Bone marrow coagulated and low-level laser therapy accelerate bone healing by enhancing angiogenesis, cell proliferation, osteoblast differentiation, and mineralization.

The present study evaluated bone marrow aspirate (BMA) and low-level laser therapy (LLLT) on bone healing. It was created critical-size defects (CSD) of 5 mm diameter in rat calvaria of 64 rats. Animals were randomly divided into four groups: Control (blood clot), BMA (coagulated BMA), LLLT (laser irradiation and blood clot), and BMA/LLLT (laser irradiation and coagulated BMA). Euthanasia was performed at 15 or 30 days postoperative. Immunohistochemical reactions were performed to identify vascular endothelial growth factor (VEGF), proliferating cell nuclear antigen (PCNA), runt-related transcription factor-2 (Runx2), bone morphogenetic protein-2 (BMP-2), osteocalcin (OCN), and osteopontin (OPN). The markers were quantified, and data were statistically analyzed. Groups BMA/LLLT and LLLT presented significantly higher VEGF expression than group control. Group BMA/LLLT presented a significantly higher expression of PCNA than all experimental groups. Groups BMA and BMA/LLLT presented significantly higher expression of BMP-2 than all experimental groups. Groups LLLT and BMA/LLLT presented significantly higher expression of OPN than groups control and BMA. Groups LLLT, BMA, and BMA/LLLT presented a significantly higher expression of OCN than group control. It can be concluded that the association of BMA and LLLT enhanced bone healing by improving expression of VEGF, PCNA, Runx2, BMP-2, OPN, and OCN.

Favourable effects of Bacillus subtilis and Bacillus licheniformis on experimental periodontitis in rats.

OBJECTIVES: The purposes of this study were to evaluate, in rats: i) the effects of Bacillus species on the development of experimental periodontitis (EP) via microtomographic, immunological and hematological assays (Experiment 1-E1); ii) the effects of Bacillus species as adjuncts to scaling and root planing (SRP) for the treatment of EP via histomorphometric and immunohistochemical analyses (Experiment 2-E2). METHODS: In E1, 24 rats were divided into groups C1 (control), PROB1, EP1 and EP-PROB1. In groups with EP, the mandibular first molar of each animal received a ligature for 14 days. In groups PROB1, animals received Bacillus species for 44 days, starting 30 days before EP induction in Group EP-PROB1. In E2, 24 rats were assigned to groups C2 (control), PROB2, EP-SRP2 and EP-SRP-PROB2. In groups with SRP, EP was induced as described in E1. The ligatures were removed after 14 days and SRP was performed. In groups PROB2, animals received Bacillus species for 15 days, starting after SRP in Group EP-SRP-PROB2. RESULTS: In E1, Group EP1 presented bone loss (BL) and eosinophil numbers greater than Group EP-PROB1 (P<0.05). In Group EP-PROB1, the receptor activator of nuclear factor-kB ligand (RANKL)/osteoprotegerin (OPG) ratio was similar to that of groups without EP. In E2, Group EP-SRP-PROB2 presented fewer TRAP-positive osteoclasts, lower immunolabeling pattern for a proinflammatory cytokine and decreased BL and attachment loss than Group EP-SRP2 (P<0.05). CONCLUSIONS: Bacillus species supplementation provided a protective effect against BL and enhanced the effects of SRP in the treatment of EP in rats.

Effect of platelet-rich plasma on the healing of mandibular defects treated with fresh frozen bone allograft: a radiographic study in dogs.

The purpose of this study was to radiographically analyze the effect of autologous platelet rich plasma (PRP) on the healing of fresh frozen bone allograft (FFBA) placed in surgically created resection defects in mandibles of dogs. Bilateral resection defects measuring 1.5 cm × 1 cm were surgically created on the inferior border of the mandible in 10 adult male dogs. The defects were randomly divided into three groups: C, FFBA, and FFBA/PRP. In Group C, the defect was filled by blood clot only. In Group FFBA, the defect was filled with particulate fresh frozen bone allograft. In Group FFBA/PRP, it was filled with particulate fresh frozen bone allograft combined with PRP. At 90 days postoperative, standardized radiographs of the mandibles were obtained and results were quantitatively evaluated. Analysis of digitized radiographs indicated that non-PRP grafts were significantly less dense than the PRP grafts. Group FFBA/PRP also presented a statistically greater mineralized tissue area than Groups C and FFBA. Within the limits of this study, it can be concluded that PRP enhanced the healing of FFBA in resection defects in mandibles of dogs.

