João César Zielak

Chlorhexidine diminishes the loss of bond strength over time under simulated pulpal pressure and thermo-mechanical stressing

OBJECTIVES The purpose of this study was to investigate the effects of chlorhexidine (CHX) digluconate at 0.2% and 2% on dentin bonding durability of etch-and-rinse and self-etch adhesive systems. METHODS In this study were used 24 extracted non-carious human third-molars. The occlusal surfaces of the molar crowns were removed with a low-speed diamond saw to expose flat dentin surfaces. The tested materials were Single-Bond (SB) (two-step etch-and-rinse adhesive) and Clearfil Tri S Bond (CTSB) (all-in-one self-etch adhesive) used in association or not with CHX at 0.2% and 2%. The bonding systems were applied according to manufacturers instructions and followed by composite application (Z250). For each condition, half of the specimens was immediately submitted to microtensile test and half of them was submitted to long-term storage of 6 months under simulated pulpal pressure and thermo-mechanical stressing before testing. The data were analyzed using Two-Way ANOVA and Tukey post hoc test (alpha=0.05). Failure patterns of the specimens were observed using scanning electron microscopy. RESULTS The falling % in bond strength over the 6-month period was: SB control-43.64%; SB/0.2%CHX-23.79%; SB/2%CHX-26.42%; CTSB control-40.94%; CTSB/0.2%CHX-37.07%; CTSB/2%CHX-22.14%. The fracture modes were predominantly adhesive, mainly in the specimens of terminal groups. CONCLUSIONS CXH digluconate at 2% was able to diminish loss of microtensile bond strength over time associated to both etch-and-rinse and self-etch adhesives. Lower concentration of CHX (0.2%) was not able to diminish the loss of bond strength over time when associated to the self-etch adhesive CTSB.

Platelet-rich plasma (PRP) impairs the craniofacial bone repair associated with its elevated TGF-β levels and modulates the co-expression between collagen III and α-smooth muscle actin.

Transforming growth factor‐β (TGF‐β) is considered the main inducer of both the α‐smooth muscle actin (α‐SMA) phenotype and collagen synthesis and deposition and plays a significant role in the tissue repair and the development of fibrosis. Since the PRP constitutes an important source of TGF‐β and its efficacy on the craniofacial bone repair remains controversy, the aim of this study was to evaluate the effect of PRP in the presence of levels of TGF‐β on PRP samples, as well as in the presence of collagen III and α‐SMA+ cells, while comparing these results by means of a histomorphometric analysis of the bone matrix and fibrous deposition on the bone repair. Four bone defects of 16 mm2 were created on the calvarium of 21 rabbits. The surgical defects were treated with either particulate autograft, particulate autograft mixed with PRP and PRP alone. Animals were euthanized at 15, 30, and 45 days postoperative. Histomorphometric and immunohistochemical analyses were performed to assess repair time, as well as the expression of collagen III, and α‐SMA. The histomorphometric results demonstrated intensive deposition of fibrous tissue while hinder bone deposition occurred in PRP groups. These results coincided with higher values of the TGF‐β on the PRP sample, also larger occurrence of diffuse collagen III deposition and higher presence of α‐SMA+ cells spread among the fibrous tissue. Thus, the higher levels of TGF‐β associated with the both expression of collagen III and α‐SMA on defect treated with PRP suggest that its biomaterial induce an effect that can be considered similarly to a fibroproliferative disorder.

Platelet-rich plasma diminishes calvarial bone repair associated with alterations in collagen matrix composition and elevated CD34+ cell prevalence

The interaction between platelets and both type I and III collagens plays an important role in modulating platelet adhesion and aggregation, also contributing to the chemotaxis of CD34+ cells. The interaction with type III collagen can maintain high levels of collagen and alter the biology of bone repair when the PRP is used. The aim of this study was to evaluate the effect of platelet-rich plasma (PRP) and autograft on the presence of type III and type I collagens, the ratio between them, as well as the presence of CD34+ progenitor cells, while comparing these results by means of a histomorphometric analysis of the bone tissue. Four bone defects (8.0mm in diameter and 2.0mm in depth) were produced on the calvarium of 23 rabbits. The surgical defects were treated with either autogenous bone grafts, autogenous bone grafts with PRP and PRP alone. Animals were euthanized at 2, 4 or 6 weeks post-surgery. Histological, histomorphometric and immunohistochemical analyses were performed to assess repair time, as well as the expression of type I and III collagens, and number of progenitor CD34+ cells. Data were analyzed using the ANOVA and Student-Newman-Keuls test (alpha=5%). An enlarged granulation and medullary tissue areas in the PRP groups were observed. The use of PRP in this study hindered bone deposition, also enhanced type III to type I collagen ratio and the chemotaxis of CD34+ progenitor cells, similarly to a thrombogenic effect.

