Regeneration of critical-sized grade II furcation using a novel injectable melatonin-loaded scaffold.

BACKGROUND: Periodontal regenerative therapy using bone-substituting materials has gained favorable clinical significance in enhancing osseous regeneration. These materials should be biocompatible, osteogenic, malleable, and biodegradable. This study assessed the periodontal regenerative capacity of a novel biodegradable bioactive hydrogel template of organic-inorganic composite loaded with melatonin. MATERIALS AND METHODS: A melatonin-loaded alginate-chitosan/beta-tricalcium phosphate composite hydrogel was successfully prepared and characterized. Thirty-six critical-sized bilateral class II furcation defects were created in six Mongrel dogs, and were randomly divided and allocated to three cohorts; sham, unloaded composite, and melatonin-loaded. Periodontal regenerative capacity was evaluated via histologic and histomorphometric analysis. RESULTS: Melatonin-treated group showed accelerated bone formation and advanced maturity, with a significant twofold increase in newly formed inter-radicular bone compared with the unloaded composite. The short-term regenerative efficacy was evident 4 weeks postoperatively as a significant increase in cementum length concurrent with reduction of entrapped epithelium. After 8 weeks, the scaffold produced a quality of newly synthesized bone similar to normal compact bone, with potent periodontal ligament attachment. CONCLUSIONS: Melatonin-loaded hydrogel template accelerated formation and enhanced quality of newly formed bone, allowing complete periodontal regeneration. Furthermore, the scaffold prevented overgrowth and entrapment of epithelial cells in furcation defects.

The effect of horizontal and vertical furcation involvement on molar survival: A retrospective study.

BACKGROUND: Periodontal involvement of the furcation area (furcation involvement, FI) has been reported to confer molars a higher risk of tooth loss. AIMS: The aim of this retrospective analysis was to assess the effect of FI on disease progression and tooth loss in molars of patients with chronic periodontitis undergoing supportive periodontal therapy (SPT) in a UK private practice setting. MATERIALS AND METHODS: Six-hundred and thirty-three molars were analysed in 100 chronic periodontitis patients treated with active periodontal therapy (APT) and followed up in SPT for at least 5 years. Molars were treated with a combination of resective, regenerative or conservative approaches, according to the different clinical needs. RESULTS: Twenty-three molars were extracted during APT and a further 23 were lost during SPT. Multivariable analysis showed that both horizontal FI and vertical furcation component were associated with increased risk of tooth loss during SPT (OR 5.26, 95% CI: 1.46-19.03, p = .012 and OR 9.83, 95% CI: 1.83-50.11, p = .006, respectively). CONCLUSION: Attention should be placed on both horizontal and vertical FI in molars, owing to their association with tooth loss during SPT.

A retrospective study on molar furcation assessment via clinical detection, intraoral radiography and cone beam computed tomography.

BACKGROUND: Accurate determination of bone loss at the molar furcation region by clinical detection and intraoral radiograph is challenging in many instances. Cone beam computed tomography (CBCT) is expected to open a new horizon in periodontal assessment. The purpose of this study was to compare and correlate accuracy of molar furcation assessment via clinical detection, intraoral radiography and CBCT images. METHODS: Eighty-three patients with chronic periodontitis who had existing CBCT scans were included. Furcation involvement was assessed on maxillary and mandibular first molars. Periodontal charts (modified Glickman's classification), intraoral (periapical and/or bitewing) radiographs (recorded as presence or absence) and axial CBCT reconstructions were used to evaluate furcation involvement on buccal and palatal/lingual sites. The correlation of furcation assessment by the three methods was evaluated by Pearson analysis. RESULTS: There were significant correlations (p < 0.05) between clinical detection and intraoral radiography, clinical detection and CBCT, as well as intraoral radiography and CBCT at all the measured sites (r values range between 0.230 to 0.644). CBCT generally exhibited higher correlation with clinical detection relative to intraoral radiography, especially at distal palatal side of maxillary first molar (p < 0.05). In addition, CBCT provided more accurate assessment, with bone loss measurement up to 2 decimals in millimeters, whereas clinical detection had 3 classes and the intraoral radiographs usually only detected the presence of furcation involvement in Glickman Class 2 and 3. CONCLUSIONS: This study validates that CBCT is a valuable tool in molar furcation assessment in addition to clinical detection and intraoral radiography.

