Lingyong Jiang

p38-MAPK signaling pathway is not involved in osteogenic differentiation during early response of mesenchymal stem cells to continuous mechanical strain

Mechanical stimuli play a significant role in the regulation of bone remodeling during orthodontic tooth movement. However, the correlation between mechanical strain and bone remodeling is still poorly understood. In this study, we used a model of continuous mechanical strain (CMS) on bone mesenchymal stem cells (BMSCs) to investigate the proliferation and osteogenic differentiation of BMSCs and the mechanism of mechano-transduction. A CMS of 10xa0% at 1xa0Hz suppressed the proliferation of BMSCs and induced early osteogenic differentiation within 48xa0h by activating Runx2 and increasing alkaline phosphatase (ALP) activity and mRNA expression of osteogenesis-related genes (ALP, collagen type I, and osteopontin). Regarding mitogen-activated protein kinase (MAPK) activation, CMS induced phased phosphorylation of p38 consisting of a rapid induction of p38 MAPK at 10xa0min and a rapid decay after 1xa0h. Furthermore, the potent p38 inhibitor SB203580 blocked the induction of p38 MAPK signaling, but had little effect on subsequent osteogenic events. These results demonstrate that mechanical strain may act as a stimulator to induce the differentiation of BMSCs into osteoblasts, which is a vital function for bone formation in orthodontic tooth movement. However, activation of the p38 signaling pathway may not be involved in this process.

Effects of hypoxia on osteogenic differentiation of rat bone marrow mesenchymal stem cells.

Bone reconstruction is essential in orthodontic treatment that caters to the correction of malocclusion by bone reconstruction. Mesenchymal stem cells (MSCs) have been demonstrated a great potency of osteogenesis. The aim of this study was to investigate the effect of hypoxia on the rat bone marrow MSCs (rBMSCs) in vitro during osteogenesis. In this study, we found that temporary exposure of rBMSCs after osteogenic induction for 7xa0days to hypoxia (2% oxygen) led to a markedxa0decrease in ALPase activity and the expression of osteocalcin and Runt related transcription factor 2/core binding factor a1 (Runx2/Cbfa1). Meanwhile, we found that exposure to hypoxia led to an early and transient increase in the level of phosphorylated ERK1/2 but had no obvious effects on mitogen-activated protein kinase (p38 MAPK) level. Based on these results, we concluded that hypoxia could inhibit osteogenic differentiation of rBMSCs possibly through MEK-ERK 1/2, while p38 MAPK may not participate in this regulation. Further exploration into the mechanisms of hypoxia on osteogenesis would surely provide reliable evidence for clinical practice.

Osteoprotegerin and ligand of receptor activator of nuclear factor : kappaB expression in ovariectomized rats during tooth movement

OBJECTIVEnTo test the null hypothesis that increased tooth displacement in ovariectomized rats is not related to differential expressions of OPG and RANKL in the periodontium.nnnMATERIALS AND METHODSnEighty-four 12-week female rats were used; half were ovariectomized and half were not. Three months later, the maxillary first molar was moved mesially. Groups of rats were sacrificed at days 0, 1, 3, 5, 7, 10, and 14 after activation. Tooth movement was measured at each time point. OPG and RANKL expressions were examined through immunohistochemistry.nnnRESULTSnOvariectomized and nonovariectomized rats showed three-phase tooth movement. In both groups, OPG expression increased at the tension area and RANKL increased at the pressure area. The OPG/RANKL ratio coincided with tooth movement, especially in the linear phase from 7 to 14 days.nnnCONCLUSIONSnThe null hypothesis is rejected. The increased rate of tooth movement in ovariectomized rats was related to differential expressions of OPG and RANKL.

Three-dimensional analysis of soft tissue changes in full-face view after surgical correction of skeletal class III malocclusion.

ObjectiveThe objective of this study was to evaluate changes in soft tissue in full-face view because of surgical correction of skeletal Class III malocclusion, using 3-dimensional (3D) laser scanning. MethodsTwenty-seven subjects with skeletal Class III malocclusion [11 males; mean age (SD), 24.0 (5.7) years] underwent bilateral sagittal split ramus osteotomy for mandibular setback combined with Lefort I osteotomy with/without maxillary advancement. Twelve patients (group 1) had mandibular setback surgery, and the other 15 (group 2) had combination surgery. Lateral cephalograms and 3D facial scan images were assessed preoperatively and postoperatively. The facial widths upon superimposition of 3D facial images were measured in the same coordinates using a Rapidform 2006 system. Paired and independent t tests were done for statistical analysis. ResultsThe midface soft tissue broadened significantly above the cheilion plane postoperatively (P < 0.05). A larger change was observed nearer to subnasale plane, and a similar trend was seen among the horizontal planes in 1- or 2-jaw surgery groups. The widths from the exocanthion plane to the subnasale plane increased more in group 2 [mean (SD), 4.45 (2.45) mm, 8.71 (2.92) mm, and 7.62 (3.13) mm] than those in group 1 [mean (SD), 1.26 (0.97) mm, 1.84 (1.06) mm, and 1.35 (0.65) mm], and this difference was significant (P < 0.05). There was a decrease below the cheilion plane with mandibular setback between groups, but this difference was not significant. ConclusionsThe measurement method used here for the shape outline of the lateral parts of the face could provide quantitative data for the clinical evaluation and objective analysis of the human face in full-face view. The midface soft tissue in subjects with skeletal Class III malocclusion exhibited a greater increase in width after bimaxillary surgery procedures than mandibular setback-only surgery.

