In vitro cultivation of canine multipotent mesenchymal stromal cells on collagen membranes treated with hyaluronic acid for cell therapy and tissue regeneration

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bone healing in critical-size defects treated with platelet-rich plasma activated by two different methods. A histologic and histometric study in rat calvaria

The purpose of this study was to analyze histologically the influence of platelet-rich plasma (PRP) coagulated with two different activators on bone healing in surgically created critical-size defects (CSD) in rat calvaria.Forty-eight rats were divided into three groups: C, PRP-C and PRP-T. An 8 mm diameter CSD was created in the calvarium of each animal. In group C, the defect was filled by a blood clot only. In groups PRP-C and PRP-T, the defect was filled with PRP activated with either calcium chloride or thromboplastin solution, respectively. Each group was divided into two subgroups (n = 8 per subgroup) and killed at either 4 or 12 weeks postoperatively. Histologic and histometric analyses were performed. The amount of new bone formed was calculated as a percentage of the total area of the original defect. Percentage data were transformed into arccosine for statistical analysis (analysis of variance, Tukey's post hoc test, p < 0.05).No defect completely regenerated with bone. Group PRP-C had a statistically greater amount of bone formation than groups C and PRP-T at both time points of analysis. No statistically significant differences were observed between groups C and PRP-T.It can be concluded that the type of activator used to initiate PRP clot formation influences its biological effect on bone healing in CSD in rat calvaria.

Mechanical removal of necrotic periodontal ligament by either Robinson bristle brush with pumice or scalpel blade. Histomorphometric analysis and scanning electron microscopy

One of the important factors accounting for successful delayed replantation of avulsed teeth is seemingly the type of root surface treatment. Removal of necrotic cemental periodontal ligament remnants may prevent the occurrence of external root resorption, which is the major cause of loss of teeth replanted in such conditions. The purpose of this study was to compare the efficacy of two mechanical techniques for removal of root-adhered periodontal ligament. Preservation or removal of the cementum layer concomitantly with these procedures was also assessed. Forty-five roots of healthy premolars extracted for orthodontic purposes were selected. After extraction, the teeth were kept dry at room temperature for 1 h and then immersed in saline for rehydration for an additional 10 min. Thereafter, the roots were assigned to three groups, as follows: group 1 (control) - the cemental periodontal ligament was preserved; group 2 - removal of the periodontal ligament by scraping root surface with a scalpel blade (SBS); group 3 - periodontal ligament remnants were removed using a Robinson bristle brush at low-speed with pumice/water slurry (RBP). The specimens were analysed histomorphometrically and examined by scanning electron microscopy. The quantitative and qualitative analyses of the results showed that the RBP technique was significantly more effective than the SBS technique for removal of the periodontal ligament remnants adhered to root surface. Both techniques preserved the cementum layer.

Periodontal Disease Decreases Insulin Sensitivity and Insulin Signaling

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Action of nicotine and ovariectomy on bone regeneration after tooth extraction in rats

Purpose: The purpose of this study was to evaluate the effects of nicotine and ovariectomy on alveolar bone regeneration after exodontias in rats.Materials and Methods: For 30 days, sham ovariectomized (OVX)/NaCl, sham OVX/nicotine, OVX/NaCl, and OVX/nicotine animals were given 2 daily injections of saline or hemisulfate of nicotine. After this period, exodontic procedures were carried out and treatment continued up to the time of euthanasia on clays 7 and 14 when the alveoli were removed for further analyses.Results: The data confirmed that nicotine significantly delays the alveolar regeneration process after dental extraction in rats and showed that the association of nicotine with ovariectomy exacerbates these results.Conclusion: These results indicate that nicotine potentiated the effect of estrogen deficiency on bone regeneration induced by ovariectomy. (c) 2010 American Association of Oral and Maxillofacial Surgeons Oral Maxillofac Surg 68:2675-2681, 2010

Dentists' level of knowledge of the treatment plans for periodontal ligament injuries after dentoalveolar trauma

This study investigated the level of knowledge held by dentists about the possible treatment plan procedures for periodontal ligament injuries after dentoalveolar trauma. A 5-item self-applied questionnaire was prepared with questions referring to the professional profile of the interviewees and to the treatment plan they would propose for periodontal ligament injuries secondary to dentoalveolar trauma. The questionnaires were filled out by 693 dentists attending the 23rd Annual Meeting of the Brazilian Society for Dental Research, and the data obtained were subjected to descriptive analysis. Either the chi-square test or Fisher's exact test was applied to assess associations among variables, at a 5% level of significance. The results revealed that dentists experienced difficulty in establishing a treatment plan for subluxation, and for extrusive, lateral and intrusive luxations. In general, holding a dental specialty degree had no influence on the knowledge about treatment plan procedures for the most severe injuries. It could be concluded that the dentists participating in this study, whether specialists or not, did not have sufficient knowledge to treat most of the periodontal ligament injuries resulting from dentoalveolar trauma adequately.

