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Preserving the marginal bone level (MBL) is essential for the long-term success of dental implant therapy, and bone remodeling around dental implants is considered to vary with time. Numerous studies comparing the platform-switching (PS) and platform-matching (PM) dental implants have indicated that PS dental implants showed a lesser reduction for the MBL, and the majority of them had a relatively short period. This study aimed to evaluate vertical and horizontal bone defects by using digital periapical radiographs to examine the changes in MBL around PM and PS dental implants over 5 years after functional loading. The vertical MBL (vMBL) was measured from the implant–abutment junction to the first bone-to-implant contact. The horizontal MBL (hMBL) was measured from the implant–abutment junction to the bone crest. All data were presented as means ± standard errors. Paired and independent t-tests with Welch’s correction were used to analyze the data. A total of 61 dental implants in 38 patients after 5 years of functional loading were evaluated. Over time, PS dental implants were more likely to gain bone; by contrast, PM dental implants were more likely to lose bone during the observation time. Changes in vMBL for PS dental implants were significantly less than those for PM dental implants at 1-year (p = 0.045), 3-year (p = 0.021), and 5-year (p = 0.010) loading. Likewise, changes in hMBL for PS dental implants were significantly smaller than in those for PM dental implants at 3-year (p = 0.021) and 5-year (p = 0.006) loading; however, the changes were minimal in both approaches. PS dental implants had a significant increment in the percentage of bone integration, whereas that for PM dental implants dropped over time, with no significance. In PS dental implants that occlude with natural teeth, vertical and horizontal bone gain was observed, and it was significant at 3 years (p = 0.023). A significant horizontal bone gain was observed in the opposing natural teeth at 3-year (p = 0.002) and 5-year loading (p = 0.002). The PS concept appears to preserve more MBL around dental implants by stabilizing the vMBL and hMBL over a 5-year period. A minimal marginal bone change was detected in both concepts. The opposing natural teeth at PS dental implants showed a favorable effect on marginal bone tissues.

Background: The maintenance of marginal bone levels around dental implants is an important criterion for evaluating the success of implants. Although computer-aided design/computer-aided manufacturing (CAD/CAM) customized abutments (CAs) provide more flexible solutions, compared with the original preformed stock abutments (PAs), there are dimensional tolerances and underlying drawbacks in the production of CAD/CAM CAs, which may change the tightness and seamless connection between fixtures and abutments set by the manufacturer and then affect the long-term stability of the abutments. This study aimed to examine the change in both mesial and distal bone levels using digital periapical radiographs to evaluate the difference between CAD/CAM CAs and original PAs.Material and methods: Radiographs were taken before delivery; after functional loading for 1 month; and after 3, 6, and 12 months; and the vertical marginal bone levels (vMBLs) of both the mesial and distal surrounding implant bones were measured. All data are presented as means ± standard errors and were analyzed using Student’s t-test. A p-value < 0.05 was judged to represent a significant difference. Results: A total of 57 implants in 50 patients were divided into 22 CAD/CAM CAs and 35 original stock abutments. The PAs appeared to have a more stable bone level. By contrast, the amount of bone level change in the CAs was higher than that in the PAs. The change in the vMBL of the CAs was significantly more than that of the PAs after functional loading for 1 month (p = 0.006), 3 months (p = 0.013), 6 months (p = 0.014), and 12 months (p = 0.002). In contrast, the distal marginal bone level was lower than the mesial marginal bone level in any period. Nevertheless, the bone levels of the CAs and PAs in any period were comparable with no significant difference. Conclusions: Significant differences were found between the mesial and distal bone levels in the PAs. The CAD/CAM CAs showed a significantly greater bone level change than the original stock abutments after functional loading.

The signaling mechanisms for Porphyromonas gingivalis lipopolysaccharide (PgLPS)-induced inflammation in human dental pulp cells are not fully clarified. This in vitro study aimed to evaluate the involvement of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in PgLPS-induced pulpal inflammation.Human dental pulp cells (HDPCs) were challenged with PgLPS with or without pretreatment and coincubation with a PI3K/Akt inhibitor (LY294002). The gene or protein levels of PI3K, Akt, interleukin (IL)-6, IL-8, alkaline phosphatase (ALP), osteocalcin and osteonectin were analyzed by reverse transcription polymerase chain reaction (PCR), real-time PCR, western blotting, and immunofluorescent staining. In addition, an enzyme-linked immunosorbent assay was used to analyze IL-6 and IL-8 levels in culture medium.In response to 5 μg/ml PgLPS, IL-6, IL-8, and PI3K, but not Akt mRNA expression of HDPCs, was upregulated. IL-6, IL-8, PI3K, and p-Akt protein levels were stimulated by 10-50 μg/ml of PgLPS in HDPCs. PgLPS also induced IL-6 and IL-8 secretion at concentrations higher than 5 μg/ml. Pretreatment and co-incubation by LY294002 attenuated PgLPS-induced IL-6 and IL-8 mRNA expression in HDPCs. The mRNA expression of ALP, but not osteocalcin and osteonectin, was inhibited by higher concentrations of PgLPS in HDPCs.P. gingivalis contributes to pulpal inflammation in HDPCs by dysregulating PI3K/Akt signaling pathway to stimulate IL-6 and IL-8 mRNA/protein expression and secretion. These results are useful for understanding the pulpal inflammation and possible biomarkers of inflamed pulp diagnosis and treatment.

Titanium is widely used in medical implants despite the release of heavy metal ions over long-term use. Zirconia is very close to the color of teeth; however, its biological inertness hinders bonding with bone tissue. Alkaline treatment and coatings of calcium phosphate can be used to enhance bone regeneration adjacent to dental implants. This study examined the effects of alkaline treatment, calcium phosphate coatings, and sintering, on the physical properties of implant material. Our analysis confirmed that the calcium phosphate species were octacalcium phosphate (OCP). The sintering of calcium phosphate was shown to create B-type HAP, which is highly conducive toward the differentiation of mesenchymal stem cells (MSCs) into osteoblasts for the facilitation of bone integration. Conclusions: This study demonstrated the room-temperature fabrication of dental implants with superhydrophilic surfaces to enhance biocompatibility.