Centrifugal forces within usually-used magnitude elicited a transitory and reversible change in proliferation and gene expression of osteoblastic cells UMR-106

Centrifugation is an important step in biochemical and molecular biological researches. But the effects of centrifugal stress on cells are still unclear. In this study, osteoblastic cells UMR-106 were subjected to a moderate centrifugal stress at 209 × g for 10xa0min. Then the cell proliferation and gene transcription after centrifugation were analyzed with flow cytometry and Real-time RT-PCR techniques, respectively. The result showed that the cell proliferation and mRNA expression of Runx2/Cbfa1, Collagen I and osteocalcin changed shortly after centrifugal loading, but recovered to pre-load levels within 24xa0h. A dose-response study of exposure cells to centrifugal force at 209, 253 and 301 × g showed that the centrifugal forces within usually-used range can rapidly influenced the mRNA expression of the osteoblast-specific genes, but no statistical differences were found among the three centrifugal magnitudes. And the fast regulation in the investigated genes was proved to be related to increased c-fos mRNA levels and subsequent activation of RTK and integrity of cytoskeleton construction. The result showed that the osteoblastic cells displayed a fast auto-regulation to usually-used centrifugal stress through multiple signal pathways.

Effect of mechanical stretch on the proliferation and differentiation of BMSCs from ovariectomized rats

Osteoporosis is characterized by a broken balance between bone formation and bone resorption. Mechanical stress has been considered to be an important factor in bone modeling and remodeling. However, biological responses of stromal cells in osteoporosis to mechanical stimuli remain unknown. To explore the correlation between mechanical stress and osteoblastic differentiation of bone mesenchymal stem cells (BMSCs) in osteoporosis, we built an osteoporosis model in ovariectomized (OVX) rats, and then investigated proliferation, alkaline phosphatase (ALP) activity, and the expression of osteoblastic genes in BMSCs under mechanical stress of 5 and 10xa0% elongation, using the Flexercell Strain system. The proliferation of BMSCs was detected using alamarBlue. The expression of osteoblastic genes was analyzed by real-time quantitative polymerase chain reaction. Protein expression was examined by Western blotting. BMSCs (OVX) and BMSCs (Sham-operated, Sham in short) proliferations were inhibited at 5 and 10xa0% elongation at day 3, compared with the un-stretched group, while BMSCs (OVX) proliferation was slower than BMSCs (Sham). ALP activity increased significantly at 10xa0% elongation in both cells, but it was less active in BMSCs (OVX) than BMSCs (Sham). At days 3 and 7, the mRNA expression of osteoblastic genes was unregulated by mechanical stretch (5 and 10xa0% elongation); however, osteoblastic gene expression in BMSCs (OVX) was less than that in BMSCs (Sham). The mRNA and protein expression of Runx2 showed similar trends in BMSCs (OVX) under mechanical stretch. These results indicate that the mechanical stretch stimulates osteoblastic differentiation of BMSCs (OVX); however, this differentiation was weaker than that of BMSCs (Sham).

Force-induced increased osteogenesis enables accelerated orthodontic tooth movement in ovariectomized rats

As the number of elderly orthodontic patients increases, the impact of postmenopausal osteoporosis on orthodontic tooth movement (OTM) has attracted a great deal of attention because OTM relies on alveolar bone remodeling. The question of whether OTM causes subsequent alveolar bone loss and is harmful to alveolar bone health under osteoporotic conditions remains to be answered. The present study aimed to clarify the influences of OTM on alveolar bone in osteoporotic rats. OTM was accelerated in ovariectomized (OVX) rats as a result of increased bone resorption in the pressure area. At the same time, anabolic bone formation was promoted in the tension area during OTM in OVX rats. Micro-CT analysis of alveolar bone revealed a decrease in BMD, BV/TV and Tb.Th. in the OTM group compared with that in non-OTM rats on day 21 of OTM, suggesting that OTM caused alveolar bone loss in OVX rats during OTM. However, the OTM-induced bone loss could be recovered 3 months after OTM in OVX rats. Thus, our findings suggest that increased osteogenesis may compensate for the increased bone resorption during and after OTM and enable effective accelerated OTM in OVX rats.

