Orientador: Rosemary Adriana Chiérici Marcantonio / Resumo: O objetivo deste estudo foi avaliar o efeito do uso de diferentes substitutos sobre o reparo ósseo de defeitos críticos de calvárias de ratos com ou sem osteoporose. Para tanto, foram divididos em três estudos. O estudo 1 avaliou o efeito de diferentes biomateriais sobre o reparo de defeitos críticos de calota (DCC) de 40 ratos saudáveis que foram aleatoriamente divididos em 5 grupos com 8 animais, de acordo com o tipo de biomaterial utilizado para preencher os DCC: Grupo COA (coágulo); Grupo AUT (osso autógeno); Grupo OBD (osso bovino desproteinizado); Grupo HA/ TCP (cerâmica bifásica composta de hidroxiapatita e β-fosfato tricálcio); Grupo TCP (β-fosfato tricálcio). Foram executadas análise microtomográfica para avaliação do comprimento linear remanescente (DLL) do DCC e o volume dos tecidos mineralizados (MT) dentro do defeito nos períodos de 3, 7, 15 e 30 dias após cirurgia. Adicionalmente, foi executado análise histométrica para avaliar a composição do tecido ósseo reparado (% Osso e % Biomaterial) no período de 30 dias. O grupo COA apresentou o menor DLL e MT dentro do DCC e maior % osso do que os outros grupos. O grupo OBD apresentou maior volume de tecidos mineralizados e maior % biomaterial do que o grupo os grupos AUT e TCP. Os grupos OBD e AUT apresentaram maior % osso que o grupo TCP. O estudo 2 avaliou o efeito do enxerto com OBD carregado de estrôncio (Sr) na cicatrização óssea em DCC em 42 ratos saudáveis. Foram feitos 2 defeitos/rato, e um destes aleatoriament... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

Nearly 8 million American adolescents participate in sports. Many sports (e.g. basketball, volleyball) require powerful muscle movements. Normally, participation declines in young adulthood. The purpose of this study was to assess longitudinal effects of interscholastic high school sport participation and muscle power on young adult bone strength. 295 young adults from the Iowa Bone Development Study participated in this study. Participants were classified into sport participation groups based on an interscholastic sport participation history questionnaire. Groups included Power Sport Participant (PSP), Other Sport Participant (OSP), and Nonparticipant (NP). Current physical activity (PA) behaviors were assessed via questionnaire. Dual x-ray absorptiometry (DXA) assessed hip areal bone mineral density (aBMD) and was used with hip structure analysis (HSA) to estimate femoral neck section modulus (FN Z) and hip cross-sectional area (CSA). Peripheral quantitative computed tomography (pQCT) provided stress-strain index (SSI) and bone strength index (BSI) at 38% and 4% cross-sectional tibial sites respectively. Vertical jump estimated muscle power at age 19. Gender-specific multiple linear regression predicted young adult bone outcomes based on sport participation groups. Mediation analysis analyzed effects of muscle power on relationships between sport participation and bone outcomes. All analyses were adjusted for current PA. For both males and females, bone outcomes for PSPs were greater than bone outcomes for NPs (P < 0.025). Bone outcomes for PSPs were also greater than OSPs in females (P < 0.025). Mean differences for PSPs and NPs differed between 6.5% to 15.7%. 14.2% to 27.5% of the effect of sport participation on bone outcomes was mediated by muscle power. These results provide evidence to say that former male power sport participants and other sport participants and female power sport participants have stronger bones than peers even when adjusting for current PA. Muscle power did not fully explain differences in all bone outcomes suggesting that sport participation has additional bone health benefits.

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.

It has been theorized that muscles generate more force on bone than body weight alone and therefore it is likely that muscle contraction drives and sustains bone adaptation (Frost 1999). Purpose: To investigate the relationship between the timing and tempo of peak growth velocities of lean tissue (LT) and bone mineral content (BMC) in boys and girls at three sites using data derived from individual growth curves. Methods: 72 boys and 70 girls were fitted with growth curves that had a distinguishable peak. Height and weight were measured for each participant and tissue assessment was performed annually using DXA. Factorial ANOVAs were completed to analyse data for differences in age, while forward regression analyses was used between LT and BMC. Results: The peak growth velocity for lean occurred significantly (P<0.05) earlier than the peak growth velocity for bone at all locations except the legs. There was a difference (P<0.001) between genders in the age of peak for both lean tissue and bone tissue at all locations with females peak growth occurring before that of males. When aligned by PHV a significant difference (P<0.05) in the timing of PBMCV was found between the arms and the legs with the peak in bone growth in the legs occurring significantly before peak bone growth in the arms. PLTV was independently associated with PBMCV at the arms (r2= .71, p<0.001), legs (r2= .53, p<0.001) and trunk (r2= .52, p<0.001). Conclusion: In conclusion, LT growth precedes BMC growth and after controlling for gender, size and maturity the magnitude of LT growth is associated with BMC growth. The findings of this study are in support the Muscle-bone Unit (Frost and Schoenau, 2000), which theorizes that localised muscle action is a driving force for bone growth. Future studies are needed to analyse bone strength as it relates to local muscle strength and usage while controlling for confounding variables.