In vitro, in vivo, and clinical studies have shown how the physicochemical and biological properties of β-tricalcium phosphate (β-TCP) work in bone regeneration. This study aimed to improve the properties of β-TCP by achieving optimum surface and bulk β-TCP chemical/physical properties through the hydrothermal addition of magnesium (Mg) and to later establish the biocompatibility of β-TCP/Mg for bone grafting and tissue engineering treatments. Multiple in vitro and in vivo analyses were used to complete β-TCP/Mg physicochemical and biological characterization. The addition of MgO brought about a modest rise in the number of β-TCP surface particles, indicating improvements in alkaline phosphatase (ALP) activity on day 21 (p < 0.05) and in the WST-1assay on all days (p < 0.05), with a corresponding increase in the upregulation of ALP and bone sialoprotein. SEM analyses stated that the surfaces of the β-TCP particles were not altered after the addition of Mg. Micro-CT and histomorphometric analysis from rabbit calvaria critical defects resulted in β-TCP/Mg managing to reform more new bone than the control defects and β-TCP control at 2, 6, and 8 weeks (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001). The hydrothermal addition of MgO to the β-TCP surfaces ameliorated its biocompatibility without altering its surface roughness resulting from the elemental composition while enhancing cell viability and proliferation, inducing more bone regeneration by osteoconduction in vivo and osteoblastic differentiation in vitro.

Camphorquinone (CQ) and resin monomers are included in dentin bonding agents (DBAs) and composite resin to restore tooth defects due to abrasion, crown fracture, or dental caries. DBAs, CQ, and bisphenol A-glycidyl methacrylate (BisGMA) applications influence the biological activities of the dental pulp. The current investigation aimed to delineate the effect of DBAs, CQ, and BisGMA on cathepsin L production/expression, lysosomal activity, and autophagy induction in human dental pulp cells (HDPCs). HDPCs were exposed to DBAs, CQ, or BisGMA with/without inhibitors for 24 h. Enzyme-linked immunosorbent assay was employed to determine the cathepsin L level in culture medium. The cell layer was utilized to measure cell viability by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl -tetrazolium bromide (MTT) assay. Real-time PCR was used to evaluate the mRNA expression. Western blotting or immunofluorescent staining was used to study protein expression. Lysosomal density was evaluated by lysotracker red staining. We found that DBAs, CQ, and BisGMA stimulated cathepsin L mRNA, protein expression, and production in HDPCs. In addition, CQ and BisGMA induced lysosomal activity, Beclin1, ATG12, LC3B, Bax, and p53 expression in HDPCs, indicating the stimulation of autophagy. Glutathione (GSH) prevented CQ- and BisGMA-induced cytotoxicity. Moreover, E64d, cathepsin L inhibitor (two cathepsin inhibitors), and Pifithrin-α (a p53 inhibitor) showed little preventive effect toward CQ- and BisGMA-induced cytotoxicity. Autophagy inhibitors (NH4Cl, Lys05) mildly enhanced the CQ- and BisGMA-induced cytotoxicity. These results indicate that DBAs stimulated cathepsin L, possibly due to their content of CQ and BisGMA that may induce cathepsin L in HDPCs. CQ and BisGMA stimulated lysosomal activity, autophagy, and apoptosis, possibly via induction of Beclin 1, ATG12, LC-3B, Bax, and p53 expression. In addition, CQ and BisGMA cytotoxicity was related to redox change and autophagy. These events are important role in pulpal changes after the restoration of tooth decay using CQ- and BisGMA-containing DBAs and resin composite.

Few studies have compared marginal bone loss (MBL) around implant-retaining overdentures (IODs) vs. implant-supported fixed prostheses (FPs). This study evaluated the mean MBL and radiographic bone-implant interface contact (r-BIIC) around IODs and implant-supported FPs. We also investigated osseointegration and MBL around non-submerged dental implants. We measured the changes between the MBL in the mesial and distal sites immediately after prosthetic delivery and after one year. The mean MBL and its changes in the IOD group were significantly higher. The mean percentage of r-BIIC was significantly higher in the FP group. MBL and its changes in males were significantly higher in the IOD group. The percentage of r-BIIC was significantly higher in the FP group. MBL in the lower site in the IOD group was significantly higher. Regarding MBL, the location of the implant was the only significant factor in the IOD group, while gender was the only significant predictor in the FP group. Regarding the r-BIIC percentage, gender was a significant factor in the FP group. We concluded that non-submerged dental implants restored with FPs and IODs maintained stable bone remodeling one year after prosthetic delivery.

A vertical root fracture (VRF) is a complex complication that usually leads to tooth extraction. The aim of this article is to review the prevalence, demography, distribution, diagnostic methods, etiology and predisposing factors, clinical features, radiographic characteristics and treatment strategies of VRFs in non-endodontically treated teeth (VRFNETT) and endodontically treated teeth (VRFETT). Search terms for each subject related to VRFNETT and VRFETT were entered into MEDLINE, PubMed and Google Scholar. Systematic reviews, retrospective cohort studies, demographic research, clinical studies, case reports and case series were reviewed. Most of the VRFs were found in patients older than 40 years old. Older populations were discovered in the non-endodontically treated VRF group when compared to the endodontically treated VRF group. Male patients were found at a greater prevalence than females in the non-endodontically treated VRF group. The initial occurrence of a VRF may accompany radiolucent lines within the root canal, unusual space between the canal wall and intracanal material, a widening of the PDL space along the periradicular surfaces, angular bony destruction, step-like bone defects, V-shaped diffuse bone defects, or root resorptions corresponding to the fracture line before the clear separation of the fractured fragment. The indicative clinical and radiographic signs of VRF included a coronally positioned sinus tract, deep-narrow periodontal defects, the displacement of a fractured fragment, periradicular radiolucent halos and the widening of the root canal space. Interestingly, VRFNETT are more often observed in the Chinese population. Some patients with multiple VRFs were observed, suggesting possible predisposing factors in genetics and tooth development. The management of a VRF usually involves a multidisciplinary approach. The common distribution and features of VRFNETT and VRFETT were elucidated to facilitate recognition and diagnosis. Besides extraction, variable therapeutic schemes, such as the repair of the VRF, root amputation and others reported in earlier literature, are available. A long-term prognosis study of the various therapeutic strategies is needed.

