INTRODUCTION: Bone marrow examination (BME) is performed in Human Immunodeficiency Virus-infected (HIV+) children with haematologic abnormalities to exclude specific disease (SD). AIMS: To describe the: (1) indications for BME , (2) utility of BME to diagnose SD, (3) patient characteristics associated with SD or non-specific disease (NSD). METHODS: Design: Retrospective review. Definitions: SD: BME positive for opportunistic infection (OI) or HIV-related malignancy. NSD: HIV-related changes only. RESULTS: Eighty six BME’s were done. Suspected SD in 56/86(65.1%) was the most common clinical indication. Bicytopaenia(n=32) and isolated cytopaenia(n=31) were the most common haematologic indications. NSD 48/86 (55.8%) was a more common finding than SD 32/86 (37.2%). Granulomas, pure red cell aplasia and malignancy were the SD identified. Pre- highly active antiretroviral therapy (HAART), advanced stage, and not being virally suppressed were significantly associated with NSD. CONCLUSION: The yield of SD (37.2%) on BME is comparable to adult studies. HAART should be instituted before BME as NSD will be the most likely finding.

Thesis (M.Sc. (Med.))--University of the Witwatersrand, Faculty of Health Sciences, 1962.

O objetivo deste trabalho foi comparar o prognóstico de sobrevida da graduação histológica após efeito da quimioterapia (graus I versus II de Huvos), visando também identificar fatores prognósticos no que diz respeito à sobrevida livre de recidiva local (SLRL), sobrevida livre de metástase (SLM) e sobrevida global (SG), em pacientes portadores de osteossarcoma primário não metastático ao diagnóstico. Vinte e quatro entre 45 pacientes admitidos no Instituto de Ortopedia e Traumatologia do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo IOT/HC/FMUSP, entre 2000 e 2004, foram eleitos para o estudo, segundo os critérios de inclusão e exclusão utilizados. As probabilidades de sobrevida acumuladas foram feitas pela técnica de Kaplan-Meier e os índices I e II de HUVOS comparados pelos testes de Log Rank. A análise multivariada foi feita pela técnica de regressão logística com modelo de risco proporcional de COX e a validade estatística estabelecida para valores de p<0,05. Os graus I e II de Huvos, quando comparados, não foram considerados de valor prognóstico em nenhuma das sobrevidas estudadas (SLRL, SLM e SG). Os fatores adversos que influenciaram o risco de recidiva local e a sobrevida global, na análise univariada foram: subtipo histológico diferente do osteoblástico (p=0,017) e o tamanho tumoral maior que 15 cm (p=0,048). Em relação à SLM o subtipo não osteoblástico (p=0,007) teve um pior prognóstico. O subtipo histológico manteve sua significância na análise multivariada em todas as sobrevidas estudadas / The purpose of this study was to compare the prognostic of survivor of histologic graduation post chemotherapy (Huvos´s grade I versus II), aiming to identify prognostic factors concerning to local recurrence free survival (LRFS), metastases free survival (MFS) and overall survival (OS) in patients with nonmetastatic primary osteosarcoma. This study included 24 patients registred in the Instituto de Ortopedia e Traumatologia do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo - Brazil, from 2000 to 2004. Survivor rates were calculed using Kaplan-Meier method. Huvos´s grade (I e II) were compared using the Log Rank test. Cox proportional hazards model was used for multifatorial analysis. Statistical significance was defined as a p value less than 0, 05. The Huvos´s grade I versus II was not significant factor for LRFS, MFS or OS. The adverse factors for LRFS and OS in univariate analysis were nonosteoblastic histologic subtypes (p=0,017) and large tumor (p=0,048). For MFS nonosteoblastic histologic subtypes (p=0,007) had worse prognostic. The histologic subtypes maintained their significance in multivariate testing on all studied survivor

