Application of Bovine Pituitary Extract and Polyglycolide/Poly(lactide‐co‐glycolide) Scaffold to the Cultivation of Bovine Knee Chondrocytes

Abstract

In vitro cultivation of primary bovine knee chondrocytes (BKCs), using bovine pituitary extract (BPE) and porous scaffolds composed of polyglycolide (PGA) and 85/15 poly(lactide-co-glycolide) (PLGA), was investigated. Here, BPE was prepared from fresh bovine pituitaries, and cylindrical PGA/PLGA scaffolds with various chemical compositions were fabricated by solvent merging/particulate leaching method. Experimental results showed that in microcarrier systems, the rate of BKC growth on PGA surfaces is faster than that on PLGA surfaces, and the decrease in the medium pH value of BKCs-adsorbed PGA particles is faster than that of BKCs-adsorbed PLGA particles. After 28-day construct cultivation, the BKC amount and the content of glycosaminoglycans and collagen per construct increased with BPE protein concentration. For a constant BPE protein concentration, a higher PGA percentage in scaffold leads to a better biological environment for the growth of BKCs and the synthesis of extracellar matrices.

Effect of genipin‐crosslinked chitin‐chitosan scaffolds with hydroxyapatite modifications on the cultivation of bovine knee chondrocytes

“…BKCs were separated from articular tissues of bovine calves by the method of enzymatic digestion, and the freshly obtained BKCs could be preserved at À858C (Sanyo, Osaka, Japan) and defrozen by the procedures explicated in the previous study (Kuo and Chung, 2005).…”

“…Seeding of BKCs on scaffolds was depicted in the previous study (Kuo and Chung, 2005) with minor modification. Briefly, after treatment by 70% ethanol for 30 min and washing by DPBS, the scaffolds were dried under ultraviolet.…”

“…In vitro BKC cultivation was achieved by the method explained in the previous study, and BKC amount, GAG content, and collagen level after 28-day cultivation were analyzed by the schemes elucidated previously (Kuo and Chung, 2005). For microtomy, fixation and dehydration of the cultivated constructs were achieved by immersion in formaldehyde over 1 h, deionized water over 30 min, 70% ethanol over 1 h, 80% ethanol over 1 h, and 95% ethanol for 2 h. The constructs were soaked in o-xylene over 3 h for transparency.…”

Application of polyethyleneimine‐modified scaffolds to the regeneration of cartilaginous tissue

“…20 Briefly, aseptic forearm knees of bovine calves were reaped at a local abattoir within 30 min of animal death and transported antiseptically to the laboratory in ice-bathed DPBS containing antibiotics. Cartilages were sliced into about 1 mm 3 cubes, digested by 0.18% type II collagenase in a CO 2 incubator at 37 C over 24 h, and centrifuged at 420g for 5 min.…”

“…20 Briefly, the scaffolds with apparent dimensions of 10 Â 10 Â 5 mm 3 were saturated with a cell suspension at a density of 8.25 Â 10 6 BKCs/construct. The seeded constructs were fastened through stainless steel wires in the middle of a spinner flask, which was stirred by two internal paddles.…”