically silent endometrial carcinoma who presented with a left tibial metastasis as the first manifestation of her disease. Ours is only the third case diagnosed by fine-needle aspiration (FNA) cytology, and the first to detail the cytomorphologic features of metastatic endometrial cancer to bone. These microscopic findings, including three-dimensional cohesive clusters with cellular overlapping and cuboidal to columnar cells exhibiting low nuclear: cytoplasmic ratios and partially vacuolated cytoplasm, differ significantly from those of endometrial carcinoma on a Papanicolaou test. The tumor bore similarity to the more commonly encountered metastatic colon cancer, but immunohistochemical staining enabled reliable distinction between these entities. A review of osseous metastases of endometrial cancer demonstrates a predilection for bones of the lower extremity and pelvis with a predominance of the endometrioid histologic subtype. In about a quarter of the cases, the bony metastasis was the first manifestation of the cancer. FNA was an effective diagnostic modality for this unusual presentation of a common malignancy. Awareness of this entity and its differential diagnosis is essential for accurate and timely diagnosis.

Aim. Out of 8 thousand species of mammals, dog is the first domesticated animal. Archaeologists studying the process of human evolution and producing excavations of ancient sites, found the bones of animals, which man ate and also bone remains of animals that lived near the sites. Wolf was the first domesticated animal of primitive tribes. This process began in the Mesolithic era 12–15 thousand years ago. Due to domestication of wolf population many indigenous, transitional and other breeds of dogs appeared. Methods. Using morphological, biochemical and genetic methods of research scientists were able to determine genome and karyotype of wild European wolf and domestic dog. Results. Domestication is a process of historical transformation of wild animals into domesticated ones. Nowadays taking into account genetics research, cytology and special experiments, researchers-cynologists came to the conclusion that only wolf is a direct ancestor of modern dogs. This is confirmed by the fact that wolf and dog have 78 chromosomes and after mating we get fertile litter. The domestication of dogs is a complicated process. While domestication of dogs to the new conditions occurred in captivity, genetic mechanisms well-known to us played an important role – mutation, recombination, hidden reserve of heredity, the direct effect of selection. Conclusions.It was proved by our research that the process of breeding in dogs which appeared as a result of domestication and microevolution of animals was accompanied by specialization and genetic differentiation of dog breeds. Кeywords: wolf, dog, domestication, karyotype, breed.

Introduction Non-small cell lung cancer presents at an advanced stage at diagnosis in two-thirds of cases. The most frequent metastatic sites are the central nervous system, adrenal glands and bones. By contrast, the thyroid gland is an extremely rare site of dissemination. Case description A 64-year-old Caucasian man previously treated with radiosurgery and brain metastasectomy followed by right middle lobectomy for a squamous cell lung carcinoma had a metachronous solitary metastasis to the thyroid gland, as confirmed by fine-needle aspiration cytology and open biopsy. He underwent curative radiotherapy, with an initial response. At 9 months’ follow-up the tumor relapsed both in the thyroid and the lung. Discussion and conclusions Review of the literature confirmed that thyroid metastasis from lung cancer is very uncommon in clinical practice. No data on the role of surgery or curative radiotherapy in thyroid metastasis are available because of the lack of prospective studies addressing the impact on survival of these treatment strategies either alone or in combination. In the case described here, radical treatment with radiotherapy allowed to obtain a modest benefit in terms of relapse-free survival. A diagnosis of metastasis to the thyroid gland should be suspected in patients who present a thyroid nodule or suggestive imaging findings when there is a history of malignancy, including lung cancer. Indeed, an early diagnosis allows to pursue radical treatment that, in selected patients, could lead to long-term survival.

