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Particulate cartilage under bioreactor-induced compression and shear.

Abstract：

PURPOSE:Our aim was to explore the effect of varying in vitro culture conditions on general chondrogenesis of minced cartilage (MC) fragments. METHODS:Minced, fibrin-associated, bovine articular cartilage fragments were cultured in vitro within polyurethane scaffold rings. Constructs were maintained either free swelling for two or four weeks (control), underwent direct mechanical knee-joint-specific bioreactor-induced dynamic compression and shear, or they were maintained free swelling for two weeks followed by two weeks of bioreactor stimulation. Samples were collected for glycosaminoglycan (GAG)/DNA quantification; collagen type I, collagen type II, aggrecan, cartilage oligomeric matrix protein (COMP), proteoglycan-4 (PRG-4) messenger RNA (mRNA) analysis; histology and immunohistochemistry. RESULTS:Cellular outgrowth and neomatrix formation was successfully accomplished among all groups. GAG/DNA and collagen type I mRNA were not different between groups; chondrogenic genes collagen type II, aggrecan and COMP revealed a significant downregulation among free-swelling constructs over time (week two through week four). Mechanical loading was able to maintain chondrogenic expression with significantly stronger expression at long-term time points (four weeks) in comparison with four-week control. Histology and immunohistochemistry revealed that bioreactor culture induced stronger cellular outgrowth than free-swelling constructs. However, weaker collagen type II and aggrecan expression with an increased collagen type I expression was noted among this outgrowth neotissue. CONCLUSIONS:The method of MC culture is feasible under in vitro free-swelling and dynamic loading conditions, simulating in vivo posttransplantation. Mechanical stimulation significantly provokes cellular outgrowth and long-term chondrogenic maturation at the mRNA level, whereas histology depicts immature neotissue where typical cartilage matrix is expected.

journal_name

Int Orthop

journal_title

International orthopaedics

authors

Wang N,Grad S,Stoddart MJ,Niemeyer P,Reising K,Schmal H,Südkamp NP,Alini M,Salzmann GM

doi

10.1007/s00264-013-2194-9

subject

Has Abstract

pub_date

2014-05-01 00:00:00

pages

1105-11

issue

eissn

0341-2695

issn

1432-5195

journal_volume

pub_type

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Particulate cartilage under bioreactor-induced compression and shear.