The sphingosine-1-phosphate receptor 1 binding molecule FTY720 inhibits osteoclast formation in rats with ligature-induced periodontitis.

BACKGROUND AND OBJECTIVE: Osteoclast precursors (OPs) re-migrate from the bone surface into blood vessels through sphingosine-1-phosphate receptor 1 (S1PR1) expression. T cells also express S1PR1, mediating their migration from the lymph nodes into blood vessels. OP and T-cell migration are one of the sequential steps related to osteoclast formation. To characterize the role of S1PR1 in osteoclast formation induced by periodontitis, we investigated the effect of S1PR1-binding molecule FTY720 (FTY) on the number of OPs and T cells in periodontal tissue and peripheral blood of rats with ligature-induced periodontitis. MATERIAL AND METHODS: Rats were divided into four groups; control (Con), FTY, periodontitis (Peri), and periodontitis+FTY (Peri+FTY) groups. Ligatures were placed around the first molars in the left and right mandibles. The rats were intraperitoneally injected with vehicle or 3 mg/kg FTY daily until they were killed. The number of osteoclasts and cluster of differentiation (CD)11b, CD3 and receptor activator of NF-κB ligand (RANKL)-positive cells in first molar furcation were counted by tartrate-resistant acid phosphatase or immunohistochemistry staining. The number of CD11b- and CD3-positive cells in peripheral blood was estimated by flow cytometry. RESULTS: The number of osteoclasts in the Peri group was higher than Con, Peri+FTY and FTY groups (p < 0.05) and CD11b, CD3 and RANKL-positive cells were also higher in the Peri group than other groups in furcation (p < 0.05). While CD11b-positive cells in furcation of the Peri+FTY group were lower than the Peri group (p < 0.05), they were higher in peripheral blood (p < 0.05). Dissimilar to CD11b-positive cells, CD3-positive cells in the Peri+FTY group were lower in peripheral blood as well as furcation than the Peri group (p < 0.05). RANKL-positive cells in furcation of the Peri+FTY group were also lower than Peri group (p < 0.05). CONCLUSION: These results indicate that FTY may facilitate re-migration of OPs from the alveolar bone surface into blood vessels, blocking T-cell migration from the lymph nodes into blood vessels and subsequently reducing osteoclast formation induced by periodontitis. This suggests that S1PR1-S1P binding may play a role in osteoclast formation of periodontitis by modulating OP and T-cell migration.

Vertical subclassification predicts survival of molars with class II furcation involvement during supportive periodontal care.

BACKGROUND: Furcation involvement is a major predictor of tooth survival. Focus has so far been on the predictive value of the horizontal component of furcation involvement. Residual periodontal support on each of the roots is likely to play a major role on retention of the furcated molar. Aim of this clinical audit study was to preliminarily assess the impact of vertical subclassification on tooth retention. METHODS: Tooth retention of class II furcated molars in 200 consecutive patients compliant with periodontal supportive care for a minimum of 10 years was retrospectively evaluated in a single practice. Randomly selected furcated molars were retrospectively diagnosed in terms of vertical subclassification (residual periodontal support on the most compromised root), and time to tooth extraction/loss was determined in clinical records. Kaplan-Meier survival curves were constructed. RESULTS: Ten-year survival of molar with class II furcation involvement was 52.5%. Survival was 91% for subclass A, 67% for subclass B and 23% for subclass C. Mean years of survival were 9.5-10.1, 8.5-9.3 and 6-7.3 for subclasses A, B and C, respectively. Tests of equality of the survival distributions showed highly significant differences in all portions of the curve (p < .001). Stratified analyses by smoking showed significant differences for the two groups (p < .001). Hazard rates for tooth extraction/loss were 4.2 and 14.7 for subclasses B and C, respectively. CONCLUSIONS: Residual periodontal support assessed as vertical subclassification of furcation involvement seems to be a good predictor of survival of molar with class II horizontal furcation. This has implication for prognosis, treatment planning and development of effective molar retention strategies.

