Ci is the concentration of nutrients, growth factors

Ri represents cell growth / decay

For Electrospun fibers

For fibers of larger size

For open pores

Spinner Flask

Here, r, t represents position and time, εσ is the volume fraction of cells within the scaffold, and can be
related to the cell diameter dcell and the cell density ρ(r, t) by

As per Michael Menten Kinetics, the reaction term in the original mass balance equation for oxygen
(nutrient) due to consumption of oxygen by the cell is given as

Monod Kinetics

Here Vmax is the maximum oxygen consumption rate, KS is the saturation, or half-rate constant, defined as the
oxygen concentration at which the consumption rate drops to half of its maximum value, µmax is the maximal
cell specific growth rate, YXS is the yield of cells per unit oxygen, and ms is the maintenance coefficient, the
minimum oxygen consumption required to keep the cells alive.
While both models are of essentially the same form, the Monod model has the advantage that the oxygen
consumption rate can be directly coupled to the cell growth rate as

Note that this equation has no inhibition term related to cell density, and is hence not self-limiting to a physically
realistic maximum cell density. However, the cell growth rate at high cell density becomes restricted by very low
oxygen diffusivity, and hence the density is limited to a physically realistic level.
Each square is called a cellular automaton that can contain a finite number of
possible cell states that evolve with time. We assume that an automaton can
contain at most a single cell. Namely a lattice square is either (1) occupied by a
cell, which is at a certain time of its life cycle, moving in a certain direction or
being stationary; or (2) empty and available for a cell to move or divide in. In
addition to the discrete automata space, time in the model is also discretized
based on the choice of the discrete time interval Δtc . The state of an automaton
is determined by its interaction with a finite number of neighboring
automata at discrete time steps.
 Cell division is considered as being modulated by the local oxygen content. The nutrient
diffuses into the scaffold from the surrounding culture medium and is consumed by cells.

C is the oxygen concentration. The nutrient uptake is described by the Michaelis–Menten

equation, where Qmax is the maximum uptake rate, and Kc the nutrient concentration at
the half uptake rate. The index ξ is a function of the spatial position x, which takes the value
of ξ = 1 if there is a cell at the local space, otherwise ξ = 0.
Boundary Condition
A quarter of the scaffold is taken as the simulational domain. Cells are confined in the

scaffold; therefore cells at boundaries 1 and 2 are not allowed to move or divide across the

border into the surrounding culture medium. Boundaries 3 and 4 are considered to be

symmetric planes at which cells undergo reflection. That is if a cell is moving or dividing

across boundaries 3 and 4 into the other part of the scaffold, then another cell from the other

part of the scaffold will move in the simulation domain, occupying the symmetric lattice in

the boundary. Since cells are not allowed to leave the scaffold, the fraction of volume

occupied by cells will gradually grow to one. Cell death has been neglected.
Cell Division
The mean value of the cell cycle, tgm, is determined according to the local nutrient
concentration by the Monod kinetics

where C is the average nutrient concentration over the local automaton that contains
the cell in question, and Rg the maximum division rate and Kg the nutrient
concentration at the half maximum division rate. Based on the mean value, the cell
cycle time is assumed to follow the Gamma distribution.
Division Counter:

Migration
Persistence Counter

The cell state index m is an integer with values from 1 through 10.
For m = 1; 2; ... ; 8, it means the cell is undergoing a persistent walk in one of the eight
directions (1 = north, 2 = east, 3 = south, 4 = west, 5 = northeast, 6 = southeast, 7 = southwest, 8 =
northwest). The persistence counter decreases by one at each time step of Δtc as the cell
migrates. When the counter reaches zero, the cell changes its moving direction by
randomly choosing one of the other seven directions provided the neighboring automaton
is available. Based on the same reason as mentioned in the division rule, the probability for the
cell to move into a neighboring corner automaton is half the edge automata to offset the
longer migration range in the oblique directions. The values of m = 9 and 10 are used to
denote the cell in one of the two stationary states (cell collision and contact inhibition
respectively), which are defined in the next.
Collision
When a cell undergoing its persistent walk collides with another cell, the two cells are
considered to stay until the collision time ta elapses, known apriori. The cell state indices
for the two cells are both reset to = 9, and the persistence time tp for each of the cells is
replaced by the collision time ta . The persistence counter for each cell is accordingly reset
When the collision time of a cell expires, i.e., the persistence counter defined
above counts down to zero, the cell resumes motility towards a randomly
selected direction by the same method defined in the migration rule.
Note that the division counter continues to work while the cell is in a stationary
phase.

Contact inhibition:
When the division counter kg of a cell counts down to zero and the cell is about to
divide, the neighborhoods of the cell are checked if there are any vacant
neighboring sites allowing for the cell to divide in. If not, the cell halts it division
at the moment, and the division counter kg is reset as one to keep the cell staying in
the G1 phase of the cell cycle. Therefore the cell is possible to resume division
provided that a vacant neighborhood is available at the next time step.
Contact-inhibition is also checked when a cell is about to change its moving
state, i.e., the persistence counter counts down to kp = 0. If the cell is found to be
contact-inhibited, it halts migration and its persistence counter kp is also reset as
one. Therefore the cell is possible to resume migration next time step. The state
index m is set to m = 10 indicating that the cell is in the contact inhibition state.

One possible Arrangement

The square size of cellular automata was taken to be ΔL = 8:5 µm based on the
dimension of chondrocytes. For the sake of symmetry, a quarter of a 1 cm wide and
0.116 cm high scaffold was taken as the computational domain which yields 588 x 68
cellular automata in a two-dimensional space. On the other hand, the grid sizes Δx and Δy of
the nutrient field were tested by randomly and uniformly distributing 20 000 cells (about a
cell volume fraction of 50%) throughout the automata. The cells were kept stationary for the
grid test without division and migration. By performing the calculation to a steady state, the
relative error for oxygen concentration was found below 0.1% when Δ x =Δy = ΔL=6.
The time step Δtc for the cellular automata simulation was determined by the specified cell
migration speed S. As the cell speeds range from 8.5 to 25.5 µm/h, the cellular time step Δtc
were set correspondingly from 1 h to 20 min. The discrete time interval Δtn for calculating
the oxygen field was adaptively chosen. It required 0.01 s for the first 100 steps to capture
the oxygen change in response to the abrupt change of automata state at each cellular time step,
and then it was switched to 1 s for the following time marching. After setting the initial
oxygen concentration, 290 cells (a volume fraction of 0.726%) were randomly distributed
throughout the automata. The initial cell cycle time for each cell was randomly assigned
based on the Gamma distribution. The initial migration direction is randomly assigned one of
the eight directions for each cell too.

Cell Distribution (Left) Normalized Nutrient Concentration (Right)