BME 430: Tissue Engineering

Tissue Fabrication Technology

Fall 2022
 What is an “ideal”
scaffold?

How can we make one?

Tissue Fabrication Technologies

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Tissue Fabrication Strategies
• Types
• Native tissues (decellularized matrices)
• Engineered tissues
• Polymers, ceramics, etc.
• Self-assembled constructs

Tissues ≈ constructs ≈ materials ≈ scaffolds

• Requirements
• Structural template
• Logistic template
• Biodegradable
• Custom designed (shape, size, chemistry, mechanical properties, in vivo and in
vitro delivery of multiple growth factors)
• Biological function
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Ideal Scaffold Properties
• Material optimization
• Ideal properties
• Processing • Crosslinkers
• High porosity with interconnected
• Co-polymers • Surface treatments
pores to allow cell infiltration and
mass/gas exchanges • Plasticizers

• Biocompatible and biodegradable • Biochemical factors

with a controlled degradation rate • Adhesion factors (e.g. RGD)
that matches new tissue formation • Cytokines or growth factors (e.g. BMPs)

• Support for cell adhesion, • Cell-based strategies

proliferation and differentiation • Precoating (e.g. endothelial cells)

• Mechanical properties that originally • Molecular biology

match the ones of the damages tissue • Plasmids
• Genetically engineered cells

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Scaffold-Free Fabrication Methods
• Relies on cell
aggregation of matrix
• Cells will reorganize
themselves in direction
of tension
• Excellent approach for
directional tissues

Not truly a
“scaffold-free”
approach… Why?

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 Tissue Organization into Contractile Muscle
• Myoblasts seeded on top of fibrin
gels produce contractile tissue
• Pins keep tissue from fully
contracting
• Note central region of construct

Huang et al. J Appl Physiol, 2005.

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 Methods to Enhance Construct Longevity
• Crosslinking gels assists in longevity of constructs
• Addition of fibroblasts aids in the health of constructs… Why?

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Khodabukus and Baar. TEC, 2009.
 Alternate Matrix Composition Affects Results
• Fibrin with Matrigel • Stem cell niches
• Note center of constructs

Khodabukus et al. Adv Healthcare Mater, 2018. Juhas et al. PNAS, 2014. 9
 “True” Scaffold Free Systems..

• Scaffold serves as anchor

• Mechanically condition
constructs (via deflection)
• Fibroblasts remodel into
“fibers”
Mubyana and Corr. TEA, 2018.
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 Cell Rings for Vascular Tissue Engineering
• Agarose posts do not facilitate cell
attachment
• Smooth muscle cells self-aggregate
to form blood vessel analogues

Strobel et al. JBMR B , 2018. 11

Gwyther et al. CTO, 2011.
Cell Sheet Fabrication
• Grow to confluence
and detach them from
culture
What material will
assist with this
process?

• Sheets can be stacked,

rolled, etc.

• Cells are making all the

matrix here

• Alternately can be
grown on restrictive
surfaces (like before)
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 Cell Sheets for Vascular Tissue Engineering
• Fibroblast sheets rolled up

L’Heureux et al. Nat Med, 2006.

L’Heureux et al. NEJM, 2007. 13
Soft Lithography and Microfluidics
• Development of
microfluidics, minute
topographies, and
embedded vascular
networks

• Expensive initial
investment

• Involved with “lab-

on-a-chip” models

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Methods of Soft Lithography
• Lithography can be used to
generate precise geometries
to develop niches for cell
growth

• After development of
photoresist, have reusable
platform

Clement et al. Acta Biomaterialia, 2013. 15

 Applications for Soft Lithography
Printing of materials to control cell
Microfluidic platforms to model cell migration
attachment

Pham et al. CMBE, 2018. Palchesko et al. PLoS ONE, 2012.

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Organ-on-a-Chip Models
• Model systems for
• Development/maturation
• Drug toxicity

• Single, or multiple compartments

• Typically include vascular

compartment (nutrient transfer)

Burdick and Vunjak-Novakovic. TEA, 2009. 17

 Organ-on-a-Chip Models
• Integrated systems will work together to combine multiple
tissue components
• Model cell interactions and fluid flow

Marturano-Kruik et al. PNAS, 2018.

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Organ-on-a-Chip Models
• Can be designed to be
parallel systems

• Typically, systems are in

series

• Ultimately will depend on

the design criteria for the
project
Yahya et al. Sensors, 2014.

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Linking Multiple Organ Systems
1. Connect outflow/inflow
• Require common media
• Metabolic wastes passed on to next organ
• Organ specific flow rates?
• Adequate O2
• Scaling? (Allometric – Biological Scaling)

Kimura et al. Drug Metab Pharmacokinet. 2018.

Linking Multiple Organ Systems
2. Functional Coupling
• Transfer of media from one
organ module to the next
• Adjustable flow rates
• Adjustable media compositions
• Can be operated at different
locations/times

Rothbauer et al. Lab Chip, 2018.

 Organ-on-a-Chip Models – Combining Everything

Multiple different cell

compartments to
observe tissue-level
interactions
Why?

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Ronaldson-Bouchard and Vunjak-Novakovic. Cell Stem Cell, 2018.
Homework
Discussion
Homework Discussion
• Stay with your group!
• 15 minutes to answer all of the questions for each paper
• For each paper discuss:
• Clinical need being addressed
• Most important results/findings of the approach
• Reasons (multiple) they did not achieve the intended goals of the
work?
• Poor experimental design
• Claims not based on data presented
• Other concerns?
• Regroup and present your answers to the class (pick a leader!
)

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