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Open Access Basic Study Issue
Experimental study of periostin promoting rapid distraction osteogenesis of the rabbit mandible
Journal of Prevention and Treatment for Stomatological Diseases 2019, 27(9): 551-556
Published: 20 September 2019
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Objective

To explore the promoting effect of periostin on rapid distraction osteogenesis of the rabbit mandible and provide experimental evidence for the clinical use of periostin to promote osteogenesis.

Methods

Twenty-four New Zealand male white rabbits underwent distraction osteogenesis, and after 3 days of retention, they were rapidly stretched at a stretch rate of 2 mm/d (total 5 d). The animals were randomly divided into group A and group B (12 per group). On the last day of the stretch, 0.5 mL of normal saline containing 40 μg of recombinant periostin was given to group B or an equal volume of normal saline was added to the control group (group A) for 8 days. At 4 weeks and 8 weeks post-stretch, 8 animals were randomly selected from each group to undergo a CT scan under general anesthesia. The bone mineral density and bone mineral content were detected by dual energy X-ray absorptiometry. Eight weeks post-stretch, all of the experimental animals were sacrificed. Six animals were randomly selected from each group for micro-CT and a histological examination, and the remaining animals were subjected to biomechanical tests.

Results

CT images showed that the new bone formation in the distraction space of group B was significantly better than that of group A at 4 and 8 weeks post-stretch. At 4 weeks and 8 weeks post-stretch, the bone mineral density in group B was (0.157 ± 0.016) g/cm2 and (0.234 ± 0.023) g/cm2, respectively, and the bone mineral content was (0.096 ± 0.010) g and (0.204 ± 0.017) g, respectively. The above four means were significantly higher in group B than in group A (P < 0.001). The micro-CT images and data suggest that the stretch gap microstructure of group B has more mature features. Histological experiments showed that the trabecular bone of group B was thick and mature, with few chondrocytes. The biomechanical test results showed that the biomechanical strength of the distraction gap in group B was (228.47 ± 39.98) N, which was 1.24 times that of group A (P = 0.045).

Conclusion

Interstitial use of periosteal protein in the distraction space of the mandible in rabbits can promote local new bone formation and mineralization.

Open Access Basic Study Issue
The role of the mTORC1 signaling pathway during osteogenic differentiation of mouse bone marrow mesenchymal cells under tension stress
Journal of Prevention and Treatment for Stomatological Diseases 2020, 28(4): 219-223
Published: 20 April 2020
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Objective

To investigate the expression of the mTORC1 signaling pathway during the osteogenic differentiation of mouse bone marrow mesenchymal cells (BMMSCs) under cyclic uniaxial tension and explore its possible role.

Methods

The BMMSCs of mice were affected by uniaxial dynamic tensile force. Western blot was used to detect the expression changes of major molecules (mTOR, Raptor, S6K) in the endogenous mTORC1 signaling pathway at 0, 1, 2, 4, and 8 hours after stretching. Chemical colorimetry, ELISA and PCR were used to detect alkaline phosphatase (ALP), osteocalcin (OCN) and Runx2 mRNA, respectively. Then, inhibition, activation and control groups were established by administration of the drugs PP242, MHY1485 and PBS, respectively. Two hours after the stress, the expression of S6K was detected by western blot, and the expression of the osteogenic signal was continuously detected by the above methods.

Results

Western blot analysis showed that the main molecules of the mTORC1 signaling pathway were all expressed within 8 hours after traction, and the highest expression was 2 hours after the stress. Compared with those in the control group, the ALP activity and OCN expression decreased and the Runx2 mRNA levels increased after the mTORC1 signal pathway was inhibited (P < 0.001); ALP activity and OCN expression increased after the mTORC1 signal pathway was activated, while the Runx2 mRNA levels decreased (P < 0.001).

Conclusion

The mTORC1 signaling pathway participates in the osteogenic differentiation of mouse BMMSCs under tension. The osteogenesis of BMMSCs under cyclic uniaxial tension would be enhanced if the mTORC1 signaling pathway was activated.

Open Access Basic Study Issue
Kaempferol promotes osteogenic differentiation of mouse bone marrow mesenchymal cells under tension stress via the mTORC1 signaling pathway
Journal of Prevention and Treatment for Stomatological Diseases 2021, 29(4): 234-240
Published: 20 April 2021
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Objective

To investigate the activation of the mammalian target of rapamycin complex 1 (mTORC1) signaling pathway molecules during the process by which kaempferol (Kae) promotes osteogenic differentiation of mouse bone marrow mesenchymal cells (BMMCs) under cyclic and uniaxial tension.

