Resveratrol Induces Mitochondrial Apoptosis and Inhibits Epithelial-Mesenchymal Transition in Oral Squamous Cell Carcinoma Cells.

OSCC is the most common malignant cancer of the head and neck. EMT is an essential cellular process critical to the morphogenesis and homeostasis of solid tissues. It is also involved in the initial stage of cancer metastasis and invasion in which cells lose epithelial characteristics. While cancer therapy protocols such as surgery, radiation, and chemotherapy are effective and useful, the drug tolerance and toxicity of OSCC patients remain a problem. Resveratrol is mainly produced in red grape skin and exhibits anti-oxidative, anti-inflammatory, anti-proliferative, and anti-cancer properties. This study was undertaken to investigate the underlying mechanisms giving rise to the induction of apoptosis by resveratrol in the human tongue squamous cell carcinoma cell line. Resveratrol treatment resulted in a time- and dose-dependent decrease in cell viability and increased the apoptotic cell ratio in CAL-27, SCC15, and SCC25 cells. Resveratrol treatment of CAL-27 cells showed that several lines of apoptotic manifestation and decreased cell migration, invasion, and EMT-inducing transcription factor. Taken together, our findings demonstrate the inhibitory effect of resveratrol in human OSCC cells via the mitochondrial pathway and that resveratrol is able to inhibit cell invasion and migration by inhibiting the EMT-inducing transcription factors.

The role of kaempferol-induced autophagy on differentiation and mineralization of osteoblastic MC3T3-E1 cells.

BACKGROUND: Kaempferol, a kind of flavonol, has been reported to possess various osteogenic biological activities, such as inhibiting bone resorption of osteoclasts and promoting the differentiation and mineralization of preosteoblasts. However, the precise cellular mechanism of action of kaempferol in osteogenesis is elusive. Autophagy is a major intracellular degradation system, which plays an important role in cell growth, survival, differentiation and homeostasis in mammals. Recent studies showed that autophagy appeared to be involved in the degradation of osteoclasts, osteoblasts and osteocytes, potentially pointing to a new pathogenic mechanism of bone homeostasis and bone marrow disease. The potential correlation between autophagy, osteogenesis and flavonoids is unclear. METHODS: The present study verified that kaempferol promoted osteogenic differentiation and mineralization and that it elevated osteogenic gene expression based on alkaline phosphatase (ALP) activity, alizarin red staining and quantitative PCR. And then we found that kaempferol induced autophagy by acridine orange (AO) and monodansylcadaverine (MDC) staining and autophagy-related protein expression. The correlation between kaempferol-induced autophagy and the osteogenic process was confirmed by the autophagy inhibitor 3-methyladenine (3-MA). RESULTS: Kaempferol promoted the proliferation, differentiation and mineralization of osteoblasts at a concentration of 10 µM. Kaempferol showed cytotoxic properties at concentrations above 50 µM. Concentrations above 10 µM decreased ALP activity, whereas those up to 10 µM increased ALP activity. Kaempferol at concentrations up to 10 µM also increased the expression of the osteoblast- activated factors RUNX-2, osterix, BMP-2 and collagen I according to RT-PCR analyses. 10 µM or less, the higher of the concentration and over time, kaempferol promoted the activity of osteoblasts. Kaempferol induced autophagy. It also increased the expression of the autophagy-related factors beclin-1, SQSTM1/p62 and the conversion of LC3-II from LC3-I. The application of 3-MA decreased the activity of ALP and the autophagy induced by kaempferol. In the RT-PCR analysis, the expression of RUNX-2, osterix, BMP-2 and collagen I was decreased. CONCLUSION: The present study showed that kaempferol stimulated the osteogenic differentiation of cultured osteoblasts by inducing autophagy.

Effect of low-level laser therapy on oral keratinocytes exposed to bisphosphonate.

