Tissue Engineering of Cartilage - Methods in Molecular Biology: Extracellular Matrix Protocols - page 288

12. Scalpel handels and blades (sizes 10, 15, and 20).

2.2. Cartilage Construct Growth
2.2.1. Preparation and Storage of Construct Scaffolds

1. Scaffold preparation. Synthetic polymer scaffolds can either be a) purchased or

b) fabricated in the laboratory. The following two scaffold types support chon-
drocyte attachment, proliferation and cartilage matrix synthesis and deposition.
They are fabricated as a) sheets or b) disks. Scaffold disks ranging from 2-3 mm
in thickness and 8-10 mm in diameter have been used routinely to generate
hyaline-like cartilage constructs.
a. Entangled, nonwoven polyglycolic acid (PGA) felt scaffolds (Albany Inter-
national Research Company, Mansfield, MA) ( see Note 2 ).
Felt, purchased in the form of 20 30 cm sheets needled to a final felt

porosity of 97% with the following characteristics, has been successfully used
in many laboratories (4 , 5) . This PGA scaffold degrades rapidly between 1 and
4 wk of culture (4) .
• 2-mm thick
• Nonheat plated
• Extruded polymer fibers
•14

×

m in diameter
• Nonwoven
b. Solvent-cast poly (lactic-co-glycolic acid) [PLGA] and polylactic acid [PLA]
foam scaffolds ( see Note 3 ). Solvent-cast, salt-leached polymer foams
with a defined pore size range and porosity can be fabricated in the labora-
tory, based on published techniques (11) . Obtain the following to fabricate
this scaffold type:
• Glass container (20 mL capacity)
• Glass pipets
• Glass Petri dish (50 mm in diameter)
• Methylene chloride (dimethyl chloride)
• PLGA; Purchase this polymer with the following characteristics: molar
ratio, between 50:50 and 85:15; inherent viscosity, 0.55-0.75
• PLA; Purchase this polymer with the following characteristics: inherent
viscosity, L-PLA 0.90-1.2, DD-PLA, 0.55-0.75
• Teflon Petri dish (50 mm in diameter)
• NaCl (common, laboratory grade)
• Sieves (to select NaCl crystals of various sizes)
• Deionized water (DI H 2 O)
• Oven (if scaffold of low crystallinity is desired)
•

µ

Orbital shaker

2. Scaffold storage.

a. Foil bags (4 mL capacity).
b. Vacuum line or dry nitrogen tank.
c. Dessicant.