Sinus floor augmentation using mineralized freeze-dried bone allograft combined with recombinant human bone morphogenetic protein-2 (rhBMP-2): A long-term retrospective study.

Background/purpose: The combination of recombinant human bone morphogenetic protein-2 (rhBMP-2) with a carrier material has not been extensively studied. This study aimed to evaluate the clinical, radiological, and histomorphometric outcomes of sinus floor augmentation using a 3:7 mixture of cancellous and cortical freeze-dried bone allografts (mixed AG) combined with rhBMP-2. Materials and methods: Mixed AG was used for sinus floor augmentation in a total of 21 patients with a residual alveolar bone height <5 mm. Among the total 47 sites, augmentation with and without rhBMP-2 was performed in 26 and 21 sites, respectively. Radiographic parameters were assessed using cone-beam computed tomography. After a six-month healing period, core biopsies were harvested for histomorphometric analysis. Results: The bone gain after healing was 13.36 ± 3.9 mm and 12.07 ± 3.8 mm in the mixed AG alone and mixed AG with rhBMP-2 groups, respectively. The survival rate of implants in both groups was 100% during the follow-up period. The proportion of newly formed bone was 24.6 ± 10.2% and 39.7 ± 18.3% in the mixed AG alone and mixed AG with rhBMP-2 groups, respectively (P < 0.05). Moreover, the percentage of residual graft material was 21.0 ± 12.2% and 9.6 ± 10.0% in the mixed AG alone and mixed AG with rhBMP-2 groups, respectively (P < 0.05). Conclusion: Mixed AG combined with rhBMP-2 could be a suitable material for sinus floor augmentation. This combination may reduce the treatment time and improve the predictability of implant placement.

A retrospective comparison of 3 approaches of vestibuloplasty around mandibular molar implants: apically positioned flap versus free gingival graft versus modified periosteal fenestration.

PURPOSE: This study aimed to clinically evaluate the efficacy of vestibuloplasty around lower molar implants using 3 different modalities: apically positioned flap alone (APF), APF with a free gingival graft (FGG), and APF with modified periosteal fenestration (mPF). METHODS: Three different vestibuloplasty procedures during second-stage implant surgery were performed at the mandibular molar area in 61 patients with a shallow vestibule and insufficient keratinized tissue (KT). The clinical measurements of KT width were recorded at baseline, immediately after surgery (T0), 6 months after surgery (T6), and 12 months after surgery (T12). Soft tissue esthetic scores were measured. RESULTS: An additional KT width gain from baseline to T12 of approximately 2 mm was obtained with FGG and mPF compared to that with APF. Shrinkage of the re-established tissue was lower with mPF and FGG than with APF, whereas the esthetic profile was better with APF and mPF than with FGG. CONCLUSIONS: Within the limitations of this study, mPF showed potential as a promising approach for vestibuloplasty around the lower molar implants compared to the traditional APF and FGG.

Sinus floor augmentation using mixture of mineralized cortical bone and cancellous bone allografts: Radiographic and histomorphometric evaluation.

BACKGROUND/PURPOSE: Due to the pneumatization of the maxillary sinus, the sinus floor augmentation is often performed to implant placement in the maxillary posterior region. The aim was to perform radiographic and histomorphometric evaluation after placement of mixed allografts (cortical freeze-dried bone allograft [FDBA] 50%:cancellous FDBA 50%) during sinus floor augmentation. MATERIALS AND METHODS: In 37 patients, anorganic bovine bone (ABB, sites = 16), mineralized cancellous bone allograft (MCBA, sites = 15), and mixed allografts (Mixed AG, sites = 20) were placed during sinus floor elevation via the lateral approach (LSFE), at total 51 sites with residual alveolar bone height (RBH) < 5 mm. Cone-beam computed tomography images were obtained before LSFE (T0), after surgery (T1), and 6 months after surgery (T2) for radiographic analysis. After a 6-month healing period, core biopsies were harvested and histomorphometric analysis was performed. RESULTS: The mean augmented bone height (ABH) of ABB, MCBA, and mixed AG groups after surgery was similar (13.86 ±â€¯4.19 mm, 13.99 ±â€¯4.07 mm, and 14.20 ±â€¯3.12 mm, respectively; P > 0.05). The mean ABH of ABB, MCBA, and mixed AG groups after 6 months was similar (13.72 ±â€¯4.55 mm, 11.83 ±â€¯3.31 mm, and 12.53 ±â€¯2.97 mm, respectively; P > 0.05). In the ABB, MCBA, and mixed AG groups, the proportion of newly formed bone (NB) was similar (36.13 ±â€¯10.01%, 39.26 ±â€¯10.72%, and 31.27 ±â€¯18.31%, respectively; P > 0.05). CONCLUSION: This result demonstrated that mixed AG led to sufficient bone augmentation and histologically comparable NB formation as compared to ABB and MCBA for sinus floor augmentation.