Background: Bone-grafting materials are required in orthopaedic surgery to treat bone defects. Bone formation assessment is required for the development of new strategies and approaches and for quality assurance and quality control of currently available materials. Approaches to the assessment of bone formation are yet to be systematically established, quantified and standardized. Aims: the overall aim of this study was to establish a set of comprehensive quantitative approaches for the assessment of bone formation and to evaluate the role of osteoblastic cells, growth factors, and scaffolds on this process. Materials & methods: both in vitro and in vivo parameters for osteoblast phenotype and bone formation were tested in osteosarcoma cell lines, Saos-2 and U2OS cells, mesenchymal cell line, C2C12 cells, primary adipose derived stromal cells (ADSCs), platelet rich plasma (PRP), and morselized bone grafts. The in vitro parameters used were measurement of alkaline phosphatase (ALP) activity, detection of bone nodules and biomineralization, and quantification of immunocytochemistry and conventional RT-PCR of osteoblast genotyping. In vivo parameters involved ectopic bone formation in nude mice and nude rats and a tibial defect model in nude rats. Histomorphometric and quantitative immunohistochemical analyses were also performed. Results: The in vitro characterization and ectopic bone formation capabiltity of Saos-2 and U2OS cells have been established. Saos-2 cell line, which presents many osteoblast genotype and phenotype, is a stable positive control for both in vitro and in vivo bone formation assessments. The measurement of ALP activity in both solid and liquid phases has been standardized. Both the genotype and phenotype of osteoblast lineage cells has been quantitatively assessed during the capability testing of ADSCs and PRP. Quantitative assessment of new bone formation and related protein markers in vivo has been successfully established through the testing of the biological properties of gamma irradiated morselized bone grafts. Conclusion: A comprehensive knowledge of the assessment of bone regeneration and formation in vitro and in vivo has been integrated and developed through years of study. A whole set of in vitro and in vivo approaches for the assessment of bone formation has been modified and standardized to best suit the different clinical applications. This thesis provides an outline of both in vitro and in vivo bone formation assessment and their clinical applications.

Le cartilage est principalement composé de collagène de type II sécrété par les chondrocytes. Nous avons montré que l'épissage alternatif de son gène Col2a1 était régulé de manière opposée par la BMP-2 et le TGF- 1 dans des chondrocytes embryonnaires de souris cultivés sur plastique. Par ailleurs, le traitement de chondrocytes à la BMP-2 nous a conduit à identifier un gène par RT-PCR différentielle, Bat (BMP-2 activated transcript), dont l'expression est stimulée par ce facteur de croissance. L'expression de ce gène favorise le phénotype des chondrocytes en culture, mais bloque leur hypertrophie. En outre, nous avons montré que la protéine Bat, très conservée au cours de l'évolution, joue un rôle dans la mise en place de l'axe dorso-ventral au cours du développement embryonnaire chez le poisson-zèbre, Danio rerio. Ce travail aura mis en évidence deux implications de la protéine Bat : dans la chondrogenèse et dans le développement précoce

Les conséquences de l’évolution démographique vers une population de plus en plus âgée ont un impact énorme sur l’économie en termes de santé. Vieillir en bonne santé est devenu un enjeu majeur et il est devenu primordial d’instaurer des politiques de prévention. La nutrition apparait comme l’une des stratégies préventives les plus prometteuses pour contrer la perte de masse osseuse liée à l’âge. Le couple Calcium/vitamine D ayant des effets limités,il est important de développer d’autres alternatives afin de préserver le capital osseux.L’étude de l’effet du collagène hydrolysé (H C) sur le tissu osseux est devenue une thématique montante ces dernières années. Plusieurs études in vivo ont permis de démontrer que l’ingestion de collagène hydrolysé améliorait le développement de la masse osseuse. L’objectif de cette thèse a été d’étudier la pertinence de l’utilisation du H C dans la mise au point de stratégies nutritionnelles pour la prévention de l’ostéoporose.Après avoir mis en évidence l’intérêt du H C sur l’activité d’ostéoblastes murins MC3T3-E1, nous avons évalué son effet in vivo sur des souris ovariectomisées : modèle d’ostéoporose post-ménopausique liée à une carence oestrogénique. Nous avons montré que le collagène hydrolysé d’origine bovine (B H C) contribue à limiter la perte osseuse induite par l'ovariectomie. Cette étude in vivo nous a permis de constater que la limitation de la perte osseuse induite est liée à un effet spécifique du B H C et dépendante de la longueur du peptide.La deuxième partie de nos travaux a consisté à mettre en lumière le rôle des métabolites de collagène sur l’activité des cellules osseuses. Ainsi, le BHC favorise la survie et la différenciation ostéoblastique et inversement pour les ostéoclastes en modulant le ratio OPG / RANKL.En résumé, nos résultats confirment le potentiel du collagène hydrolysé dans la prise en charge de la perte osseuse liée à l’âge et soulignent l’intérêt du collagène hydrolysé dans la conception de stratégies novatrices pour le management des troubles ostéoporotiques