Background/purpose: Interleukin 1 beta (IL-1β) is a pro-inflammatory cytokine involved in the acute and chronic inflammatory processes of dental pulp. Intercellular adhesion molecule-1 (ICAM-1) and IL-8 are two major inflammatory mediators. However, the role of interleukin-1 receptor-associated kinases (IRAKs) signaling pathways in responsible for the inflammatory effects of IL-1β on dental pulp cells is not clear. Methods: Cultured human dental pulp cells were exposed to IL-1β with/without pretreatment and co-incubation with IRAK1/4 inhibitor or SB203580 (p38 inhibitor). IRAK-1 phosphorylation was evaluated by immunno fluorescent staining. The protein expression of ICAM-1 and IL-8 were tested by western blotting. The secretion of soluble ICAM-1 (sICAM-1) and IL-8 was measured by enzyme-linked immunosorbant assay (ELISA). Results: IL-1β stimulated IRAK-1 phosphorylation of pulp cells within 120 min of exposure. IRAK1/4 inhibitor attenuated the IL-1β-induced ICAM-1, but not IL-8 protein expression. IRAK1/4 inhibitor also prevented the IL-1β-induced sICAM-1, but not IL-8 secretion. SB203580 showed little effect on IL-1β-induced sICAM-1 secretion, but effectively inhibited its induction of IL-8 secretion in pulp cells. Conclusion: The Results reveal the important role of IL-1β in pulpal inflammatory responses via stimulation of IL-8 and ICAM-1 expression and secretion. Moreover, IL-1β-induced effects on IL-8 and ICAM-1 are differentially regulated by IRAK1/4 and p38 signaling in dental pulp cells. Blocking of IRAKs and p38 signaling may have potential to control inflammation of dental pulp in the future. Keywords: Dental pulp, Inflammation, ICAM-1, Interleukin-1β, IL-8, Signal transduction, IRAKs

Background: In mixed dentition analysis, estimation of the mesiodistal width of unerupted permanent canines and premolars is essential for successful diagnosis and treatment planning. The present study aimed to develop a simple linear equation to predict permanent tooth sizes from mixed dentition analysis for Taiwanese people. Methods: The sample comprised 200 dental casts, derived from Taiwanese patients (100 males and 100 females, age, 12–35 years). Mesial distal tooth widths were measured in dental casts with a digital caliper. A student’s t-test was conducted to detect tooth size correlations with gender-specific differences, as well as intra-arch counterparts. Standard linear regression was conducted to develop a simple equation representing predictions of canine-premolar relationships. Results: All teeth were not significantly different between the left and right sides, regardless of gender and upper or lower arches. In terms of types of teeth, males had larger tooth dimensions in both arches than females. New regression equations for estimating the dimensions of the unerupted canines and premolars in the Taiwanese population were developed. Conclusions: Using a sample of Taiwanese people, new models derived for females and males separately were developed, which should provide highly accurate predictions for unerupted canines and premolars in the Taiwanese population.

AIM To investigate the effects of butyric acid (BA), a metabolic product generated by pulp and root canal pathogens, on the viability and intercellular adhesion molecule-1 (ICAM-1) production of endothelial cells, which are crucial to angiogenesis and pulpal/periapical wound healing. METHODOLOGY Endothelial cells were exposed to butyrate with/without inhibitors. Cell viability, apoptosis and reactive oxygen species (ROS) were evaluated using an MTT assay, PI/annexin V and DCF fluorescence flow cytometry respectively. RNA and protein expression was determined using a polymerase chain reaction assay and Western blotting or immunofluorescent staining. Soluble ICAM-1 (sICAM-1) was measured using an enzyme-linked immunosorbent assay. The quantitative results were expressed as mean ± standard error (SE) of the mean. The data were analysed using a paired Student's t-test where necessary. A p-value ≤0.05 was considered to indicate a statistically significant difference between groups. RESULTS Butyrate (>4 mM) inhibited cell viability and induced cellular apoptosis and necrosis. It inhibited cyclin B1 but stimulated p21 and p27 expression. Butyrate stimulated ROS production and hemeoxygenase-1 (HO-1) expression as well as activated the Ac-H3, p-ATM, p-ATR, p-Chk1, p-Chk2, p-p38 and p-Akt expression of endothelial cells. Butyrate stimulated ICAM-1 mRNA/protein expression and significant sICAM-1 production (p

A major number of studies have demonstrated Beta-tricalcium phosphate (β-TCP) biocompatibility, bioactivity, and osteoconductivity characteristics in bone regeneration. The aim of this research was to enhance β-TCP's biocompatibility, and evaluate its physicochemical properties by argon glow discharge plasma (GDP) plasma surface treatment without modifying its surface. Treated β-TCP was analyzed by scanning electron microscopy (SEM), energy-dispersive spectrometry, X-ray photoelectron spectroscopy (XPS), X-ray diffraction analysis, and Fourier transform infrared spectroscopy characterization. To evaluate treated β-TCP biocompatibility and osteoblastic differentiation, water-soluble tetrazolium salts-1 (WST-1), immunofluorescence, alkaline phosphatase (ALP) assay, and quantitative real-time polymerase chain reaction (QPCR) were done using human mesenchymal stem cells (hMSCs). The results indicated a slight enhancement of the β-TCP by GDP sputtering, which resulted in a higher Ca/P ratio (2.05) than the control. Furthermore, when compared with control β-TCP, we observed an improvement of WST-1 on all days (p p p

Betel quid (BQ) chewing increased the risk of oral cancer and oral submucous fibrosis (OSMF), an oral premalignant disorder (OPMD) with malignant transformation potential. BQ components such as areca nut (AN), trauma by coarse AN fiber, catechin, copper, alkaloids, stimulated reactive oxygen species (ROS), inflammation and cytotoxicity are suggested to be the contributing factors. They may induce tissue inflammation, proliferation of fibroblasts and collagen deposition, myofibroblast differentiation and contraction, collagen cross-links and inhibit collagen phagocytosis, finally leading to the development of OSMF and oral cancer. These events are mediated by BQ components-induced changes of extracellular matrix (ECM) turnover via regulation of TGF-β1, plasminogen activator inhibitor-1 (PAI-1), cystatin, lysyl oxidase (LOX) and tissue inhibitors of metalloproteinases (TIMPs) and metalloproteinases (MMPs). Genetic susceptibility is also involved in these disease processes. Further understanding the molecular mechanisms of BQ-induced OSMF and oral cancer can be helpful for future disease prevention and treatment.