Clinical concepts for regenerative therapy in furcations.

Furcation involvements present one of the greatest challenges in periodontal therapy because furcation-involved molar teeth respond less favorably to conventional periodontal therapy compared with noninvolved molar or nonmolar teeth. Various regenerative procedures have been proposed and applied with the aim of eliminating the furcation defect or reducing the furcation depth. An abundance of studies and several systematic reviews have established the effectiveness of membrane therapy (guided tissue regeneration) for buccal Class II furcation involvement of mandibular and maxillary molars compared with open flap surgery. Bone grafts/substitutes may enhance the results of guided tissue regeneration. However, complete furcation closure is not a predictable outcome. Limited data and no meta-analyses are available on the effects of enamel matrix proteins for furcation regeneration. Enamel matrix protein therapy has demonstrated clinical improvements in the treatment of buccal Class II furcation defects in mandibular molars; however, complete closure of the furcation lesion is achieved only in a minority of cases. Neither guided tissue regeneration nor enamel matrix protein therapy have demonstrated predictable results for approximal Class II and for Class III furcations. Promising preclinical data from furcation regeneration studies in experimental animals is available for growth factor- and differentiation factor-based technologies, but very limited data are available from human clinical studies. Although cell-based therapies have received considerable attention in regenerative medicine, their experimental evaluation in the treatment of periodontal furcation lesions is at a very early stage of development. In summary, the indications and the limitations for currently available treatment modalities for furcation defects are well established. New regenerative treatments are clearly needed to improve the predictability of a complete resolution of furcation defects.

Localization of RANK, RANKL and osteoprotegerin during healing of surgically created periodontal defects in sheep.

BACKGROUND AND OBJECTIVE: Modeling of periodontal bone regeneration in a large animal enables better examination of the spatial and temporal regulation of osteogenesis and the remodeling of the healing defect. RANK, RANKL and osteoprotegerin (OPG) are known to be important regulators of bone healing. The aim of this study was to create periodontal defects surgically in a large animal model and to examine bone regeneration and the expression of RANK, RANKL and OPG proteins in the defect site during bone regeneration. MATERIAL AND METHODS: Periodontal defects were made in the furcation of the second mandibular premolar of sheep. Wound healing was examined 6 h, and 1, 4 and 6 wk after surgery and in control tissue. The teeth and defect region were decalcified and paraffin embedded. Immunohistochemistry for RANK, RANKL and OPG was conducted. Osteoclasts were identified using TRAP staining. RESULTS: The defects were examined at different time points after surgery and by 6 wk the defect region had fully regenerated with new bone, albeit less dense than that in the unwounded controls. RANK-positive osteoclasts were present at the edge of the wound from week 1 and were found within the defect at week 6, corresponding to osteoclast activation and bone remodeling. RANKL staining increased from week 1 compared with unwounded tissue, and peaked at 4 and 6 wk, as the osteoblast numbers increased. At the same time, OPG immunostaining was high in controls and at week 6, suggesting that it may act to block RANKL and control the bone remodeling within the defect. CONCLUSION: Distinctive temporal and spatial expression patterns for RANK, RANKL and OPG proteins were observed during healing of surgically created periodontal wounds in a sheep model. The research identifies possible therapeutic approaches to periodontal bone repair via modulation of these members of the tumor necrosis factor family.

Cigarette smoke inhalation increases the alveolar bone loss caused by primary occlusal trauma in a rat model.

