The Journal of Bone and Joint Surgery, Vol 76, Issue 7 1042-1051, Copyright © 1994 by Journal of Bone and Joint Surgery, Inc
Chondrogenesis in periosteal explants. An organ culture model for in vitro study
SW O'Driscoll, AD Recklies and AR Poole
Cartilage and Connective Tissue Research Laboratory, St. Michael's Hospital, University of Toronto, Canada.
Periosteal grafts have chondrogenic potential and have been used to repair
defects in articular cartilage. We studied the effects of the culture
conditions and of transforming growth factor-beta 1 on chondrogenesis in
rabbit periosteal explants that were cultured in vitro. A total of 390
periosteal explants were obtained from the anteromedial sides of the
proximal parts of the tibiae of eleven rabbits that were two weeks, two
months, or six months old. The culture medium (alpha minimum essential
medium or Dulbecco minimum essential medium) contained fetal calf serum,
with or without transforming growth factor-beta 1, at a concentration of
one or ten nanograms per milliliter for the first two weeks of culture.
Three hundred and twenty-one explants were submerged in liquid medium and
sixty-nine were suspended in an agarose gel; they were then evaluated
histochemically, histomorphometrically, and by collagen-typing. In the
media without agarose, in the presence of ten nanograms of transforming
growth factor-beta 1 per milliliter, chondrogenesis was commonly seen after
two to four weeks with use of safranin-O staining and histomorphometry. In
the agarose gels, chondrogenesis from the periosteum was observed at four
and six weeks and was enhanced by the presence of one or ten nanograms of
transforming growth factor-beta 1 per milliliter. The combination of
agarose with transforming growth factor-beta 1 most favored the formation
of cartilage, which was maximum at six weeks in the presence of ten
nanograms of transforming growth factor-beta 1 per milliliter. Under these
conditions, chondrogenesis occurred in almost every explant, with 50 +/- 30
per cent of the tissue being composed of cartilage. Type-II collagen was
present in the explants that had undergone chondrogenesis.