Peri-implantitis is the pathological condition of connective tissue inflammation and the progressive loss of supporting bone around dental implants. One of the primary causes of peri mucositis evolving into peri-implantitis is bacterial infection, including infection from Porphyromonas gingivalis. Enhancing the surface smoothness of implants helps to prevent P. gingivalis adhesion to the implant&rsquo, s surface. Interaction analyses between bacteria and the surface roughness of zirconia (Zr) discs subjected to a glow discharge plasma (GDP) treatment compared with non-plasma-treated autoclaved control Zr discs were done. Examinations of the material prosperities revealed that the GDP-treated Zr group had a smoother surface for a better wettability. The GDP-treated Zr discs improved the proliferation of the osteoblast-like cells MG-63, and the osteoblastic differentiation was assessed through alkaline phosphatase detection and marker gene bone sialoprotein (Bsp) and osteocalcin (OC) induction. Scanning electron microscopy demonstrated a relatively low P. gingivalis adhesion on GDP-treated Zr disks, as well as lower colonization of P. gingivalis compared with the control. Our findings confirmed that the GDP treatment of Zr discs resulted in a significant reduction of P. gingivalis adhesion and growth, demonstrating a positive correlation between surface roughness and bacteria adhesion. Therefore, the GDP treatment of Zr dental implants can provide a method for reducing the risk of peri-implantitis.

Background/purpose: Interleukin 1 beta (IL-1β) is a pro-inflammatory cytokine involved in the inflammatory processes of dental pulp. IL-8 and urokinase plasminogen activator (uPA) are two inflammatory mediators. However, the role of transforming growth factor beta-activated kinase-1 (TAK1) and mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling pathways in responsible for the effects of IL-1β on IL-8 and uPA expression/secretion of dental pulp cells are not clear. Methods: Human dental pulp cells were exposed to IL-1β with/without pretreatment with 5z-7-oxozeaneaeol (a TAK1 inhibitor) or U0126 (a MEK/ERK inhibitor). TAK1 activation was determined by immunofluorescent staining. The protein expression of IL-8 was tested by western blot. The expression of IL-8 and uPA mRNA was studied by reverse transcriptase-polymerase chain reaction (RT-PCR). The secretion of IL-8 and uPA was measured by enzyme-linked immunosorbent assay. Results: Exposure of dental pulp cells to IL-1β (0.1–10 ng/ml) stimulated IL-8 and uPA expression. IL-1β also induced IL-8 and uPA secretion of dental pulp cells. IL-1β stimulated p-TAK1 activation of pulp cells. Pretreatment and co-incubation of pulp cells by 5z-7oxozeaenol (1 and 2.5 μM) and U0126 (10 and 20 μM) prevented the IL-1β-induced IL-8 and uPA expression. 5z-7oxozeaenol and U0126 also attenuated the IL-1β-induced IL-8 and uPA secretion. Conclusion: IL-1β is important in the pathogenesis of pulpal inflammatory diseases and repair via stimulation of IL-8 and uPA expression and secretion. These events are associated with TAK1 and MEK/ERK signaling. Blocking of TAK1 and MEK/ERK signaling has potential to control inflammation of dental pulp. Keywords: Dental pulp, Healing, Inflammation, Interleukin-1β, MEK/ERK, Signal transduction, TAK1, Urokinase plasminogen activator

Background: The objective of this study was to evaluate the effects of opposite tooth conditions on change in marginal bone level (MBL) around submerged dental implants. Materials and methods: The study included healthy patients with one or two implants. Structures opposite implants were either natural teeth (NT) or fixed restorations (FRs). MBLs were measured on digital periapical radiographs at the mesial and distal aspects of each implant. Results: Sixty implants were inserted by the 3-year follow-up. Mean MBLs for NT were 0.21 ± 0.33 mm before prosthetic loading and 0.30 ± 0.41 mm 3 years later (p = 0.001). Mean MBLs with FRs were 0.36 ± 0.45 mm before loading and 0.53 ± 0.50 mm 3 years later (p &lt, 0.001). Changes in mean MBL from the 6-month follow-up to the 1- and 3-year follow-ups were statistically significant (p &lt, 0.01) for implants opposite NT. However, changes in mean MBL from the 6-month follow-up to the 1-year (p = 0.161) and 3-year follow-ups (p = 1.000) were not significant for implants opposite FRs. Between baseline and the 3-year follow-up, MBL change was relatively small and did not differ regarding NT and FRs. Conclusion: Bone loss was greater if submerged dental implants were opposed by FRs. MBLs around submerged implants continued to change after 3 years if NT opposed implants.

Absorbable porcine collagen membrane with a bovine bone graft can be considered for regenerative treatment in periodontal class II furcation defects. We evaluated the clinical efficacy of guided tissue regeneration (GTR) treatment with bovine bone xenograft and a porcine collagen membrane in molars with class II furcations. Probing depth (PD), clinical attachment level (CAL), and bone level (BL) were recorded at baseline and at 3, 6, and 9 months postoperatively. Thirty class II furcation defects from the lower and upper molars were assessed. Significant improvements in PD and CAL were observed from baseline to 9 months in all groups (p &lt, 0.01). BL improved in all groups except group A in the upper molars in radiographic assessment (p &lt, 0.05). The lower and upper molars showed PD reduction of 50.5% ± 7.44% and 46.2% ± 11.2%, respectively, at 9 months (p = 0.044). In furcations of 1–3 mm, the lower and upper molars showed PD reductions of 51.2% ± 4.49% and 36.5% ± 16.14%, respectively (p = 0.035). The lower and upper molars showed a CAL gain of 51.1% ± 4.64% and 33.6% ± 18.8%, respectively (p = 0.037). Thus, GTR with bovine bone graft and porcine collagen membrane yielded good results in class II furcations, with better results in the lower than in the upper molars.

The most commonly used cryoprotectant for cells and tissues is dimethyl sulfoxide (DMSO). However, the cytotoxic effect of DMSO is a concern when high concentrations are used. The aim of this study was to assess the cryoprotective effects of static magnetic fields (SMFs) on human red blood cells (RBCs) during cryopreservation. Before the freezing process, RBCs were suspended in media containing 0, 7.5, or 15% DMSO. Then, samples were frozen at −80 °C with a 0.4-T SMF for 24 h. After the cells were thawed, the survival rate, morphology, and mechanical stability of cellular membranes were examined. The results show that SMFs exhibit the largest cryoprotective efficiency when DMSO was not present in the freezing medium. In addition, cell morphology and membrane stability of the frozen-thawed RBCs were not changed after 0.4-T SMF treatment. These results demonstrate that SMFs increase the survival rate of thawed RBCs frozen in a DMSO-free medium. Accordingly, SMF exposure during the freezing process improved the cryopreservation efficiency of RBCs.

