Seed Funded Projects

The Center for Advanced Biomanufacturing encourages collaboration with faculty working toward similar goals in tissue engineering and biomanufacturing. Please contact us to talk about how your research aligns with our mission of discovering new ways to help people heal from muscle injuries and defects. 

Engineering Morphogen Signaling Centers

Design microparticles of polymer containing the instructing morphogens (BMP4 and/or BMP2) that will be used to induce embryonic development in aggregates of embryonic stem cells. Control the size of particles, concentration of proteins delivered, and timing of instructive release to induce controlled and reproducible embryonic-like development in vitro. Design microparticles of polymer containing a combination of morphogen antagonists (Dkk1, Lefty1, Cer1) to modulate the gradient of Nodal and Wnt such that the developing anterior domain is protected from these two posteriorizing factors, allowing the formation of the anterior head domain. A pilot experiment showed an antagonistic effect of Dkk1 on the Wnt gradient we induced in our embryoids resulted in the expression of telencephalic markers at the anterior end. We will assess the results by analyzing the characteristic morphogenesis (live, immunostaining) and its reproducibility as a function of the parameters used and against controls (empty particles).

Equipment/PI: Lampe Electrospinner

Soft Hydrogel Microfluidics for In Vitro Vasculature Models Containing Aligned Smooth Muscle Cells

Vasculatures are composed of endothelial cells that are surrounded by smooth muscle cells, with an intervening fibrous basement membrane between the two cell types. Endothelial cells are longitudinally aligned by shear stress along the direction of blood flow, while smooth muscle cells are aligned circumferentially, orthogonal to the direction of blood flow, under radial cyclic strain. While there are a number of in vitro models of vasculatures to mimic endothelial cells aligned in a microfluidic channel, there are few models that include orthogonal arrangement between endothelial and smooth muscle cells. The availability of such an in vitro model would enable more realistic studies of vasculatures, including angiogenesis, invasion, intravascular survival and extravasation of tumor cells. The key theme of our innovation is use of structural and microfluidic cues to hydrogel microchannels so that fluid flow coupled to the well-controlled mechanical stiffness properties of hydrogels can transmit shear and compressive cues to patterned cells at neighboring interfaces for creating orthogonally aligned endothelial and smooth muscle cell layers that mimic in vivo arrangements. In the first approach, cell-laden hydrogels will be patterned and cultured under compressive strain due to fluid flow in the micropatterned structure. In the second and third approaches aimed at implantation in vivo, hydrogel channels will be created by transfer pattern from PDMS or by 3D bioprinting sacrificial layers for subsequent cell culture under fluidic compressive strain.

Equipment/PI: Nathan Swami

3D Printed Antioxidative Nanofullerene Scaffolds for Bone Repair

With the advancement of three-dimensional (3D) printing as a vital tool in tissue engineering, there is an ever-growing need to develop new biomaterial scaffolds that can be 3D printed with significant bioactivities. Tissue engineering, consisting of scaffolds, stem cells, and various growth factors (GFs), has been extensively investigated for bone fracture repair, but results were not satisfactory. In scaffolds for tissue engineering, GFs provide powerful biological signals to improve the performance of stem cells. However, they are generally expensive with a short in-vivo half-life that prevents a maximal effect. On the other hand, overloaded reactive oxygen species (ROS) have been considered as a crucial player prohibiting tissue regeneration. In the past decade, fullerene (C60), a nano chemical molecule with outstanding ROS scavenging activity, has been implicated to have a strong potential in treatment of several bone diseases, such as intervertebral disc degeneration, osteonecrosis, and osteoporosis. Therefore, we hypothesize that nano-fullerenes can serve as alternatives to GFs and will enhance osteogenesis leading to effective bone repair. This hypothesis is supported by our in vitro and in vivo pilot study showing that a fullerene derivative C60 pyrrolidine tris-acid (abbreviated as FTA) is able to induce osteogenesis of a bone marrow stem cell line D1. Our ultimate goal is to explore a novel 3D printed antioxidative biomaterial scaffold with FTA which can be combined with bone marrow stem cells for repair of bone fractures.

Equipment/PI: 3D Bioprinters and Rheometer

Instructional Microporous Scaffolds for Treatment of Volumetric Muscle Loss

Hydrogels for regeneration of large volumes of tissue has yet to be translated to the
clinic. The elusive balance of material degradation/rearrangement with tissue ingrowth
has been a major research hurdle limiting the progress of hydrogel biomaterials (a
material class capable of mimicking the physiological physical/chemical environment)
from translation to the clinic. The use of injectable microporous annealed particle
(MAP) scaffolds that achieve a seamless material-tissue boundary while allowing for
degradation-free cellular infiltration provides a potential shortcut around this hurdle.
Early results have shown a clear integrative benefit to this approach. However, the MAP
platform has presently only been approached as an inert scaffold and lacks the
instructive capability to promote an organized and directed tissue response over large
distances (>1 cm). To start to grow a toolbox capable of achieving this goal, we
propose to incorporate both temporary and permanent instructive elements throughout
the MAP matrix of microgels.

Spatially Patterned 3D Cultures of T Cells and B Cells Towards a Lymph-Node-On-a-Chip
There is a nationwide effort towards developing so-called “organs on chip” to provide
insight into the cellular and biochemical mechanisms of disease and as a platform for
drug screening. So far, there has been no on-chip model of a lymph node beyond
simple co-cultures of various cells, despite the centrality of this organ to the immune
response. We have identified two significant obstacles to developing a functional
lymph-node-on-a-chip: (i) the ability to pattern T cells, B cells, and dendritic cells to
recapitulate the organization of the node, and (ii) providing cells a matrix network on
which to migrate realistically. This seed grant proposal addresses the former.
The overarching project goal is to pattern 3D cultures of lymphocytes to recapitulate
the critical features of the lymph node’s organized structure and show that the cultures
retain basic function. As a starting point, we will pattern small circular regions of B
cells to mimic B cell follicles, within a larger structure of T cells mixed with dendritic
cells. The soft nanoimprint lithography method will be used because of its ability to
pattern cell-laden gels in close proximity, with excellent spatial resolution and fidelity to
the original pattern and its potential for use in future multi-layer patterns.