BACKGROUND AND OBJECTIVE: Occlusal trauma (OT) and smoking are both factors that alter alveolar bone metabolism and therefore could synergistically act on alveolar bone loss. The aim of this experimental study was to evaluate the influence of short-term cigarette smoke inhalation (CSI) on inter-radicular alveolar bone loss promoted by primary OT in a rat model. MATERIAL AND METHODS: Forty-eight animals were randomly assigned to one of three groups based on treatment type: OT + CSI (n = 16), animals were exposed to CSI three times per day, for 8 min per exposure, and they concomitantly received unilateral vertical augmentation creating an occlusal interference inducing experimental OT; OT (n = 16), animals received only unilateral vertical augmentation; negative control (NC; n = 16), animals maintained for equal periods to achieve periodontal baseline values of periodontal ligament dimension. Each group was divided into two subgroups (n = 8) based on treatment length: 7 or 14 d. RESULTS: After 7 d, the OT + CSI group exhibited significantly higher bone loss compared to the NC group (p = 0.0022). After 14 d, the OT (p < 0.0001) and OT + CSI (p < 0.0001) groups presented significantly higher bone loss compared to the NC group, and OT + CSI resulted in significantly higher bone loss than OT alone (p = 0.0241). The number of tartrate-resistant acid phosphatase-positive cells on the linear surface of the bone crest after 7 d was significantly higher in the OT + CSI group as compared to the NC and OT groups (p < 0.0001 and p = 0.0045, respectively) and remained significantly higher in the OT + CSI group after 14 d, compared to the OT group (p < 0.0001). CONCLUSION: Short-term CSI increases early bone loss in association with OT after 7 d, and this worsens in severity after 14 d of exposure.

Regeneration of the periodontium using enamel matrix derivative in combination with an injectable bone cement.

OBJECTIVES: Enamel matrix derivative (EMD) has proven to enhance periodontal regeneration; however, its effect is mainly restricted to the soft periodontal tissues. Therefore, to stimulate not only the soft tissues, but also the hard tissues, in this study EMD is combined with an injectable calcium phosphate cement (CaP; bone graft material). The aim was to evaluate histologically the healing of a macroporous CaP in combination with EMD. MATERIALS AND METHODS: Intrabony, three-wall periodontal defects (2 × 2 × 1.7 mm) were created mesial of the first upper molar in 15 rats (30 defects). Defects were randomly treated according to one of the three following strategies: EMD, calcium phosphate cement and EMD, or left empty. The animals were killed after 12 weeks, and retrieved samples were processed for histology and histomorphometry. RESULTS: Empty defects showed a reparative type of healing without periodontal ligament or bone regeneration. As measured with on a histological grading scale for periodontal regeneration, the experimental groups (EMD and CaP/EMD) scored equally, both threefold higher compared with empty defects. However, most bone formation was measured in the CaP/EMD group; addition of CAP to EMD significantly enhanced bone formation with 50 % compared with EMD alone. CONCLUSIONS: Within the limits of this animal study, the adjunctive use of EMD in combination with an injectable cement, although it did not affect epithelial downgrowth, appeared to be a promising treatment modality for regeneration of bone and ligament tissues in the periodontium. CLINICAL RELEVANCE: The adjunctive use of EMD in combination with an injectable cement appears to be a promising treatment modality for regeneration of the bone and ligament tissues in the periodontium.

Application of collagen hydrogel/sponge scaffold facilitates periodontal wound healing in class II furcation defects in beagle dogs.