Due to demographic changes healthy aging has become a major issue and interest in prevention has raised accordingly. In this context, nutrition has become one of the most promising strategies to counter age-related complications, including bone loss. Effects of hydrolyzed collagen (HC) on the bone tissue have focused attention in recent years. Several in vivo studies have demonstrated that hydrolyzed collagen ingestion may preserve bone mass. The aim of this thesis was to study the relevance of HC use in the development of nutritional strategies for osteoporosis prevention. The interest of HC was first investigated on the activity of murine osteoblasts MC3T3-E1. Then, we evaluated itseffect on ovariectomized mice: a postmenopausal osteoporosis model related to estrogenic deficiency. We have shownthat bovine hydrolyzed collagen (BHC) helps reducing bone loss induced by ovariectomy and that observed effect wasspecific and not related to protein content and that it was dependent on the mean molecular weight of the peptide. The second part of our work emphasizes the role of collagen metabolites on bone cell activity. We demonstrated that enreiched serum in BHC promotes osteoblast survival and differentiation, conversely inhibits osteoclastogenesis andmodulates the OPG / RANKL ratio. In summary, our results confirm the benefit of hydrolyzed collagen on bone and point out the value of hydrolyzed collagen in the design of innovative strategies for the management of osteoporotic disorders

Bone marrow mesenchymal stem cells (BM-MSCs) are pluripotent cells that can differentiate into a variety of non-hematopoietic tissues. They also maintain healthy heamatopoiesis by providing supportive cellular microenvironment into BM. In this thesis, MSCs were characterized in terms of their morphological, immunophenotypical and differentiation properties. Then, they were examined by attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy together with hierarchical clustering, and FTIR microspectroscopy. In the first part of this study, global structural and compositional changes in BM-MSCs during beta thallasemia major (

Indiana University-Purdue University Indianapolis (IUPUI)
Communication between bone cells is important for the maintenance of bone mass. Although osteocytes are deeply embedded within the mineralized matrix, they are essential for the regulation of osteoblast and osteoclast functions. However, the intracellular proteins that control the morphology and function of osteocytes, and their ability to communicate with other bone cells are still unknown. Kalirin is a novel multi-domain GTP exchange factor (GEF) protein that activates the RhoGTPases. Recently, we found that 14 week old female Kalirin knockout (Kal-KO) mice exhibit a 45% decrease in trabecular bone density and have significantly lower cortical area, perimeter, thickness and polar cross-sectional moment of inertia (-12.6%, -7.2%, -7.6% and -21.9%, respectively) than WT mice. Kalirin was found to be expressed in osteoclasts and osteoblasts but its expression and function in osteocytes is currently unclear. We examined the role of Kalirin on the morphology and function of osteocytes. Primary osteocytes were isolated by sequential collagenase digestions from long bones (femurs and tibias) of 10-week old WT and Kal-KO mice. Immunofluorescent staining revealed Kalirin was localized to the perinuclear region of primary osteocytes and MLO-Y4 cells, and was detected along the cytoplasmic processes of primary osteocytes. We also examined primary osteocytes isolated from the long bones of Kal-KO and WT mice for changes in the length and number of cytoplasmic processes. Kal-KO osteocytes were found to express significantly fewer cytoplasmic processes per cell (3.3±0.21) than WT osteocytes (4.7±0.3). In addition, the cytoplasmic processes of Kal-KO osteocytes were shorter (79.5±4.6 µm) than those observed for WT osteocytes (85.4±3.6 µm) (p <0.01). Quantitative PCR revealed the expression of mRNA for the three major Kalirin isoforms (Kal-7, Kal-9, Kal-12) in primary osteocytes and in MLO-Y4 cells. Moreover, the mRNA levels of osteoprotegerin (OPG) and SOST, which are important for controlling osteoclast differentiation and Wnt signaling leading to bone formation, respectively, were reduced in Kal-KO osteocytes. Next, the role of Kalirin in osteocyte morphology and function was further examined. Treatment of MLO-Y4 cells for 5 days with nerve growth factor, which is known to activate Kalirin in neurons, or over-expression of the Ser-Thr kinase domain of Kal-12, promoted cytoplasmic process elongation and upregulated phosphorylated ERK and RhoA levels. Together, these results suggest that Kalirin controls osteocyte morphology and function in part by regulating cytoskeletal remodeling and the activity of ERK and RhoA. Furthermore, Kalirin may control the bone remodeling cycle by regulating osteocyte signaling to osteoclasts and osteoblasts.

