| Objective Under the guidance of the theory of bone bonding, dental implant technology is developing rapidly,however, the consequent peri-implantitis and how to maintain the normal bone mass and bone mineral density around the implant has been concerned by the scholars. Osteoblasts play an important role in bone formation, bone morphogenetic protein -2 is the most important factor in the regulation of bone formation,recent studies have found that BMP-2 participates in the damage repair of bones and teeth, so it has been used in oral tissue-engineered. Cinnamaldehyde (CA)has no toxic effect on a certain drug concentration range, the effect of antibacterial and anti-inflammatory on periodontal disease has a certain therapeutic significance,and CA has an positive effect on the proliferation and osteogenesis of osteoblasts. But the mechanism of promoting osteogenesis has not been reported. This experiment using the different concentration of CA on osteoblasts, and to evaluate the osteogenetic ability, discusses the mechanism of action of promote osteogenesis,provide preliminary theoretical basis for clinical application.Methods 1 .Cultivation of MC3T3 cells in vitro, according to the experimental design MC3T3 cells were divided into 7 groups,including a blank control group and six different drug concentration groups. In 1,3, 5, 7 days each group of cells was determined by MTT method to measuring the wavelength of 570 nm OD value by MTT method. 2.MC3T3 cells were cultured in serum medium containing osteogenic inducing medium and passaged for 2 times,in 3, 5, 7 days according to ALP test kit requirements for operation and easuring the wavelength of 560nm OD value. 3.With containing the CA concentration respectively 0,10,20 ?g/mL of medicated serum culture medium to group,extract total RNA from MC3T3 cells by Trizol method,then reverse transcription-polymerase chain reaction ( RT-PCR) to examine three groups of BMP-2 mRNA gene expression, in order to study groups with different concentrations of CA on osteoblast of bone function.Results 1. Compared with the control group, 10,20 ?g/mL CA cell morphology was no significant change,similar size and contacting with neighboring cells by the way of peripheral processes.40,80,160,320 ?g/mL CA cell morphology changes gradually,the cell becomes round and smaller,some cell appears to take off the wall floating phenomenon.2.The effect of 10,20 ?g/mL CA on MC3T3, compared with the control group,the osteoblast proliferation value decreased slightly but there were no obvious difference between them , alkaline phosphatase activity rised significantly, and the alkaline phosphatase activity of osteoblasts in the 10 ?g/mL group was higher than that in the 20 ?g/mL group.Compared with the blank group, the osteoblast proliferation decreased clearly when the CA was 40,80,160,320 ?g/mL, ALP activity was relieved in different degree.At the same time, a negative correlation between the concentration of CA and osteoblast proliferation level was found, the amount of cell proliferation and drug intervention time were positively correlated.3. Compared with the blank group in the same period of time, CA increased the mRNA expression of BMP-2 at the concentration of 10,20 pg/mL. Compared to the low concentration groups,the mRNA BMP-2 expression in the 10 pg/mL group was higher than 20 ?g/mL group,but there were no obvious difference between them.Conclusions In conclusion, the low concentration group (10,20 ?g/mL) of CA on osteoblast has a rise in a certain effect, this promotion effect is related to CA significantly promote the expression of osteoblasts in BMP-2 mRNA. |
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