Date of Award

Summer 8-2-2019

Degree Type

Thesis

Degree Name

MS Biology

Department

Biology

Advisor

Jessica Cottrell, Ph.D.

Committee Member

Angela Klaus, Ph.D.

Committee Member

Constantine Bitsaktsis, Ph.D.

Keywords

bone, fracture, healing, insulin-mimetic, vanadium, osteoblast

Abstract

Osteoblastogenesis is an essential part of the bone healing process. Insulin has been shown to improve bone healing in both normal and diabetic bone healing models. In addition, insulin mimetic compounds such as Zinc chloride (ZnCl2) and Vanadyl acetylacetonate (VAC) have also been shown to improve bone healing in these models as well. The purpose of this study was to determine the effects of vanadium compounds VAC and Vanadium (II) sulfate (VSO4) in osteoblast proliferation and function. In addition the mechanisms by which growth and function are facilitated by these Vanadium compounds were also evaluated. In this study, we determined if treatment with VAC and VSO4 could induce osteoblast proliferation in the MC3T3-E1 mouse cell line. MC3T3-E1 cells were treated with 50ug/ml insulin, 50ug/ml ascorbic acid, 0µM, 5µM, 10µM, 15µM, 25µM, 50µM, or 100µM, of VAC or VSO4 and harvested at the following time points 6hrs, 24hrs, 4 days, 7 days, 10 days, 14 days, 17 days, and 21 days. MC3T3-E1 proliferation, function, and mechanisms of growth were evaluated via MTT Assays, Alizarin Red staining/Alcian blue staining, and immunoblotting respectively. Our results demonstrate that both VAC and VSO4 are capable of inducing MC3T3-E1 proliferation as shown by the MTT assay. Treatment with VAC and VSO4 can also induce calcium and proteoglycan deposition as shown by Alizarin red and Alcian blue staining in MC3T3-E1 cells. VAC and VSO4 treatment also increased activation of protein kinase B (Akt) to its activated form p-Akt over time. Our results coincide with other studies that show that Vanadium compounds can stimulate osteoblastogenesis. Also that this is achieved through utilization of insulin pathways and promoting proper function of osteoblast cells. Our data suggests that VAC and VSO4 may be useful as medications for promoting bone growth during the fracture healing process.

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