前言：間充質幹細胞容易擴增並且能分化為成骨細胞、軟骨細胞和脂肪細胞，並且能對炎症、感染和損傷做出反應，並且遷移到相應的組織部位。這些特性使間充質幹細胞成為骨骼組織工程學中非常重要的細胞來源。外周血間充質幹細胞是一種存在於血液中的間充質幹細胞，而主要的間充質幹細胞存在與骨髓中，被稱之為骨髓間充質幹細胞。在我們實驗室之前的研究中通過DNA微陣列發現外周血間充質幹細胞中很多基因的表達與骨髓間充質幹細胞有很大區別。這其中的一些基因可能參與調控間充質幹細胞的分化和歸巢，我們從中挑選了三個變化比較明顯的基因--CRBP1, N-cadherin和 SOX11做進一步研究。本研究的目的在於研究CRBP1, N-cadherin和 SOX11在骨髓間充質幹細胞分化和遷移中的作用及相關機理。
方法：培養的骨髓間充質幹細胞來源於6-8周大小的SD大鼠。細胞的表型經過多分化潛能測試（成骨分化，成脂分化和成軟骨分化）和流式細胞儀檢驗。克隆大鼠的CRBP1, N-cadherin和SOX11基因到慢病毒載體。而且還設計了針對CRBP1和 N-cadherin的shRNA及非特異性對照shRNA。慢病毒由暫態轉染293FT細胞產生。細胞遷移實驗採用了BD Falcon的細胞遷移系統（cell culture insert）。實驗採用了定量PCR、免疫共沉澱、western雜交和雙螢光報告檢驗。對於體內實驗，細胞經感染帶有不同基因的病毒後，種植到Si-TCP材料並移植到裸鼠皮下。8周後，收集樣品進行組織學和免疫組織學分析。最後，我們建立了大鼠的股骨開放式骨折模型，並在4天后將SOX11基因修飾的間充質幹細胞通過心臟注射打到大鼠體內。4周後，收集股骨骨折樣品並進行microCT、力學測試和組織學分析。
結果：CRBP1過表達能夠促進骨髓間充質幹細胞的成骨分化潛能，並能抑制其成脂分化。進一步的機理研究表明CRBP1可以通過與RXRα的蛋白相互作用抑制RXRα誘導的β-catenin降解，從而維持β-catenin和磷酸化-ERK1/2在較高的水準，導致間充質幹細胞成骨能力增強；N-cadherin過表達可以促進間充質幹細胞的遷移，但是卻通過下調β-catenin和磷酸化ERK1/2抑制其成骨分化。過表達SOX11可以通過增強BMP信號通路促進三系分化。SOX11還可以通過啟動CXCR4的表達來促進細胞遷移。最後，在大鼠的股骨開放骨折模型上通過系統注射，我們證明穩定過表達SOX11的間充質幹細胞遷移到骨折部位的數量明顯增加。這些細胞到達骨折部位以後可以起始骨痂的鈣化，促進骨折的修復。
結論：本研究證明CRBP1, N-cadherin 和SOX11具有調節骨髓間充質幹細胞遷移和/或分化的功能。這些基因也許會成為幹細胞治療的新靶點。系統注射SOX11基因修飾的骨髓間充質幹細胞對於骨折修復可能具有較好的療效。本研究初步研究了CRBP1, N-cadherin 和SOX11在間充質幹細胞中的作用，為探討以間充質幹細胞為基礎的組織工程的某些新臨床應用提供了一些線索。
Introduction: Mesenchymal stem cells (MSCs) can be easily harvested, expanded, and have the capability of differentiating into osteoblasts, chondrocytes and adipocytes, and they can home to various tissues in response to stimuli such as inflammation, infection and injuries. MSCs are therefore valuable cell source for musculoskeletal tissue engineering. Peripheral blood-derived MSCs (PB-MSCs) are one kind of MSCs that reside in peripheral blood, whereas the main source of MSCs is bone marrow-derived MSCs (BM-MSCs). In our previous study, we found many genes were differentially expressed in the PB-MSCs compared to their counterpart BM-MSCs demonstrated by microarray analysis, among which the effects of CRBP1, SOX11 and N-cadherin on MSCs in terms of migration and differentiation are studied.
