Date of Award

Fall 1991

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biological Sciences

Program/Concentration

Biology

Committee Director

Lioyd Wolfinbarger, Jr.

Committee Member

Keith A. Carson

Committee Member

Edgar Stillwell

Call Number for Print

Special Collections LD4331.B46 Z44

Abstract

Acid demineralized bone (DMB) implanted in extraosseous sites induces bone formation. In vivo studies have shown that DMB stimulates the differentiation of mesenchymal cells into osteogenic cells. In this study an in vitro system was developed to demonstrate the osteoinductivity of DMB quantitatively. This in vitro working system was used to assess the osteoinductivity of variously processed DMB products as a means to evaluate their processing protocols. A human dermal fibroblastic (HDF) cell line was initiated from tissue explants as a working model, along with a human periosteal (HPO) cell line as a control. Ground DMB processed by LifeNet Transplant Services was added to HDF cell cultures and changes in cell growth and functional characteristics were monitored over a 7-day incubation period. Cell proliferation rate decreased, as indicated by decreased total cell number and decreased rate of [3H]-thymidine incorporation. Cell size decreased, using the amount of protein content per cell as a measure. Alkaline phosphatase (AP) activity also increased. These characteristics of stimulated HDF cells listed above are more typical of HPO cells rather than HDF cells used as a control, suggesting that DMB had stimulated HDF cells to display phenotypic characteristics typical of osteogenic cells.

AP activity was chosen as a quantitative measure for osteoinductivity of various kinds of DMB products because it has been reported to be an exclusive indicator of osteoinducing cells and has been widely used as a marker for osteogenic cells. AP activities of HDF cells showed no significant difference when cultured with demineralized bone and ethanol-pretreated demineralized bone, indicating that ethanol pretreatment does not affect the osteoinductive potential of DMB.

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DOI

10.25777/6bbd-tj45

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