Effect of antisense oligonucleotide against mouse dentine matrix protein 1 on mineralization ability and calcium ions metabolism in odontoblast-like cell line MDPC-23.
OBJECTIVE
To study the mineralization ability and the dynamic changes of intracellular and extracellular concentrations of calcium ions in the odontoblast-like cell line MDPC-23 affected by antisense oligonucleotide (AS-ODN) against mouse dentine matrix protein 1 (DMP1).


METHODS
The expression of DMP1 in MDPC-23 cells was detected by an immunohistochemical method and its blocking outcome by the Western blot method. The alkaline phosphatase (ALP) activity, size and number of mineralized nodules, and the intracellular free ([Ca2+]if), total ([Ca2+]it) and the extracellular ([Ca2+]e) calcium ion concentrations in MDPC-23 cells in the experimental group affected with AS-ODN were compared with those in the control group (paired-samples t-test).


RESULTS
Dentine matrix protein 1 was stably expressed in a stable way in MDPC-23 cells; the expression was only just detectable at 12 h and became negative after 24 h affected by AS-ODN. Compared with the control groups, ALP activity of MDPC-23 cells in the AS-ODN group was decreased (P < 0.05), and both the number and size of mineralized nodules were smaller than those in the control group. [Ca2+]if in the AS-ODN group increased and then decreased after 24 h. [Ca2+]it dropped substantially to the lowest point at 24 h (P < 0.01). [Ca2+]e increased before treatment for 24 h and then dropped, however, it was still higher than that of the control group.


CONCLUSIONS
Antisense oligonucleotide against DMP1 could decrease mineralization ability and affect the intracellular and extracellular concentrations of calcium ions in MDPC-23 cells. This would indicate that DMP1 regulates the metabolism and transportation of calcium ions in odontoblasts, and thus boosts dentine mineralization.
Pang
J L
JL
Department of Operative Dentistry and Endodontics, Qindu Stomatological Hospital, The Fourth Military Medical University, Xi'an City, China.
Wu
B L
BL
He
W X
WX
Zhang
Y Q
YQ
Zhao
H P
HP
Xie
Z H
ZH
D000469:Alkaline Phosphatase / Q000032:analysis / Q000187:drug effects; D000818:Animals; D015153:Blotting, Western; D002113:Calcification, Physiologic / Q000187:drug effects* / Q000235:genetics; D002118:Calcium / Q000032:analysis / Q000378:metabolism*; D002460:Cell Line; D016326:Extracellular Matrix Proteins / Q000187:drug effects* / Q000235:genetics; D005110:Extracellular Space / Q000378:metabolism; D007150:Immunohistochemistry; D042541:Intracellular Space / Q000378:metabolism; D051379:Mice; D009804:Odontoblasts / Q000187:drug effects* / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*; D010750:Phosphoproteins / Q000187:drug effects* / Q000235:genetics; D013997:Time Factors
C513308:Dmp1 protein, mouse; D016326:Extracellular Matrix Proteins; D016376:Oligonucleotides, Antisense; D010750:Phosphoproteins; D000469:Alkaline Phosphatase; D002118:Calcium