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Objective:To investigate the partial delivery ability in vivo of oligonucleotide MT01 mediated by polyethyleneimine(PEI) derivative PEN, and to illuminate its effect on the alveolar bone reconstruction and the biological safety in the rats with experimental tooth movement. Methods:Forty-eight Wistar male rats were randomly divided into PEN,MT01, sulfuration-modified MT01 (MT01s), and PEN/MT01 groups. The rat experimental tooth model was established, and the above four drugs were injected into the mesial buccal mucosa of the first molar in the left sides of the rats as drug intervention sides,and the PBS were injected into the mesial buccal mucosa of the first molar in the right sides as PBS control sides;once every 3 d. After 14 d, the rats were sacrificed. HE staining was used to detect the histopathological changes of important organs of the rats;the photos and X-ray films were applied to measure the distance of tooth movement, and RT-PCR was used to detect the expression levels of the osteogenic marker genes Runt-related transcription factor 2(Runx2),special protein 7(SP7) and osteocalcin(OCN) mRNA. Results:After local injection of the above drugs, no obvious inflammatory cell infiltration and structural differences were found in the main organs of the rats and no obvious abnormalities were found in all organs tissues. Compared with the PBS control sides, the movement distance of the first molars in the drug intervention sides in MT01, MT01s, and PEN/MT01 groups was reduced (P0.05). The data of the distance between the two teeth of the rats in various groups was PEN/MT01 group > MT01s group > MT01 group>PEN group, and the difference between PEN/MT01 group and MT01s group was statistically significant (P0.05). Conclusion:PEI derivative PEN can transfer MT01 into the cell safely and efficiently; it can increase the expression levels of Runx2, SP7 and OCN mRNA in the periodontium tissue and decrease the tooth movement distance in the experimental tooth movement rats.
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Objective:To investigate the changes of expression levels of TLR9 and IL-6 in the periodontal tissue during the experimental tooth movement of the rats, and the effecst of the MT01 on the expression of TLR9,TRAF6 and IL-6 in periodontal tissue,and to clarify its related mechanisms.Methods:Thirty Wistar rats were randomly divided into MT01 intervention group(n=6) and non-MT01 group(n=24).Force of 0.49 N was applied to move the upper first molars mesially. The rats in Non-MT01 intervention group were sacrificed on the days 3,7,14 and 21, and the rats in MT01 intervention group were all sacrificed on the day 7. RT-qPCR was used to detect the expression levels of TLR9,TRAF6 and IL-6 mRNA in maxillary first molar alveolar bone in each group.Results:The expression levels of IL-6 and TLR9 mRNA in loaded side were higher than those in control side(P<0.05), and reached the maximum levels on the day 7(P<0.01);with the interpose of MT01, the expression levels of TLR9 and TRAF6 mRNA were lower than control side(P<0.01).Conclusion: MT01 could down-regulate the expression levels of TLR9 and TRAF6 during orthodontic tooth movement and eventually resists the inflammation during the tooth movement.
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Objective:To investigate the effect of MT01 on the differentiation of osteoblasts under infected condition through determining the expression level of collagen Ⅰ (Col Ⅰ )mRNA in MG63 cells treated with Porphyromonas gingivalis (Pg).Methods:The cultured MG63 cells were divided into blank control,MT01,Pg, and MT01+Pg groups.MT01 at a concentration of 1 mg·L-1 was added into the MG63 cells,and the cells were incubated for 3 h.The cells treated with PBS (1 mg·L-1 )were used as control group.Then Pg (MOI=100∶1) was added.Real-time PCR was used to detect the expression levels of ColⅠ mRNA in MG63 cells at 2,4,6,8, 12 and 24 h after incubation.Results:Compared with blank control group,the levles of ColⅠ mRNA in the MG63 cells in MT01 and MT01 + Pg groups had no significant changes at 2 and 4 h (P > 0.05);the Col Ⅰ mRNA expression levels in MT01 group at 8,12 and 24 h were increased (P < 0.05 or P < 0.01).Compared with Pg group,the expression levels of ColⅠ mRNA in MT01+ Pg at 2 and 4 h were decreased,but the expression levels of ColⅠ mRNA were increased at 6,8,12 and 24 h (P <0.05 or P <0.01).Conclusion:MT01 can up-regulate the expression level of Col Ⅰ mRNA in the infected MG63 cells; MT01 could promot the differentiation of osteoblasts under infected condition.
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Objective To observe the distribution of oligodexynucleotide (ODN)MT01 in main organ tissues of the rats at different time points and to discuss the regularity of the distribution of MT01 preliminarily. Methods 60 male Wistar rats were randomly divided into experimental group(n=30)and control group(n=30). The rats in experimental group was locally injected with Cy5 labeled MT01 in gingival mucosa,whereas the rats in control group were injected with MTO1.The samples of rat lung,liver spleen,kidney,heart,and brain tissues were collected at 15 min,1 h,4 h,8 h,16 h,1 d,2 d,3 d,4 d,and 5 d after injection,and the distribution of MT01 fluorescence was observed by laser scanning confocal microscope.The ratio of fluorescence positive cells indicated the amount of MT01 that had been taken up by different organs.Results No positive fluorescence cells were observed in control group.Whereas,in experimental group ,the positive fluorescence cells were detected in the tissue samples of lung,liver,spleen and kidney but not in the tissue samples of heart and brain.The positive fluorescence cells distributed focally in kidney tissue and presented primarily in the cytoplasm of renal tubular epithelial cells.The ratios of positive fluorescence cells changed regularly with time in liver, spleen and kidney tissues and the highest level was detected at 4,3 and 4 d after injection.No distinct regularity of the ratio of positive fluorescence cells was observed in lung tissue.Conclusion MT01 can be taken up by liver,spleen and lung tissue and primarily by kidney with regularity in distribution.