Due to autogenous bone limitations, some substitute bone grafts were developed. Collagenated porcine graft (CPG) is able to regenerate new bone, although the number of studies is insufficient, highlighting the need for future studies to better understand the biomaterial. In order to understand better CPG&prime, s possible dental guided bone regeneration indications, the aim of this work was to determine CPG&prime, s biological capacity to induce osteoblast differentiation in vitro and guided bone regeneration in vivo, whilst being compared with commercial hydroxyapatite and beta tricalcium phosphate (HA/&beta, TCP) and porcine graft alone. Cell cytotoxicity (WST-1), alkaline phosphatase activity (ALP), and real-time polymerase chain reaction (qPCR) were assessed in vitro. Critical size defects of New Zealand white rabbits were used for the in vivo part, with critical size defect closures and histological analyses. WST-1 and ALP indicated that CPG directly stimulated a greater proliferation and confluency of cells with osteoblastic differentiation in vitro. Gene sequencing indicated stable bone formation markers, decreased resorption makers, and bone remodeling coupling factors, making the transition from osteoclast to osteoblast expression at the end of seven days. CPG resulted in the highest new bone regeneration by osteoconduction in critical size defects of rabbit calvaria at eight weeks. Nonetheless, all biomaterials achieved nearly complete calvaria defect closure. CPG was found to be osteoconductive, like porcine graft and HA/&beta, TCP, but with higher new bone formation in critical size defects of rabbit calvaria at eight weeks. CPG can be used for different dental guided bone regeneration procedures, however, further studies are necessary.

Background: There are 200-600 million betel quid (BQ) chewers in the world. BQ increases oral cancer risk. Matrix metalloproteinase-9 (MMP-9) is responsible for matrix degradation, cancer invasion and metastasis. Whether areca nut extract (ANE), a BQ component, stimulates MMP-9 secretion, and the related signaling pathways awaits investigation. Results: ANE (but not arecoline) stimulated MMP-9 production of gingival keratinocytes and SAS cancer epithelial cells. ANE stimulated TGF-β1, p-Smad2, and p-TAK1 protein expression. ANE-induced MMP-9 production/expression in SAS cells can be attenuated by SB431542 (ALK5/Smad2 inhibitor), 5Z-7-Oxozeaenol (TAK1 inhibitor), catalase, PD153035 (EGFR tyrosine kinase inhibitor), AG490 (JAK inhibitor), U0126 (MEK/ERK inhibitor), LY294002 (PI3K/Akt inhibitor), betel leaf (PBL) extract, and hydroxychavicol (HC, a PBL component), and melatonin, but not by aspirin. Conclusions: AN components contribute to oral carcinogenesis by stimulating MMP-9 secretion, thus enhancing tumor invasion/metastasis. These events are related to reactive oxygen species, TGF-β1, Smad2-dependent and –independent signaling, but not COX. These signaling molecules can be biomarkers of BQ carcinogenesis. PBL, HC and melatonin and other targeting therapy can be used for oral cancer treatment. Methods: ANE-induced MMP-9 expression/secretion of oral epithelial cells and related TGF-β1, Smad-dependent and –independent signaling were studied by MTT assay, RT-PCR, western blotting, immunofluorescent staining, and ELISA.

After tooth extraction, alveolar ridge loss due to resorption is almost inevitable. Most of this bone loss occurs during the first six months after the extraction procedure. Many studies have indicated that applying socket-filling biomaterials after extraction can effectively reduce the resorption rate of the alveolar ridge. The purpose of this study was to investigate the clinical efficacy of the application of a hydroxyapatite/&beta, tricalcium plus collagen (HA/&beta, TCP + collagen) dental bone graft in dental sockets immediately after tooth extraction, so as to prevent socket resorption. The study was conducted on 57 extraction sockets located in the mandible and maxilla posterior regions in 51 patients. HA/&beta, TCP + collagen was inserted into all of the dental sockets immediately after extraction, and was covered with a flap. Follow-up was performed for three months after extraction, using radiographs and stents for the vertical and horizontal alveolar ridge measurements. A minimal alveolar bone width reduction of 1.03 &plusmn, 2.43 mm (p &lt, 0.05) was observed. The height reduction showed a slight decrease to 0.62 &plusmn, 1.46 mm (p &lt, 0.05). Radiographically, the bone height was maintained after three months, indicating a good HA/&beta, TCP + collagen graft performance in preserving alveolar bone. In conclusion, the HA/&beta, TCP + collagen graft demonstrated adequate safety and efficacy in dental socket preservation following tooth extraction.

Peri-implantitis has become a common complication, accompanied by soft tissue inflammation. Porphyromonas gingivalis infection is the major cause of inflammation and progressive bone loss in the jaws. The surface property of titanium implants is a key factor in the alteration of osseointegration and P. gingivalis adhesion. However, the interplay between P. gingivalis and the surface properties of implants, subjected to different treatments, is not well described. Therefore, we focused on the surface properties of titanium implants, titanium disks that were autoclaved alone were used as controls. Those that were autoclaved and then subjected to low-temperature plasma (LTP) at 85 W and 13.56 MHz and with 100 mTorr of argon gas at room temperature for 15 min formed the experimental group. LTP-treated disks had smoother surfaces than the control group disks. The physical properties, such as scanning electron microscope (SEM), energy dispersive X-ray spectrometer (EDX), and X-ray photoelectron spectroscopy (XPS), demonstrated the surface composition was changed after LTP treatment. Further, osteoblastic cell proliferation enhancement was observed in the LTP-treated titanium surfaces. The results also revealed relatively less P. gingivalis adhesion to the LTP-treated disks than on the control disks on spectrophotometry and SEM. These findings clarified that P. gingivalis adhesion is reduced in implants subjected to LTP treatment. Thus, LTP treatment of peri-implantitis with the settings used in the present study is an option that needs further investigation.