Bisphosphonate-related osteonecrosis of the jaw (BRONJ) is a side effect of bisphosphonate therapy. However, its pathophysiology is not yet fully elucidated, and effective treatment of BRONJ remains unclear. The aim of this study is to investigate the effects of alendronate on oral keratinocytes and of low-level laser therapy (LLLT) on alendronate-treated keratinocytes, specifically by evaluating their viability, apoptosis, and wound healing function after irradiation. Oral keratinocyte cells (HaCaT) were exposed to 25 µM alendronate. Then, laser irradiation was performed with a low-level Ga-Al-As laser (λ = 808 ± 3 nm, 80 mW, and 80 mA; NDLux, Seoul, Korea) using 1.2 J/cm(2) energy dose. Viability was analyzed using MTT assay. Apoptosis was measured by Hoechst staining, caspase assay. Changes in secretion of IL-8, VEGF, and collagen type I were studied by ELISA and immunofluorescence microscopy. Scratch wound assays were also performed to measure cellular migration. Our results show that alendronate inhibits keratinocyte viability, expression of IL-8, VEGF, and collagen type I which are intimately related to healing events and cell migration while promoting apoptosis. Our results serve to demonstrate the utility of LLLT in partially overcoming the inhibitory effects of this bisphosphonate. From these results, the authors believe that the present study will provide an experimental basis for a fuller explanation of the clinical effects of LLLT as a BRONJ treatment modality.

Transient ipsilateral mydriasis during correction of left blowout fracture.

Mydriasis, either bilateral or unilateral, seldom occurs during reconstruction of periorbital fracture. Anisocoria, a unilateral mydriasis, requires more urgent assessment than bilateral mydriasis does. Pharmacologic agents, local anesthetic infiltration, as well as direct or indirect oculomotor nerve damage are possible causes of unilateral mydriasis. Few cases have been reported about intraoperative temporary ipsilateral mydriasis during correction of blowout fracture. We have experienced an unusual case of anisocoria and report the case with literature reviews.

Low-level laser therapy induces the expressions of BMP-2, osteocalcin, and TGF-ß1 in hypoxic-cultured human osteoblasts.

The aim of this study was to examine the effect of low-level laser therapy (LLLT) on the cell viability and the expression of hypoxia-inducible factor-1s (HIF-1s), bone morphogenic protein-2 (BMP-2), osteocalcin, type I collagen, transforming growth factor-ß1 (TGF-ß1), and Akt in hypoxic-cultured human osteoblasts. Human fetal osteoblast cells (cell line 1.19) were cultured under 1 % oxygen tension for 72 h. Cell cultures were divided into two groups. At the experimental side, low-level laser (808 nm, GaAlAs diode) was applied at 0, 24, and 48 h. After irradiation, each cell culture was incubated 24 h more under hypoxia. Total energy was 1.2, 2.4, and 3.6 J/cm(2), respectively. Non-irradiated cultures served as controls. Comparisons between the two groups were analyzed by t test; a p value <0.05 was considered statistically significant. Hypoxia resulted in a decrease in the expression of type I collagen, osteocalcin, and TGF-ß1 (p < 0.001, p < 0.001, and p < 0.01, respectively). Cell viability and BMP-2 expression were not decreased by hypoxic condition. On the other hand, LLLT on hypoxic-cultured osteoblast promoted the expression of BMP-2, osteocalcin, and TGF-ß1 (p < 0.05, p < 0.01, and p < 0.001, respectively). Cell proliferation was also increased time-dependently. However, hypoxia decreased in type I collagen expression (p < 0.001), and LLLT did not affect type I collagen expression in hypoxic-cultured osteoblasts. Furthermore, LLLT inhibited HIF-1 and Akt expression in hypoxic conditioned osteoblasts. We concluded that LLLT induces the expression of BMP-2, osteocalcin, and TGF- ß1 in 1 % hypoxic-cultured human osteoblasts.

Decreasing Effect of Lidocaine·HCl on the Thickness of the Neuronal and Model Membrane.