Novel Use of Silicone Sheets for Immediate Implant Placement in Fresh Molar Extraction Sockets.

In the recent years, the immediate placement of dental implants into fresh extraction sites has become an acceptable treatment approach. However, immediate molar implant placement presents specific challenges because of the anatomical and physiologic limitations. Such implant surgeries commonly require procedures that use a barrier membrane to generate bone and soft tissue or one that seals the molar extraction socket through a coronally advanced flap. Here, as an alternative, we report a method for treating molar extraction socket wounds in the hard and soft tissues after immediate placement of an implant using a silicone sheet.

Effects of platelet-rich plasma on tooth replantation in dogs: a histologic and histomorphometric analysis.

PURPOSE: The purpose of this study was to evaluate the effects of platelet-rich plasma (PRP) on periodontal healing of replanted root surfaces in dogs histologically and histomorphometrically. METHODS: A total of 36 roots of mandibular incisors and premolars from 6 mongrel dogs were used. The roots were randomly divided into 3 groups: 1) a positive control group (n=12), in which the periodontal ligament (PDL) and cementum were retained and the roots were soaked in saline; 2) a negative control group (n=12), in which the PDL and cementum were removed and the roots were soaked in saline; and 3) an experimental group (n=12), in which the PDL and cementum were removed and the roots were soaked in PRP. After soaking the root surfaces, the extracted roots were replanted into the extraction sockets. The roots were covered using a coronally repositioned flap. RESULTS: Histologically, irregular-thickness PDL-like and cementum-like tissues were observed in the 4-week experimental group and the positive control group. PDL-like tissue and cementum-like tissue with a more uniform thickness were observed at 8 weeks. In the negative control group, PDL-like tissue and cementum-like tissue were rarely found, and root resorption and ankylosis were observed. In the cross-sectional histomorphometric analysis, the experimental group demonstrated a higher rate of formation of cementum-like tissue and a lower tooth ankylosis rate than the positive and negative control groups at 4 and 8 weeks. Although there was a significant difference in the tooth ankylosis rate and the formation of cementum-like tissue across the 3 groups (P<0.05), no statistical significance was observed between any pair of groups (P>0.017). CONCLUSIONS: Applying PRP to root surfaces during tooth replantation in dogs can reduce tooth ankylosis and increase PDL-like and cementum-like tissue formation.

Effects of 1,25-dihydroxyvitamin D3 on the differentiation of MC3T3-E1 osteoblast-like cells.

PURPOSE: The purpose of this study was to evaluate the effects of 1,25-dihydroxyvitamin D3 on the proliferation, differentiation, and matrix mineralization of MC3T3-E1 osteoblast-like cells in vitro. METHODS: MC3T3-E1 osteoblastic cells and 1,25-dihydroxyvitamin D3 were prepared. Cytotoxic effects and osteogenic differentiation were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, alkaline phosphatase (ALP) activity assay, ALP staining, alizarin red S staining, and reverse transcription-polymerase chain reaction (RT-PCR) for osteogenic differentiation markers such as ALP, collagen type I (Col-I), osteocalcin (OCN), vitamin D receptor (VDR), and glyceraldehyde 3-phosphate dehydrogenase. RESULTS: The MTT assay showed that 1,25-dihydroxyvitamin D3 did not inhibit cell growth and that the rate of cell proliferation was higher than in the positive control group at all concentrations. ALP activity was also higher than in the positive control group at low concentrations of 1,25-dihydroxyvitamin D3 (10-10, 10-12, and 10-14 M). RT-PCR showed that the gene expression levels of ALP, Col-I, OCN, and vitamin D receptor (VDR) were higher at a low concentration of 1,25-dihydroxyvitamin D3 (10-12 M). Alizarin red S staining after treatment with 1,25-dihydroxyvitamin D3 (10-12 M) showed no significant differences in the overall degree of calcification. In contrast to the positive control group, formation of bone nodules was induced in the early stages of cell differentiation. CONCLUSIONS: We suggest that 1,25-dihydroxyvitamin D3 positively affects cell differentiation and matrix mineralization. Therefore, it may function as a stimulating factor in osteoblastic bone formation and can be used as an additive in bone regeneration treatment.