Platelet-rich plasma derived from bone marrow aspirate promotes new cementum formation.

BACKGROUND: This study evaluates the influence of platelet-rich plasma derived from bone marrow aspirate (PRP-BMA) on the healing of periodontal fenestration defects in rats. METHODS: Periodontal fenestration defects were surgically created in the mandibles of 40 rats. The animals were randomly divided into two groups, control and PRP-BMA, in which defects were filled with blood clot or PRP-bma, respectively. Animals were euthanized at either 10 or 30 days post-surgery. Histologic, histometric, and immunohistochemical analyses were performed. Percentage of new bone area (NBA), area of bone trabeculae (ABT), new cementum (NC), and extension of remaining defect were histometrically evaluated. Proliferating cell nuclear antigen (PCNA), bone sialoprotein (BSP), osteocalcin (OCN), and tartrate-resistant acid phosphatase (TRAP) immunohistochemical staining were performed. Immunolabeled cells were quantified. Data were statistically analyzed (analysis of variance; Tukey, P <0.05). RESULTS: At 10 days, control and PRP-BMA groups presented similar amounts of NBA and ABT; NC formation was not observed. At 30 days, control and PRP-BMA groups presented similar amounts of NBA and ABT; the PRP-BMA group showed NC formation with collagen fibers inserted obliquely or perpendicularly to the root surface. NC formation was not observed in any control group specimen. PRP- BMA presented higher numbers of PCNA-positive and BSP-positive cells than control at 10 and 30 days post-surgery. No significant differences in the number of either OCN-positive or TRAP-positive cells were observed between groups at 10 or 30 days. CONCLUSION: PRP-BMA promoted NC formation with a functional periodontal ligament when applied at experimental periodontal fenestration defects.

Platelet-rich plasma, low-level laser therapy, or their combination promotes periodontal regeneration in fenestration defects: a preliminary in vivo study.

BACKGROUND: This study histomorphometrically analyzes the influence of platelet-rich plasma (PRP), low-level laser therapy (LLLT), or their combination on the healing of periodontal fenestration defects (PFDs) in rats. METHODS: PFDs were surgically created in the mandibles of 80 rats. The animals were randomly divided into four groups: 1) C (control) and 2) PRP, defects were filled with blood clot or PRP, respectively; 3) LLLT and 4) PRP/LLLT, defects received laser irradiation, were filled with blood clot or PRP, respectively, and then irradiated again. Animals were euthanized at either 10 or 30 days post-surgery. Percentage of new bone (NB), density of newly formed bone (DNB), new cementum (NC), and extension of remaining defect (ERD) were histomorphometrically evaluated. Data were statistically analyzed (analysis of variance; Tukey test, P <0.05). RESULTS: At 10 days, group PRP presented ERD significantly lower than group C. At 30 days, group PRP presented NB and DNB significantly greater than group C. Groups LLLT, PRP, and PRP/LLLT showed significant NC formation at 30 days, with collagen fibers inserted obliquely or perpendicularly to the root surface. NC formation was not observed in any group C specimen. CONCLUSIONS: LLLT, PRP, or their combination all promoted NC formation with a functional periodontal ligament. The combination PRP/LLLT did not show additional positive effects compared to the use of either therapy alone.

Bone marrow aspirate combined with low-level laser therapy: a new therapeutic approach to enhance bone healing.

