And my soul hungered; and

I kneeled down before my Maker, and

I cried unto him in mighty prayer and
supplication for mine own soul; and all
the day long did I cry unto him; yea,
and when the night came I did still raise
my voice high that it reached the
heavens.

Enos 1:4
 Reading Questions 19
1. What limits growth of cells? Nutrient limitations, oxygen limits
2. What is the cell lifeline number? The relative importance of
consumption of an essential nutrient, and of diffusion of the
nutrient to the cell.
3. For what kinds of tissue would a zero order model most likely
apply? Not many, cartilage, outer layer of skin.
4. For what kinds of tissue would a first order model most likely
apply? Most tissues that are metabolically active.
5. What transport mechanisms will determine oxygen transport
within pores of a scaffold covered with growing cells?
Consumption of the oxygen by the cells will require Fick’s
second law of concentration changing with time, and diffusion.
 Transport in Tissue Engineering
A. Cells from healthy tissue usually don't
proliferate indefinitely as do tumor cells.
B. Cells are very picky about their growth
conditions.
1. Their default mode is not to grow.
2. Nutrients (glucose, O2, etc.), growth factors, and
temperature must be just right.
3. Waste products (lactic acid, CO2, etc.) must be
removed
C. In vitro growth/survival is limited by nutrient
transport through the tissue
1. Nutrient concentrations decrease as they transport
into the “tissue”
a. A concentration gradient drives the diffusion
b. Nutrients are partially consumed as they pass
through the cells
2. O2 is often the limiting nutrient in reconstructed
tissues
a. Monolayer of confluent hepatocytes, the limiting
distance of unstirred water between the O2 and the
cells is 0.95 mm.
b. In a cylinder of hepatocytes, the radius must be
< 181 µm to maintain internal cells
3. The critical length can be estimated using the
cell lifeline number (S),
a. relative importance of consumption of an
essential nutrient, and of diffusion of the nutrient to
the cell:

where RA is the rate of consumption (mole/cm3· s),

l is the diffusion length (cm), D is the diffusivity
(cm2/s), and co is the nutrient concentration
(mol/cm3) at or near the surface.
b. When S = O(1), then l is the critical length that the
cells can grow from the surface.
 Flat Film Models
1. Tissue-cultured skin
2. Tissue-cultured retina
3. Tissue-cultured myocardial patches

No motion in
the film layer
 Class Exercise 1
0 0 0 0

Reduce this to an equation for steady-state

diffusion through the tissue of a flat film, where
Rox (rate of oxygen consumption) = -RA and is
assumed to be constant (zero-order kinetics).
Assume no flow within the film layer.
𝜕𝜕2 𝑐𝑐𝐴𝐴
0= 𝐷𝐷𝐴𝐴𝐴𝐴 − 𝑅𝑅𝑂𝑂𝑂𝑂
𝜕𝜕𝑦𝑦 2
 Class Exercise 1 (cont.)
a. For what kinds of tissue would this zero-order
model most likely apply?
Not many, only when diffusion is faster than
consumption of oxygen. Examples where it may
apply are slow growing tissue like cartilage and
the dead skin layer that is not highly vascularized.
b. For what kinds of tissue would this zero-order
model most likely NOT apply?
Most tissues with blood supply but that are not
actively consuming oxygen (e.g. deep skin). Active
tissues (e.g. liver, brain, heart) may have ROX = kCOX
 Class Exercise 2
2. Derive the concentration profile for oxygen
through the film of growing cells, where the
boundary conditions are assumed to be:
a. BC 1: @y=0, cox=c0 (assuming much faster
transfer in the flowing nutrient solution)
b. BC 2: @y=L, dcox/dy=0 (assuming the oxygen
concentration will stop changing when the
oxygen drops to a critical concentration)
c. Assume that Rox is zero order.
 Class Exercise 2 Solution