Effect of preameloblast-conditioned medium and CPNE7 on root surfaces in dogs: a histologic and histomorphometric evaluation.

Preameloblast-conditioned medium (PACM) has been reported as a potent dentin regenerative material, but its effects as a mixture on periodontal regeneration and the role of CPNE7 in PACM are not known. The purpose of this study is to evaluate the histologic and histomorphometric effects of preameloblast-conditioned medium (PACM) and CPNE7 on periodontal tissue healing in dogs. Seventy-two mandibular premolar roots from ten dogs were extracted and randomly divided into six groups (n = 12 each): (1) positive control group; (2) negative control group; (3) cementum-removed and PACM-treated group; (4) cementum-preserved and PACM-treated group; (5) CPNE7-inactivated PACM-treated group; and (6) recombinant CPNE7-treated group. The extracted roots were replanted into extraction sockets for 4 and 8 weeks and analyzed histologically. Most of the root surfaces in the negative control group showed ankylosis; and those in the experimental groups showed newly formed PDL-like and cementum-like tissues. Histomorphometric analysis of horizontal sections showed that the mean length of the PDL on the roots of the positive controls was similar to those in cementum-removed or -preserved and PACM-treated group at 8 weeks (p = 1.08). Sagittal sections showed that the mean length of the new cementum on the roots in cementum-removed and PACM-treated group was significantly greater than that in CPNE7-inactivated PACM-treated group (p = 0.037). The mean length of the newly formed PDL on the roots in CPNE7- inactivated PACM-treated and rCPNE7-treated groups was significantly greater than that in the negative controls at 8 weeks (p = 0.037, p = 0.036). The use of PACM and CPNE7 in tooth replantation resulted in increased PDL and cementum formation, suggesting the beneficial role of PACM and CPNE7 in periodontal tissue healing.

Licochalcone-E induces caspase-dependent death of human pharyngeal squamous carcinoma cells through the extrinsic and intrinsic apoptotic signaling pathways.

The aim of the present study was to investigate licochalcone-E (Lico-E)-induced apoptosis and the associated apoptotic signaling pathway in FaDu cells, a human pharyngeal squamous carcinoma cell line. Treatment with Lico-E exhibited significant cytotoxicity on FaDu cells in a concentration-dependent manner. The IC50 value of Lico-E in FaDu cells was ~50 µM. Treatment with Lico-E increased the number of dead FaDu cells. Furthermore, chromatin condensation, which is associated with apoptotic cell death, was observed in FaDu cells treated with Lico-E for 24 h. By contrast, Lico-E did not produce cytotoxicity or increase the number of dead cells when applied to human normal oral keratinocytes (hNOKs). Furthermore, chromatin condensation was not observed in hNOKs treated with Lico-E. Treatment with Lico-E increased the expression of Fas ligand and the cleaved form of caspase-8 in FaDu cells. Furthermore, treatment with Lico-E increased the expression of pro-apoptotic factors, including apoptosis regulator BAX, Bcl-2-associated agonist of cell death, apoptotic protease-activating factor 1, caspase-9 and tumor suppressor p53, while decreasing the expression of anti-apoptotic factors, including apoptosis regulator Bcl-2 and Bcl-2-like protein 1 in FaDu cells. The expression of cleaved caspases-3 and poly (ADP-ribose) polymerase was significantly upregulated following treatment with Lico-E in FaDu cells, while Lico-E-induced apoptotic FaDu cell death was partially suppressed by treatment with Z-VAD-FMK, a pan caspase inhibitor. Therefore, Lico-E-induced oral cancer (OC) cell-specific apoptosis is mediated by the death receptor-dependent extrinsic and mitochondrial-dependent intrinsic apoptotic signaling pathways. In conclusion, these data suggested that Lico-E exhibits potential chemopreventive effects and warrants further developed as a chemotherapeutic agent against OC.