BACKGROUND AND OBJECTIVE: A three-dimensional scaffold may play an important role in periodontal tissue engineering. We prepared bio-safe collagen hydrogel, which exhibits properties similar to those of native extracellular matrix. The aim of this study was to examine the effect of implantation of collagen hydrogel/sponge scaffold on periodontal wound healing in class II furcation defects in dogs. MATERIAL AND METHODS: The collagen hydrogel/sponge scaffold was prepared by injecting collagen hydrogel, cross-linked to the ascorbate-copper ion system, into a collagen sponge. Class II furcation defects (of 5 mm depth and 3 mm width) were surgically created in beagle dogs. The exposed root surface was planed and demineralized with EDTA. In the experimental group, the defect was filled with collagen hydrogel/sponge scaffold. In the control group, no implantation was performed. Histometric parameters were evaluated 2 and 4 wk after surgery. RESULTS: At 2 wk, the collagen hydrogel/sponge scaffold displayed high biocompatibility and biodegradability with numerous cells infiltrating the scaffold. In the experimental group, reconstruction of alveolar bone and cementum was frequently observed 4 wk after surgery. Periodontal ligament tissue was also re-established between alveolar bone and cementum. Volumes of new bone, new cementum and new periodontal ligament were significantly greater in the experimental group than in the control group. In addition, epithelial down-growth was suppressed by application of collagen hydrogel. CONCLUSION: The collagen hydrogel/sponge scaffold possessed high tissue compatibility and degradability. Implantation of the scaffold facilitated periodontal wound healing in class II furcation defects in beagle dogs.

Comparison of mesenchymal stem cells and autogenous cortical bone graft in the treatment of class II furcation defects in dogs.

The purpose of this study was to compare the effectiveness of mesenchymal stem cells (MSCs) with platelet-rich plasma (PRP) as scaffold and autogenous cortical bone (ACB) graft with and without PRP in the regenerative treatment of class II furcation defects in dogs. The mandibular second, third, and fourth premolars (P2, P3, P4) and maxillary P3 and P4 of both sides in three dogs were selected for experimentation. Class II furcation defects (5 mm in height and 2 mm in depth) were surgically created. Five weeks after the first operation, scaling + root planning (group 1), PRP (group 2), ACB (group 3), combination of ACB/PRP (group 4), and combination of MSCs/PRP (group 5) treatments were performed during open flap debridement. The percentage of cementum and alveolar bone formation was evaluated by histomorphometric analysis after a healing period of 8 weeks. There was new cementum along with periodontal ligament and coronal growth of alveolar bone in all groups. Cementum formation was significantly higher in groups 3, 4, and 5 compared to the control group (P < 0.05) with no significant difference between groups 2, 3, 4, and 5. Alveolar bone formation was similar in all groups (P > 0.05). It can be concluded that periodontal regeneration with complete filling of class II furcation defects with cementum, alveolar bone, and periodontal ligament is obtained 8 weeks after ACB, ACB/PRP, and MSCs/PRP treatments; however, efficacy of none is higher than another.

Wound healing following regenerative procedures in furcation degree III defects: histomorphometric outcomes.

Degree III furcation involvements were surgically created at four first molars in each of three monkeys. Following 6 weeks of healing, full-thickness flaps were elevated. Following 24% EDTA gel conditioning, the defects were treated with one of the following: (1) enamel matrix proteins (EMD), (2) guided tissue regeneration (GTR) or (3) a combination EMD and GTR. The control defects did not receive any treatment. After 5 months of healing, the animals were sacrificed. Three 8 µm thick histological central sections, 100 µm apart, were used for histomorphometric analysis in six zones of each tooth either within the furcation area or on the pristine external surface of the root. In all specimens, new cementum with inserting collagen fibres was formed. Following GTR or GTR + EMD, cementum was formed up to and including the furcation fornix indicating complete regeneration on the defect periphery. Periodontal ligament fibres were less in all four modalities compared to pristine tissues. In the teeth treated with GTR and GTR + EMD a higher volume of bone and periodontal ligament tissues was observed compared to EMD. After 5 months of healing, regenerated tissues presented quantitative differences from the pristine tissues. In the two modalities where GTR alone or combined with EMD was used, the regenerated tissues differed in quantity from the EMD-treated sites.

Accuracy and Reliability of CBCT Compared to Clinical Probing in Detection of Trifurcation Defects: An In Vivo Study.

