Distribution of hyaluronan in the epiphysial growth plate: turnover by CD44-expressing osteoprogenitor cells.
In the present study, we have examined the distribution of both hyaluronan and its receptor, CD44, during the process of endochondral ossification in the mouse tibia. Histochemical staining revealed that a large amount of hyaluronan was present in the lacunae located in the zone of hypertrophy, but it was greatly reduced or absent from the zone of erosion. In addition, hyaluronan was present in the cytoplasm of osteoprogenitor cells located in the zone of erosion. These cells also expressed CD44 on their surfaces, as revealed by double-label immunohistochemistry. These results suggested that the osteoprogenitor cells may use CD44 to bind and internalize hyaluronan, and subsequently degrade it with lysosomal enzymes. To test this possibility, we examined the human cell line, MG-63, which closely resembles osteoprogenitor cells. These cells produced several different forms of CD44, as determined by western blotting (85, 116 and 150 kDa). In addition, the binding of isotopically labeled hyaluronan to detergent extracts of these cells was blocked by a monoclonal antibody to CD44. Similarly, the degradation of hyaluronan by these cultured cells was also inhibited by a monoclonal antibody to CD44. To determine if these cells could remove hyaluronan from the growth plate, the cells were cultured directly on top of thin sections of the epiphysial region of long bone. After 16 hours, the sections were stained for hyaluronan. The MG-63 cells removed significant amounts of hyaluronan present in the zone of hypertrophy, and this effect was blocked by an excess of soluble hyaluronan and by a monoclonal antibody to CD44.(ABSTRACT TRUNCATED AT 250 WORDS)
Pavasant
P
P
Department of Cell Biology, Georgetown University Medical Center, Washington, DC 20007.
Shizari
T M
TM
Underhill
C B
CB
D000818:Animals; D002352:Carrier Proteins / Q000032:analysis*; D006132:Growth Plate / Q000737:chemistry*; D006801:Humans; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid / Q000032:analysis*; D015576:Hyperostosis / Q000378:metabolism / Q000473:pathology; D007150:Immunohistochemistry; D051379:Mice; D010006:Osteoblasts / Q000276:immunology / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000032:analysis*; D016201:Receptors, Lymphocyte Homing / Q000032:analysis*; D013977:Tibia / Q000166:cytology; D014407:Tumor Cells, Cultured
D002352:Carrier Proteins; D018960:Hyaluronan Receptors; D011956:Receptors, Cell Surface; D016201:Receptors, Lymphocyte Homing; D006820:Hyaluronic Acid