Biomedical Sciences ETDs

Publication Date

8-1-2023

Abstract

Development of a cell culture based bone model can contribute to biomedical research by producing a reproducible model system that mimics a bone remodeling disorder and reduces the use of animals in research studies. Synthetic biology was used to engineer osteoblast cell lines to alter a key protein in the RANKL pathway in efforts to control the bone remodeling process. Osteoblast cell lines, K7M2 and MC3T3-E1 Subclone 4 were engineered to express osteoprotegerin (OPG) and inhibit OPG expression, respectively. The upregulation of OPG expression was confirmed with Real Time PCR and Quantikine® Immunoassay. To identify the functionality of the engineered K7M2-OPG-Myc cell line, a resorption pit assay using calcium phosphate as synthetic biomimetic material was used. RAW cells were differentiated to osteoclasts using varying concentrations of RANKL in conditioned medium from engineered K7M2-OPG-Myc cell line in the calcium phosphate assay. Conditioned Medium from K7M2-OPG-Myc cells inhibited resorption activity of formed osteoclasts. This thesis work was the first step taken to enable the development of a cell culture based bone model capable of remodeling on its own.

Keywords

Osteoblasts, Osteoclasts, Osteocytes, Bone Remodeling Model

Sponsors

The Laboratory Directed Research and Development program at Sandia National Laboratories, a multimission laboratory managed and operated by National Technology and Engineering Solutions of Sandia LLC, a wholly owned subsidiary of Honeywell International Inc. for the U.S. Department of Energy’s National Nuclear Security Administration under contract DE-NA0003525

Document Type

Thesis

Language

English

Degree Name

Biomedical Sciences

Level of Degree

Masters

Department Name

Biomedical Sciences Graduate Program

First Committee Member (Chair)

Laura V. Gonzalez-Bosc

Second Committee Member

Christina Salas

Third Committee Member

Jerilyn A. Timlin

Download

Share

COinS