Fibroblast seeding and culture in biodegradable porous substrates

Abstract

A natural poly(hydroxybutyrate‐co‐9% hydroxyvalerate) copolyester was processed into a three‐dimensional porous foam structure by salt leaching/solvent casting with previously sieved sodium chloride salts. Laboratory‐built P(HB‐9% HV) foams and commercial collagen sponges were cut into small rectangular specimens, sterilized, and prewetted using ethanol, rinsed with Dulbecco's minimum essential medium + 10% serum culture media, and seeded with fibroblasts isolated from canine anterior cruciate ligaments. The fibroblast cultures into such porous substrates were performed from 0 to 35 days by incubation (5% CO₂) at 37°C. It demonstrated that the P(HB‐HV) sustained a cell proliferation rate similar to that observed in collagen sponges, up to at least 35 days, with a maximal cell density on the day 28 in culture. On the other hand, the P(HB‐HV) materials kept their structural integrity during the culture period while the collagen foams contracted greatly. Further, the total protein production after 4 weeks in culture was found to be twice as high (190 ± 10%) in the P(HB‐9% HV) foam than in the collagen foam. Porous P(HB‐HV) materials appear to be adequate polymeric substrates for cell cultures. However, further evaluations are still required to confirm such preliminary results. © 1995 John Wiley & Sons, Inc.