Targeting P38 Pathway Regulates Bony Formation via MSC Recruitment during Mandibular Distraction Osteogenesis in Rats.

Distraction osteogenesis (DO) is a widely used self-tissue engineering. However, complications and discomfort due to the long treatment period are still the bottleneck of DO. Novel strategies to accelerate bone formation in DO are still needed. P38 is capable of regulating the osteogenic differentiation of both mesenchymal stem cells (MSCs) and osteoblasts, which are crucial to bone regeneration. However, it is not clear whether targeting p38 could regulate bony formation in DO. The purpose of the current work was to investigate the effects of local application of either p38 agonist anisomycin or p38 inhibitor SB203580 in a rat model of DO. 30 adult rats were randomly divided into 3 groups: (A) rats injected with DMSO served as the control group; (B) rats injected with p38 agonist anisomycin; (C) rats injected with p38 inhibitor SB203580. All the rats were subjected to mandibular distraction and the injection was performed daily during this period. The distracted mandibles were harvested on days 15 and 30 after surgery and subjected to the following analysis. Micro-computed tomography and histological evaluation results showed that local application of p38 agonist anisomycin increased new bone formation in DO, whereas p38 inhibitor SB203580 decreased it. Immunohistochemical analysis suggested that anisomycin promoted MSC recruitment in the distraction gap. In conclusion, this study demonstrated that local application of p38 agonist anisomycin can increase new bone formation during DO. This study may lead to a novel cell-based strategy for the improvement of bone regeneration.

Calcitonin gene-related peptide enhances osteogenic differentiation and recruitment of bone marrow mesenchymal stem cells in rats.

The present study evaluated the effects of calcitonin gene-related peptide (CGRP) on bone marrow mesenchymal stem cells (BMMSCs) in vitro and in a rat model of mandibular distraction osteogenesis (MDO). Rat BMMSCs were isolated then treated with CGRP or CGRP antagonist (CGRP8-37). The proliferation and migration ability of BMMSCs was determined using 5-bromo-2'-deoxyuridine and Transwell assays, respectively. Osteogenic-related gene expression was analyzed with reverse transcription-quantitative polymerase chain reaction. For the in vivo analysis, thirty MDO rats were randomly assigned to control, CGRP or CGRP8-37 groups. To evaluate the mobilization of BMMSCs, nestin and stromal cell-derived factor 1 (SDF-1) were detected by immunohistochemistry and ELISA. Rats were sacrificed following 14 days and new bone formation was assessed by histological and micro-computed tomography analysis. In the in vitro results, the CGRP group demonstrated significantly higher migration and proliferation, as well as enhanced alkaline phosphatase and runt-related transcription factor 2 expression compared with the control. In the in vivo experiments, bone mineral density of the newly formed bone in the CGRP group was significantly higher than controls. The nestin and SDF-1 expression in the CGRP group was also significantly upregulated. In conclusion, the present study demonstrated that CGRP administration increased new bone formation, possibly via enhancing BMMSC migration and differentiation in MDO rats.

The mechanically activated p38/MMP-2 signaling pathway promotes bone marrow mesenchymal stem cell migration in rats.

OBJECTIVE: The aim of the present study was to investigate the effect of static strain on bone marrow mesenchymal stem cell (BMMSC) migration and whether the p38/matrix metalloproteinase-2 (MMP-2) axis plays a role in induction of BMMSC migration under mechanical strain. DESIGN: Both in vivo and in vitro investigations were performed. Twelve adult male Sprague-Dawley rats were randomly divided into 2 groups (n=6 per group). Rats in the experimental group underwent right mandibular distraction osteogenesis, whereas rats in the control group were subjected to osteotomy in the mandible without distraction. Immunohistochemistry and immunofluorescence were performed to evaluate phospho-p38 (p-p38) and Nestin expression. BMMSCs were isolated from rat mandibles. BMMSCs in the experimental group were subjected to static mechanical strain for 2h, whereas those in the control group underwent no strain. The biological roles of static strain and the p38/MMP-2 axis in BMMSC migration were evaluated by Transwell assays and western blotting by inhibiting p38 phosphorylation. RESULTS: There were significantly more Nestin+ cells in the bone calluses of the experimental group than in those of the control group. In addition, Nestin+/p-p38+ cell numbers were significantly higher in the experimental group than in the control group, indicating that static strain activated p38 signaling in BMMSCs in vivo. In accordance with in vivo results, static strain in vitro stimulated phosphorylation of p38 in BMMSCs. Furthermore, expression of MMP-2 was elevated in BMMSCs under static strain compared with the control, and strain-induced MMP-2 expression was abolished by inhibition of p38 phosphorylation in BMMSCs. Moreover, Transwell assay results showed that static strain promoted BMMSC migration, which was abolished by inhibition of p38 phosphorylation. CONCLUSIONS: The present study demonstrated that static strain can promote the migration ability of BMMSCs via p38/MMP-2 signaling. To the best of our knowledge, this study is the first report demonstrating that the p38/MMP-2 axis governs BMMSC migration under static mechanical strain.