Coumestrol Counteracts Interleukin-1ß-Induced Catabolic Effects by Suppressing Inflammation in Primary Rat Chondrocytes.

In the present study, we investigated the anti-catabolic effects of coumestrol, a phytoestrogen derived from herbal plants, against interleukin-1ß-induced cartilage degeneration in primary rat chondrocytes and articular cartilage. Coumestrol did not affect the viability of human normal oral keratinocytes and primary rat chondrocytes treated for 24 h and 21 days, respectively. Although coumestrol did not significantly increase the proteoglycan contents in long-term culture, it abolished the interleukin-1ß-induced loss of proteoglycans in primary rat chondrocytes and knee articular cartilage. Furthermore, coumestrol suppressed the expression of matrix-degrading enzymes such as matrix metalloproteinase-13, -3, and -1 in primary rat chondrocytes stimulated with interleukin-1ß. Moreover, the expression of catabolic factors such as nitric oxide synthase, cyclooxygenase-2, prostaglandin E2, and inflammatory cytokines in interleukin-1ß-stimulated primary rat chondrocytes was suppressed by coumestrol. In summary, these results indicate that coumestrol counteracts the catabolic effects induced by interleukin-1ß through the suppression of inflammation. Therefore, based on its biological activity and safety profile, coumestrol could be used as a potential anti-catabolic biomaterial for osteoarthritis.

The association between radiographic embrasure morphology and interdental papilla reconstruction using injectable hyaluronic acid gel.

PURPOSE: The purpose of this study was to evaluate the clinical efficacy of enhancing deficient interdental papilla with hyaluronic acid gel injection by assessing the radiographic anatomical factors affecting the reconstruction of the interdental papilla. METHODS: Fifty-seven treated sites from 13 patients (6 males and 7 females) were included. Patients had papillary deficiency in the upper anterior area. Prior to treatment, photographic and periapical radiographic standardization devices were designed for each patient. A 30-gauge needle was used with an injection-assistance device to inject a hyaluronic acid gel to the involved papilla. This treatment was repeated up to 5 times every 3 weeks. Patients were followed up for 6 months after the initial gel application. Clinical photographic measurements of the black triangle area (BTA), height (BTH), and width (BTW) and periapical radiographic measurements of the contact point and the bone crest (CP-BC) and the interproximal distance between roots (IDR) were undertaken using computer software. The interdental papilla reconstruction rate (IPRR) was calculated to determine the percentage change of BTA between the initial and final examination and the association between radiographic factors and the reconstruction of the interdental papilla by means of injectable hyaluronic acid gel were evaluated. RESULTS: All sites showed improvement between treatment examinations. Thirty-six sites had complete interdental papilla reconstruction and 21 sites showed improvement ranging from 19% to 96%. The CP-BC correlated with the IPRR. More specifically, when the CP-BC reached 6 mm, virtually complete interdental papilla reconstruction via injectable hyaluronic acid gel was achieved. CONCLUSIONS: These results suggest that the CP-BC is closely related to the efficacy of hyaluronic acid gel injection for interdental papilla reconstruction.

Biochanin-A antagonizes the interleukin-1ß-induced catabolic inflammation through the modulation of NFκB cellular signaling in primary rat chondrocytes.