Objective: To describe remodeling of the mesial and distal marginal bone level around platform-switched (PS) and platform-matched (PM) dental implants that were sandblasted with large grit and etched with acid over a three-year period. Materials and Methods: Digital periapical radiographs were obtained at the following time-points: during Stage I of the surgical placement of dental implants, before loading, immediately after loading (baseline), and one, three, six, 12, and 36 months after loading for measuring the horizontal and vertical marginal bone levels. Results: Sixty implants were successfully osseointegrated during the overall observation period. Vertical marginal bone levels for the PS and PM dental implants were 0.78 &plusmn, 0.77 and 0.98 &plusmn, 0.81 mm, respectively, whereas the horizontal marginal bone levels for the PS and PM implants were 0.84 &plusmn, 0.45 and 0.98 &plusmn, 0.68 mm, respectively. During the time leading up to the procedure until 36 months after the procedure, the average vertical marginal bone level resulted in less bone loss for the PS and PM groups&mdash, 0.96 &plusmn, 1.28 and 0.30 &plusmn, 1.15 mm, respectively (p &lt, 0.05). The mean levels of the horizontal marginal bone also showed increases of 0.48 &plusmn, 1.01 mm in the PS and 0.37 &plusmn, 0.77 mm in the PM groups from the time before loading until 36 months after the procedure. However, these increases were not statistically significant (p &gt, 0.05). Conclusion: PS dental implants appeared to be more effective than PM implants for minimizing the mean marginal vertical and horizontal marginal bone loss during the three-year period. Regardless of which abutment connection was used, the dental implant in the present retrospective investigation exhibited minimal marginal bone remodeling, thus indicating long-term stability.

The aim of this study was to incorporate nanoscale Fe3O4 particles into a poly-L-lactide (PLLA) matrix to fabricate a magnetic and biodegradable composite. The physical and osteogenic functions of this material were tested. Injection molding was used to fabricate four nano-Fe3O4/PLLA composites with Fe3O4 mix ratios of 0%, 20%, 30%, and 40% (w/w). X-ray diffraction and hysteresis loop tests were performed to evaluate the physical properties of the nano-Fe3O4/PLLA composites. Tensile strength tests showed that the progressive addition of nano-Fe3O4 particles to the PLLA matrix results in higher elastic modulus and lower tensile strength. Images from scanning electron microscopy demonstrated that osteoblasts cultured on the 20% nano-Fe3O4/PLLA surface exhibited abundant filaments, which are a morphologic characteristic of osteoblastic differentiation. These results suggest that the 20% nano-Fe3O4/PLLA composite used in this study has the potential for future tissue engineering applications. POLYM. COMPOS., 2016. © 2016 Society of Plastics Engineers

Melanogenesis is a complex process that can lead to pigmentation defects. Various chemical skin-lightening products have been developed to treat pigmentation disorders. However, these chemical products can cause harmful adverse effects. Therefore, the development of safer, natural bleaching ingredients is a trend for sustainability. It has been reported that unsaturated fatty acids exhibit significant antimelanogenic effects. Sapindus mukorossi seed oils contain abundant unsaturated fatty acids; however, these have not yet been investigated for beneficial effects on skin tone evenness. In this study, we tested the possibility of using S. mukorossi oil for the treatment of hyperpigmentation in an in vitro model. Free fatty acid compositions and β-sitosterol were determined by gas chromatography-mass spectrometry and high-pressure liquid chromatography, respectively. The effect of S. mukorossi oil on melanoma B16F10 cell viability was detected using the 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-tetrazolium bromide assay. The inhibitive effects of fatty acids and β-sitosterol in S. mukorossi oil on α-melanocyte-stimulating hormone (MSH)-induced melanogenesis was evaluated by detecting melanin formation and tyrosinase activity. Our results showed that S. mukorossi oil produced no significant cytotoxicity in B16F10 cells at various concentrations compared with the control group. The enhancement of melanin formation induced by α-MSH was reduced by S. mukorossi oil. We also found that the primary fatty acid contributing to the antimelanogenesis effect was eicosenoic acid. These results suggest that S. mukorossi seed oil can effectively inhibit melanogenesis and has the potential for future development as a de-hyperpigmentation product within a waste utilization context.

Platelets play crucial roles in thrombosis and hemostasis through platelet activation and aggregation that are crucial in cardiovascular diseases. Hydroquinone (HQ) and its derivatives are present in many dermatological creams, paints, motor fuels, air, microorganisms, and plant products like wheat bread, fruit, coffee, and red wine. The effect of HQ on humans is not clear. In this study, we found that HQ (>25 μM) inhibited arachidonic acid (AA)-induced platelet aggregation. HQ suppressed AA-induced thromboxane B2 production of platelets. HQ (>10 μM) also attenuated ex vivo platelet-rich plasma aggregation. HQ prevented the interleukin (IL)-1β-induced 8-isoprostane, and PGE2 production, but not IL-8 production of pulp cells. These results indicate that HQ may have an antiplatelet effect via inhibition of thromboxane production. HQ has antioxidative and anti-inflammatory effects, and possible inhibition of COX. Exposure and consumption of HQ-containing products, food or drugs may have antiplatelet, antioxidative, and anti-inflammatory effects.