Background: Furcation defects are areas of pathological bone resorption in multirooted teeth. The aim of the study was to compare the measurements of trifurcation bone loss, measured using CBCT, versus clinical measurements in order to evaluate the efficacy of CBCT as an adjunctive diagnostic tool. Material and Methods. The included patients had both CBCT scans for maxillary molars and completed periodontal charts. Clinical examination consisted of probing and detection of vertical and horizontal furcation defects. These were measured and recorded. CBCT measurements were then evaluated using the linear measuring tool in Carestream imaging software (Carestream, Rochester, USA) and iCAT (Imaging Sciences, Hatfield, USA). These measurements of the CBCT images were then documented and compared to clinical findings. The two examiners were blinded to each other's measurements. Results: The most common tooth with a detected furcation defect was tooth #2 (31.7%), followed by tooth #15 (26.8%) and #3 (21.9%). The least common tooth with a detected furcation defect was #14 (19.5%). The mean values of buccal furcation for clinical and CBCT measurements were 3.01 mm and 2.6 mm, respectively. The measurements of mesial furcation were 2.5 mm and 2.2 mm for CBCT. The distal measurement of clinical examination was 2.7 mm and for CBCT was 2.44 mm. Conclusion: CBCT can be used as an adjunct to clinical furcation measurements and adds useful diagnostic information to assess trifurcation defects. In addition, CBCT limited field of view (FOV) can provide relatively high-resolution images at a reduced dose that is comparable to two-dimensional imaging.

Guided tissue regeneration: a decision-making model.

BACKGROUND: Guided tissue regeneration (GTR) has the potential to promote periodontal regeneration, which is one of the goals of periodontal surgery. While many successful reports of periodontal regeneration using barrier membranes exist in the literature, considerable heterogeneity of GTR outcomes is more typical of current reports. The reasons for this variability are numerous, but could be attributed to differences in surgical skills and case selection. There is a need for a current analysis of the factors affecting success and the formation of evidence-based treatment guidelines for GTR. METHODS: Available English literature pertaining to guided tissue regeneration was reviewed. Sources included peer-reviewed journal publications, online resources, and textbooks. Specific consideration was made to factors affecting GTR outcomes, especially in the context of systematic reviews and meta-analyses. RESULTS: Factors, including patient systemic conditions and compliance, defect features, local factors and surgical techniques and materials, that influence GTR outcomes were analyzed and entered into a decision-making model. CONCLUSION: A decision-making model was formulated based upon current evidence regarding factors that influence guided tissue regeneration outcomes. Meticulous case selection based upon known influential variables may help to minimize inconsistency in GTR outcomes.

Auto-transplanted mesenchymal stromal cell fate in periodontal tissue of beagle dogs.

BACKGROUND AIMS: Mesenchymal stromal cells (MSC) possess multilineage differentiation potential and characteristics of self-renewal. It has been reported that MSC can acquire characteristics of cells in the periodontal ligament (PDL) in vitro. Moreover, the transplantation of MSC has been shown to be a promising strategy for treating periodontal defects. However, little is known about the fate of MSC in periodontal tissue in vivo. The aim of this study was to trace the paths of MSC after transplantation into periodontal tissues in vivo. METHODS: MSC labeled with bromodeoxyuridine (BrdU) were transplanted into periodontal defects of beagle dogs. Six weeks after surgery, the animals were killed and decalcified specimens were prepared. Migration and differentiation of MSC were detected by single/double immunohistochemistry and a combination of immunohistochemistry and in situ hybridization. RESULTS: BrdU-labeled MSC were observed distributing into periodontal tissue that included alveolar bone, PDL, cementum and blood vessels and expressing surface markers typical of osteoblasts and fibroblasts. CONCLUSIONS: Cumulatively, our data suggest that MSC migrate throughout periodontal tissue and differentiate into osteoblasts and fibroblasts after transplantation into periodontal defects at 6 weeks in vivo, and have the potential to regenerate periodontal tissue.

Accuracy of three-dimensional imaging in assessing maxillary molar furcation involvement.