Com o propósito de contribuir com os estudos do comportamento do tecido ósseo e auxiliá-los em uma situação próxima da real, que é a de comportamento dinâmico, foi construí da uma maquina de ensaio de fadiga (MEF) para corpos de prova de ossos. Utilizaram-se corpos de prova de mesma dimensão retirados de um mesmo animal (boi), no qual os corpos de prova da tíbia esquerda foram submetidos a 42.000 ciclos na máquina de ensaio de fadiga e, posteriormente, submetidos à flexão na máquina universal de ensaio e os corpos de prova da tíbia direita foram levados aos ensaios de flexão sem terem passados por fadiga. Os resultados obtidos para os CDPs, com e sem fadiga, demonstraram que as propriedades mecânicas dos CDPs tiveram sensível índice de variança, caracterizando assim a eficiência da MEF. Além disso, a MEF é capaz de executar ensaios com valores de amplitude de 0,5 mm e de 1,0 mm, bastando para isso a troca do excêntrico por um destes que já foram confeccionados. / A fatigue\'s testing machine (MEF) was built to be used in bone samples, with the purpose of giving aid and contribution to the study of bone tissue\'s behavior in a situation close to a real one, that is the dynamic behavior. Samples all with the same dimension taken from only one animal (an ox) were used. From those samples some were from the left shaft (diaphysis) and were submitted to 42.000 cycles in the fatigue\'s testing machine and after that submitted to bending test in the universal machine. Meanwhile, the right shinbone samples were submitted to bending tests without going through previous fatigue. Results were obtained from bone samples (also called CDPs) which went through fatigue and from those that didn\'t. The results showed that mechanical properties from the CDPs had a sensible variant rate, which then characterized how efficient the fatigue testing machine is. Besides, the MEF is capable of making tests with the values of amplitude of 0,5 mm and 1,0 mm, just by switching the eccentric by one of those which were already pre-fabricated.

Li, Cheuk Wun. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 121-135). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

by Pang Sai Yau. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves [87-90]). / Chapter chapter one： --- introduction / Chapter 1.1 --- General Introduction --- p.1 / Chapter 1.2 --- biology of cancellous bone grafts --- p.2 / Chapter 1.2.1 --- Biology of bone graft incorporation --- p.2 / Chapter 1.2.1.1 --- Osteogenesis --- p.2 / Chapter 1.2.1.2 --- Vascularization --- p.3 / Chapter 1.2.1.3 --- Osteoinduction --- p.3 / Chapter 1.2.1.4 --- Osteoconduction --- p.4 / Chapter 1.2.2 --- Histological changes of bone grafts after bone transplantation --- p.4 / Chapter 1.2.2.1 --- Histologic pictures of cancellous autograft --- p.4 / Chapter 1.2.2.2 --- Histologic pictures of cancellous bone allograft --- p.5 / Chapter 1.2.2.3 --- Summary of the histologic changes of bone grafts --- p.5 / Chapter 1.3 --- application of cancellous bone grafts --- p.6 / Chapter 1.3.1 --- Principles of graft incorporation --- p.6 / Chapter 1.3.1.1 --- Operative site --- p.6 / Chapter 1.3.1.2 --- Graft material --- p.7 / Chapter 1.3.1.2.1 --- Autogenic cancellous bone --- p.8 / Chapter 1.3.1.2.2 --- Autogenic cortical bone --- p.9 / Chapter 1.3.2.2.3 --- Vascularized autogenic bone grafts --- p.9 / Chapter 1.3.2.2.4 --- Bone allografts --- p.10 / Chapter 1.3.2.2.5 --- Graft adjuncts and substitutes --- p.11 / Chapter 1.3.2.3 --- Systemic factors influencing gaft incorporation --- p.13 / Chapter 1.3.2.4 --- Local factors influencing graft incorporation --- p.13 / Chapter 1.3.3 --- Bone graft complications --- p.13 / Chapter 1.3.4 --- Placement of a graft --- p.14 / Chapter 1.3.5 --- Bone graft harvesting --- p.15 / Chapter 1.3.5.1 --- Iliac bone graft --- p.15 / Chapter 1.3.5.2 --- Femoral head bone allograft --- p.16 / Chapter 1.4 --- Application of flow conductance concept in a cancellous bone graft --- p.17 / Chapter 1.4.1 --- Physical structure of cancellous bone --- p.17 / Chapter 1.4.2 --- Porosity of cancellous bone --- p.17 / Chapter 1.4.3 --- Flow conductance concept --- p.18 / Chapter chapter two： --- material and method / Chapter 2.1 --- Transplantation of cancellous bone graft - Rabbit model --- p.19 / Chapter 2.1.1 --- Preparation of porcine cancellous bone graft --- p.19 / Chapter 2.1.1.1 --- Bone drilling --- p.19 / Chapter 2.1.1.2 --- Defat and freeze-dry --- p.20 / Chapter 2.1.2 --- Flow conductance measurement --- p.21 / Chapter 2.1.2.1 --- Porosity measurement --- p.21 / Chapter 2.1.2.2 --- Conductance measurement --- p.24 / Chapter 2.1.3 --- Rabbit model --- p.26 / Chapter 2.1.4 --- Methods of assessment --- p.29 / Chapter 2.1.4.1 --- Intraosseous pressure measurement --- p.29 / Chapter 2.1.4.2 --- Histologic study --- p.30 / Chapter 2.1.4.3 --- Blood flow study - use of tracer microspheres --- p.30 / Chapter 2.2 --- Flow conductance measurement of human cancellous bone --- p.34 / Chapter chapter three： --- results / Chapter 3.1 --- Results of the effects of various conductance of the grafts on bone healing in animal model --- p.38 / Chapter 3.1.1 --- Intraosseous pressure measurement --- p.38 / Chapter 3.1.2 --- Histological study --- p.40 / Chapter 3.1.3 --- Blood flow study of cancellous bone grafts --- p.52 / Chapter 3.2 --- Human specimens study --- p.62 / Chapter chapter four： --- discussion / Chapter 4.1 --- Discussion of the results in vivo study --- p.66 / Chapter 4.1.1 --- Intraosseous pressure measurement - a baseline study --- p.66 / Chapter 4.1.2 --- Effects of flow conductance of porcine cancellous grafts on bone regeneration --- p.67 / Chapter 4.1.2.1. --- Threshold conductance --- p.67 / Chapter 4.1.2.2. --- Histological score --- p.68 / Chapter 4.1.3 --- Discussion of graft healing from the blood flow study --- p.70 / Chapter 4.1.3.1 --- Tibia blood supply in relation to bone healing --- p.70 / Chapter 4.1.3.2 --- Effect of different flow conductance on blood flow changes in the tibia-graft structure --- p.72 / Chapter 4.1.4 --- "Comparison of length, porosity and conductance as the parameter on graft healing" --- p.74 / Chapter 4.2 --- Discussion on human bone specimens study --- p.76 / Chapter 4.3 --- General discussion --- p.78 / Chapter 4.3.1 --- The limitation of the animal model --- p.78 / Chapter 4.3.2 --- Some problems related to the clinical aspects --- p.79 / Chapter chapter five： --- conclusion --- p.81

Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease which predominantly affects the synovial joints. Local gene therapy represents an approach to produce therapeuticmolecules (i.e. soluble TNF receptor (sTNFR)-Fc and interleukin-1 receptor antagonist (IL-1Ra)) directly in arthritic joints. Gene therapy could be designed to link the level of therapeutic gene expression directly to disease activity, through the use of transcriptional and posttranscriptional regulatory elements. The experiments in this thesis describe the construction of multi-responsive, composite synthetic promoters, comprised of the binding sites for an array of transcription factors activated in arthritic joints. Optimal spatial arrangements of binding sites in relation to each other and to the TATA box were determined by Assembly PCR cloning and the functionality of the resulting synthetic promoters revealed additive or synergistic induction of luciferase reporter gene expression in response to combined stimulation. Candidate synthetic promoters were cloned into a lentiviral vector between insulator elements and displayed significantly enhanced induction, in excess of 1,500 fold in response to combined stimulation. Inflammation-specific activation of lentiviral synthetic promoters was confirmed in a carrageenan-induced paw inflammation mouse model, which demonstrated the strong correlation between local luciferase gene expression and paw inflammation. Post-transcriptional gene regulation was also investigated by exploiting the differential expression of endogenous miR-23b during inflammation. Insertion of miR-23b target sites into the 3’UTR of the luciferase gene subjected luciferase mRNA to regulation by miR-23b. Experiments demonstrated that high basal gene expression driven by constitutive and inducible promoters was significantly downregulated by miR-23b without significantly impairing high gene expression upon stimulation. Overall, the experiments in this thesis have confirmed the induction of inflammation-specific gene expression, regulated by inflammationresponsive endogenous transcriptional and post-transcriptional elements.

Hydroxyapatite (HA) is a highly biocompatible calcium phosphate which closely resembles the mineral component of bone. Porous structures composed of biocompatible materials are believed to enhance fixation (and integration) as they encourage the ingrowth of bone into the implant. Consequently there is great interest in the potential of porous HA as an alternative to bone homo grafts and auto grafts. However, despite the interest in the field, there has been no thorough investigation into the physical and structural properties of porous HA and their effects on bone ingrowth. The material used in this study was a carbonated apatite (containing < 0.9 % levels of trace element impurities) with a trabecular macrostructure, that had been converted from bovine cancellous bone. The apparent density of the material ranged from 0.35 -1.45 g.cm-3 and the macrostructural morphology varied from an open equiaxed foam to a columnar honeycomb-like structure. The ultimate compressive stress was strongly related (r = 0.9) to the square of the apparent density, while compressive modulus was influenced by both apparent density and macrostructural morphology. Transmission electron microscopy of human osteoblast-like cells cultured on the material demonstrated that cells were closely associated with the surface. Specimens with densities of 0.6.0.9 and 1.2 g.cm-3 were then implanted in a lapine cancellous site for periods of 10 days, 3, 5, 13 and 26 weeks. After implantation all specimens elicited a highly biocompatible response, with active areas of bone deposition, remodelling and revascularization and no fibrous encapsulation. The amount of bone ingrowth within the implant (25-10%) after.5 weeks was found to vary with apparent density (0.6-1.2 g.cm-3) indicating that osseointegration was a function of macrostructural morphology. Pushout testing of retrieved spedmens indicated that all implants were securely fixed by 5 weeks (2-3 MPa). Compression testing demonstrated that after 5 weeks low density implants were sufficiently reinforced by bone ingrowth to equal the compressive strength of the host tissue (6 MPa) which increased to approximately 20 MPa at 3 and 6 months.