Butyric acid as a histone deacetylase (HDAC) inhibitor is produced by a number of periodontal and root canal microorganisms (such as Porphyromonas, Fusobacterium, etc.). Butyric acid may affect the biological activities of periodontal/periapical cells such as osteoblasts, periodontal ligament cells, etc., and thus affect periodontal/periapical tissue destruction and healing. The purposes of this study were to study the toxic effects of butyrate on the matrix and mineralization marker expression in MG-63 osteoblasts. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Cellular apoptosis and necrosis were analyzed by propidium iodide/annexin V flow cytometry. The protein and mRNA expression of osteoprotegerin (OPG) and receptor activator of nuclear factor kappa-B ligand (RANKL) were analyzed by Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR). OPG, soluble RANKL (sRANKL), 8-isoprostane, pro-collagen I, matrix metalloproteinase-2 (MMP-2), osteonectin (SPARC), osteocalcin and osteopontin (OPN) secretion into culture medium were measured by enzyme-linked immunosorbant assay. Alkaline phosphatase (ALP) activity was checked by ALP staining. Histone H3 acetylation levels were evaluated by immunofluorescent staining (IF) and Western blot. We found that butyrate activated the histone H3 acetylation of MG-63 cells. Exposure of MG-63 cells to butyrate partly decreased cell viability with no marked increase in apoptosis and necrosis. Twenty-four hours of exposure to butyrate stimulated RANKL protein expression, whereas it inhibited OPG protein expression. Butyrate also inhibited the secretion of OPG in MG-63 cells, whereas the sRANKL level was below the detection limit. However, 3 days of exposure to butyrate (1 to 8 mM) or other HDAC inhibitors such as phenylbutyrate, valproic acid and trichostatin stimulated OPG secretion. Butyrate stimulated 8-isoprostane, MMP-2 and OPN secretion, but not procollagen I, or osteocalcin in MG-63 cells. Exposure to butyrate (2&ndash, 4 mM) for 3 days markedly stimulated osteonectin secretion and ALP activity. In conclusion, higher concentrations of butyric acid generated by periodontal and root canal microorganisms may potentially induce bone destruction and impair bone repair by the alteration of OPG/RANKL expression/secretion, 8-isoprostane, MMP-2 and OPN secretion, and affect cell viability. However, lower concentrations of butyrate (1&ndash, 4 mM) may stimulate ALP, osteonectin and OPG. These effects are possibly related to increased histone acetylation. These events are important in the pathogenesis and repair of periodontal and periapical destruction.

Butyric acid as a histone deacetylase (HDAC) inhibitor was produced by a number of periodontal and root canal microorganisms (such as Porphyromonas, Fusobacterium etc.). Butyric acid may affect the biological activities of periodontal/periapical cells such as osteoblasts, periodontal ligament cells etc., and thus affect periodontal/periapical tissue destruction and healing. The purposes of this study were to study the toxic effects of butyrate on matrix and mineralization markers’ expression of MG-63 osteoblasts. Cell viability and proliferation were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Cellular apoptosis and necrosis were analyzed by propidium iodide/Annexin V flow cytometry. Protein and mRNA expression of OPG, and RANKL were analyzed by western blotting and RT-PCR. OPG, soluble RANKL (sRANKL), 8-isoprostane, pro-collagen I, MMP-2, osteonectin (SPARC), osteocalcin and osteopontin secretion into culture medium were measured by enzyme-linked immunosorbant assay. Histone H3 acetylation levels were evaluated by immunofluorescent staining (IF) and western blot. We found that butyrate induced morphologic changes of growing MG-63 cells, with bigger and flattened in appearance. Butyrate activated histone H3 acetylation of MG-63 cells. Exposure of MG-63 cells to butyrate partly decreased cell number with no marked increase in apoptosis and necrosis. Butyrate stimulated RANKL protein expression, whereas it inhibited OPG protein expression. Butyrate also inhibited the secretion of OPG in MG-63 cells, whereas sRANKL level was below detection limit. Butyrate stimulated 8-isoprostane, MMP-2 and osteopontin secretion, but not procollagen I, osteonectin, osteocalcin in MG-63 cells. In conclusion, butyric acid generated by periodontal and root canal microorganisms may potentially induce bony destruction and impair bone repair by alteration of OPG/RANKL expression/secretion, 8-isoprostane, MMP-2, and osteopontin secretion, and affect cell proliferation. These effects are possibly related to increased histone acetylation. These events are important in the pathogenesis of periodontal and periapical destruction.

Background/purpose: Implant-abutment connection is the component responsible for the transmitting of occlusal force from the crown down to the implant fixture. Different connection geometric structure will lead to different mechanical performance. The purpose of this study was to compare the stability of internal hex Implant -abutment connection with internal hex with Morse taper implant-abutment connection by testing their compressive strength. Materials and methods: This was an in vitro study. The test group and the control group had 8 specimens separately. The test group was internal hex combined with Morse taper implant connection design, and the control group was internal hex connection design. Static force was applied to the specimens at a 30° angle until failure. The testing protocol was designed according to ISO14801 regulations. We compared the compressive strength of both groups. Results: The control group showed significantly higher compressive strength than the test group (p

The biocharacteristics of xenogeneic grafts make them a possible substitute for autogenous bone grafts in dental bone graft procedures. This study aimed to develop a novel porcine graft with collagen capable of generating new bone in bone defects via osteoconduction over 8 weeks of healing and to compare it with a porcine graft. The porcine collagen graft was made to undergo a cell viability test (MTT) and alkaline phosphatase assay (ALP). The surgical procedure was performed in 20 male adult New Zealand white rabbits. Four calvarial critical-size defects of 6 mm in diameter were prepared in each rabbit. The upper left defect was filled with a porcine graft of 500–1000 μm, the upper right with a porcine collagen graft, the lower left with hydroxyapatite/beta-tricalcium phosphate and the lower right served as the control without any filling material. The rabbits were divided and sacrificed at 2, 4, 6 and 8 weeks after surgery. Histological and micro-CT scan results showed that the performance of the porcine collagen graft is superior for regenerating new bone. Porcine collagen graft showed cell viability and osteoblast-like cell differentiation in vitro. The results indicate that porcine collagen graft is a potential bone substitute for clinical application.

Background/purpose This study investigated the cumulative survival rate of the mandibular implant-supported overdentures in patients treated from 2005 to 2014 at the Chang Gung Memorial Hospital Implant Center. Materials and methods Mandibular implant-supported overdentures were placed in 61 patients (32 males and 29 females) during the period from 2005 to 2014. These patients were categorized into two groups according to the type of attachment system used, namely, Group A and Group B. Group A included 31 patients (17 males and 14 females who received a total of 124 implants) whose denture was retained by a Hader bar and cast ERA attachments, whereas Group B included 30 patients (15 males and 15 females who received a total of 120 implants) whose denture was retained by a Hader bar and bilateral, extension cantilevers with clips. Results At the end of the follow-up period, 238 implants remained. Among the failed implants, two implants were in Group A (failure rate 2/124 = 1.6%), whereas four implants were in Group B (failure rate 4/120 = 3.3%). Fifty percent (3/6) of the failed implants were placed in the distal anterior mandible and 50% (3/6) were placed in the middle anterior mandible. The condition of the opposing arch was also analyzed in relation to the survival rate. The failure rate among patients with maxillary complete dentures was only 1.6%, whereas those wearing maxillary removable partial dentures had the highest implant failure rate (3/61 = 4.9%). Conclusion Our results indicate that implant-supported overdentures provide a reliable and effective alternative to conventional complete mandibular dentures. Overdentures with Hader bars and bilateral, cast ERA attachments resulted in fewer prosthetic complications.