Methods

BMMCs isolated and cultured in vitro were subjected to uniaxial dynamic tension with a 10% shape variable. The appropriate concentration of Kae was selected by cytotoxicity testing. The endogenous mTOR signal was inhibited by pp242. Four hours after traction, alkaline phosphatase (ALP) and osteocalcin (OCN) were detected by chemical colorimetry and ELISA, and the relative concentration of intracellular calcium was detected by flow cytometry. Phosphorylation of mTOR, 4E/BP1, and ribosomal protein S6 kinases (S6K), which are the main molecules of the endogenous mTORC1 signaling pathway, and expression of osteogenic transcription factors (Runx2 and Osterix) were detected by western blotting (WB), and mRNA expression levels of the above factors were detected by qRT-PCR.

Results

The cytotoxicity test showed that 10 μmol/L Kae had little inhibitory effect on cell proliferation but had the strongest osteogenic ability. Four hours after stretching, Kae effectively promoted the osteogenic differentiation of BMMCs. The expression of ALP was (153.04 ± 18.72) U/mg, the expression of OCN was (1.64 ± 0.25) U. The mRNA and protein levels of Runx2 and Osterix were upregulated, and the intracellular calcium content was decreased. The mRNA and protein phosphorylation of mTOR and S6K was upregulated, and the opposite effect was observed with 4E/BP1. After pp242 was added to inhibit mTOR signaling, mTOR and S6K mRNA and protein phosphorylation were downregulated, but 4E/BP1 mRNA and protein phosphorylation was upregulated. The osteogenic differentiation of BMMCs was also significantly inhibited, mRNA and protein expression of Runx2 and Osterix were significantly downregulated, ALP and OCN expression were downregulated, and intracellular calcium content was increased.

Conclusion

Kae promotes osteogenic differentiation of mouse BMMCs under uniaxial dynamic tension through the mTORC1 signaling pathway.

Open Access Issue
Clinical study of the treatment effects of direct current iontophoresis of triamcinolone acetonide on oral submucous fibrosis
Journal of Prevention and Treatment for Stomatological Diseases 2019, 27(10): 638-641
Published: 20 October 2019
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Objective

To observe the clinical effects of direct current iontophoresis (DCI) of triamcinolone acetonide (TA) on treatment of oral submucous fibrosis.

Methods

Fifty-one patients were randomly divided in group A (n=25), which underwent local submucous injection with TA (20 mg for each side) once a week for 8 weeks, and group B (n=26), which underwent local TA administration by DCI (20 mg for each side) for 8 weeks, follow-up was performed 12 weeks after treatment. The distances between the upper and lower central incisors were recorded. The visual analogue scale (VAS) was used to evaluate the pain caused by the two treatments. The Oral Health Impact Profile (OHIP)-14 was used to assess the entire effect of treatments.

Results

At 8 weeks of treatment, the mouth opening was in-creased by 3.25 ± 0.77 mm in group A and 3.76 ± 0.88 mm in group B (P < 0.05). At 20 weeks after treatment, the mouth opening was increased by 2.61 ± 0.62 mm in group A and 2.53 ± 0.52 mm in group B (P>0.05). After the first treatment, the VAS score of group A was 7.88 ± 0.80, and the VAS score of group B was 2.47 ± 0.64, resulting in a statistically significant difference (P < 0.001). After the last treatment, the VAS score was 7.29 ± 0.53 in group A and 1.77 ± 0.48 in group B, resulting in a statistically significant difference (P < 0.001). The VAS scores of the two groups were decreased after treatment, and the difference between groups was statistically significant (P < 0.001). The OHIP-14 score of patients in group A before treatment was 31.44 ± 2.55, while that in group B was 32.04 ± 2.20 (P>0.05). At 20 weeks after treatment, the score in group A was 13.52 ± 3.31 and that in group was 12.04 ± 2.84 (P>0.05). The OHIP-14 scores of the two groups were significantly decreased before and after treatment, and the difference between groups was statistically significant (P < 0.001).

Conclusion

The results suggest that local TA administration by DCI might be a promising method for treatment in the early and middle disease periods in OSF patients.

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