C0
2
𝜕𝜕 𝑐𝑐𝐴𝐴 y=0
0 = 𝐷𝐷𝐴𝐴𝐴𝐴 2
− 𝑅𝑅𝑂𝑂𝑂𝑂
𝜕𝜕𝑦𝑦 y=L

𝑅𝑅𝑂𝑂𝑂𝑂 𝜕𝜕 2 𝑐𝑐𝑂𝑂𝑂𝑂 𝑅𝑅𝑂𝑂𝑂𝑂 𝑑𝑑𝑐𝑐𝑂𝑂𝑂𝑂

= � 𝑑𝑑𝑑𝑑 = � 𝑑𝑑
𝐷𝐷𝐴𝐴𝐴𝐴 𝜕𝜕𝑦𝑦 2 𝐷𝐷𝐴𝐴𝐴𝐴 𝑑𝑑𝑑𝑑

𝑅𝑅𝑂𝑂𝑂𝑂 𝑑𝑑𝑐𝑐𝑂𝑂𝑂𝑂 𝑑𝑑𝑐𝑐𝑂𝑂𝑂𝑂

𝑦𝑦 + 𝐶𝐶1 = BC 2: at y=L, =0
𝐷𝐷𝐴𝐴𝐴𝐴 𝑑𝑑𝑑𝑑 𝑑𝑑𝑑𝑑

𝑅𝑅𝑂𝑂𝑂𝑂 𝑅𝑅𝑂𝑂𝑂𝑂 𝑅𝑅𝑂𝑂𝑂𝑂 𝑑𝑑𝑐𝑐𝑂𝑂𝑂𝑂

𝐿𝐿 + 𝐶𝐶1 = 0 - 𝐿𝐿 = 𝐶𝐶1 so (𝑦𝑦 − 𝐿𝐿) =
𝐷𝐷𝐴𝐴𝐴𝐴 𝐷𝐷𝐴𝐴𝐴𝐴 𝐷𝐷𝐴𝐴𝐴𝐴 𝑑𝑑𝑑𝑑
Class Exercise 2 Solution (cont.)
𝑅𝑅𝑂𝑂𝑂𝑂 𝑑𝑑𝑐𝑐𝑂𝑂𝑂𝑂 𝑅𝑅𝑂𝑂𝑂𝑂
(𝑦𝑦 − 𝐿𝐿) = ∫ 𝐷𝐷 𝑦𝑦 − 𝐿𝐿 𝑑𝑑𝑑𝑑 = 𝑑𝑑𝐶𝐶𝑂𝑂𝑂𝑂
𝐷𝐷𝐴𝐴𝐴𝐴 𝑑𝑑𝑑𝑑 𝐴𝐴𝐴𝐴
0
𝑅𝑅𝑂𝑂𝑂𝑂 𝑦𝑦 2
− 𝐿𝐿𝐿𝐿 + 𝐶𝐶2 = 𝐶𝐶𝑂𝑂𝑂𝑂 BC 1: at y=0, 𝐶𝐶𝑂𝑂𝑂𝑂 = 𝐶𝐶0
𝐷𝐷𝐴𝐴𝐴𝐴 2

𝐶𝐶2 = 𝐶𝐶0

𝑅𝑅𝑂𝑂𝑂𝑂 𝑦𝑦 2
𝐶𝐶𝑂𝑂𝑂𝑂 = − 𝐿𝐿𝐿𝐿 + 𝐶𝐶0
𝐷𝐷𝐴𝐴𝐴𝐴 2
 Class Exercise 3
3. Using the concentration gradient expression
from Class Exercise 2:

a. impose the following: @y=L, cox= ccrit

b. derive an expression for the length (L) for

known values of D, Rox, c0, and ccrit
 Special Problem 6
Repeat the derivation from our class in which the
concentration profile of oxygen in a flat layer
of cultured cells was obtained from the species
continuity equation. Then take that solution
further by deriving an expression for the critical
thickness of the cell layer at which the oxygen
concentration would drop to ccrit (called “L” in II. C. 3
in the notes).
 Class Exercise 4
4. If we had assumed first-order kinetics (Rox=kcox),

a. How would you do the problem differently?

b. What form would the solution take?

