Proliferation and morphology of pre-osteoblast cells on 25 – 1000 micron diameter patterned features on polyacrylamide hydrogels

Bone formation, also known as osteogenesis, occurs in three distinct phases: proliferation, during which cells multiply, form nodules, and begin expressing early stage osteogenic markers; extracellular matrix development and maturation, during which mature cells in nodules build a collagen matrix and secrete the building blocks for mineralization; and mineralization, during which bone mineral crystals nucleate and grow. While it is known that cell proliferation, maturation, morphology, and density all play a role of in osteogenesis, most studies of controlled cell differentiation either focus proliferation and maturation in large cell colonies or morphology of single cells. However, large cell colonies are difficult to examine because of the presence of multiple bone nodules and mineral nucleation sites, and the single cell differentiation experiments ignore the importance of cell proliferation, cell maturation, and cell-cell contact/interaction. In this work, microcontact printing is used to pattern 19 kPa hydrogel substrates with 1000, 500, 250, 100, 50, and 25 micron diameter circles of fibronectin and collagen. Experiments were conducted in which cell proliferation of D1 ORL UVA cells was measured as the cells filled the patterns. Cell seeding density and cell density at confluence do not depend on media type or feature size, as long as cells are not confined to a small area (25 micron). The fold increase is significantly higher for larger features, where the cells have ample area to rapidly proliferate, but not significant for smaller features where the cells do not rapidly proliferate. Proliferating cells have an elongated morphology and mature cells at confluence are cuboidal. In future work, quantitative cell shape, proliferation, and early osteogenic and adipogenic marker data will be combined into a simulation to further study osteogenesis.

Name thesis:degree_name

M.S.

Level thesis:degree_level

Thesis

Discipline thesis:degree_discipline

Mechanical Engineering

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2016

Author dc:creator

• Berent, Zachary T

Contributors dc:contributor

• Wagoner Johnson, Amy J.

• osteogenesis
• microcontact printing

Statement dc:rights

• © 2016 Zachary Taylor Berent

Language dc:language

en

Handle dc:identifier

http://hdl.handle.net/2142/90964

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/90964