Cell-Scaffold Interaction

Types of Interactions

• Scaffold influences cell viability, growth, function and motility.

• Types of cellular interaction under influence of scaffold
1. Adhesion
2. Migration
3. Aggregation
 1. Adhesion
• Most tissue derived cells require attachment to a solid surface for viability
and growth.
• Cell adhesion to a surface is critical because it is followed by other
important phenomena like cell spreading, migration and differentiated cell
function.
• Phenomena
i. Cell attachment
ii. Cell spreading
iii. Focal adhesion

• Techniques used to determine cell adhesion are-

1. Sedimentation-detachment assay
2. Centrifugation assay
3. Fluid-flow chambers
i. Cell attachment- cells attach to the surface of
the scaffold and form monolayer on the
scaffold
ii. Cell spreading- surface attached cells divide
and proliferate to cover the surface of the
scaffold. The cells also penetrate inside the
interconnected pores of scaffold.
iii. Focal adhesion- Focal adhesions are large,
dynamic protein complexes through which the
cytoskeleton (protein present in the cell outside
the cytoplasm e.g intigrin, actin, myosin) of a
cell connects to the extracellular matrix
(scaffold).
 Techniques to determine cell adhesion

• Sedimentation-detachment assay
i) sedimentation of cells onto a surface
ii) incubation of the sedimented cells in
culture medium for some period of time
iii) detachment of loosely adherent cells by
removal of the culture medium and
repeated washing

• The extent of adhesion is determined by

the number of cells that remain
associated with the surface or the number
of cells that were extracted with washes.
 Techniques to determine cell adhesion

• Centrifugation assay
i) Seeding of cells onto a scaffold surface
ii) incubation of the cells in culture medium
for some period of time
iii) the plate is inverted and subjected to a
controlled detachment force by
centrifugation.
The extent of cell attachment is then
quantified
 Techniques to determine cell adhesion
• Fluid-flow chambers
Fluid mechanical forces are utilized to produce cell detachment in a
well-controlled and quantifiable manner.
i) Cell suspension is injected into the chamber, and the cells are
permitted to settle onto the surface of scaffold and adhere.
ii) After incubation the fluid is forced between two parallel plates and
non adherent cells are removed with the flow of fluid, while adherent
cells remain on the surface, which can be quantified.
 2. Migration
• Migration of individual cells within a tissue is critical for formation
of the architecture of organs.

• In tissue engineering, the ability of cells to move, in association

with scaffold surface or through other cells, will be an essential part
of new tissue formation or regeneration.

• Techniques used to determine cell migration are-

1. Under agarose test
2. Filter assaying
3. Direct visualization
 Techniques to determine cell migration
• Under agarose test
i) a cell suspension is placed in a well of
semisolid agarose
ii) motile cells crawl on the solid substrate
underneath agarose.
• Filter Assay
i) cell suspension is placed on a filter with small
pores
ii) motile cells crawl through the pores of the filter
material to the other side, where they are
detected.
• Direct visualization assays
- the paths of movement of many individual cells are
directly observed for cells migrating on
surfaces and within solid gels
 3. Aggregation
• Important in tissue development
• It correlates cell-cell interaction with cell differentiation, viability
and migration for subsequent tissue formation.
• Aggregate morphology allows re-establishment of cell-cell contact
in tissues, thus cell function and survival rate are enhanced in
aggregate culture.
• Formation of aggregates-
by incubating cells in suspension and adding serum proteins to
promote cell aggregation.
• Techniques used to determine aggregation are-
1. Direct visualization
2. Electronic particle counter
3. Aggregometers
 Techniques to determine aggregation

1. Direct visualization –
Monitoring aggregate size to determine extent of aggregation

2. Electronic particle counter -

Invented by Moscona, determines kinetics of aggregation by
measuring aggregate size distribution over time.
This procedure utilizes computer image analysis to follow
disappearance of single cells with time.

3. Aggregometers -
Small angle light scattering through rotating sample cuvettes are
used to produce continuous record of aggregate growth.