Methods: BM-MSCs and PB-MSCs were cultured from 6-8 weeks SD rats. The phenotypes of MSCs were characterized by tri-lineage (adipo-, osteo- and chondrogenic) differentiation and flow cytometry analysis. The genes encoding rat CRBP1, SOX11 and N-cadherin were cloned into lentiviral vectors respectively. shRNAs targeting CRBP1, N-cadherin, and one nonspecific shRNA were designed. Pseudo-lentivirus was produced by transient transfection of 293FT cells. Cell migration was examined using transwell insert culture system. Quantitative RT-PCR, CO-IP, western blot and dual-luciferase assay were employed in the studies. For in vivo study, MSCs transduced with different genes were seeded on Si-TCP scaffolds and implanted subcutaneously in nude mice. 8 weeks later, the samples were collected for histological and immunohistological analysis. Finally, an open femoral fracture model was established in 8-week old SD rats, SOX11-modified MSCs were injected at four days after fracture. At 4-week after MSCs injection, the femurs were collected for microCT, mechanical test and histological analysis.
Results: For CRBP1gene, our results showed that CRBP1 overexpression promoted osteogenic differentiation of BM-MSCs, while inhibited their adipogenic differentiation. We demonstrated that CRBP1 promoted osteogenic differentiation by inhibiting RXRα-induced β-catenin degradation through physical interactions, and maintaining β-catenin and pERK1/2 at higher levels. For N-cadherin gene, we found that N-cadherin overexpression promoted MSCs migration, and suppressed osteogenic potential of MSCs through inhibiting ERK and β-catenin signaling pathways. For SOX11 gene, we demonstrated that SOX11 overexpression enhanced the adipo-, osteo- and chondrogenic differentiation of BM-MSCs, through enhancing BMP signaling pathways. The migration capacity of BM-MSCs was also enhanced when Sox-11 was overexpressed, through activating CXCR4 expression. Finally, in the open femur fracture model we demonstrated that a larger number of SOX11-overexpressing BM-MSCs migrated to the fracture site, initiated earlier callus ossification and improved bone fracture healing quality.
Conclusions: This study demonstrated that CRBP1, N-cadherin and SOX11 gene can regulate the migration and/or differentiation potentials of BM-MSCs. These genes may become new therapeutic targets in stem cell therapy applications. Systemic administration of genetically modified SOX11-overexpressing BM-MSCs may be useful in promoting fracture healing. Overall, this study defined some unknown functions of CRBP1, N-cadherin and SOX11 in MSCs and shed the lights on some novel therapeutic implications for MSCs-based tissue engineering.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Xu, Liangliang.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2012.
Includes bibliographical references (leaves 128-144).
Abstract also in Chinese.