This study evaluated the influence of bone marrow aspirate (BMA), low-level laser therapy (LLLT) and their combination on bone healing in surgically created critical-size defects (CSDs) in rat calvaria. 40 rats were divided into four groups: C (control), BMA, LLLT and BMA/LLLT. A 5mmdiameter CSD was created in the calvarium of each animal. In Group C, the defect was filled by blood clot only. In Group BMA, the defect was filled with BMA. In groups LLLT and BMA/LLLT, the defect received laser irradiation (InGaAlP laser), was filled with blood clot or BMA respectively, and irradiated again. Animals were euthanized 30 days postoperatively. Histomorphometric and immunohistochemical analyses were performed. Newly formed bone area (NFBA) was calculated as percentage of the total area of the original defect. Proliferating cell nuclear antigen (PCNA), runt-related transcription factor 2 (Runx2) and osteocalcin (OCN) immunohistochemical staining were performed. PCNA-positive, Runx2-positive and OCN-positive cells were quantified. Data were statistically analyzed. Group BMA/LLLT had significantly greater NFBA than groups C, BMA or LLLT. Group BMA presented significantly greater NFBA than control, while group LLLT did not. Group BMA/LLLT presented a significantly higher number of PCNA-positive and OCN-positive cells than any of the other groups. Groups BMA/LLLT and BMA showed a significantly lower number of Runx2-positive cells than groups C or LLLT. The combination of BMA/LLLT yielded significantly greater bone formation in surgically created CSD in rat calvaria when compared to control, or either treatment alone.

Effect of platelet-rich plasma on bone healing of fresh frozen bone allograft in mandibular defects: a histomorphometric study in dogs.

OBJECTIVE: This study histologically analyzed the healing of fresh frozen bone allograft (FFBA) with or without platelet-rich plasma (PRP) in bony defects surgically created in the mandible of dogs. MATERIAL AND METHODS: Bilateral resection defects were surgically created on the inferior border of the mandible in 10 dogs. The defects were randomly divided into three groups: C (the defect was filled by blood clot only), FFBA (the defect was filled with particulate FFBA), and FFBA/PRP (the defect was filled with particulate FFBA combined with PRP). The animals were euthanized at 12 weeks post-surgery. Histomorphometric and histological analyses were performed. Data were statistically analyzed using Friedman test and Dunn test (P < 0.05). RESULTS: Group FFBA/PRP presented a statistically greater mineralized bone area and a smaller marrow bone area than Groups FFBA and C. CONCLUSIONS: The use of PRP improved the incorporation of FFBA, increasing the density of newly formed bone at 12 weeks post-surgery.

Effectiveness of two methods for preparation of autologous platelet-rich plasma: an experimental study in rabbits.

OBJECTIVES: The purpose of this study was to compare the quantity and quality of platelets in platelet-rich plasma (PRP) samples prepared using either the single- or the double-centrifugation protocol. METHODS: Ten adult white New Zealand rabbits were used. Ten ml of blood were drawn from each animal via cardiac puncture. Each blood sample was divided into two equal parts for PRP preparation: 5 ml of blood were centrifuged according to a single-centrifugation protocol (Group I), and 5 ml were centrifuged according to a double-centrifugation protocol (Group II). Manual platelet counts were performed on the whole blood and PRP samples of each group. Smears were also done on all samples in order to see the morphology of the platelets. The data obtained in the manual platelet count were submitted to statistical analysis (repeated measures ANOVA, Tukey, P<.05). RESULTS: The average whole blood platelet count was 446,389/µl. The PRP samples in Group II presented an average platelet amount significantly higher than that of Group I (1,986,875 ± 685,020/µl and 781,875 ± 217,693/µl, respectively). The PRP smears from Group II were the only one to present platelets with altered morphology (75% of the smears). A few lymphocytes with increased cytoplasm were observed in the PRP smears of both Groups I (25% of the smears) and II (62.5% of the smears). CONCLUSIONS: Within the limits of this study, it can be concluded that the double-centrifugation protocol resulted in higher platelet concentrations than did the single-centrifugation protocol. However, the double-centrifugation protocol caused alterations in platelet morphology and was more sensitive to small processing errors.

Bone healing in critical-size defects treated with new bioactive glass/calcium sulfate: a histologic and histometric study in rat calvaria.

