Thesis
The effects of bioprinting materials on HEPM cell proliferation and cytokine release
University of Iowa
Master of Science (MS), University of Iowa
Spring 2018
DOI: 10.17077/etd.bfewys08
Abstract
<p>Objectives: Three-dimensional (3D) bioprinting is a manufacturing process that incorporates viable cells into a 3D matrix by adding layer upon layer of material. The objectives of this study are to characterize a novel matrix of collagen and hydroxyapatite and to assess the effects of the 3D bioprinting process on cytotoxicity, proliferation rate, and cytokine expression of Homo sapiens palatal mesenchyme (HEPM) cells.</p>
<p>Methods: For this, we prepared a 3D matrix of collagen and hydroxyapatite without and with cells. We used light microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM) to characterize the structure and arrangement of the collagen fibers. We then incubated the matrix with known standards of cytokines to measure adsorption. We assessed the proliferation and viability of HEPM cells in the presence of the 3D construct and after 3D bioprinting. Finally, we assessed the cytotoxicity of this matrix for HEPM cells and assessed its effect on the production of chemokines and cytokines. A one-way fixed effect ANOVA was fit to concentrations of cytokines and pairwise group comparisons were conducted using Tukey’s Honest Significant Differences test (p< 0.05).</p>
<p>Results: The matrix was found to contain interwoven strands of collagen and some hydroxyapatite crystals that did not absorb cytokines except for MIP-1a (p< 0.05). The matrix was found to be non-cytotoxic using an AlamarBlue® assay. We found that the cell proliferation rate was lower when seeded on the 3D construct than in 2D culture. We also found that the proliferation rate was very low for the HEPM cells in the 3D bioprinted constructs. In the presence of the 3D construct, the HEPM cells had similar expression profiles of the cytokines measured (P > 0.05 for GM-CSF, IL-6, IL-8, and RANTES).</p>
<p>Conclusion: 3D-bioprinting has the potential to be used in dentistry as a novel osteogenic bone grafting material. Here we show that a novel matrix of collagen and hydroxyapatite is non-cytotoxic to HEPM cells and does not induce a proinflammatory response.</p>
Details
- Title: Subtitle
- The effects of bioprinting materials on HEPM cell proliferation and cytokine release
- Creators
- Robert David Swenson - University of Iowa
- Contributors
- Kim A. Brogden (Advisor)Satheesh Elangovan (Committee Member)Liu Hong (Committee Member)
- Resource Type
- Thesis
- Degree Awarded
- Master of Science (MS), University of Iowa
- Degree in
- Oral Science
- Date degree season
- Spring 2018
- DOI
- 10.17077/etd.bfewys08
- Publisher
- University of Iowa
- Number of pages
- x, 77 pages
- Copyright
- Copyright © 2018 Robert David Swenson
- Language
- English
- Description illustrations
- color illustrations
- Description bibliographic
- Includes bibliographical references (pages 63-69).
- Public Abstract (ETD)
Objectives: Three-dimensional (3D) printing is a method of making something by adding one layer of material at a time. 3D bioprinting is a special type of 3D printing where the material used contains living cells and the material is chosen to keep the cells alive until they are used to treat a patient. We call the 3D bioprinted material a 3D construct. One potential use of these constructs is for bone grafting. The 3D bioprinting process puts stress on the cells. If the cells are too stressed during the printing process they will not be able to function as bone producing cells. In this set of experiments, we measure the health of the cells after they have gone through the process, their ability to grow in the material, and the amount of proteins called cytokines that the cells produce.
Methods: We made 3D constructs with and without cells. We used a variety of microscopes that can magnify up to several hundred thousand times to look at the structure of the material. We took samples of the material with cells and measured the cytokines expressed by the cells. We used a specialized test to look for cells that were damaged during the process.
Results: The 3D bioprinting process and the 3D construct we developed was well tolerated by the cells. The cells grow at the same rate with the material as without the material present. The cells did not increase the amount of cytokines that they produced after going through the 3D bioprinting process.
Conclusion: This new 3D construct and process has potential to be used as a bone graft. Future studies will measure other important factors such as its safety, cost, and ability to grow new bone.
- Academic Unit
- Oral Pathology, Radiology and Medicine; Craniofacial Anomalies Research Center
- Record Identifier
- 9983777004902771
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