Leukocyte-Platelet-Rich Plasma (L-PRP) Induces an Abnormal Histophenotype in Craniofacial Bone Repair Associated with Changes in the Immunopositivity of the Hematopoietic Clusters of Differentiation, Osteoproteins, and TGF-β1

BACKGROUND Leukocyte-platelet-rich plasma (L-PRP) is considered an important source of growth factors, especially Transforming growth factor β 1 (TGF-β1), which modulates the proliferation and regulation of mesenchymal cells, and also exerts an influence on the hematopoiesis, osteogenesis, and adipogenesis in bone microenvironment. Thus, the aim of this study was to evaluate the effect of L-PRP on the calvarial bone repair and compare its results on the presence of TGF-β1, CD34, CD45, bone morphogenetic protein 2 (BMP2), BMPR1B, and Runx2 proteins detected by immunohistochemistry. MATERIAL AND METHODS Four bone defects were created on the calvaria of 23 rabbits. The defects were treated with autograft, L-PRP alone, and L-PRP mixed with autograft. The animals were euthanized at 2, 4, and 6 weeks post-surgery. RESULTS Unlike autograft and sham groups, the defects treated with L-PRP demonstrated significant positivity to TGF-β1, while the BMP2 was scarce. These results coincided with the lower bone matrix deposited and larger medullary area, which were composed of fibrosis, when treated with only L-PRP, or intense adiposity on defects filled with L-PRP mixed with autograft. The fibrosis that occurred was associated with a minor percentage of osteoproteins, intense presence of CD34(+) CD45(-) cells, and significant expression of TGF-β1 in all time periods analyzed. The adiposity occurred from the major presence of osteoprogenitor BMPR1B (+) Runx2(+) cells simultaneously to BMP2(-) TGF-β1(+) and CD34(+) CD45(+/-) expressions predominantly on the earlier period. CONCLUSION From this study, it can be concluded that the L-PRP used alone or mixed to autograft hindered the osteoneogenesis due to suppression of immunoexpression of BMP2, while the immunopositivity of TGF-β1 was intense. When used alone, the L-PRP induced a fibrotic condition associated with TGF-β1 presence and lack of osteoproteins, but when L-PRP was mixed to autograft, it induced the presence of the osteolineage cells (BMPR1B (+) Runx2(+) ), but also inhibited the terminal osteoblastic maturation associated with the lack of BMP2 and the presence of TGF-β1(+) , a fact that contributed to cellular transdifferentiation into fat cells.

Influence of protaper finishing files and sodium hypochlorite on cleaning and shaping of mandibuldar central incisors - a histological analysis

Objective: This study investigated the influence of the last apical instrument of the ProTaper system with and without 2.5% sodium hypochlorite for cleaning mandibular central incisors. Material and Methods: Thirty two mandibular central incisors were divided into six study groups: Group I – F1 instrument with 2.5% sodium hypochlorite; Group II – F1 and F2 with 2.5% sodium hypochlorite; Group III – F1, F2 and F3 with 2.5% sodium hypochlorite; Group IV – F1 with distilled water; Group V – F1 and F2 with distilled water; Group VI – F1, F2 and F3 with distilled water. The two remaining teeth comprised the negative control group. The specimens were prepared following the principles of the technique suggested by the manufacturer and then submitted to histological preparation and morphometric analysis. Data were analyzed statistically by the Kruskal Wallis test at 1% significance level. Results: There was statistically significant difference (p<0.01) between all study groups, except between Groups I and VI. Conclusions: It was concluded that no technique allowed complete cleaning of the root canals. However, the technique of finishing preparation of the apical third with the F3 instrument with 2.5% sodium hypochlorite irrigation was the most effective.

Dental infection simulating skin lesion

Orocutaneous fistulas or cutaneous sinus, a tract of dental origin, is an uncommon but well-documented condition that usually requires emergency treatment. Such condition may be misdiagnosed by physicians and dentists and may sometimes be confused with bone and skin tumor, osteomyelitis, congenital fistula, salivary gland fistula, pyogenic granuloma, infected cyst, deep mycotic infection, and other pathologies. A case of facial sinus tract that was initially misdiagnosed by a physician as a nonodontogenic lesion is presented. Nonsurgical endodontic therapy was the treatment of choice for this case. Facial cutaneous sinus tracts must be considered of dental origin. Early diagnosis and prompt treatment minimize patient discomfort and esthetic problems, reducing the possibility of further complications such as sepsis and osteomyelitis.

Nonprocessed adipose tissue graft in the treatment of dehiscence bone defects in rabbit tibiae: a pilot study.

Objective:To evaluate the bone repair of surgically created dehiscence-type defects (3 × 5 mm) around dental implants in rabbit tibia using nonprocessed adipose tissue graft or autogenous bone graft. Materials and Methods:The bone defects were randomly assigned to 3 groups: blood clot (C), autogenous bone (AB), and nonprocessed adipose tissue (AT). After 3 months, the animals were euthanized. Histomorphometric analyses were performed, and the results were analyzed with analysis of variance and Tukey tests (P ⩽ 0.05). Statistics were performed for the percentage of the bone-to-implant contact (BIC) and bone area (BA) within the limits of the threads. Results:The results for BIC in the AT (37.75% ± 28.03%) and C (40.57 ± 13.71%) groups were statistically similar, whereas the AB group had the greatest percentage of BIC (83.37% ± 11.85%). For all groups, the BA percentage was similar (61.48% ± 30.89% in AT; 72.90% ± 14.10% in C; 84.23% ± 11.96% in AB), with no statistically significant differences. Conclusion:Nonprocessed adipose tissue is not a comparable substitute for autogenous bone in the treatment of dehiscence bone defects around titanium dental implants.