Declaration --- p.i
Abstract --- p.ii
摘要 --- p.v
Acknowledgements --- p.vii
Chapter 1 --- p.1
Introduction --- p.1
Chapter 1.1 --- Mesenchymal stem cells --- p.2
Chapter 1.1.1 --- Characteristics of mesenchymal stem cells --- p.2
Chapter 1.1.2 --- Bone marrow- and peripheral blood-derived MSCs --- p.4
Chapter 1.1.3 --- Other tissue-derived MSCs --- p.5
Chapter 1.2 --- Adipogenesis of MSCs --- p.6
Chapter 1.3 --- Chondrogenesis of MSCs --- p.7
Chapter 1.4 --- Osteogenesis of MSCs --- p.8
Chapter 1.4.1 --- Regulators of osteogenesis --- p.9
Chapter 1.4.2 --- Stratergies for improving bone tissue engineering --- p.11
Chapter 1.5 --- Signaling pathways involved in osteogenesis --- p.13
Chapter 1.5.1 --- ERK signaling pathway --- p.14
Chapter 1.5.2 --- Wnt signaling pathway --- p.15
Chapter 1.5.3 --- BMP signaling pathway --- p.17
Chapter 1.6 --- Migration of MSCs --- p.20
Chapter 1.7 --- Fracture healing --- p.22
Chapter 1.8 --- Clinical application of MSCs --- p.23
Chapter 1.8.1 --- BM-MSCs vs. PB-MSCs --- p.24
Chapter 1.8.2 --- Autologous vs. Allogeneic MSCs transplantation --- p.25
Chapter 1.9 --- Scope of the present study --- p.26
Chapter 1.9.1 --- CRBP1 --- p.26
Chapter 1.9.2 --- N-cadherin --- p.27
Chapter 1.9.3 --- SOX11 --- p.27
Chapter 1.10 --- Experimental scheme --- p.29
Chapter 2 --- p.31
Comparison between PB-MSCs and BM-MSCs --- p.31
Chapter 2.1 --- Chapter introduction --- p.32
Chapter 2.2 --- Materials and methods --- p.33
Chapter 2.2.1 --- Cell culture --- p.33
Chapter 2.2.2 --- Flow cytometry --- p.33
Chapter 2.2.3 --- Adipogenic differentiation --- p.34
Chapter 2.2.4 --- Osteogenic differentiation --- p.34
Chapter 2.2.5 --- RNA Extraction and Real-time PCR --- p.34
Chapter 2.3 --- Results --- p.35
Chapter 2.3.1 --- Morphology of PB-MSCs --- p.35
Chapter 2.3.2 --- Cellular surface markers of BM-MSCs and PB-MSCs --- p.36
Chapter 2.3.3 --- Multi-differentiation potential of BM-MSCs and PB-MSCs --- p.38
Chapter 2.3.4 --- Target genes expression in BM-MSCs and PB-MSCs --- p.39
Chapter 2.4 --- Discussion and future work --- p.40
Chapter 3 --- p.41
Role of CRBP1 in Differentiation and Migration of MSCs --- p.41
Chapter 3.1 --- Chapter introduction --- p.42
Chapter 3.2 --- Materials and methods --- p.46
Chapter 3.2.1 --- Chemicals --- p.46
Chapter 3.2.2 --- Isolation and culture of BM-MSCs --- p.46
Chapter 3.2.3 --- RNA Extraction and Real-time PCR --- p.47
Chapter 3.2.4 --- Plasmid construction, transfection, production of lentivirus and infection --- p.48
Chapter 3.2.5 --- Osteogenic differentiation --- p.50
Chapter 3.2.6 --- Adipogenic differentiation --- p.50
Chapter 3.2.7 --- Western blot --- p.51
Chapter 3.2.8 --- Immunofluorescence labeling and fluorescence microscopy --- p.52
Chapter 3.2.9 --- Cell migration assay --- p.52
Chapter 3.2.10 --- Ectopic bone formation assay --- p.52
Chapter 3.2.11 --- Statistical analysis --- p.53
Chapter 3.3 --- Results --- p.53
Chapter 3.3.1 --- Transducing BM-MSCs with lentivirus carrying CRBP1 or shRNAs --- p.53
Chapter 3.3.2 --- CRBP1 accelerates osteogenesis of BM-MSCs via enhancing ERK1/2 and β-catenin pathways --- p.56
Chapter 3.3.3 --- CRBP1 stabilizes β-catenin by inhibiting RXRα-induced degradation --- p.58
Chapter 3.3.4 --- CRBP1 inhibits adipogenesis of BM-MSCs --- p.61