This study examined the mechanism of action of a local anesthetic, lidocaine·HCl. Energy transfer between the surface fluorescent probe, 1-anilinonaphthalene-8-sulfonic acid, and the hydrophobic fluorescent probe, 1,3-di(1-pyrenyl) propane, was used to determine the effect of lidocaine·HCl on the thickness (D) of the synaptosomal plasma membrane vesicles (SPMV) isolated from the bovine cerebral cortex, and liposomes of the total lipids (SPMVTL) and phospholipids (SPMVPL) extracted from the SPMV. The thickness (D) of the intact SPMV, SPMVTL and SPMVPL were 1.044±0.008, 0.914±0.005 and 0.890±0.003 (arbitrary units, n=5) at 37℃ (pH 7.4), respectively. Lidocaine·HCl decreased the thickness of the neuronal and model membrane lipid bilayers in a dose-dependent manner with a significant decrease in the thickness, even at 0.1 mM. The decreasing effect of lidocaine·HCl on the membrane thickness might be responsible for some, but not all of its anesthetic action.

The Effect of Lidocaine · HCl on the Fluidity of Native and Model Membrane Lipid Bilayers.

The purpose of this study is to investigated the mechanism of pharmacological action of local anesthetic and provide the basic information about the development of new effective local anesthetics. Fluorescent probe techniques were used to evaluate the effect of lidocaine·HCl on the physical properties (transbilayer asymmetric lateral and rotational mobility, annular lipid fluidity and protein distribution) of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex, and liposomes of total lipids (SPMVTL) and phospholipids (SPMVPL) extracted from the SPMV. An experimental procedure was used based on selective quenching of 1,3-di(1-pyrenyl)propane (Py-3-Py) and 1,6-diphenyl-1,3,5-hexatriene (DPH) by trinitrophenyl groups, and radiationless energy transfer from the tryptophans of membrane proteins to Py-3-Py. Lidocaine·HCl increased the bulk lateral and rotational mobility of neuronal and model membrane lipid bilayes, and had a greater fluidizing effect on the inner monolayer than the outer monolayer. Lidocaine·HCl increased annular lipid fluidity in SPMV lipid bilayers. It also caused membrane proteins to cluster. The most important finding of this study is that there is far greater increase in annular lipid fluidity than that in lateral and rotational mobilities by lidocaine·HCl. Lidocaine·HCl alters the stereo or dynamics of the proteins in the lipid bilayers by combining with lipids, especially with the annular lipids. In conclusion, the present data suggest that lidocaine, in addition to its direct interaction with proteins, concurrently interacts with membrane lipids, fluidizing the membrane, and thus inducing conformational changes of proteins known to be intimately associated with membrane lipid.

Expression of receptor activator of nuclear factor -kappaB ligand, receptor activator of nuclear factor -kappaB, and osteoprotegerin, following low-level laser treatment on deproteinized bovine bone graft in rats.

The aim of this study was to investigate by immunohistochemistry the effects of low-level laser (LLL) irradiation on the expression of the receptor activator of nuclear factor -kappaB ligand (RANKL), osteoprotegerin (OPG), and the receptor activator of nuclear factor -kappaB (RANK) in deproteinized bovine bone grafts in rats. Twenty-four male Sprague-Dawley rats aged 15 weeks were allocated to either an experimental group that underwent LLL irradiation during bone healing at the bone graft sites of the rats' calvarial bone defects or a control group. In the experimental group, gallium-aluminum-arsenide (Ga-Al-As) diode LLL (wavelength 808 nm; output 96 mW) was used to irradiate three areas on and around bone defects. The radiation was administered by the contact method for 10 s at 8.3 J/cm(2), once a day for 7 days. The total dose over the complete schedule was 40.32 J. The animals were killed on days 7, 14 or 21. The results of immunohistochemical analysis showed that the expression of RANKL (P = 0.199), OPG (P = 0.035), and RANK (P = 0.020) in the experimental group significantly increased from day 7, with a more even distribution than in the control group, and that this difference prevailed until the end of the experiment. Bone density of the experimental group after trichrome staining was also higher than in the control group. These results suggest that LLL irradiation facilitates bone metabolism during bone healing at the sites of deproteinized bovine bone grafts in rats.

