Background: Osteoporosis is a serious condition in which bone of the skeleton deteriorates substantially, resulting from an imbalance of the bone remodeling process. Affecting more than 40 million Americans, osteoporosis leads to major fractures of the hip, spine. Although many forms of treatment are widely used, including anabolic and anti-resorptive agents, the vast majority of prescribed medications have minor to severe adverse effects. Thus, many sufferers of osteoporosis seek natural forms of treatment. Estrogen replacement therapy is effective in preventing and treating ovarian hormone-deficiency bone loss, yet the risks far exceed the benefits. Similar to soy isoflavonic compounds, which have been shown to exude beneficial effects on bone metabolism, ferutinin, a daucane extract of the giant fennel (Ferula communis) are believed to have estrogen-like activity. Purpose: To elucidate the effects of ferutinin on osteoblastic differentiation and mineralization. Methods: MC3T3-E1 preosteoblast-like cells were treated with ferutinin (10-12, 10-10, 10-8, 10-6, 10-5, 10-4 M), 17ß-estradiol (10-8 M), or no treatment (control). Cells cultured in an osteogenic medium (50 μg/ml ascorbic acid & 10 mM ß-glycerophosphate) contatining 5% charcoal-dextran treated FBS for up to nine, fifteen, or twenty-eight days for the assessment of the following: cell viability using an MTT assay, alkaline phosphatase activity measured colorimetrically, calcium nodule formation and mineralization using 40 mM alizarin red stain which was lifted and absorbance measured, and osteocalcin concentrations. Results: Cell viability remained stable in cells treated with 10-12, 10-10, 10-8, and 10-6 M of ferutinin. Alkaline phosphatase activity was enhanced but not significantly (13.4% greater than control) in cells treated with 10-10 M ferutinin. Osteocalcin levels and the absorbance of retained stain ferutinin-treated cells were not significant. Conclusions: Ferutinin at a dose of 10-10 M appeared to have a mild augmenting effect on osteoblastic differentiation; however, further experimentation with other in vitro and vivo models is needed to fully elucidate its effects. / A Thesis submitted to the Department of Nutrition, Food and Exercise Sciences in partial fulfillment of the requirements for the degree of Master of Science. / Summer Semester, 2011. / June 29, 2011. / Ferutinin, MC3T3-E1 cells, differentiation, alkaline phosphatase / Includes bibliographical references. / Bahram H. Arjmandi, Professor Directing Thesis; Maria Spicer, Committee Member; Cathy W. Levenson, Committee Member.

Bone fractures are one of the most common injuries in the United States, encompassing 6.2 million incidences, and costing the healthcare system roughly $20 billion annually. The majority of this cost falls upon a unique type of fracture known as a non-union fracture, defined by incomplete healing after 9 months. The economic burden in combination with the frequency by which these incidences occur offer a unique opportunity for research and improvement in the healthcare field. Previous research on the fracture repair process showed that dietary deficiency led to delayed healing producing a rachitic-like effect on the endochondral bone formation process that occurs during fracture healing. This research will build off the understanding of hypophosphatemia on bone fracture repair utilizing a unique temporal clustering approach to assess changes in the transcriptomic expression within callus tissues of control fed and dietary phosphate restricted animals. Using a temporal cluster modeling technique developed by our group (Lu et al. 2019), twelve clusters were generated for the gene expression data extracted from the callus tissues of B6 strain mice at time points 3, 5, 7, 10, 14, 18-, 21-, 28-, and 35-days post fracture, in, control and phosphate restricted dietary groups. Groupings of clusters were used to establish the temporal expression patterns over the time course of healing and identify the movement of genes that changed their temporal expression patterns between the control and phosphate restricted diets. Biological process categories were established for each cluster, grouping using both Ingenuity Pathway Analysis (IPA) and the NIH DAVID Bioinformatics Database ontology assessment programs. Genes based on their association with four key tissue developmental processes in fracture repair, skeletogenesis, myogenesis, vasculogenesis, and neurogenesis were analyzed. The analysis showed shifts to later peak expression times for all four categories. Further analysis illuminated three specific regulatory pathways that were significantly impacted by hypophosphatemia, Hippo and WNT signaling pathways and the circadian rhythm pathway while oxidative phosphorylation was both shifted and showed reduced expression. The shifts in expression time and level of these pathways demonstrate their importance to bone fracture repair and their impacts on mesenchymal stem cell differentiation. From the data analysis it is clear that limiting dietary phosphate results in impaired mesenchymal stem cell differentiation caused by delayed Hippo and WNT signaling. Further it is evident that the processes of skeletogenesis, myogenesis, vasculogenesis, and neurogenesis are heavily interconnected, often showing overlapping genes through all four processes. Based on these shifts and impairments in specific signaling we identified novel mechanisms by which hypophosphatemia can impair fracture callus growth and development and delay healing.

