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Microscale Additive Manufacturing of Collagen Cell Culture Scaffolds

Bell, Alex E.
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1448037063

Abstract Details

2015, PhD, University of Cincinnati, Engineering and Applied Science: Biomedical Engineering.
Communication between cells and their environment is a key influencing factor on cell fate in tissues. Through integrin receptors, cells can sense and respond to structural features of the extracellular matrix. Signaling-factor receptors can detect gradient changes in concentration of a multitude of soluble ligands. Both mechanical and biochemical signals are presented to cells in defined spatial patterns. One of the most important sets of signals for developing or regenerating tissue are those guiding the formation of a vascular supply. Vascular development is crucial to recapitulation of further tissue function. Replicating these 3D patterns in vitro is a considerable challenge for the long-term goal of engineering functional tissue replacements. Multiphoton crosslinking (MPC) is an additive manufacturing process using light-activated chemistry to produce crosslinking by the absorption of multiple photons. MPC is quadratically dependent on light intensity, allowing isolation of the reaction to the focal plane. With diffraction-limited optics, this provides 1 µm resolution. MPC has the potential to give unlimited 3D control of the structural and biochemical cues imparted by functional proteins and peptides at a sub-cellular scale. Potential benefits of MPC include both the ability to spatially isolate presentation of a given factor in a concentration-controlled manner and sequestering the factor to provide a sustained dose over an extended period of cell culture and tissue regeneration. The MPC technique is essentially applicable to any protein, as several amino acids can react with an excited radical species. MPC of protein substrates has not yet been utilized toward functional tissue engineering applications. This is due to multiple factors, including the need for the substrate protein to be soluble, form a stable 3D structure, and have the unexposed portion readily removed. Thus, MPC has been limited in use primarily to highly soluble proteins such as albumin, which is not effective as a tissue scaffold material. Patterning of functional proteins onto scaffolds using MPC has also been limited by the need for the unexposed materials to be removed from the scaffold following the patterning process. This has meant that the technique, thus far, is largely employed on synthetic hydrogels with peptide fragments. This dissertation encompasses studies that established new additive manufacturing methods for cellular scaffolds in dermal wound healing. Completion of this work has expanded the capability of MPC to allow 3D fabrication of type I collagen scaffolds with spatial control of local structural and biochemical signals at a sub-cellular scale. Initial studies were performed to assess the effectiveness of these scaffolds patterned with VEGF for the induction of vasculogenesis by spatially defining the presentation of vasculogenic signals. These materials were used in a form as near as possible to their native state in an attempt to present a signaling environment more similar to a native ECM. These studies were directed toward implementation in vasculogenic dermal wound scaffolds, but the methods used are generally applicable to creation of structural and chemical patterning in any TE/RM application.
Vasille Nistor, Ph.D. (Committee Chair)
J. Matthew Kofron, Ph.D. (Committee Member)
Jason Shearn, Ph.D. (Committee Member)
Lilit Yeghiazarian, Ph.D. (Committee Member)
118 p.
Biomedical Research
Multiphoton fabrication; 3D printing; tissue engineering; collagen scaffolds; additive manufacturing; vasculogenesis angiogenesis

Recommended Citations

Citations

  • Bell, A. E. (2015). Microscale Additive Manufacturing of Collagen Cell Culture Scaffolds [Doctoral dissertation, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1448037063

    APA Style (7th edition)

  • Bell, Alex. Microscale Additive Manufacturing of Collagen Cell Culture Scaffolds. 2015. University of Cincinnati, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1448037063.

    MLA Style (8th edition)

  • Bell, Alex. "Microscale Additive Manufacturing of Collagen Cell Culture Scaffolds." Doctoral dissertation, University of Cincinnati, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1448037063

    Chicago Manual of Style (17th edition)

ucin1448037063
641
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This open access ETD is published by University of Cincinnati and OhioLINK.