Chapter 3.3.5 --- CRBP1 overexpression has no effect on MSCs migration potential --- p.63
Chapter 3.3.6 --- CRBP1 promotes ectopic bone formation in vivo --- p.64
Chapter 3.4 --- Discussion --- p.66
Chapter 3.5 --- Future work --- p.73
Chapter 4 --- p.74
Role of N-cadherin in Differentiation and Migration of MSCs --- p.74
Chapter 4.1 --- Chapter introduction --- p.75
Chapter 4.2 --- Materials and methods --- p.78
Chapter 4.2.1 --- Chemicals --- p.78
Chapter 4.2.2 --- Isolation and culture of BM-MSCs --- p.78
Chapter 4.2.3 --- Plasmid construction, transfection, production of lentivirus and infection --- p.79
Chapter 4.2.4 --- Osteogenic differentiation and ALP activity assay --- p.81
Chapter 4.2.5 --- Western blot --- p.81
Chapter 4.2.6 --- Ectopic bone formation assay --- p.82
Chapter 4.2.7 --- Statistical analysis --- p.82
Chapter 4.3 --- Results --- p.83
Chapter 4.3.1 --- Expression of N-cadherin during osteogenesis in MSCs --- p.83
Chapter 4.3.2 --- N-cadherin overexpression inhibits osteogenesis through suppressing β-catein and ERK1/2 signaling pathways --- p.84
Chapter 4.3.3 --- N-cadherin silencing increases osteogenesis through enhancing β-catenin and ERK1/2 signaling pathways --- p.86
Chapter 4.3.4 --- N-cadherin promotes migration of MSCs --- p.87
Chapter 4.3.5 --- Cellular surface markers of SV40-immortalized MSCs --- p.89
Chapter 4.3.6 --- N-cadherin inhibits ectopic bone formation in vivo --- p.89
Chapter 4.4 --- Discussion --- p.91
Chapter 4.5 --- Future work --- p.94
Chapter 5 --- p.96
Role of SOX11 in Differentiation and Migration of MSCs --- p.96
Chapter 5.1 --- Chapter introduction --- p.97
Chapter 5.2 --- Materials and methods --- p.105
Chapter 5.2.1 --- Plasmid construction, transfection, production of lentivirus and infection --- p.105
Chapter 5.2.2 --- Cell culture --- p.106
Chapter 5.2.3 --- Luciferase reporter gene assay --- p.106
Chapter 5.2.4 --- Osteogenic differentiation and ALP activity assay --- p.106
Chapter 5.2.5 --- Adipogenic differentiation --- p.107
Chapter 5.2.5 --- Chondrogenic diffferentiation --- p.107
Chapter 5.2.6 --- Western blot --- p.108
Chapter 5.2.7 --- RNA Extraction and Real-time PCR --- p.108
Chapter 5.2.8 --- Cell migration --- p.110
Chapter 5.2.9 --- Ectopic bone formation --- p.110
Chapter 5.2.10 --- Fracture healing model and analysis --- p.111
Chapter 5.2.11 --- Statistical Analysis --- p.112
Chapter 5.3 --- Results --- p.112
Chapter 5.3.1 --- SOX11 is upregulated during osteogenesis of BM-MSCs --- p.112
Chapter 5.3.2 --- SOX11 promotes adipogenesis in BM-MSCs --- p.113
Chapter 5.3.3 --- SOX11 promotes migration of BM-MSCs --- p.114
Chapter 5.3.4 --- SOX11 promotes osteogenesis in BM-MSCs --- p.115
Chapter 5.3.5 --- SOX11 promotes chondrogenesis of MSCs --- p.117
Chapter 5.3.6 --- Mechanisms of how SOX11 regulates differentiation and migration of MSCs --- p.118
Chapter 5.3.7 --- SOX11-modified MSCs promote bone fracture healing in an open femur fracture rat model --- p.122
Chapter 5.4 --- Discussion --- p.126
Chapter 5.5 --- Future work --- p.131
Appendix --- p.153

Meng, Fanbiao.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2013.
Includes bibliographical references (leaves 114-138).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.