Bone Morphogenetic Proteins (BMPs) belong to the Transforming Growth Factor Beta superfamily of growth factors. While BMP signaling has been shown to induce skeletogenic differentiation of mesenchymal stem cells and be involved in the formation of ectopic bone and ossification, BMPs also are involved in the differentiation of many other tissues including neurogenic tissues. Prior studies from our laboratory showed that BMP-2 induction of chondrogenic differentiation of the C3H10T1/2 murine mesenchymal stem cell line was associated with increased oxidative metabolism. This study was performed to test the hypothesis that BMP-2 promotes increased oxidative metabolism during the differentiation of other types of cells. Using the neurogenic cell line, SH-SY5Y, this study examined whether cellular differentiation induced by BMP-2 also was associated with increased oxidative phosphorylation in non-mesenchymal stem cells. SH-SY5Y cells were grown in growth medium (DMEM/F12 with 10% FBS, 1% Penicillin/Streptomycin, L-glutamine, and non-essential amino acids). Cells were plated appropriately at differing seeding densities (Day 0), treated one time with ± 200ng/mL BMP-2 two days after plating and analyzed on Day 4. Oxidative metabolism was measured using a Seahorse XF Analyzer that measures oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). BMP-2 induction of neural cell differentiation was assayed by quantifying dendrite communication between neurons and decreased proliferative capacity as assessed by overall culture DNA contents. Mitochondrial density after BMP-2 treatment was examined using vital mitochondrial labeling. Groups treated with BMP-2 contained significantly less DNA content than control groups (p=0.006). BMP-2 treated cells had on average more dendritic interactions with one or two and more processes than control groups (p=0.396; p=0.872), while there were a larger percentage of cells not treated with BMP-2 that had zero dendritic interactions (p=0.470). All parameters of oxidative metabolism were increased in cells treated with BMP-2. More specifically, basal respiration and ATP production were significantly increased in BMP-2 treated cells (p=0.031; p=0.010). SH-SY5Y cells were significantly affected by BMP-2 treatment in both DNA content and oxidative phosphorylation. The diminished DNA content with BMP-2 treatment is consistent with the known decrease in cellular proliferation that is associated with neural cell differentiation. While there were small increases in dendritic interactions these were not significant and are inclusive for demonstrating BMP-2’s effect on neural differentiation. Upregulation of oxidative phosphorylation indicated after BMP-2 treatment was validated by the substantial increases in metabolic parameters associated with BMP-2 treatment. Through the utilization of the neural cell line, SH-SY5Y, this research suggests a more generalized functionality of BMP-2 in upregulation of oxidative metabolism as well as differentiation non-specific to bone and chondrocyte cell lineage.

Thesis (M.Ost.)--Unitec New Zealand, 2008. / Includes bibliographical references (leaves 119-123).

Thesis (M.Ost.)--Unitec New Zealand, 2009. / Includes bibliographical references (leaves 88-92).

Thesis (M.Ost.)--Unitec New Zealand. / Coda (electronic version) title-page has 2009 date. Includes bibliographical references (leaves 57-58).

The primary purpose of this research was to compare quantifiable structural asymmetry to changes visualized in center of pressure (CoP) and/or postural sway by the use of an Isobalance forceplate in order to introduce principles of osteopathic medicine to a Latin America culture. Osteopathic manual medicine (OMM) was used to correct structural dysfunction found in the study participants at the Baxter Institute in Tegucigalpa, Honduras. Study participants were students and staff members ranging in age from 18 to 35 and consisting of 24 males and two females. The examination period lasted a total of five weeks. During weeks one, three, and five, baseline measurements were taken prior to manipulation using .05 alpha to test significance. Measurements were repeated post OMM. An educational video was provided during each session. Pretest and posttest results demonstrated an improvement in understanding of OMM materials provided to participants. A bivariable chi-square test found that, when manipulated, those with sacrum dysfunction have an association with a positive improvement in postural sway (TIC 1 & 2) (p<.05). Also, the multivariable logistic regression model found that individuals who had no initial change in postural sway (TIC 1 & 2) were more likely to move to a positive improvement of time in the center of the premeasured diameter calculated by the Isobalance forceplate, than a decrease in time spent in the center (i.e. negative improvement) throughout the time of the study (p<.05). / Ph. D.

Thesis (M.Ost.)--Unitec New Zealand, 2006. / Includes bibliographical references (leaves 50-53).

Thesis (M.Ost.)--Unitec New Zealand, 2008. / Coda (electronic version) title page has 2009 as date of publication. Includes bibliographical references (leaves 43-50).

Thesis (M.Ost.)--Unitec New Zealand, 2009. / Includes bibliographical references (leaves 57-63).