Date of Award

8-2007

Document Type

Thesis

Degree Name

Master of Science (MS)

Legacy Department

Bioengineering

Advisor

Burg, Karen J.L.

Committee Member

Webb , Ken

Committee Member

Kellam , James F.

Abstract

Over 500,000 bone graft procedures are performed each year in the United States. Bone grafting involves a surgical procedure to replace missing bone. Problems can arise with donor and defect sites during and after surgery, sometimes resulting in poor clinical results. The development and optimization of bone graft substitutes via a tissue engineering approach could markedly improve bone graft surgical outcome. Demineralized bone matrix (DBM), a bone graft material, is currently used in a clinical setting but has variable success rates.
The primary objective of the research presented in this thesis was to assess the cellular activity of D1 mouse stromal cells seeded on either partially demineralized bone matrix (PDBM) substrates or completely demineralized bone matrix substrates (CDBM). Before performing a cell-based experiment involving the varying levels of DBM, a study was performed to determine the optimal media conditions for osteoblast formation. Additives β-glycerophosphate and L-ascorbic acid were added to α-MEM media and cultured with D1 cells on DBM to assess the effects of these additives on the rate of cellular differentiation. Significant differences in osteoblast activity were not noted between the two medium conditions.
The final study evaluated cellular activity at four specific time points over a 36 day period after seeding D1 cells on DBM of varying demineralization levels. Based on the assays performed at the four time points, it appears that the D1 cells on the PDBM fragments differentiated toward the osteoblast phenotype and the cellular fragments began to mineralize between 24 and 36 days after initiation of culture, while the cellular CDBM showed minimal potential for mineralization over the course of the study. Additional studies with more frequent time points are necessary to gain a better understanding of the cellular activity of the D1 cells on the DBM.

Share

COinS