This study analyzed histologically the influence of new spherical bioactive glass (NBG) particles with or without a calcium sulfate (CS) barrier on bone healing in surgically created critical-size defects (CSD) in rat calvaria. A CSD was made in each calvarium of 60 rats, which were divided into three groups: C (control): the defect was filled with blood clot only; NBG: the defect was filled with NBG only; and NBG/CS: the defect was filled with NBG covered by CS barrier. Subgroups were euthanized at 4 or 12 weeks. Amounts of new bone and remnants of implanted materials were calculated as percentages of total area of the original defect. Data were statistically analyzed. In contrast to Group C, thickness throughout defects in Groups NBG and NBG/CS was similar to the original calvarium. At 4 weeks, Group C had significantly more bone formation than Group NBG/CS. No significant differences were found between Group NBG and either Group C or Group NBG/CS. At 12 weeks, Group C had significantly more bone formation than Group NBG or NBG/CS. NBG particles, used with or without a CS barrier, maintained volume and contour of area grafted in CSD. Presence of remaining NBG particles might have accounted for smaller amount of new bone in Groups NBG and NBG/CS at 12 weeks post-operative.

Treatment of gingival recessions associated to cervical abrasion lesions with subepithelial connective tissue graft: a case report.

Extensive gingival recessions associated with cervical abrasions are common among the population. Several different surgical and/or restorative therapies have been proposed to correct these lesions. This manuscript reports the treatment of multiple gingival recessions associated to cervical abrasions. The procedure involved the utilization of subepithelial connective tissue graft (SCTG) combined with coronally advanced flap onto a previously restored root surface. At the postoperative follow-up visits, the success of the restorative/surgical approach was confirmed by the absence of bleeding to probing and periodontal pockets as well as presence of gingival tissue with normal color, texture and contouring. After 18 months of follow-up, the clinical conditions are stable with satisfactory root coverage and periodontal health. An excellent esthetical outcome was achieved and the patient is satisfied with case resolution.

Effect of platelet-rich plasma on bone healing of autogenous bone grafts in critical-size defects.

AIM: This study histologically analysed the effect of autogenous platelet-rich plasma (PRP), prepared according to a new semiautomatic system, on healing of autogenous bone (AB) grafts placed in surgically created critical-size defects (CSD) in rabbit calvaria. MATERIAL AND METHODS: Sixty rabbits were divided into three groups: C, AB and AB/PRP. A CSD was created in the calvarium of each animal. In Group C (control), the defect was filled by blood clot only. In Group AB (autogenous bone graft), the defect was filled with particulate autogenous bone. In Group AB/PRP (autogenous bone graft with platelet-rich plasma), it was filled with particulate autogenous bone combined with PRP. All groups were divided into subgroups (n=10) and euthanized at 4 or 12 weeks post-operatively. Histometric and histologic analyses were performed. Data were statistically analysed (anova, t-test, p<0.05). RESULTS: Group C presented significantly less bone formation compared with Group AB and AB/PRP in both periods of analysis (p<0.001). At 4 weeks, Group AB/PRP showed a statistically greater amount of bone formation than Group AB (64.44 +/- 15.0%versus 46.88 +/- 14.15%; p=0.0181). At 12 weeks, no statistically significant differences were observed between Groups AB and AB/PRP (75.0 +/- 8.11%versus 77.90 +/- 8.13%; p>0.05). It is notable that the amount of new bone formation in Group AB/PRP at 4 weeks was similar to that of Group AB at 12 weeks (p>0.05). CONCLUSION: Within its limitation, the present study has indicated that (i) AB and AB/PRP significantly improved bone formation and (ii) a beneficial effect of PRP was limited to an initial healing period of 4 weeks.

Periodontal tissue response to coverage of root cavities restored with resin materials: a histomorphometric study in dogs.