Effects of dimyristoylphosphatidylethanol and ethanol on thickness of neuronal membrane lipid bilayers.

The aim of this study was to provide a basis for examining the molecular mechanism for the pharmacological action of ethanol. Energy transfer between the surface fluorescent probe 1-anilinonaphthalene-8-sulfonic acid and hydrophobic fluorescent probe 1,3-di(1-pyrenyl)propane was used to examine the effect of both dimyristoylphosphatidylethanol (DMPEt) and ethanol on the thickness (D) of the synaptosomal plasma membrane vesicles (SPMV) isolated from the bovine cerebral cortex. The thickness (D) of the intact SPMV was 1.044 +/- 0.008 (arbitrary units, n=5) at 37 degrees C (pH 7.4). Both DMPEt and ethanol decreased the thickness of the SPMV lipid bilayer in a dose-dependent manner with a significant decrease in thickness observed at 5 microM and 25 mM, respectively. It was assumed that both ethanol and DMPEt cause interdigitation in the SPMV lipid bilayers. The effects of ethanol on the neuronal membranes were attributed to its direct and indirect actions. The indirect action of ethanol refers to the action of phosphatidylethanol, which is an ethanol abnormal metabolite, on the neuronal membranes. The decrease in membrane thickness by both DMPEt and ethanol might be responsible for some, but not all of its anesthetic actions.

The effect of lead on calcium release activated calcium influx in primary cultures of human osteoblast-like cells.

To further understand the significance of bone as a target tissues of lead toxicity, as well as a reservoir of systemic lead, it is necessary to define the effect of lead on the calcium release activated calcium influx (CRACI) in primary cultures of human osteoblast-like cells (OLC). Pb2+ inhibited the immediate CRACI dose-dependent manner. Influx of Pb2+ into human OLC was increased dose-dependent manner. The present study demonstrates that the interference of Pb2+ with CRACI of human OLC is at least twofold: (1) the initiation of CRACI, i.e., the measurable influx of Ca2+ upon Ca2+ readdition, is partially inhibited by Pb2+ and (2) the influx of Pb2+ was enhanced after CRACI had been induced.

Amphiphilic effects of local anesthetics on rotational mobility in neuronal and model membranes.

To provide a basis for studying the molecular mechanism of pharmacological action of local anesthetics, we carried out a study of the membrane actions of tetracaine, bupivacaine, lidocaine, prilocaine and procaine. Fluorescence polarization of 12-(9-anthroyloxy)stearic acid (12-AS) and 2-(9-anthroyloxy)stearic acid (2-AS) were used to examine the effects of local anesthetics on differential rotational mobility between polar region and hydrocarbon interior of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex, and liposomes of total lipids (SPMVTL) and phospholipids (SPMVPL) extracted from the SPMV. The two membrane components differed with respect to 2 and 12 anthroyloxy stearate (2-AS, 12-AS) probes, indicating that a difference in the membrane fluidity may be present. In a dose-dependent manner, tetracaine, bupivacaine, lidocaine, prilocaine and procaine decreased anisotropy of 12-AS in the hydrocarbon interior of the SPMV, SPMVTL and SPMVPL, but tetracaine, bupivacaine, lidocaine and prilocaine increased anisotropy of 2-AS in the membrane interface. These results indicate that local anesthetics have significant disordering effects on hydrocarbon interior of the SPMV, SPMVTL and SPMVPL, but have significant ordering effects on the membrane interface, and thus they could affect the transport of Na(+) and K(+) in nerve membranes, leading to anesthetic action.

Clinical changes of TMD and condyle stability after two jaw surgery with and without preceding TMD treatments in class III patients.

