Recombinant human midkine stimulates proliferation of articular chondrocytes.
OBJECTIVE
Midkine, a heparin-binding growth factor, promotes population growth, survival and migration of several cell types, but its effect on articular chondrocytes remains unknown. The aim of this study was to investigate its role on proliferation of articular chondrocytes in vitro and in vivo.


METHODS
Bromodeoxyuridine incorporation and MTT assays were performed to examine the proliferative effect of recombinant human midkine (rhMK) on primary articular chondrocytes. Activation of extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3-kinase (PI3K) was analysed using western blot analysis. Systemic and local delivery of rhMK into mice and rats was preformed to investigate the proliferative effect of rhMK in vivo, respectively. Histological evaluation, including measurement of articular cartilage thickness, cell density, matrix staining and immunostaining of proliferating cell nuclear antigen was carried out.


RESULTS
rhMK promoted proliferation of articular chondrocytes cultured in a monolayer, which was mediated by activation of ERK and PI3K. The proliferative role of rhMK was not coupled to dedifferentiation of culture-expanded cells. Consistent with its action in vitro, rhMK stimulated proliferation of articular chondrocytes in vivo when it was administered subcutaneously and intra-articularly in mice and rats, respectively.


CONCLUSIONS
Our results demonstrate that rhMK stimulates proliferation of primary articular chondrocytes in vitro and in vivo. The results of this study warrant further examination of rhMK for treatment of animal models of articular cartilage defects.
Zhang
Z H
ZH
Laboratory of Regeneromics, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.
Li
H X
HX
Qi
Y P
YP
Du
L J
LJ
Zhu
S Y
SY
Wu
M Y
MY
Lu
H L
HL
Yu
Y
Y
Han
W
W
D000818:Animals; D001973:Bromodeoxyuridine / Q000378:metabolism; D002358:Cartilage, Articular / Q000166:cytology* / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D004396:Coloring Agents / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D000078792:Midkine; D009414:Nerve Growth Factors / Q000235:genetics / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000494:pharmacology; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
D004396:Coloring Agents; C066254:MDK protein, human; D009414:Nerve Growth Factors; D011994:Recombinant Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D000078792:Midkine; C022616:thiazolyl blue; D001973:Bromodeoxyuridine
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