Hypoxia suppresses osteogenesis of bone mesenchymal stem cells via the extracellular signal‑regulated 1/2 and p38‑mitogen activated protein kinase signaling pathways

There is a growing body of evidence indicating an association between osteoporosis and vascular diseases, which are associated with reduced blood supply. Decreased vascular flow results in a hypoxic gradient in the local microenvironment, affecting local bone remodeling. Bone mesenchymal stem cells (BMSCs) have been demonstrated to be the key to bone remodeling. To elucidate the molecular mechanisms involved in vascular supply and osteoporosis, the present study investigated the effect of hypoxia on BMSCs inxa0vitro during osteogenesis. The BMSC osteogenesis process was evaluated by alkaline phosphatase (ALP) activity assay and the mRNA expression of the osteogenic markers runt‑related transcription factor 2 (Runx2), ALP and osteocalcin. The function of extracellular signal‑regulated kinase (ERK)1/2 and p38 kinase were studied under hypoxia using specific inhibitors. The results demonstrated that hypoxia reduces the osteogenic differentiation of BMSCs by inactivating Runx2, followed by decreased ALP activity and mRNA expression levels of ALP, collagen type I and osteocalcin. Furthermore, these data suggested that the ERK1/2 and p38‑mitogen activated protein kinase signaling pathways might participate in hypoxia‑induced differentiation of BMSCs toward the osteogenic phenotype. Compared with ERK1/2, the p38‑Runx2 signaling pathway might exert a relatively more prominent effect in the above process. These findings may help to elucidate the pathophysiology of osteoporosis caused by decreased vascular supply.

Mechanical strain promotes osteogenic differentiation of bone mesenchymal stem cells from ovariectomized rats via the phosphoinositide 3‑kinase/Akt signaling pathway

Osteoporosis has become an overwhelming public health problem worldwide. As an elementary physiological factor to regulate bone formation and regeneration, mechanical strain may be used as a non‑invasive intervention in osteoporosis prevention and treatment. However, little is known regarding the underlying mechanism. The aim of the current study was to investigate the effect of continuous mechanical strain (CMS) on osteogenic differentiation of bone mesenchymal stem cells (BMSCs) from ovariectomized rats (OVX BMSCs). In addition, involvement of the phosphatidylinositolxa03‑kinase (PI3K)/Akt signaling pathway in biomechanical signal transduction and its function were evaluated. The results demonstrated that OVX BMSCs subjected to CMS exhibited higher alkaline phosphatase (ALP) activity and deeper staining at 24 and 48xa0h. In addition, CMS upregulated the mRNA expression levels of ALP, collagen type I, runt related transcription factor 2 (Runx2), as well as the protein expression level of Runx2 in a time‑dependent manner. The PI3K/Akt signaling pathway was rapidly activated by CMS, with its phosphorylation level reaching its maximum in a short duration and a large quantity of phosphorylated‑Akt remaining in the nucleus. Pre‑treatment with a selective blocker significantly blocked the strain‑induced activation of PI3K/Akt and reduced the commitment of OVX BMSCs into osteoblasts, demonstrating a dominated regulative effect of PI3K/Akt signaling in strain‑induced osteogenesis. These results indicated that CMS induced the early differentiation of OVX BMSCs towards an osteogenic phenotype by activating the PI3K/Akt signaling pathway.

Temporomandibular joint changes after activator appliance therapy: a prospective magnetic resonance imaging study.

AbstractThe aim of this prospective clinical and magnetic resonance imaging study was to analyze the effect of 1-year Activator (Yi-fan Dental Co., Shanghai, China) treatment in internal anatomical relationships of the temporomandibular joint (TMJ) complex, including the condyle-disc relationship, condyle-fossa relationship, condylar height change, disc length change, and morphologic change of the glenoid fossa. The study was composed of patients with class II division 1 malocclusion (11 girls and 13 boys) who underwent 1-year Activator treatment. All the patients were in the acceleration or peak phase of the pubertal growth spurt. Magnetic resonance imaging in closed-mouth position and lateral cephalometric radiographs before and after 1 year of Activator treatment were analyzed metrically. Overall, condylar height showed a significant increase (P < 0.001), and the eminence angle decreased (P = 0.037). TMJ disc length has no statistically significant change before and after treatment. A slight advancement (P = 0.041) was found in the sagittal condylar position. A significant backward movement (P = 0.04) was shown in the sagittal disc position. Our results showed that the disc is not impaired by Activator therapy; it seems possible that adaptive remodeling, including a shallower glenoid fossa and increased condylar height, was seen after treatment.