AIM: To assess the accuracy of cone beam computed tomography (CBCT) in detecting furcation involvement (FI) in maxillary molars. MATERIAL AND METHODS: Fourteen patients with generalized advanced chronic periodontitis were consecutively recruited and treated non-surgically. In maxillary molars considered for furcation surgery due to increased FI and/or increased probing pocket depths during re-evaluation, CBCT was performed and the degree of FI was evaluated from the CBCT images. Furcation surgery was performed in 25 maxillary molars. Intra-surgical FI assessments were compared with data derived from CBCT images. RESULTS: Overall, 84% of the CBCT data were confirmed by the intra-surgical findings (weighted kappa=0.926, 95% confidence interval: 0.681-1.0). While 14.7% (11 sites) were underestimated (CBCT less than intra-surgical value), in only 1.3% (one site) did the CBCT data lead to an overestimation compared with the intra-surgical analysis. The agreement between both assessments was the highest in distopalatal furcation entrances, followed by buccal and mesiopalatal. CONCLUSIONS: CBCT images demonstrate a high accuracy in assessing the loss of periodontal tissue and classifying the degree of FI in maxillary molars.

Histologic Evaluation of Artificial Floors Under MTA and Nano-Filled Resin-Modified Glass Ionomer Used to Repair Furcation Perforations in Dogs.

Objective: This study aimed to compare the tissue reaction of two repair materials for furcation perforations, nano-filled resin modified glass ionomer (Nano-FRMGI) and mineral trioxide aggregate (MTA), used with or without an artificial floor. Methods: A total of 96 teeth in 6 dogs were used for this study. After access cavities, root canals were prepared and obturated with gutta percha using cold lateral condensation technique. Perforations were then created on the floors of the pulp chambers. The perforations divided into four groups n=24/group that were sealed with MTA alone, MTA with calcium sulphate artificial floor (CSAF), FRMGI alone and Nano-FRMGI with CSAF. All access cavities were filled with composite resin. Two dogs were sacrificed at 1, 3, and 6 month. The experimental tooth along with the surrounding alveolar bone were cut in block sections and histologically evaluated for tissue response. Data were analyzed by Chi-square (P≤0.05). Results: MTA and MTA with CSAF showed more bone and cementum apposition when compared to Nano-FRMGI at 6-month interval. MTA and MTA with CSAF showed less bone resorption, epithelium proliferation and inflammation compared to Nano-FRMGI at 6-month interval. Conclusion: MTA with CSAF or MTA-alone show better outcomes in the repair of pulp chamber floor perforation.

Research on promoting periodontal regeneration with human basic fibroblast growth factor-modified bone marrow mesenchymal stromal cell gene therapy.

BACKGROUND AIMS: Recently, it has been found that effective periodontal regeneration can be induced by bone marrow mesenchymal stromal cell (BMSC) transplantation or local application of basic fibroblast growth factor (bFGF). The aim of the present study was to assess, in dogs, the efficacy of periodontal regeneration via the delivery of BMSC transfected with bFGF to repair destruction of periodontal tissue. METHODS: BMSC from dogs were isolated, cultured and purified via density-gradient centrifugation. Polymerase chain reaction (PCR) was employed to clone bFGF cDNA from human periodontal cells, and the product was then ligated into the eukaryotic expression vector pDC316-IREs-EGFP. BMSC transfected with pDC316bFGF-IREs-EGFP were transplanted into root furcation defects of beagle dogs. After 6 weeks, regeneration in defects was assessed via clinical examination, X-ray, histologic observation and micro-CT analysis. RESULTS: DNA sequence analysis showed that the bFGF sequence of recombinant plasmid pDC316bFGF-IREs-EGFP was consistent with that reported by GeneBank. bFGF expression was detected with Western blotting, and active bFGF in supernatant was also observed. Our animal experiment proved that the regenerating speed of periodontal bone tissue in groups transplanted with BMSC containing the modified bFGF gene was higher than in those transplanted with BMSC alone. CONCLUSIONS: A successfully constructed eukaryotic expression vector containing human bFGF in pDC316bFGF-IREs-EGFP could produce bioactive bFGF in vitro. bFGF overexpression mediated by the recombinant plasmid pDC316bFGF- IREs-EGFP accelerated periodontal regeneration.