BACKGROUND: This study are to identify the symptomatic changes and condylar stability after 2 jaw surgery without preceding treatments for Temporomandibular joints(TMJ) in class III patients with the TMJ symptoms; and to assess therapeutic effect of 2 jaw surgery and the necessity of preceding treatment for alleviation of TMJ symptoms. METHODS: 30 prognathic patients with preexisting TMJ symptoms were divided into 2 groups according to presence or absence of preceding treatments before the surgery. We evaluated symptomatic changes on both TMJ by questionnaires and clinical examinations. And we reconstructed 3D cone beam computed tomography images before 2 jaw surgery, immediately after the surgery, and 6 months or more after the surgery with SimPlant software, and analyzed the stability of condylar position on 3D reconstruction model. Significances were assessed by the Wilcoxon signed rank test on SPSS ver. 20.0. RESULTS: Both groups had favorable changes of TMJ symptoms after orthognathic surgery. And postoperative position of condyle had good stability during follow-up period. CONCLUSION: 2 jaw surgery without preceding treatments for TMD can have therapeutic effect for TMD patients with class III malocclusion.

Facial Bone Fracture Patients Visiting Pusan National University Hospital in Busan and Yangsan: Trends and Risks.

PURPOSE: This study examined patients with facial bone fracture visiting Pusan National University Dental Hospital to understand the trends, and to enhance appropriate care and treatment for patients with facial bone fracture. METHODS: We investigated 531 patients presenting with facial bone fracture in Yangsan and 802 patients in Busan from January 2010 to December 2013. We divided the patients by year, month, gender, age, site, and cause to compare with historic data and other studies. RESULTS: The gender ratio was 3.58:1 in Yangsan and 4.31:1 in Busan. Patients aged in their 20s had the highest number of facial bone fractures in both Yangsan and Busan. The most frequent fracture site was the mandible, and the most frequent cause was slip down in both Yangsan and Busan. CONCLUSION: The investigation and comparison of patients with facial bone fracture who visited Pusan National University Hospital located at Yangsan and Busan from 2010 to 2013 found a difference in the total number of patients at each hospital, but the trends were not significantly different.

The impact factors on 5-year survival rate in patients operated with oral cancer.

OBJECTIVES: The purpose of this study is to analyze clinical impact factors on the survival rate, and to acquire basic clinical data for the diagnosis of oral cancer, for a determination of the treatment plan with long-term survival in oral cancer patients. MATERIALS AND METHODS: Through a retrospective review of the medical records, the factors for long-term survival rate were analyzed. Thirty-seven patients, among patient database with oral cancer treated in the Department of Oral and Maxillofacial Surgery at Pusan National University Hospital within a period from March 1998 to March 2008, were selected within the study criteria and were followed-up for more than 5 years. The analyzed factors were gender, age, drinking, smoking, primary tumor site, type of cancer, TNM stage, recurrence of affected region, and metastasis of cervical lymph node. The 5-year survival rate on the impact factors was calculated statistically using the Kaplan-Meier method. RESULTS: By classification of clinical TNM at the 1st visit, there were 11 (29.7%) cases for stage I, 11 (29.7%) cases for stage II, 3 (8.1%) cases for stage III, and 12 (32.5%) cases for stage IV. The 5-year survival rate of total oral cancer patients after the operation were 75.7%, pathological TNM stage related 5-year survival rate were as follows: stage I 90.0%, stage II 81.8%, stage III 100% and stage IV 45.5%; in which the survival rate difference by each stage was significantly observed. The recurrence of cervical lymph node was the significant impact factor for the survival rate, because only 30.0% the survival rate in recurrent cases existed. During the follow-up, there were 15 (40.5%) patients with confirmed recurrence, and the 5-year survival rate of these patients was decreased as 46.7%. CONCLUSION: The classification of clinical and pathological TNM stage, local recurrence after surgery, and metastasis of cervical lymph node after surgery were analyzed as the 3 most significant factors.

A comparative study of the effectiveness of sinus bone grafting with recombinant human bone morphogenetic protein 2-coated tricalcium phosphate and platelet-rich fibrin-mixed tricalcium phosphate in rabbits.