 Class Exercise 4 (Solution)
𝜕𝜕 2 𝐶𝐶𝑂𝑂𝑂𝑂 𝜕𝜕 2 𝐶𝐶𝑂𝑂𝑂𝑂 𝑘𝑘𝑘𝑘𝑂𝑂𝑂𝑂
0 = 𝐷𝐷𝐴𝐴𝐴𝐴 2
− 𝑘𝑘𝑘𝑘𝑂𝑂𝑂𝑂 =
𝜕𝜕𝑦𝑦 𝜕𝜕𝑦𝑦 2 𝐷𝐷𝐴𝐴𝐴𝐴

𝑘𝑘 𝑘𝑘
Solution: 𝐶𝐶𝑂𝑂𝑂𝑂 = 𝐴𝐴 sinh + 𝐵𝐵 cosh
𝐷𝐷𝐴𝐴𝐴𝐴 𝐷𝐷𝐴𝐴𝐴𝐴

Apply Boundary Conditions:

𝑘𝑘 𝑘𝑘
𝐶𝐶𝑂𝑂𝑂𝑂 = 𝐷𝐷1 exp(
𝐷𝐷𝐴𝐴𝐴𝐴
𝑦𝑦) + 𝐷𝐷2 exp(−
𝐷𝐷𝐴𝐴𝐴𝐴
y)

See page 427 in Chapter 10

 Thicker Geometries
A. Guiding the cell growth
1. Micropatterning of a surface with hydrophilic
and hydrophobic patterns using lithography
technology. Example: Nerve tissue…
a. Smooth-walled channels produce long straight
organized nerve axons
b. Textured surfaces make scattered nerve junctions
2. Scaffolds – made from biodegradable
polymers so they go away
Want the polymer to degrade and disappear at
the same rate as the cells grow
td
= O(1)
th
where td = characteristic time for degradation
th = characteristic time for healing
 Pores
1. What transport mechanisms will determine
oxygen concentrations within the pores?

where k = mass-transfer coefficient from the pore liquid to the cell

boundary
 Pores (continued)
@ z=0, cpore = c0
Boundary conditions:
@ z=L,
a. By writing the differential equation as shown
above, what assumption is being made about
relative transport rates across the pore and into
the cells?
Transport in the r-direction into the growing
cell layer will be slow, so there is no gradient in
r-direction. Only gradient is in the z-direction.
Assuming cpore>>ccells, the differential equation becomes

which has the general solution:

where

Finally, applying the boundary conditions and solving gives

where and mL is called the Thiele modulus.

3. What other models are possible besides
ccells = ~0

Gradient in the r-direction because cells consume

O2 fast
𝜕𝜕𝐶𝐶𝑂𝑂𝑂𝑂 𝜕𝜕𝐶𝐶𝑂𝑂𝑂𝑂
( 𝐷𝐷𝑂𝑂𝑂𝑂 −𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝 = 𝐷𝐷𝑂𝑂𝑂𝑂 −𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐 ) at r=R
𝜕𝜕𝑟𝑟 𝜕𝜕𝑟𝑟
4. In the growing cell layer….
a. Given the general species continuity
equation in cylindrical coordinates….

What differential equation would describe the

oxygen transfer in the cell layer? At SS, no
velocities:
1 𝜕𝜕 𝐶𝐶𝑂𝑂𝑂𝑂
0 = 𝐷𝐷𝑂𝑂𝑂𝑂−𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐 𝑟𝑟 + 𝑅𝑅𝑂𝑂𝑂𝑂
𝑟𝑟 𝜕𝜕𝜕𝜕 𝜕𝜕𝜕𝜕
b. What boundary conditions?
At r=R, Cox = C0 and r = R+δ, dCox/dr = 0
c. How must we treat the concentration outside
the cell layer differently than we did in the case
for the flat plate on pp.1-2?
Concentration changes in z-direction.

d. What effect does the answer to part c above

have on the direction(s) of diffusion inside the
cell layer?
If COX changes in z-direction outside cells,
then it changes in z-direction also inside cell
layer. Some O2 will diffuse in cells in z-
direction.
5. How would you begin your efforts to obtain a
solution to this problem?
Look at the limiting cases: fast and slow O2
consumption. Superposition of linear
solutions and BCs. Numerical methods.

6. What is the heat transfer analogy to this

problem?
Heat transfer in pores coated with material
that generates heat, like uranium or metal
heaters.
 Special Problem 7
In our class, we discussed the problem of steady-state
oxygen transport in a long, thin cylindrical pore (no
convection) lined with cultured cells (see III.B of the
notes). We considered whether it was appropriate to
model such a problem using “film theory.” By not
making the “film theory” assumption, formulate the
governing differential equation by eliminating
irrelevant/negligible terms from the species continuity
equation. Also write the appropriate boundary
conditions.
Note: you are NOT being asked to solve this problem to
obtain a concentration profile.