Thorough documentation of the accidental aspiration and ingestion of foreign objects during dental procedure is necessary: review and analysis of 617 cases.

OBJECTIVES: To review the cases of accidental aspiration and ingestion of foreign objects during dental procedure, and to emphasize the importance of thorough documentation of the accidents. METHODS: A comprehensive search on (dental procedure/treatment/practice), (aspiration/inhalation), and (ingestion/swallow) was performed for all years before 1st October 2014 available. The statistic analysis was made on the variables including journals and reported year, patients' age, gender, general conditions, dental procedure and location for procedure, foreign objects, site of involvement, possible causes, anesthesia during procedure and treatment, symptoms, treatment time and treatment modality, follow-up, and so on. RESULTS: A total of 617 cases reported by 45 articles from 37 kinds of journals were included and analyzed. Most reports made detailed record. While some important variables were recorded incompletely, including patient's general conditions, location for procedure, clinical experience of the involving dentists, tooth position of procedure, possible causes, and anesthesia during procedure and treatment for the accident. CONCLUSIONS: Aspiration and ingestion of foreign objects are rare and risky complication during dental procedure. Each accident should have thorough documentation so as to provide enough information for the treatment and prevention.

Immobilization of chitosan film containing semaphorin 3A onto a microarc oxidized titanium implant surface via silane reaction to improve MG63 osteogenic differentiation.

Improving osseointegration of extensively used titanium (Ti) implants still remains a main theme in implantology. Recently, grafting biomolecules onto a Ti surface has attracted more attention due to their direct participation in the osseointegration process around the implant. Semaphorin 3A (Sema3A) is a new proven osteoprotection molecule and is considered to be a promising therapeutic agent in bone diseases, but how to immobilize the protein onto a Ti surface to acquire a long-term effect is poorly defined. In our study, we tried to use chitosan to wrap Sema3A (CS/Sema) and connect to the microarc oxidized Ti surface via silane glutaraldehyde coupling. The microarc oxidization could formulate porous topography on a Ti surface, and the covalently bonded coating was homogeneously covered on the ridges between the pores without significant influence on the original topography. A burst release of Sema3A was observed in the first few days in phosphate-buffered saline and could be maintained for >2 weeks. Coating in phosphate-buffered saline containing lysozyme was similar, but the release rate was much more rapid. The coating did not significantly affect cellular adhesion, viability, or cytoskeleton arrangement, but the osteogenic-related gene expression was dramatically increased and calcium deposition was also abundantly detected. In conclusion, covalent bonding of CS/Sema could strongly improve osteogenic differentiation of osteoblasts and might be applied for Ti implant surface biofunctionalization.

Local injection of substance P increases bony formation during mandibular distraction osteogenesis in rats.

Substance P is a neuropeptide that is distributed in those sensory nerve fibres that innervate the medullary tissues of bone. It is a potent accelerator of proliferation and differentiation of osteoblasts in vitro. However, its capacity for promoting repair of mandibular defects is not known. We have investigated the osteogenic effects of local injections of substance P during mandibular distraction osteogenesis in rats. Twenty Sprague-Dawley rats were randomly assigned to 2 groups (n = 10 in each): substance P 10(-7) mmol/l in normal saline 0.2ml was injected into the experimental group, and saline alone into the controls. The mandibular distraction rate was 0.2mm every 12hours for 10 days. Daily injections of substance P or saline were given during the distraction period. Regeneration of bone was assessed quantitatively on days 15 and 29 using microcomputed tomography (microCT), and histological analysis. The rate of bony union in the group treated with substance P was significantly higher than that in the saline alone group on day 29 (p=0.001) The microCT images and quantitation showed more callus and more mature cortical bone when substance P was given than with control. Histological examination showed that cartilaginous tissues had formed in the middle of the distraction gaps in both groups. Bony bridges were seen only in the substance P group at the final time point (day 29). Injection of substance P into the gap of a rat mandible during mandibular distraction improved formation of good-quality bone and accelerated bony union.

Application of internal fixation of steel-wire limited loop in early Achilles tendon rupture.