Treatment of Experimental Periodontal Disease by Photodynamic Therapy in Rats With Diabetes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Evaluation of tissue response to periodontal dressings: Histological study in tooth sockets of rats

Although the use of periodontal dressings is currently limited, there are some indications for their use. Selection of any material that will have direct contact with live tissues, such as periodontal dressings, should be careful in order to allow surgical wound healing. The aim of this study was to evaluate the intensity of inflammatory response and bone formation in tooth sockets of rats after implantation of three periodontal dressings. After removal of the right maxillary incisors of 84 male rats, each tooth socket received implantation of a polyethylene tube, 63 of which were filled with non-eugenol periodontal dressing and the remaining 21 tubes remained empty (control group). Histological evaluation assessed the intensity of inflammatory response and presence and location of bone tissue formation at postoperative periods of 7, 14 and 28 days. Statistical analysis was performed by the Kruskal-Wallis test at 5% significance level. Regarding the inflammatory infiltrate, at 28 days, there was statistically significant difference between one of periodontal dressings and control group (p < 0.05). Analysis of postoperative periods, showed that the control group presented statistically significant reduction in the inflammatory infiltrate comparing the 14- and 28-day periods (p < 0.05). Regarding bone tissue formation, there was difference in control group between the 7- and 28-day periods (p < 0.05). Within the experimental conditions, it may be concluded that no differences were found in the inflammatory response among the groups at 7 and 14 days and that Voco pac (TM) dressing induced a more intensive inflammatory reaction at 28 days.

Treatment of experimental periodontal disease by photodynamic therapy in immunosuppressed rats

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Experimental periodontal disease treatment by subgingival irrigation with tetracycline hydrochloride in rats

Objective: The aim of this study was to compare subgingival irrigation with tetracycline hydrochloride (TTC-HCL) as adjunctive treatment to scaling and root planning (SRP) on induced periodontitis in rats. Material and Methods: In 60 rats, periodontal disease was ligature-induced at the mandibular left first molar. After 7 days, the ligature was removed and all animals were submitted to SRP, and divided into 2 groups according to the following treatment: C (n=30) - subgingival irrigation with 1 mL of saline; T (n=30) - subgingival irrigation with 1 mL of TTC-HCL (50 mg/mL). Ten animals in each group were euthanized at 7, 15 and 30 days posttreatment. The histometric values were statistically analyzed (p<0.05). Results: In the histometric analysis, at 7, 15 and 30 days, Group T (0.72+/-0.05 mm(2), 0.57+/-0.14 mm(2), 0.62+/-0.07 mm(2)), showed less bone loss (p<0.05) than Group C (1.35+/-0.25 mm(2); 1.40+/-0.31 mm(2); 1.29+/-0.27 mm(2)), respectively. Conclusions: Subgingival irrigation with TTC-HCL was an effective adjunctive treatment for periodontal disease induced in rats.

Evaluation of Centrifuged Bone Marrow on Bone Regeneration Around Implants in Rabbit Tibia

Objective: To evaluate the bone regeneration of cervical defects produced around titanium implants filled with blood clot and filled with centrifuged bone marrow (CBM) by means of histomorphometric analysis.Materials and Methods: Twelve rabbits received 2 titanium implants in each right tibia, with the upper cortical prepared with a 5-mm drill and the lower cortex with a 3-mm-diameter drill. Euthanasia was performed to allow analysis at 7, 21, and 60 days after operation. The samples were embedded in light curing resin, cut and stained with alizarin red and Stevenel blue for a histomorphometric analysis of the bone-to-implant contact (BIC) and the bone area around implant (BA). The values obtained were statistically analyzed using the nonparametric Kruskal-Wallis test (P = 0.05).Results: At 60 days postoperation, the groups had their cervical defects completely filled by neo-formed bone tissue. There was no statistically significant difference between the groups regarding BIC and BA during the analyzed periods.Conclusion: There was no difference in the bone repair of periimplant cervical defects with or without the use of CBM. (Implant Dent 2012;21:481-485)

Bone regeneration in surgically created defects filled with autogenous bone: an epifluorescence microscopy analysis in rats

Although the search for the ideal bone substitute has been the focus of a large number of studies, autogenous bone is still the gold standard for the filling of defects caused by pathologies and traumas, and mainly, for alveolar ridge reconstruction, allowing the titanium implants installation. Objectives: The aim of this study was to evaluate the dynamics of autogenous bone graft incorporation process to surgically created defects in rat calvaria, using epifluorescence microscopy. Material and methods: Five adult male rats weighing 200-300 g were used. The animals received two 5-mm-diameter bone defects bilaterally in each parietal bone with a trephine bur under general anesthesia. Two groups of defects were formed: a control group (n=5), in which the defects were filled with blood clot, and a graft group (n=5), in which the defects were filled with autogenous bone block, removed from the contralateral defect. The fluorochromes calcein and alizarin were applied at the 7th and 30th postoperative days, respectively. The animals were killed at 35 days. Results: The mineralization process was more intense in the graft group (32.09%) and occurred mainly between 7 and 30 days, the period labeled by calcein (24.66%). Conclusions: The fluorochromes showed to be appropriate to label mineralization areas. The interfacial areas between fluorochrome labels are important sources of information about the bone regeneration dynamics.

