Michel Reis Messora

Bone healing in critical-size defects treated with platelet-rich plasma: a histologic and histometric study in rat calvaria

BACKGROUND AND OBJECTIVE The purpose of this study was to analyze histologically the influence of autologous platelet-rich plasma on bone healing in surgically created critical-size defects in rat calvaria. MATERIAL AND METHODS Thirty-two rats were divided into two groups: the control group (group C) and the platelet-rich plasma group. An 8-mm-diameter critical-size defect was created in the calvarium of each animal. In group C the defect was filled by a blood clot only. In the platelet-rich plasma group, 0.35 mL of platelet-rich plasma was placed in the defect and covered by 0.35 mL of platelet-poor plasma. Both groups were divided into subgroups (n = 8) and killed at either 4 or 12 wk postoperatively. Histometric (using image-analysis software) and histologic 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, p < 0.05). RESULTS No defect completely regenerated with bone. The platelet-rich plasma group had a statistically greater amount of bone formation than group C at both 4 wk (17.68% vs. 7.20%, respectively) and 12 wk (24.69% vs. 11.65%, respectively) postoperatively. CONCLUSION Within the limits of this study, it can be concluded that platelet-rich plasma placed in the defects and covered by platelet-poor plasma significantly enhanced bone healing in critical-size defects in rat calvaria.

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

BACKGROUND AND OBJECTIVE 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. MATERIAL AND METHODS 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, Tukeys post hoc test, p < 0.05). RESULTS 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. CONCLUSION 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.

Influence of the proportion of particulate autogenous bone graft/platelet-rich plasma on bone healing in critical-size defects: An immunohistochemical analysis in rat calvaria

This immunohistochemical analysis evaluated the influence of the proportion of particulate autogenous bone (AB) graft/platelet-rich plasma (PRP) on bone healing in surgically created critical-size defects (CSD) in rat calvaria. 25 rats were divided into 5 groups: C (control), AB, AB/PRP-50, AB/PRP-100 and AB/PRP-150. A 5 mm diameter critical-size defect was created in the calvarium of each animal. In Group C, defect was filled by blood clot only. In Group AB, defect was filled with AB graft. In Groups AB/PRP-50, AB/PRP-100 and AB/PRP-150, defects were filled with AB graft combined with 50, 100 and 150 microl of PRP, respectively. Animals were euthanized at 30 days post-operative. Osteocalcin (OC) and osteopontin (OP) immunohistochemical staining were performed. OP-positive and OC-positive cells were quantified. The amount of vital bone (VBA) and remaining bone graft particles (RPA) were calculated as percentage of the total area of the original defect. Data were statistically analyzed (ANOVA, Tukey, p<0.05). Group AB/PRP-100 presented a significantly higher number of OP-positive cells than groups C, AB/PRP-50 and AB/PRP-150 (p<0.05) and also a significantly higher number of OC-positive cells than all of the other groups (p<0.05). The number of OP-positive cells in Group AB/PRP-150 was significantly lower than in Group AB (p<0.05). Statistically significant differences were not found between groups with regard to either VBA or RPA (p>0.05). However, clear trends were indicated by an increase in VBA in Group AB/PRP-100 when compared to groups C, AB and AB/PRP-150 and an increase in RPA in Group AB/PRP-150 when compared to groups AB, AB/PRP-50 and AB/PRP-100. The proportion AB graft/PRP seems to influence bone healing in CSD in rat calvaria. The highest expression of osteocalcin and osteopontin, and thus the most developed healing, was found in Group AB/PRP-100 indicating that there may be an optimal proportion beyond which increasing the concentration PRP is deleterious.

Antimicrobial Photodynamic Therapy as an Adjunct to Non-Surgical Treatment of Aggressive Periodontitis: A Split-Mouth Randomized Controlled Trial

BACKGROUND The management of aggressive periodontitis (AgP) represents a challenge for clinicians because there are no standardized protocols for an efficient control of the disease. This randomized controlled clinical trial evaluated the effects of repeated applications of antimicrobial photodynamic therapy (aPDT) adjunctive to scaling and root planing (SRP) in patients with AgP. METHODS Using a split-mouth design, 20 patients with generalized AgP were treated with aPDT + SRP (test group) or SRP only (control group). aPDT was applied at four periods. All patients were monitored for 90 days. Clinical, microbiologic, and immunologic parameters were statistically analyzed. RESULTS In deep periodontal pocket analysis (probing depth [PD] ≥ 7 mm at baseline), the test group presented a decrease in PD and a clinical attachment gain significantly higher than the control group at 90 days (P < 0.05). The test group also demonstrated significantly less periodontal pathogens of red and orange complexes and a lower interleukin-1β/interleukin-10 ratio than the control group (P < 0.05). CONCLUSION The application of four sessions of aPDT, adjunctive to SRP, promotes additional clinical, microbiologic, and immunologic benefits in the treatment of deep periodontal pockets in single-rooted teeth in patients with AgP.

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.