OBJECTIVES: The objective of this histologic study was to evaluate platelet-rich fibrin (PRF)-mixed tricalcium phosphate (TCP) and recombinant human bone morphogenic protein 2 (rhBMP-2)-coated TCP in their potential to enhance bone regeneration in sinus elevation in rabbits as well as in their inflammatory features. STUDY DESIGN: Bilateral round-shaped defects (diameter 8.0 mm) were formed in the maxillary anterior sinus walls of 36 New Zealand white rabbits. The defects were grafted with TCP only (control group), with rhBMP-2-coated TCP (experimental group A) and with PRF-mixed TCP (experimental group B). Each group included 12 rabbits. The animals were killed at 3 days, 1 week, 2 weeks, 4 weeks, 6 weeks, and 8 weeks. The specimens underwent decalcification and were stained for histologic analysis. RESULTS: There were no significant differences in inflammatory features among the groups at 3 days or the first week after operation. In a histomorphometric analysis, the new bone formation ratio showed significant differentiation between groups A and B. The TCP-only control group showed a relatively lower bone formation ratio rather than the experimental groups. The PRF-mixed TCP group showed a larger bone formation area, compared with both the control group and group A. CONCLUSIONS: In the results of the histologic evaluation (hematoxylin-eosin, Masson trichrome stain), the experimental groups A and B showed rapid bone formation, remodeling, and calcification in the second week. Moreover, there was a significant difference between those experimental groups and the control group in the new bone formation area at the fourth, sixth, and eighth weeks. The PRF-mixed TCP showed more rapid bone healing than the rhBMP-2-coated TCP or the TCP-only control.

The effect of methanol on the structural parameters of neuronal membrane lipid bilayers.

The structures of the intact synaptosomal plasma membrane vesicles (SPMVs) isolated from bovine cerebral cortexs, and the outer and the inner monolayer separately, were evaluated with 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1,3-di(1-pyrenyl)propane (Py-3-Py) as fluorescent reporters and trinitrophenyl groups as quenching agents. The methanol increased bulk rotational and lateral mobilities of SPMVs lipid bilayers. The methanol increased the rotational and lateral mobilities of the outer monolayers more than of the inner monolayers. n-(9-Anthroyloxy)stearic acid (n-AS) were used to evaluate the effect of the methanol on the rotational mobility at the 16, 12, 9, 6, and 2 position of aliphatic chains present in phospholipids of the SPMVs outer monolayers. The methanol decreased the anisotropy of the 16-(9-anthroyloxy)palmitic acid (16-AP), 12-(9-anthroyloxy)stearic acid (12-AS), 9-(9-anthroyloxy)stearic acid (9-AS), and 6-(9-anthroyloxy)stearic acid (6-AS) in the SPMVs outer monolayer but it increased the anisotropy of 2-(9-anthroyloxy)stearic acid (2-AS) in the monolayers. The magnitude of the increased rotational mobility by the methanol was in the order at the position of 16, 12, 9, and 6 of aliphatic chains in phospholipids of the outer monolayers. Furthermore, the methanol increased annular lipid fluidity and also caused membrane proteins to cluster. The important finding is that was far greater increase by methanol in annular lipid fluidity than increase in lateral and rotational mobilities by the methanol. Methanol alters the stereo or dynamics of the proteins in the lipid bilayers by combining with lipids, especially with the annular lipids. In conclusion, the present data suggest that methanol, in additions to its direct interaction with proteins, concurrently interacts with membrane lipids, fluidizing the membrane, and thus inducing conformational changes of proteins known to be intimately associated with membranes lipids.

Mutually opposite effects of dopamine.HCl and chlorpromazine. HCl on the thickness of liposomal lipid bilayers.

