2

Materials

Cell type and culture techniques should be chosen according to the

application. The scaffold material can include natural or synthetic

biomaterials. The method described refers to parameters tested

in a modified polyurethane for approaches in cartilage tissue

engineering.

2.1

Bioreactor

System

All components used in the bioreactor system are included in the

equipment. The dual real-time compressive and fluid flow modes

were mounted as described in this section.

1. ElectroForce^® BioDynamic 5200^® (Bose/TA Instruments)

bioreactor, connected to a peristaltic pump and assembled to

22N load cells. The bioreactor possesses four force modules,

each one connected to a chamber (Fig. 3) (see Note 1).

2. 180 mL of Dulbecco’s Modified Eagle’s (DMEM) supplemen-

ted with fetal bovine serum 10% (FBS) and antibiotics (penicil-

lin 10.000 units, streptomycin 10 mg, and amphotericin B

25 μg per mL), for each bioreactor chamber (see Note 2).

3. 3-Stop PVC Pump tubings hermetically mounted on the

chambers (see Note 3).

4. Three way stopcocks connected to the pump tubings (see Note

4).

5. 0.22 μM syringe filters.

6. A cell culture incubator large enough to have the entire biore-

actor system housed in it, connected to a CO2 gas supply.

Fig. 2 Laser scanning confocal microscopy images of aggrecan labeling under static conditions versus

dynamic-mechanical bioreactor, under fluid flow and mechanical compression. Indirect labeling for aggrecan

is identified in red by Alexa Fluor 647. Bars 50 μm

Optimized Dynamics in Bioreactors System

129