Project Page - Microengineered microvascular network for 3D tissue construct

DATE CREATED: Dec. 22 2008

ACTION PLAN

PROBLEMS IDENTIFIED (PI) / OUTSIDE SKILL REQUIRED (OSR) / RESOLVED (R)

PAPER TITLE : Fabrication and assembly of endothelialized vascular network

A) Background

Controlling microenvironment of cells have potential advantages of predefined structures and functions in tissue engineering field. Cell behavior and biochemical, mechanical functionality of tissue can be tailored by petterning with microfabrication technology. Harmonized cell extracellular matrice, microstructure, mechanophysiological environments are **** to cell survival, proliferation and differentiation.

Tissue engineering with porous scaffold has been limited to single or two types of cells without predetermined vascular structure.

Here, we suggest 3D vascular tissue structure with defined structures for tissue engineering.

Hyaluronic acid and collagen were used as ECM materials.

B) Hypothesis

HA and collagen IPN is suitable for microengineered tissue engineering and can be integrated to 3D structures

B-1) Specific Aims

Make micropatterned vascular structure show feasibility of hydrogel micropattern for 3D tissue engineering.

Stack vascular network sheets to show 3D integrated structure.

C) General Experimental Approach (Design etc)

- Mechanical stability/ Control of degradability(HA)/ Vascular Lumen formation

- Multilayer formation, burst pressure test

- Perfusion culture, cell alignment

C-1) Materials and Methods(GENERAL EXPERIMENTAL APPROACH)

1. Gel materials preparation and gelation (fig. 1)

2. Mechanical evaluation (fig.2)

3. Patterning of microvascular structure(fig. 3)

4. Cell patterning and Immunostaining (fig. 4, 5)

5. Multilayer culture and evaluation

D) Results and Discussion

D-1) Conclusion

E) Potential Figures

Figure 1 : Schematic of fabrication

Figure 2. Mechanical Properties of HA-Colagen IPN

a) Compressive modulus : Jason Nicol

b) Microstructure(SEM image) : Mark Brigham

Figure 3. Cytocompatibility of IPN surface with HUVEC

a) HUVEC cell morphology on the surface of HA-Col IPN

b) HUVEC cell adhesion density after 6 hours of seeding and cells' area on the surface after 6 hours of seeding

Figure 4. Bonding two channels and lumen formation

a) Characterization of lumen formation

b) Secondary Crosslinking - Binding and Leaking Test

Figure 5. Characterization of the lumen comprising endothelized-channels

Figure 6. Building 3D multilayer Vascularized Cardiac Tissue

a) Section staining with using Anti CD 31 specific marker of HUVECs

Figure to be added like this : confocal microscope

b) Viability for day-3 with HUVEC-HL 1 coculture in HA-collagen IPN

Figure 7. Perfusion of microvascular structure

- Endothelial cell actin microfilament, static vs perfusion

Figure 8. Cell morphology along the vascular structure

- Cell morphology around bifurcation vs straight vessel

F) Future Directions