In vivo effect of photodynamic therapy on periodontal bone loss in dental furcations

Background: The purpose of this study was to histometrically evaluate the influence of photodynamic therapy on bone loss in furcation areas in rats with experimentally induced periodontal disease.Methods: Ligatures were placed on the first mandibular molar in rats. Then the animals were divided into four groups: control group = no treatment; methylene blue group (MB) = treated topically with methylene blue (100 mu g/ml); laser group (LLLT) = treated with low-level laser therapy; and photodynamic therapy group (PDT) = treated topically with MB followed by LLLT (4.5 J/cm(2)). Rats from all groups were sacrificed at 7, 15, or 30 days postoperatively. The area of bone loss in the furcation region of the first molar was histometrically analyzed. Data were analyzed statistically (analysis of variance and Bonferroni tests; P<0.05).Results: The PDT group demonstrated less bone loss compared to the other groups at 7 days (1.986 +/- 0.417 mm(2)); at 15 days, the PDT (1.641 +/- 0.115 mm(2)) and MB groups (1.991 +/- 0.294 mm(2)) demonstrated less bone loss compared to the control (4.062 +/- 0.416 mm(2)) and LLLT (2.641 +/- 0.849 mm(2)) groups.Conclusion: Within the parameters used in this study, PDT may be an effective alternative for control of bone loss in furcation areas in periodontitis.

Nonsurgical Periodontal Therapy Combined with Laser and Photodynamic Therapies for Periodontal Disease in Immunosuppressed Rats

Background. Periodontal disease is often associated with systemic diseases and is characterized by destruction of the tissues supporting the teeth. Patients using immunosuppressive drugs such as tacrolimus are among those who suffer from tissue destruction. Objective. We sought to evaluate the effects of laser and photodynamic therapies (PDT; nonsurgical) as an adjunct to scaling and rootplaning (SRP) in the treatment of corona-induced periodontitis in rats immunosuppressed with tacrolimus (Prograf).Materials and Methods. The animals were divided into 5 groups. Each groups had 6 rats. Group I, the control group, received only saline solution throughout the study period of 42 days and did not receive periodontal treatment; group II received saline solution and SRP; group III received tacrolimus (1 mg/kg per day) and was treated with SRP; group IV animals were treated identically to group III and then administered laser treatment; and in group V, the animals were treated identically to group III and then administered PDT.Results. Statistical analysis indicated decreased bone loss with the progression of time (P = .035). There was no difference between the bone loss associated with the types of treatment administered to groups I, II, and III (P > .9) or groups IV and V (P > .6). The analysis also indicated that immunosuppression was not a bone loss-determining factor.Conclusion. Laser and PDT therapies were effective as an adjunctive treatment to SRP in reducing bone loss caused by experimental periodontitis induced in animals being treated systemically with tacrolimus.

Periodontal and Prosthetic Biologic Considerations to Restore Biological Width in Posterior Teeth

\ The biologic width is an essential dental space that always needs to be maintained to ensure periodontal health in any dental prosthetic restorations. An iatrogenic partial fixed prosthesis constructed in lower posterior teeth predisposed the development of subgingival caries, which induced violation of the biologic width in involved teeth, resulting in an uncontrolled inflammatory process and periodontal tissue destruction. This clinical report describes a periodontal surgical technique to recover a violated biologic width in lower posterior teeth, by crown lengthening procedure associated with free gingival graft procedure, to ensure the possibility to place a modified partial fixed prosthesis in treated area. The procedure applied to recover the biologic width was crown lengthening with some modifications, associated with modified partial fixed prosthesis to achieve health in treated area. The modified techniques in both surgical and prosthetic procedures were applied to compensate the contraindications to recover biologic width by osteotomy in lower posterior teeth. The result, after 4 years under periodic control, seems to achieve the projected goal. Treating a dental diseased area is necessary to diagnose, eliminate, or control all etiologic factors involved in the process. When the traditional methods are not effective to recover destructed tissues, an alternative, compensatory, and adaptive procedure may be applied to restore the sequelae of the disease, applying a restorative method that respects the biology of involved tissues.