BACKGROUND: The purpose of this study was to histomorphometrically evaluate the response of periodontal tissues covering Class V resin restorations in dogs. METHODS: After raising a mucoperiosteal flap, bony defects measuring 5 x 5 mm were created on the buccal aspect of the canines of five dogs followed by cavity preparations on the root surface measuring 3 x 3 x 1 mm. Before repositioning the flap to cover the bone defect, the cavities were restored with composite resin (CR) or resin-modified glass ionomer cement (RMGIC) or were left unrestored as control (C). The dogs were euthanized 90 days after surgery. Specimens comprising the tooth and periodontal tissues were removed, processed routinely, cut into longitudinal serial sections in the buccolingual direction, and stained with hematoxylin and eosin (H&E) or Masson's trichrome. The most central sections were selected for histomorphometric analysis. RESULTS: Histomorphometric analysis revealed apical migration of epithelial tissue onto the restorative materials (RMGIC and CR). The C group presented significantly longer connective tissue attachment (P <0.05) than the RMGIC and CR groups and significantly higher bone regeneration (P <0.05) compared to the RMGIC group. Histologically, the cervical third (CT) of all groups had the most marked chronic inflammatory infiltrate. CONCLUSIONS: Within the limits of this study, it can be concluded that the restorative materials used exhibit biocompatibility; however, both materials interfered with the development of new bone and the connective tissue attachment process.

Bone healing in critical-size defects treated with bioactive glass/calcium sulfate: a histologic and histometric study in rat calvaria.

OBJECTIVE: The purpose of this study was to analyze histologically the influence of bioactive glass (BG) with or without a calcium sulfate (CS) barrier on bone healing in surgically created critical-size defects (CSD) in rat calvaria. MATERIAL AND METHODS: A CSD was made in each calvarium of 48 rats. They were divided into three groups: C (control): blood clot only; BG: defect filled with BG; and BG/CS: defect filled with BG covered by a CS barrier. Animals were euthanized at 4 or 12 weeks. Formation of new bone was evaluated histomorphometrically. RESULTS: No defect completely regenerated with bone. BG particles were observed in Groups BG and BG/CS at both periods of analysis. The thickness throughout the healing area in Groups BG and BG/CS was similar to the original calvarium, while Group C presented a thin connective tissue in the center of the defect in both periods of analysis. At 4 weeks, Groups C and BG/CS presented significantly more bone formation than Group BG. No significant differences were found between Groups C and BG/CS. At 12 weeks, no significant differences in the amount of bone formation were observed among the three groups. When comparing 4 and 12 weeks, there was a significant increase in new bone formation within groups BG and BG/CS, but not C. CONCLUSION: BG particles, used with or without a CS barrier, maintained the volume and contour of the area grafted in CSD. However, they did not lead to a significant difference in bone formation when compared with control at 12 weeks post-operative.

Paradental cyst: report of two cases.

BACKGROUND: The paradental cyst is an odontogenic lesion of inflammatory origin that has few clinical signs and symptoms apart from recurring acute episodes. A well-defined radiolucency associated with the roots or distal to the crown may be seen radiographically. The purpose of this article is to report on different aspects of two cases involving paradental cysts. In the first case, the patient complained about recurring pericoronitis. A semilunar-shaped radiolucency on the distal aspect of the mandibular third molar was noted on the periapical radiograph. In the second case, the patient's main complaint was chronic trauma of the overlying mucosa. Radiographs revealed an enlarged pericoronal space. METHODS: In both cases, the mandibular third molar was extracted due to a lack of space. Lesional samples were sent for histopathologic analysis. RESULTS: In the first case, the drainage of cystic fluid and a regular concavity were found during tooth removal. In the second case, a nodular lesion was found adhering to the disto-buccal surface of the tooth arising from the distal wall of a periodontal pocket. The histopathologic analysis revealed a hyperplastic stratified squamous epithelium with arcading lining a fibrous capsule with inflammatory infiltrate, resulting in a final diagnosis of a paradental cyst. CONCLUSIONS: The presence of a paradental cyst should be considered when recurrent inflammatory periodontal processes are associated with partially erupted vital teeth, even when characteristic radiographic findings are absent. Definitive diagnosis requires a clinicopathologic correlation incorporating surgical, radiographic, and histologic findings.

Autogenous bone graft with or without a calcium sulfate barrier in the treatment of Class II furcation defects: a histologic and histometric study in dogs.