Biochanin-A, a phytoestrogen derived from herbal plants, protected from the IL-1ß-induced loss of proteoglycans through the suppression of matrix degrading enzymes such as matrix metalloproteinase (MMP)-13, MMP-3, MMP-1, and ADAMTS-5 in primary rat chondrocytes and the knee articular cartilage. It also suppressed the expression of IL-1ß-induced catabolic factors such as nitric oxide synthase 2, cyclooxygenase-2, prostaglandin E2, and inflammatory cytokines. Furthermore, biochanin-A suppressed the IL-1ß-induced phosphorylation of NFκB, and inhibited its nuclear translocation in primary rat chondrocytes. These results indicate that biochanin-A antagonizes the IL-1ß-induced catabolic effects through its anti-inflammatory activity that involves the modulation of NFκB signaling.

Six Month Clinical Evaluation of Interdental Papilla Reconstruction with Injectable Hyaluronic Acid Gel Using an Image Analysis System.

PURPOSE: Obtaining predictable and aesthetically pleasing interdental papilla is challenging in dental reconstruction. Hyaluronic acid gel has been successfully used to reduce facial creases and similar abnormalities. The purpose of this study was to clinically assess the efficiency of interdental papilla reconstruction with injectable hyaluronic acid gel. MATERIALS AND METHODS: Ten patients with 43 treated sites in the maxillary anterior region were studied. Photographic standardization devices were designed for image analysis before treatment. This treatment was repeated up to five times during 3-week intervals. Patients were followed 6 months after initial gel application. RESULTS: Twenty-nine sites had complete papilla reconstruction and 14 sites improved from 39 to 96% of interdental papilla reconstruction rate. Complete interdental papilla reconstruction was performed when black triangle at initial examination had area of ≤0.25 mm(2) , height of ≤1mm, or width of ≤0.5mm. CONCLUSION: Injectable hyaluronic acid gel may be a promising treatment for enhancing papillary esthetics. CLINICAL SIGNIFICANCE: Interdental papilla reconstruction with injectable hyaluronic acid gel can be suggested as a minimally invasive treatment option for interdental papilla deficiencies in small areas. However, long-term results of this treatment beyond 6 months are not yet known. (J Esthet Restor Dent 28:221-230, 2016).

Berberine-induced anticancer activities in FaDu head and neck squamous cell carcinoma cells.

In the present study, we investigated berberine­induced apoptosis and the signaling pathways underlying its activity in FaDu head and neck squamous cell carcinoma cells. Berberine did not affect the viability of primary human normal oral keratinocytes. In contrast, the cytotoxicity of berberine was significantly increased in FaDu cells stimulated with berberine for 24 h. Furthermore, berberine increased nuclear condensation and apoptosis rates in FaDu cells than those in untreated control cells. Berberine also induced the upregulation of apoptotic ligands, such as FasL and TNF-related apoptosis-inducing ligand, and triggered the activation of caspase-8, -7 and -3, and poly(ADP ribose) polymerase, characteristic of death receptor-dependent extrinsic apoptosis. Moreover, berberine activated the mitochondria­dependent apoptotic signaling pathway by upregulating pro-apoptotic factors, such as Bax, Bad, Apaf-1, and the active form of caspase-9, and downregulating anti-apoptotic factors, such as Bcl-2 and Bcl-xL. In addition, berberine increased the expression of the tumor suppressor p53 in FaDu cells. The pan-caspase inhibitor Z-VAD-fmk suppressed the activation of caspase-3 and prevented cytotoxicity in FaDu cells treated with berberine. Interestingly, berberine suppressed cell migration through downregulation of vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2, and MMP-9. Moreover, the phosphorylation of extracellular signal-regulated kinase (ERK1/2) and p38, components of the mitogen-activated protein kinase pathway that are associated with the expression of MMP and VEGF, was suppressed in FaDu cells treated with berberine for 24 h. Therefore, these data suggested that berberine exerted anticancer effects in FaDu cells through induction of apoptosis and suppression of migration. Berberine may have potential applications as a chemotherapeutic agent for the management of head and neck squamous carcinoma.

Biological Effects of the Herbal Plant-Derived Phytoestrogen Bavachin in Primary Rat Chondrocytes.