The influence of thyroid hormones on periodontitis-related bone loss and tooth-supporting alveolar bone: a histological study in rats.

BACKGROUND AND OBJECTIVE: Recent studies have pointed to potentially periodontal risk indicators, however no information is available on the impact of changes in thyroid hormone levels on the progression of periodontitis and on the quality of alveolar bone. Thus, the aim of the present study was to evaluate histologically, in rats, the influence of thyroid hormones on the rate of periodontal bone loss resulting from ligature placement and on the quality of tooth-supporting alveolar bone. MATERIAL AND METHODS: Thirty-six male Wistar rats were randomly assigned to the following groups: healthy (control, n = 12), hypothyroidism (n = 12) and hyperthyroidism (n = 12). Once alterations were confirmed by total serum levels of triiodothyronine and thyroxine, ligatures were randomly placed around one of the first mandibular molars. Thirty days later, the animals were killed and specimens routinely processed for serial decalcified sections. The parameters assessed were periodontitis-related bone loss, quality of tooth-supporting alveolar bone and the number of cells positive for tartrate-resistant acid phosphatase (TRAP), a marker of bone resorption. RESULTS: At the ligated sites, intergroup analysis revealed that hypothyroidism significantly increased the bone loss resulting from ligature-induced periodontitis (p = 0.02) and the number of TRAP-positive cells on the linear surface of bone crest (p = 0.01). In addition, no significant differences were detected regarding the quality of the bone (p = 0.24) or the number of TRAP-positive cells in the area of the interradicular bone for ligated teeth among the groups (p = 0.17). CONCLUSION: It may be concluded that decreased serum levels of thyroid hormones may enhance periodontitis-related bone loss, as a function of an increased number of resorbing cells, whereas the tooth-supporting alveolar bone seems to be less sensitive to alterations in hormone levels.

Induction of cementogenesis and periodontal ligament regeneration by recombinant human transforming growth factor-beta3 in Matrigel with rectus abdominis responding cells.

BACKGROUND AND OBJECTIVE: In primates and in primates only, the transforming growth factor-b proteins induce endochondral bone formation. Transforming growth factor-b3 also induces periodontal tissue regeneration. Two regenerative treatments using human recombinant transforming growth factor-b3 were examined after implantation in mandibular furcation defects of the nonhuman primate, Papio ursinus. MATERIAL AND METHODS: Class III furcation defects were surgically created bilaterally in the mandibular first and second molars of two adult Chacma baboons (P. ursinus). Different doses of recombinant transforming growth factor-beta3 reconstituted with Matrigel matrix were implanted in the rectus abdominis muscle to induce heterotopic ossicles for subsequent transplantation to selected furcation defects. Twenty days after heterotopic implantation, periodontal defects were re-exposed, further debrided and implanted with minced fragments of induced heterotopic ossicles. Contralateral class III furcation defects were implanted directly with recombinant transforming growth factor-beta3 in Matrigel matrix with the addition of minced fragments of autogenous rectus abdominis muscle. Treated quadrants were not subjected to oral hygiene procedures so as to study the effect of the direct application of the recombinant morphogen in Matrigel on periodontal healing. Histomorphometric analyses on undecalcified sections cut from specimen blocks harvested on day 60 measured the area of newly formed alveolar bone and the coronal extension of the newly formed cementum along the exposed root surfaces. RESULTS: Morphometric analyses showed greater alveolar bone regeneration and cementogenesis in furcation defects implanted directly with 75 microg of transforming growth factor-beta3 in Matrigel matrix with the addition of minced muscle tissue. CONCLUSION: Matrigel matrix is an optimal delivery system for the osteogenic proteins of the transforming growth factor-beta superfamily, including the mammalian transforming growth factor-beta3 isoform. The addition of minced fragments of rectus abdominis muscle provides responding stem cells for further tissue induction and morphogenesis by the transforming growth factor-beta3 protein.