BACKGROUND: The purpose of this study was to histologically evaluate the healing of surgically created Class II furcation defects treated using an autogenous bone (AB) graft with or without a calcium sulfate (CS) barrier. METHODS: The second, third, and fourth mandibular premolars (P2, P3, and P4) of six mongrel dogs were used in this study. Class II furcation defects (5 mm in height x 2 mm in depth) were surgically created and immediately treated. Teeth were randomly divided into three groups: group C (control), in which the defect was filled with blood clot; group AB, in which the defect was filled with AB graft; and group AB/CS, in which the defect was filled with AB graft and covered by a CS barrier. Flaps were repositioned to cover all defects. The animals were euthanized 90 days post-surgery. Mesio-distal serial sections were obtained and stained with either hematoxylin and eosin or Masson's trichrome. Histometric, using image-analysis software, and histologic analyses were performed. Linear and area measurements of periodontal healing were evaluated and calculated as a percentage of the original defect. Percentage data were transformed into arccosine for statistical analysis (analysis of variance; P <0.05). RESULTS: Periodontal regeneration in the three groups was similar. Regeneration of bone and connective tissue in the furcation defects was incomplete in most of the specimens. Statistically significant differences were not found in any of the evaluated parameters among the groups. CONCLUSION: Periodontal healing was similar using surgical debridement alone, AB graft, or AB graft with a CS barrier in the treatment of Class II furcation defects.

Bone healing in surgically created defects treated with either bioactive glass particles, a calcium sulfate barrier, or a combination of both materials. A histological and histometric study in rat tibias.

OBJECTIVE: The purpose of this study was to histologically analyze the influence of bioactive glass and/or a calcium sulfate barrier on bone healing in surgically created defects in rat tibias. MATERIAL AND METHODS: Sixty-four rats were divided into 4 groups: C (control), CS (calcium sulfate), BG (bioactive glass), and BG/CS (bioactive glass/calcium sulfate). A surgical defect was created in the tibia of each animal. In Group CS, a calcium sulfate barrier was placed to cover the defect. In Group BG the defect was filled with bioactive glass. In Group BG/CS, it was filled with bioactive glass and protected by a barrier of calcium sulfate. Animals were sacrificed at 10 or 30 days post-operative. The formation of new bone in the cortical area of the defect was evaluated histomorphometrically. RESULTS: At 10 days post-operative, Group C presented significantly more bone formation than Groups CS, BG, or BG/CS. No statistically significant differences were found between the experimental groups. At 30 days post-operative, Group C demonstrated significantly more bone formation than the experimental groups. Groups CS and BG/CS showed significantly more bone formation than Group BG. No statistically significant differences were found between Group CS and BG/CS. CONCLUSIONS: (a) the control groups had significantly more bone formation than the experimental groups; (b) at 10 days post-operative, no significant differences were found between any of the experimental groups; and (c) at 30 days post-operative, the groups with a calcium sulfate barrier had significantly more bone formation than the group that used bioactive glass only.

Healing of dehiscence defects following root surface demineralization with tetracycline: a histologic study in monkeys.

BACKGROUND: The purpose of this study was to histologically evaluate the healing of experimental dehiscence defects after surface demineralization with tetracycline hydrochloride. METHODS: Six adult male monkeys (Cebus apella) were used in this study. Dehiscence defects were surgically created on the buccal aspect of the mandibular lateral incisors in all animals. The root surfaces were debrided and planed. In a split-mouth design, a 10% tetracycline hydrochloride solution was applied to one tooth for 4 minutes (T group), followed by irrigation with saline. The contralateral tooth served as a control (C group). The flaps were repositioned and sutured. The animals were sacrificed at 6 months postoperatively and histological sections were processed. Computer-assisted histomorphometric analysis was used to evaluate the formation of new cementum, new bone, new connective tissue attachment, and length of the epithelium (junctional and sulcular). RESULTS: Bone regeneration was similar in both groups (1.5 +/- 0.3 mm for the T group and 1.5 +/- 0.6 mm for the C group). The C group showed more new cementum than the T group (2.3 +/- 0.3 mm versus 2.2 +/- 0.3 mm) as well as a longer epithelium (1.0 +/- 0.3 mm versus 0.9 +/- 0.2 mm). The T group presented more new connective tissue attachment (3.1 +/- 0.2 mm) than the C group (2.9 +/- 0.6 mm). However, no statistically significant differences were detected between the two groups. CONCLUSIONS: The amount of new attachment was similar in both groups. Root conditioning with 10% tetracycline solution did not produce any additional new attachment in comparison to the controls.