The aim of this study was to examine the anabolic and anticatabolic functions of bavachin in primary rat chondrocytes. With bavachin treatment, chondrocytes survived for 21 d without cell proliferation, and the proteoglycan content and extracellular matrix increased. Short-term monolayer culture of chondrocytes showed that gene induction of both aggrecan and collagen type II, major extracellular matrix components, was significantly upregulated by bavachin. The expression and activities of cartilage-degrading enzymes such as matrix metalloproteinases and a disintegrin and metalloproteinase with thrombospondin motifs were inhibited significantly by bavachin, while tissue inhibitors of metalloprotease were significantly upregulated. Bavachin inhibits the expression of inducible nitric oxide synthase, a representative catabolic factor, and downregulated the expression of nitric oxide, cyclooxygenase-2, and prostaglandin E2 in a dose-dependent manner in chondrocytes. Our results suggest that the bavachin has anabolic and potent anticatabolic biological effects on chondrocytes, which may have considerable promise in treating articular cartilage degeneration in the future.

Effect of fibroblast growth factor on injured periodontal ligament and cementum after tooth replantation in dogs.

PURPOSE: The purpose of this animal study was to perform a histological and histomorphometric analysis in order to elucidate the effect of fibroblast growth factor-2 (FGF-2) on injured periodontal ligament (PDL) and cementum after tooth replantation in dogs. METHODS: The roots of 36 mandibular premolars from six mongrel dogs were used in this study. The roots were randomly divided into three groups: (1) a positive control group (n=12), in which the PDL was retained; (2) a negative control group (n=12), in which the PDL and the cementum between the notches were removed; and (3) an experimental group (n=12), in which the PDL and the cementum between the notches were removed and the roots were soaked in an FGF-2 solution (30 µg/0.1 mL). After treating the root surfaces, the extracted roots were replanted into extraction sockets. The animals were sacrificed four and eight weeks after surgery for histologic and histomorphometric evaluation. RESULTS: At four and eight weeks, normal PDLs covered the roots in the positive control group. In the negative control group, most replanted roots showed signs of replacement resorption. In the experimental group, new PDL-like tissue and cementum-like tissue were observed to partially occupy the region between the root surfaces and the newly formed bone. Histomorphometric analysis showed that the mean length of the newly formed cementum-like tissue on the roots treated with FGF-2 was significantly greater than that of the tissue on the roots in the negative control group (four weeks, P=0.008; eight weeks, P=0.042). However, no significant differences were observed between the roots treated with FGF-2 and the negative control roots with respect to newly formed PDL-like tissue. CONCLUSIONS: The results of this study suggest that use of FGF-2 on injured root surfaces promotes cementogenesis after tooth replacement in dogs.

Clinical evaluation of a biphasic calcium phosphate grafting material in the treatment of human periodontal intrabony defects.

PURPOSE: The aim of this study was to compare the clinical outcome of open flap debridement (OFD) with a biphasic calcium phosphate (BCP) graft to that of OFD without BCP graft for the treatment of intrabony periodontal defects (IBDs). METHODS: The study included 25 subjects that had at least one intrabony defect of 2- or 3-wall morphology and an intrabony component≥4 mm as detected radiographically. Subjects were randomly assigned to treatment with (BCP group, n=14) or without BCP (OFD group, n=11). Clinical parameters were recorded at baseline and 6 months after surgery and included the plaque index, gingival index, probing depth (PD), clinical attachment level (CAL), and gingival recession (REC). A stringent plaque control regimen was enforced for all of the patients during the 6-month observation period. RESULTS: In all of the treatment groups, significant PD reductions and CAL gains occurred during the study period (P<0.01). At 6 months, patients in the BCP group exhibited a mean PD reduction of 3.7±1.2 mm and a mean CAL gain of 3.0±1.1 mm compared to the baseline. Corresponding values for the patients treated with OFD were 2.5±0.8 mm and 1.4±1.0 mm, respectively. Compared to OFD group, the additional CAL gain was significantly greater in the patients in BCP group (P=0.028). The additional PD reduction was significant for the BCP group (P=0.048). The REC showed a significant increase in both groups, and the amount of recession was significantly smaller in the BCP group than OFD group (P=0.023). In radiographic evaluation, the height of the bone fill in the BCP group was significantly greater than OFD group. CONCLUSIONS: The clinical benefits of BCP found in this study indicate that BCP may be an appropriate alternative to conventional graft materials.