The aim of this study was to provide a basis for examining the molecular mechanism for the pharmacological action of dopamine.HCl (DA) and chlorpromazin.HCl (CPZ). Radiationless energy transfer from the surface fluorescent probe, 1-anilinonaphthalene-8-sulfonic acid, to the hydrophobic fluorescent probe, bispyrenylpropane (Py-Py), was used to examine the effects of DA and CPZ on the thickness (D) of the liposomal lipid bilayers prepared with total lipids (SPMVTL) and phospholipids (SPMVPL) extracted from neuronal membranes. The thickness (D) of intact SPMVTL and SPMVPL (37 degrees C, pH 7.4) were 0.914 +/- 0.010 and 0.886 +/- 0.009 (arbitrary units, n = 5), respectively. DA decreased the thickness of both SPMVTL and SPMVPL in a dose-dependent manner with a significant decrease in thickness observed even at 40 x 10(-9) M and 40 x 10(-9) M, respectively. On the other hand, CPZ increased the thickness of both SPMVTL and SPMVPL in a dose-dependent manner with a significant increase in thickness observed even at 35 x 10(-5) M and 35 x 10(-5) M, respectively. The sensitivities to the decreasing and increasing effect of the membrane lipid bilayers thickness by DA and CPZ, respectively, differed according to the liposomes in descending order of SPMVPL and SPMVTL. The decreasing and increasing action of DA and CPZ, respectively, on the membrane thickness had many effects that may be responsible for the dopaminergic receptor-DA and -CPZ interaction as well as the protein-lipid interaction.

Effect of low-level laser treatment after installation of dental titanium implant-immunohistochemical study of RANKL, RANK, OPG: an experimental study in rats.

BACKGROUND AND OBJECTIVES: This experiment using an animal experimental model was conducted in order to investigate the effect of low-level laser therapy (LLLT) on the healing of the dental titanium implant. STUDY DESIGN/MATERIALS AND METHODS: The experimental group received LLLT for a week and the control group did not. Each group consisted of 10 rats. Two rats from the groups were euthenized on the days 1, 3, 7, 14, and 21 of the experiment. The expression of receptor activator of nuclear factor kB ligand (RANKL), osteoprotegerin (OPG), and receptor activator of nuclear factor kB (RANK) were investigated. RESULTS: The expression of RANKL was observed from the initial stage of the installation of the implant for both the experimental and control groups. However, the degree of expression was higher in the experimental group. The degree of expression of OPG increased remarkably in the experimental group, while in the control group the degree of expression increased only slightly. In the experimental group, the expression of RANK was observed from the first day, but in the control group, it was weakly observed after day 3. The overall expression within the bone was slight on day 7 in the control group, while an active expression was observed in the experimental group. Bone density after installation of dental titanium implant during osseointegration in the experimental group was higher than the control group. The surface and structure of the titanium implant was not damaged by low-level laser (LLL). CONCLUSIONS: From the above results, the expression of OPG, RANKL, and RANK during the osseointegration of the dental titanium implant was observed within bone tissue. The application of the LLL influenced the expression of OPG, RANKL, and RANK, and resulted in the expansion of metabolic bone activity and increased the activity of bone tissue cells.

Bone regeneration in mandibular distraction osteogenesis combined with compression stimulation.

PURPOSE: This study compared modified distraction osteogenesis (DO) protocol with conventional DO protocol on healing bone formation. Computer simulation was performed to understand the mechanical environment of modified DO protocol, which applies compression during the consolidation period. MATERIALS AND METHODS: Fifty rats were used in this study. Twenty-five rats in the conventional DO (control) group were sacrificed at postoperative days 11, 21, 28, 35, and 49 after osteotomy. In the modified DO (experimental) group, compression was applied on day 7 after distraction (day 18 postoperatively) for 4 days during the early consolidation period and 25 rats were sacrificed on postoperative day 19, 28, 39, 46, and 53. The histologic and radiographic findings were used to compare the 2 groups. Further, computer simulation was used to predict the mechanical environment of healing bone under conventional and modified DO protocol. RESULTS: Radiographic findings showed that the experimental group resulted in denser and wider healing bone. Histologically, the experimental group yielded more mature lamellar bone than the control group. Computer simulation showed that absolute values of tissue strains were nearly double in the control group because of the softer healing tissues. Both the experimental and control groups showed high strains at the ridge crest. Concentrated tensile strain along the distraction direction at the ridge crest might hinder bone formation at the interface, while compressive strain could facilitate the process. CONCLUSION: This study proposed a modified DO protocol of adding compression during the early consolidation period of conventional DO protocol. This new technique appears to provide faster and denser bone regeneration.