It has been theorized that muscles generate more force on bone than body weight alone and therefore it is likely that muscle contraction drives and sustains bone adaptation (Frost 1999). Purpose: To investigate the relationship between the timing and tempo of peak growth velocities of lean tissue (LT) and bone mineral content (BMC) in boys and girls at three sites using data derived from individual growth curves. Methods: 72 boys and 70 girls were fitted with growth curves that had a distinguishable peak. Height and weight were measured for each participant and tissue assessment was performed annually using DXA. Factorial ANOVAs were completed to analyse data for differences in age, while forward regression analyses was used between LT and BMC. Results: The peak growth velocity for lean occurred significantly (P<0.05) earlier than the peak growth velocity for bone at all locations except the legs. There was a difference (P<0.001) between genders in the age of peak for both lean tissue and bone tissue at all locations with females peak growth occurring before that of males. When aligned by PHV a significant difference (P<0.05) in the timing of PBMCV was found between the arms and the legs with the peak in bone growth in the legs occurring significantly before peak bone growth in the arms. PLTV was independently associated with PBMCV at the arms (r2= .71, p<0.001), legs (r2= .53, p<0.001) and trunk (r2= .52, p<0.001). Conclusion: In conclusion, LT growth precedes BMC growth and after controlling for gender, size and maturity the magnitude of LT growth is associated with BMC growth. The findings of this study are in support the Muscle-bone Unit (Frost and Schoenau, 2000), which theorizes that localised muscle action is a driving force for bone growth. Future studies are needed to analyse bone strength as it relates to local muscle strength and usage while controlling for confounding variables.

Bone grafts are used to treat more than 300,000 fracture patients yearly, as well as patients with congenital defects, bone tumors, and those undergoing spinal fusion. Given the established limitations of autograft and allograft bone, there is a substantial need for bone graft substitutes. Tissue engineering strategies employing the addition of osteogenic cells and/or osteoinductive factors to porous scaffolds represent a promising alternative to traditional bone grafts. While many bone defects are in load-bearing sites, very little is known about the response of bone grafts and their substitutes to mechanical loading, despite vast documentation on the ability of normal bone to adapt to its mechanical environment. The goal of this research was to quantify the effects of controlled in vivo mechanical stimulation on bone graft repair and bone graft substitutes and identify the local stress/strain environment associated with load-induced changes in bone formation. The global hypothesis that cyclic in vivo mechanical loading improves mineralized matrix formation within bone grafts and bone graft substitutes was addressed in this work using orthotopic and ectopic models specifically designed to facilitate modeling of local stresses and strains. In the first study, a bone defect repair model utilizing an orthotopic implant capable of supplying a controlled mechanical stimulus to a trabecular allograft showed a significant reduction in new bone formation with controlled in vivo mechanical loading. Although the reason remains unclear, loading conditions may not have been ideal for increased bone formation or potential micromotion may have influenced the results. A second study demonstrated for the first time that controlled in vivo mechanical stimulation enhances mineralized matrix production on a mesenchymal stem cell-seeded polymeric construct using a novel subcutaneous implant system. In addition, the local stresses and strains associated with this adaptive response were predicted. The novel subcutaneous implant represents technology which may be adapted for the preparation of tissue-engineered bone constructs, capitalizing on the benefits of mechanical loading and a vascularized in vivo environment. Such an approach may produce larger, stronger, and more homogeneous constructs than could be developed in a static culture system subject to diffusional limitations.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2010. / Härtill 4 uppsatser.

published_or_final_version / Orthopaedics and Traumatology / Doctoral / Doctor of Philosophy

Recombinant human bone morphogenetic proteins (rhBMPs) are increasingly used for reconstructing bony defects, fracture non-unions, and augmenting existing bone volumes. For the numerous patients taking bisphosphonates, the impact of prior bisphosphonate treatment on rhBMP bone induction is not well understood. Objective: to evaluate the effect of the prior treatment with zoledronate on rhBMP induced bone formation in mice. Methods: 42 mice were pre-treated with 0, 2, and 20 µg of zoledronate/mouse. The osteoinductive activity of a bioimplant, containing rhBMP-2, was assessed using the mouse muscle pouch assay, and analyzed with micro CT and histology. Results: micro CT demonstrated that BMP bioimplants placed in mice pretreated with 20 µg of zoledronate, formed bony ossicles of greater volume but reduced bone density compared to controls. Histologically, the heterotopic ossicles from the 20 µg group consisted of more immature bone than those from the other groups. Conclusion: bone induced by rhBMP-2 in mice pre-treated with a high concentration of zoledronate was immature as evidenced by radiographic and histologic appearance.

Non-invasive estimates of bone quality are of great interest as they could potentially improve the diagnosis and treatments for bone disorders, such as osteoporosis, and could aid in preoperative planning of surgical interventions, improving patient outcomes in orthopaedic trauma. Quantitative computed tomography (QCT) is currently being investigated as a method of estimating structural bone mineral content (sBMC) and, thus, bone quality. However, the reported correlations between QCT and sBMC have been variable and the relationships reported between sBMC and the apparent elastic modulus (E*) of bone have been even more variable. A series of investigations are presented, relating QCT, sBMC, apparent density and the E* of bone. These studies were performed with the ultimate objective of determining the appropriateness of QCT as a predictor of the E* of bone and whether the E* of bone could be predicted from information regarding the sBMC and apparent density. / Thesis (Master, Mechanical and Materials Engineering) -- Queen's University, 2008-01-31 22:00:08.77 / This work was funded by the Canadian Institutes of Health Research.