Radiographic and histological evaluation of ectopic application of deproteinized bovine bone matrix.

Objective: To evaluate, through radiographic and histological analysis, the tissue reaction induced by a biomaterial based on deproteinized bovine bone matrix (DBBM) in the muscle of sheep. sMaterials and Methods: Sixteen sheep were used. The animals underwent surgery to insert polyethylene tubes containing the biomaterial in the muscle of the lower back (ectopic site) and were euthanized after 3 and 6 months. Each sheep received three tubes: Group 1 - sham group (negative control - tube without biomaterial), Group 2 - particulate autogenous bone (positive control), and Group 3 - DBBM biomaterial (GenOx Inorg). The material removed was evaluated by radiographic, macroscopic, and microscopic analysis, descriptively. Results: Macroscopic analysis showed that Group 3 had a greater tissue volume maintenance. Microscopic analysis indicated that Group 1 had a higher concentration of dense, thin collagen fibers (3 and 6 months); in Group 2, there was a decrease in the inflammatory process and the deposition of dense, thin collagen fibers (3 and 6 months); in Group 3, the presence of a dense connective tissue was noted, in which the DBBM particles (3 months) were found. On the periphery of these particles, a deposition of basophilic material was found, indicating the formation of mineral particles and the formation of tissues with osteoid characteristics (6 months). Conclusion: Based on the results obtained, it can be concluded that the biomaterial based on DBBM led to the formation of tissue with similar characteristics to an osteoid matrix in a postoperative period of 6 months. However, none of the groups evaluated showed ectopic bone neoformation.

Leukocyte-platelet-rich plasma diminishes bone matrix deposition in rat calvaria treated with autograft due to simultaneous increase in immunohistochemical expression of Indian Hedgehog, transforming growth factor-β, and parathyroid-1 receptor

PURPOSE This study evaluated the immunohistochemical presence of Indian Hedgehog (IHH), transforming growth factor-β (TGF-β), and parathyroid-1 receptor (PTH1R) in calvaria bone repair, and compared these results with the histological bone matrix features in defects treated with autograft in the presence or absence of L-PRP. MATERIAL AND METHODS An artificial bone defect measuring 5 × 1 mm was produced in the calvaria of 28 Wistar rats. Randomly the defects were treated with autograft and autograft mixed with L-PRP. The animals were euthanized at 15 and 40 days post-surgery. Data were analyzed by Student-Newman-Keuls test (p ≤ .05) for immunohistochemical interpretation. RESULTS The results revealed that the histological characteristic of bone matrix deposited in the defect was different in the defects treated with L-PRP. The group that received only the autograft demonstrated larger haversian bone matrix deposited, whereas the group that received autograft mixed with L-PRP revealed trabecular bone deposition. These results coincided with significantly higher immunopositivity for IHH, TGF-β1, and PTH1R in the L-PRP group. CONCLUSION These results suggest that L-PRP altered the biological characteristic of the autograft, increasing the bone cells IHH+ but inducing a trabecular bone associated with intense quantities of TGF-β and PTH1R.

Immunoexpression of PPAR-? and Osteocalcin Proteins for Bone Repair of Critical-Size Defects Treated with Fragmented Autogenous Abdominal Adipose Tissue Graft

Immunoexpression of PPAR-γ and osteocalcin proteins was evaluated for bone repair of critical-size defects (CSDs), created in rat calvaria (n=42) and treated with fragmented abdominal autogenous adipose tissue graft. Three groups (n=14) were formed: C (control - blood clot), AB (autogenous bone) and AT (fragmented adipose tissue). The groups were divided into subgroups (n=7) for euthanasia at 30 and 90 days. Histological and immunohistochemical analyses were performed. Data were subjected to descriptive statistics (mode). A complete bone closure was observed in Group AB 90 days after surgery. In Group C, repair was achieved by the formation of collagen fiber bundles oriented parallel to the wound surface at both post-surgery periods. In Group AT the type of healing was characterized by dense connective tissue containing collagen fiber bundles arranged amidst the remaining adipose tissue, with rare heterotopic bone formation associated with fibrosis and different types of tissue necrosis. Immunostaining of PPAR-γ was not observed in any specimen from Groups C and AB. In Group AT, the immunostaining of PPAR-γ was more evident 30 days after surgery. Immunostaining of osteocalcin was present in all groups and at both postoperative periods. The fragmented autogenous abdominal adipose tissue graft did not favor the repair of critical-size bone defects created surgically in rat calvaria as evidenced by the positive immunostaining of PPAR-γ protein and the negative immunostaining of osteocalcin in the osteoblast-like cells and bone matrix.