Hydroxyapatite (HA) is a biologically active ceramic used in surgery to replace bone. While HA promotes bone growth, it suffers from weak mechanical properties and does not possess any antibacterial property. Multi walled carbon nanotubes (MWCNTs), as one of the strongest and stiffest materials, have the potential to strengthen and toughen HA, thus expanding the range of clinical uses for the material. Furthermore, Silver nanoparticles (Ag NPs) can be decorated to sidewalls of the MWCNTs which could be released over a period of time to prevent infection following surgery. This work sought to develop and characterise Ag NPs- MWCNTs – HA composites in four main areas: 1) production and characterisation of the composite, 2) evaluation of mechanical properties, 3) investigation of antimicrobial property and 4) assessment of biological response to in vitro cell culture. Pristine (p-MWCNTs) and acid treated MWCNTs (f-MWCNTs) were decorated with Ag NPs. In the presence of 0.5 wt % Ag NPs-MWCNTs, HA was precipitated by the wet precipitation method in the presence of either poly vinyl alcohol (PVA) or Hexadecyl trimethyl ammonium bromide (HTAB) as the surfactant. Composites were characterised using various techniques and the diameteral tensile strength and compressive strength of the composites were measured. The antibacterial effect of these composites was investigated against clinically relevant microbe, Staphylococcus aureus. To determine the ability of the HOB cells to differentiate and mineralize in the presence of the composite, HOB cells were cultured on the composites for 21 days. Gene expression studies was performed along with the biochemical assays and scanning electron microscopy was used for qualitative analysis. Pure HA was used as control in all the studies. The study revealed that both the MWCNTs and surfactants play a crucial role in the nucleation and growth of the HA. XRD and FTIR characterisation revealed that HA was the primary phase in all the synthesised powders. Composites made with f-MWCNTs were found to have better dispersion and better interaction with the HA compared to composites with p-MWCNTs. Although mechanical strength was improved in all the composites, p-MWCNTs composites exhibiting maximum strength. Antibacterial studies showed 80% bacterial reduction in the treatment composites compared to pure HA. The biocompatibility study showed reduced activity of the HOB cells, however, no significant difference was observed between the control and the treatments. This systematic study of the synthesis and properties of the Ag NPs- MWCNTs-HA composites has resulted in improved understanding of the production and processing of these materials and the effect of MWCNTs and silver nanoparticles on primary human osteoblast cells. Additionally, it has yielded interesting biocompatibility result favouring the use of MWCNTs in the development of implants. There is potential to translate Ag NPs-MWCNTs-HA composites into clinically approved product.

Hydroxyapatite (HA) bioceramics are commercially available as bone graft substitute materials. The aim of the current research was to characterise the electrical properties of hydroxyapatite-barium titanate (HABT) composites and to assess in vitro biological responses to the composites in order to investigate their potential use as bone graft substitutes. A range of HABT ceramics of different compositions was manufactured and their electrical properties were measured. The microstructure and piezoelectric properties of the ceramics were both dependent on the proportion of barium titanate (BT) present. Composites containing more than 70% BT displayed piezoelectric charge coefficients (d33) of up to 86.3±7.9pCN-1 (95% BT). The ferroelectric nature of the 90 and 95% BT materials was confirmed by assessment of their ferroelectric hysteresis loops. The highest piezoelectric voltage coefficient (g33) recorded was 14x10-3Vm-1Pa-1 (90% BT). Following the assessment of the electrical properties, the HABT ceramic containing 90% BT was selected for the assessment of biological responses to the composites. The proliferation, viability, activity and morphology of human osteoblast-like cells cultured on HABT were comparable to those cultured on hydroxyapatite (HA) up to 7 days after seeding. The remnant polarisation of poled HABT induced an increase in cell attachment. This influence was independent of the nature (positive or negative) of the polarisation. Poling was not found to influence cell morphology, activity or differentiation in the first 7 days of incubation. At 14 days after seeding, results were inconsistent, indicating some variations in cell population and differentiation depending on the composition and poling of the ceramics respectively. This study has substantially defined the electrical properties of a range of HABT ceramics. It indicates their in vitro biocompatibility and thus their potential for use as bone graft substitutes. These results provide a benchmark against which future work investigating the influence of mechanical loading and longer term studies may be measured.