The aim of this study was to fabricate biodegradable poly-l-lactic acid (PLLA) bone screws containing iron oxide (Fe3O4) nanoparticles, which are radiopaque and 3D-printable. The PLLA composites were fabricated by loading 20%, 30%, and 40% Fe3O4 nanoparticles into the PLLA. The physical properties, including elastic modulus, thermal properties, and biocompatibility of the composites were tested. The 20% nano-Fe3O4/PLLA composite was used as the material for fabricating the 3D-printed bone screws. The mechanical performance of the nano-Fe3O4/PLLA bone screws was evaluated by anti-bending and anti-torque strength tests. The tissue response and radiopacity of the nano-Fe3O4/PLLA bone screws were assessed by histologic and CT imaging studies using an animal model. The addition of nano-Fe3O4 increased the crystallization of the PLLA composites. Furthermore, the 20% nano-Fe3O4/PLLA composite exhibited the highest thermal stability compared to the other Fe3O4 proportions. The 3D-printed bone screws using the 20% nano-Fe3O4/PLLA composite provided excellent local tissue response. In addition, the radiopacity of the 20% nano-Fe3O4/PLLA screw was significantly better compared with the neat PLLA screw.

Background/purpose The purpose of this study was to determine fracture loads of human maxillary first premolars restored with condensable composite resin (Surefil, Dentsply) or leucite-reinforced ceramic inlays (ProCAD, Ivoclar Vivadent) in mesial occlusal distal cavity preparations of differing pulpal depths and isthmus widths. Materials and methods Ninety freshly extracted intact maxillary first premolars were divided into nine groups of 10. One group was an unprepared control. Eight groups of teeth were prepared to create different widths and depths of mesial occlusal distal cavities. Subsequently, teeth were restored with either condensable composite resin or ceramic inlay. Specimens were then subjected to axial loading with a 4.82-mm diameter steel ball in a universal testing machine at a crosshead speed of 0.5 mm/minute. Loads were applied against tooth structure, and the steel ball did not contact restorative material. Data were analyzed with multiple analysis of variance and Tukey post hoc testing. Results Mean fracture load of teeth restored with Surefil and a 3-mm cavity width was greater than in groups with a 2-mm cavity width (P = 0.0315). Teeth restored with ProCAD were observed to have significantly higher mean cusp fracture load compared to teeth restored with composite resin (P = 0.0003). Conclusion A bonded ceramic restoration restores the fracture load of a tooth comparable to an intact tooth independent of the examined design parameters, whereas the fracture loads of composite-resin-restored teeth were dependent on cavity widths. Cavity pulpal floor depth is not a significant factor of cusp fracture resistance in a tooth restored with either a ceramic inlay or composite resin.

Background/purpose The present study analyzed the subjective responses of patients to surgical placement of dental implants and the subsequent functioning of mandibular implant-retained overdentures as compared with the functioning of conventional dentures. This study sought to ascertain patient satisfaction and long-term effectiveness of tooth replacement with mandibular implant-retained overdentures. Materials and methods Patients (n = 86) unable to adapt to a conventional mandibular complete denture were treated with four one-stage titanium plasma sprayed or sandblasted, large-grit, and acid-etched threaded implants supporting an overdenture retained by a cast bar and extracoronal attachments. These patients were evaluated clinically during periods ranging from 6 months to 12 years. Eighty-three patients provided answers to queries regarding their impressions and feelings both prior to and after treatment for the implant-retained overdentures. Results During the study period, none of the implants or restorations failed. Ninety-eight percent of patients responding to the questionnaire were satisfied with their new overdentures. Retaining and supporting mandibular overdentures with implants yielded improvements in comfort, providing the patients with not only effective oral rehabilitation but also greater self-confidence in social situations. Conclusion The results indicate that mandibular arch rehabilitation by using implant-retained overdenture therapy serves as a predictable and effective treatment protocol.

For years, in order to improve bone regeneration and prevent the need of a second stage surgery to remove non-resorbable membranes, biological absorbable membranes have gradually been developed and applied in guided tissue regeneration (GTR). The present study’s main objective was to achieve space maintenance and bone regeneration using a new freeze-dried developed porcine collagen membrane, and compare it with an already commercial collagen membrane, when both were used with a bovine xenograft in prepared alveolar ridge bone defects. Prior to surgery, the membrane’s vitality analysis showed statistically significant higher cell proliferation in the test membrane over the commercial one. In six beagle dogs, commercial bone xenograft was packed in lateral ridge bone defects prepared in the left and right side and then covered with test porcine collagen membrane or commercial collagen membrane. Alveolar height changes were measured. Histomorphometric results, in vitro and in vivo properties indicated that the new porcine collagen membrane is biocompatible, enhances bone xenograft osteoconduction, and reduces the alveolar ridge height reabsorption rate.

Modification of the physiochemical properties of titanium surfaces using glow discharge plasma (GDP) and fibronectin coating has been shown to enhance the surface hydrophilicity, surface roughness, cell adhesion, migration, and proliferation. Thisin vivostudy aimed to evaluate the bone integration efficacy of a biologically modified implant surface. Two different surface-modified implants (Ar-GDP and GDP-fib) were placed in the mandibular premolar area of six beagle dogs for 2–8 weeks. Three techniques [histologic evaluation, resonance frequency analysis (RFA), and microcomputed tomography (micro-CT) evaluation] were used to detect the implant stability and bone-implant contact. The implant stability quotient values of GDP-fib implants were significantly greater than the Ar-GDP implants at 2 and 4 weeks (P<0.01). The bone volume/total volume ratio of GDP-fib implants was greater than the Ar-GDP implants in micro-CT evaluation. A high positive correlation was observed between RFA and micro-CT measurements. At 2 weeks, osteoblasts were seen to line the implant surface, and multinuclear osteoclasts could be seen on the surface of old parent bone. After 8 weeks, a majority of the space in the wound chamber appeared to be replaced by bone. Enhancement of the stability of biologically modified implants was proved by the results of RFA, micro-CT, and histological analysis. This enhanced stability may help fasten treatment and be clinically beneficial.