OBJECTIVE: To explore the clinical effect and safety of internal fixation of steel-wire limited loop in early Achilles tendon rupture. METHODS: Seventy-six patients respectively with early transected and avulsed types of Achilles tendon rupture were selected and treated with internal fixation of steel-wire limited loop. The patients began to take exercise for their lower limbs through continous passive motion as early as possible after surgical repair, and the loops were removed after 3-5 months. Six months later, the condition of complications including Achilles tendon re-rupture, wound fistula, wound infection and skin necrosis, cutaneous sensation in sural nerve dominance region, time back to preinjury work or learning as well as time to physical activities were observed. One year later, the therapeutic effect was evaluated, and the maximum circumferences of bilateral legs and ruptured plane circumferences of Achilles tendon were measured. RESULTS: The wound of all patients healed well, no complications like Achilles tendon re-rupture, wound fistula, wound infection and skin necrosis occured, and the cutaneous sensation in sural nerve dominance region was normal. The mean time back to preinjury work or learning as well as to pysical activities of all patients were respectively 10 and 22 weeks. Seventy out of 76 patients (92.1%) achieved an excellent effect, and 6 (7.9%) good effect. The excellent and good rate came up to 100%. The maximum circumference in the affected leg decreased to 2 mm averagely compared with the offside, while the ruptured plane circumferences of Achilles tendon in the affected side increased to 2.2 mm compared with the offside. CONCLUSIONS: For early Achilles tendon rupture, internal fixation of steel-wire limited loop can recover the ankle function better, return to the preinjury state in the shortest time, and has few complications.

[Clinical efficacy of immediate implants restoration in anterior zone].

PURPOSE: To elevate the clinical efficacy of immediate loading in anterior implants. METHODS: Forty eight implants were placed in 33 patients and the implants were immediately loaded to provide support for fixed provisional prosthesis within 3-6 months. Then, metal-ceramic crowns were restored. The patients were revisited every 12 months after restoration. RESULTS: The follow-up period was 3 to 50 months. All cases achieved good clinical efficacy. CONCLUSION: Clinical studies on the placement of immediate restoration in anterior teeth have revealed predictable results.

Histologic analysis of acellular dermal matrix in the treatment of anal fistula in an animal model.

BACKGROUND: Human acellular dermal matrix (ADM) has been used successfully for the treatment of severe burns, ureter support, and abdominal wall reconstruction. This study was designed to evaluate the mechanism of ADM in the closure of anal fistula in an experimental porcine model. STUDY DESIGN: The fistula-in-ano model was created in the porcine model and treated with ADM in 14 animals. Fistula specimens were obtained at hours 12 and 24 and on days 3, 7, 14, 28, 60. Hematoxylin and eosin staining, Masson trichrome staining, and immunohistochemical staining for alpha smooth muscle actin and matrix metalloproteinase 9 were performed. RESULTS: The cell density increased from hour 12 to day 7 and decreased from day 7 to day 28 (p < 0.001). Mature vessels stained with alpha smooth muscle actin were identified at day 7. Alpha smooth muscle actin-positive myofibroblasts were found in clusters at the edge of the ADM at day 7. The density of vessels (p < 0.001) and myofibroblasts (p < 0.001) increased from day 7 to day 14. The density of matrix metalloproteinase 9 increased from hour 12 to day 7 and decreased from day 14 to day 60 (p < 0.001). Partially organized bundles of muscle were found by day 60. CONCLUSIONS: We suggest that ADM is a reasonable new option for closure of anal fistulas. Anal fistulas begin to heal as early as 12 hours, and day 7 may be an important time point to judge whether the fistula healed preliminarily or not. The ability of ADM to become vascularized and remodeled by autologous cells may be advantageous for anal fistula healing.

[Clinical study of maxillary sinus floor elevation with simultaneous placement of implants from the top of alveoli].

PURPOSE: To evaluate the clinical results of maxillary sinus floor elevation technique with bone condensers on implantation. METHODS: 104 patients underwent maxillary sinus floor elevation and 126 implants were inserted and restored. Clinical examination and radiographs were conducted. Spiral CT scanning and three-dimensional reconstruction was performed among 30 cases during follow-up. RESULTS: All the patients with 9.16 mm (5-11 mm) height of residual bone in the posterior maxilla underwent sinus floor elevation with simultaneous implant placement using bone condensers. The elevation height was 2-6 mm. The follow-up time was 16-82 months. No sinus complication was observed during the follow-up period. The survival rate was 100%. Spiral CT scanning and three-dimensional reconstruction showed that dental implants were submerged in the alveoli around, their tops were covered with bone. CONCLUSIONS: With bone condensers, maxillary sinus floor elevation with simultaneous placement of implants from the top of alveoli was a predictable and safe technique. There were no implants loose or lost, no clinical complaint of maxillary sinus area, and X-ray examination showed well osseointegration.

[Maxillary sinus lifting and simultaneously placement of implants from the top of alveoli].

OBJECTIVE: To study the effect of maxillary sinus floor elevation by the Frialit-2 Bone Condenser for implantation. METHODS: 11 patients underwent sinus floor lift by The Frialit-2 Bone Condenser and were inserted 14 implants. The time of following up was 10 - 21 months. RESULTS: There were no implant loose or lost, no clinical complaint of maxillary sinus area, and X-ray exam showed well osseointegration. CONCLUSIONS: The Frialit-2 bone condenser can be used for lifting sinus floor, and the sinus elevation without lateral access allows the insertion of implants with no additional surgical stress for the patients.