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.

Topical sodium alendronate combined or not with photodynamic therapy as an adjunct to scaling and root planing: Histochemical and immunohistochemical study in rats.

OBJECTIVE: The purpose of this study was to evaluate influence of topical sodium alendronate (ALN), photodynamic therapy (aPDT), or a combination thereof as adjuvant to scaling and root planing (SRP) in the treatment of experimental periodontitis in rats. BACKGROUND: Therapeutic protocols to control periodontitis progression that aim to equalize bacterial action and load with tissue immune response are well addressed in current scientific research. METHODS: Experimental periodontitis was induced in 96 rats with a ligature around the mandibular left first molar. After 7 days, ligature was removed and animals were treated according to the following experimental groups (n = 8): control-SRP plus saline solution; ALN-SRP plus ALN; aPDT-SRP plus methylene blue irrigation, followed by low-level laser therapy (LLLT); and ALN/aPDT-SRP plus ALN and methylene blue irrigation followed by LLLT. The animals were euthanized at 7, 15, and 30 days after treatments. Collagen maturation (picrosirius red staining) and immunohistochemical analyses (TRAP, RANKL and osteoprotegerin [OPG]) were performed. Data were submitted to statistical analysis (P < .05). RESULTS: At 7 days, group ALN presented a significantly higher number of TRAP-positive cells and percentage of immature collagen fibers than group ALN/aPDT, while group ALN/aPDT presented a significantly higher percentage of mature collagen fibers than group ALN. At 30 days, group ALN presented significantly lower percentage of immature collagen fibers and higher percentage of mature collagen fibers than control. CONCLUSION: It can be concluded that topical use of ALN coadjutant to SRP, alone or combined with aPDT, enhanced collagen maturation and reduced osteoclastogenesis during the healing of experimental periodontitis.

Effects of the prebiotic mannan oligosaccharide on the experimental periodontitis in rats.

AIM: To evaluate the effect of the prebiotic (PREB) mannan oligosaccharide (MOS) on the progression of the experimental periodontitis (EP) and intestinal morphology in rats. MATERIALS AND METHODS: Forty rats were randomly allocated into groups (n = 10): C (control), PREB, EP and EP-PREB. Groups PREB and EP-PREB received MOS incorporated into the feed daily. After 30 days, groups EP and EP-PREB received a cotton ligature around their mandibular first molars, kept for 14 days. Morphometrical, histomorphometrical, microcomputed tomography, gene expression analyses and immunohistochemistry were performed. Data were statistically analysed (p < 0.05). RESULTS: Group EP-PREB showed less interproximal bone loss, area without bone in the furcation and bone porosity, and greater bone mineral density than group EP (p < 0.05). It was also observed a significant decrease in IL-10 and IFN-γ gene expression, besides a decrease in TNF-α and IL-1ß and an increase in TGF-ß immunolabeling score for group EP-PREB. Group EP-PREB also presented villous height and crept depth values similar to group C, while group EP presented reduced values (p < 0.05). CONCLUSION: It can be concluded that the oral administration of MOS promotes a protective effect against alveolar bone loss caused by EP in rats, modifying histologic and immune-inflammatory parameters, in addition to protecting the intestine.

Long-term therapy with intravenous zoledronate increases the number of nonattached osteoclasts.

The purpose of this study was to investigate the influence of long-term therapy with intravenous zoledronate (ZA) on the healing of extraction sockets in rats. Forty rats, divided into groups C (Control) and Z (Zoledronate), received intravenous injections of either saline solution or ZA for 24 weeks. Their right maxillary incisor was extracted. Euthanasia was performed at 7 or 28 days postoperative. Histomorphometric (Newly Formed Bone Area) and immunohistochemical (RANKL, OPG and TRAP) analyses were performed. Data were statistically analyzed (ANOVA, Tukey's test and Kruskal-Wallis, Dunn's Multiple Comparison test).Groups C and Z showed similar new bone area, RANKL and OPG immunolabeling. The number of TRAP-positive multinucleated cells was significantly higher in Group Z than in Group C at 28 days. A significantly higher proportion of nonattached osteoclasts were seen in Group Z than in Group C at both periods of analysis. Long-term therapy with intravenous ZA stimulated nonattached osteoclast formation in extraction sockets in rats, thus decreasing local bone resorption. However, it did not influence bone formation by osteoblasts.

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.