Probiotic Therapy Reduces Periodontal Tissue Destruction and Improves the Intestinal Morphology in Rats With Ligature-Induced Periodontitis

BACKGROUND With increase in the incidence of resistance to antibiotics, probiotics are emerging as a promising adjunctive periodontal therapy. The authors of this study evaluate the influence of probiotic (PROB) supplementation on ligature-induced periodontitis (LIP) and intestinal morphology in rats. METHODS Thirty-two rats were randomly divided into four groups: control (C), LIP, PROB, and LIP/PROB. In groups PROB and LIP/PROB, the PROB was administered orally by addition to the drinking water of the animals for 44 days. In groups LIP and LIP/PROB, the mandibular right first molar of the animals received a cotton ligature that was left in the same position for 14 days. All animals were euthanized 44 days after the start of the PROB supplementation. The jaws were resected and histomorphometric analyses were performed. The measurements included evaluation of attachment loss (AL) and alveolar bone level (ABL) on the distal root of the mandibular first molar. Samples of the duodenum, jejunum, and ileum were also dissected from each animal to evaluate the villous height (VH) and crypt depth (CD). The data obtained were subjected to statistical analyses (analysis of variance, Tukey; P <0.05). RESULTS Mean values of AL and ABL were significantly higher in group LIP compared with group LIP/PROB (AL: 3.05 ± 0.57 mm and 1.78 ± 0.63 mm, respectively; ABL: 4.21 ± 0.42 mm and 3.38 ± 0.17 mm, respectively). In group LIP/PROB, the mean values of VH and CD of the jejunum were significantly higher than the ones from group LIP (VH: 672.1 ± 83.3 µm and 528.0 ± 51.7 µm, respectively; CD: 463.8 ± 100.9 µm and 269.0 ± 48.4 µm, respectively). CONCLUSIONS It can be concluded that PROB supplementation 1) reduces AL and alveolar bone loss in rats with LIP and 2) can protect the small intestine from reactive changes induced by LIP.

Influence of the ratio of particulate autogenous bone graft/platelet-rich plasma on bone healing in critical-size defects: a histologic and histometric study in rat calvaria.

The purpose of this study was to analyze histomorphometrically the influence of the ratio of particulate autogenous bone (AB) graft/platelet‐rich plasma (PRP) on bone healing in surgically created critical‐size defects (CSD) in rat calvaria. Fifty rats were divided into five groups: Group C (control), Group AB, Group AB/PRP‐50, Group AB/PRP‐100, and Group AB/PRP‐150. A 5‐mm diameter critical‐size defect was created in the calvarium of each animal. In Group C, the defect was filled by blood clot only. In Group AB, the defect was filled with 0.01 mL of AB graft. In Groups AB/PRP‐50, AB/PRP‐100, and AB/PRP‐150, the defects were filled with 0.01 mL of AB graft combined with 50, 100, and 150 µL of PRP, respectively. All animals were euthanized at 30 days postoperative. Histomorphometry, using image analysis software, and histology analyses were performed. New Bone Area (NBA) and the remaining bone graft particles area (RPA) were calculated as a percentage of the total area of the original defect. Percentage data were transformed into arccosine for analysis. No defect completely regenerated with bone. Group AB/PRP‐50 (41.78 ± 13.48%) had a significantly greater NBA than Groups C (19.29 ± 5.11%), AB (27.43 ± 10.90%) or AB/PRP‐150 (19.17 ± 8.45%) (p < 0.05). No significant differences were observed between groups AB/PRP‐50 and AB/PRP‐100 or among groups AB, AB/PRP‐100, and AB/PRP‐150 with regard to NBA (p > 0.05). Group AB/PRP‐150 (31.59 ± 3.22%) had a significantly greater RPA than Groups AB (19.09 ± 5.21%), AB/PRP‐50 (17.33 ± 4.43%), and AB/PRP‐100 (19.72 ± 3.62%) (p < 0.001). No significant differences were observed among groups AB, AB/PRP‐50, and AB/PRP‐100 with regard to RPA (p > 0.05). The ratio AB graft/PRP influences bone healing in surgically created CSD in rat calvaria.

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.

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.

Effects of Probiotic Therapy on Metabolic and Inflammatory Parameters of Rats With Ligature-Induced Periodontitis Associated With Restraint Stress

BACKGROUND This study evaluates the effects of probiotic therapy (PT) in rats with ligature-induced periodontitis associated with restraint stress. METHODS Sixty-four rats were divided into control, stress (STR), probiotic (PROB), periodontal disease (PD), STR-PROB, STR-PD, STR-PROB-PD, and PROB-PD groups. The probiotic was added to the drinking water for 44 days. PD was induced by a ligature. In STR groups, the animals were subjected to restraint stress for 2.5 hours per day for 30 days. RESULTS Rats with PD exhibited increased alveolar bone loss (P <0.05), as well as increased levels of cyclooxygenase-2, serum C-terminal telopeptide (CTX), p38 mitogen-activated protein kinase (p38), and receptor activator of nuclear factor-κB ligand and decreased levels of osteoprotegerin (OPG). Stressed rats presented high levels of C-peptide, corticosterone, and glucose (P <0.05). In general, the presence of stress reduced the expression of CTX and p38 (P <0.05). PT reduced alveolar bone loss in unstressed animals. It also decreased expression of CTX and induced increased expression of OPG in unstressed animals with PD. However, PT was not effective in preventing bone loss or altering the expression of inflammatory markers in stressed animals. PT decreased the number of inflammatory cells in the periodontal tissue (P <0.05). Groups with stress and PD showed decreased villous height and crypt depth. Stress seemed to prevent part of the probiotic beneficial effects on the small intestine. CONCLUSIONS Based on the methodology used, PT may reduce tissue breakdown resulting from PD in unstressed rats. The protocol used for restraint stress influenced the immunomodulatory effects of PT in intestinal and periodontal tissues.