Investigation of adipose-derived mesenchymal stem cells interaction with electrospun demineralized bone matrix nanofiber scaffolds

Nanofiber demineralized bone matrix (DBM) scaffolds were fabricated by electrospinning, and their ability to support cell adhesion and cell viability of murine adipose-derived mesenchymal stem cells (AD-MSCs) for short-term in culture media was investigated. Poly (ε-caprolactone) (PCL) scaffolds were used as control surfaces. Live cell stain calcein-AM and CellTiter 96® Non-Radioactive Cell Proliferation assays were used for cell adhesion and cell proliferation, respectively. DBM scaffolds supported greater cell adhesion compared to PCL nanofiber scaffolds. For cell viability, the two types of scaffolds behaved similarly. The results led to further research on DBM scaffolds. The ability to support osteoblastic differentiation of AD-MSCs for long-term (three weeks) in osteogenic differentiation media was also investigated. Both PCL scaffolds and DBM scaffolds seeded with no cells were used as control surfaces. The total protein content of viable AD-MSCs on the scaffolds was assessed by bicinchoninic acid (BCA) assay. Nanofiber scaffolds displayed increased levels of alkaline phosphatase (ALP) activity for the first week for all cases. ALP activity dropped after one week. Scanning electron microscopy (SEM) and alizarin calcium staining techniques were used to examine mineralization patterns qualitatively on DBM and PCL nanofiber scaffolds. DBM scaffolds deposited more calcium mineral than PCL scaffolds during three-week experiments. Mineralization was quantified by energy-dispersive X-ray spectroscope (EDS). After three weeks of culture, EDS revealed high calcium and phosphorus deposition on DBM scaffolds compared to PCL controls. The DBM scaffolds exhibited increased mineralization over three weeks, both with and without cells. These results demonstrate that the adhesion, proliferation, and osteogenic differentiation of AD-MSCs were influenced by DBM scaffolds.

Name thesis:degree_name

Master of Science (M.S.)

Level thesis:degree_level

Masters

Discipline thesis:degree_discipline

Chemical and Biological Engineering

Grantor dc:publisher

Colorado State University. Libraries

Year dc:date.issued

2016

Authors dc:creator

• Yaprak Akgul, Selin, author
• Kipper, Matt, advisor
• Popat, Ketul, advisor
• Bailey, Travis, committee member
• Ehrhart, Nicole, committee member

Statement dc:rights

• Copyright and other restrictions may apply. User is responsible for compliance with all applicable laws. For information about copyright law, please see https://libguides.colostate.edu/copyright.

Language dc:language.iso

eng, English

Identifier URI

https://doi.org/10.25675/3.018968

OAI identifier oai:identifier

oai:mountainscholar.org:10217/173527