Photobiomodulation Activates Microglia/Astrocytes and Relieves Neuropathic Pain in Inferior Alveolar Nerve Injury.

Objective: This study aimed to determine microglial/astrocyte changes and their associated analgesic effect in inferior alveolar nerve injury (IANI) model rats treated with photobiomodulation therapy (PBMT) using a 940-nm diode laser. Background: Very few basic studies have investigated microglial/astrocyte dynamics following PBMT aimed at relieving neuropathic pain caused by IANI. Methods: Rats were divided into an IANI-PBM group, IANI+PBM group, and sham+PBM group. Observations were made on the day before IANI or the sham operation and on postoperative days 3, 5, 7, 14, and 28. PBMT was delivered for 7 consecutive days, with an energy density of 8 J/cm2. Behavioral analysis was performed to determine pain thresholds, and immunohistological staining was performed for the microglia marker Iba1 and astrocyte marker glial fibrillary acidic protein, which are observed in the spinal trigeminal nucleus. Results: Behavioral analysis showed that the pain threshold returned to the preoperative level on postoperative day 14 in the IANI+PBM group, but decreased starting from postoperative day 1 and did not improve thereafter in the IANI-PBM group (p ≤ 0.001). Immunological analysis showed that microglial and astrocyte cell counts were similar in the IANI+PBM group and IANI-PBM group shortly after IANI (day 3), but the expression area was larger (p ≤ 0.001) and hypertrophy of microglia and astrocyte cell bodies and end-feet extension (i.e., indicators of activation) were more prominent in the IANI+PBM group. Conclusions: PBMT after IANI prevented hyperalgesia and allodynia by promoting glial cell activation shortly after injury.

CO2 Laser for Esthetic Healing of Injuries and Surgical Wounds with Small Parenchymal Defects in Oral Soft Tissues.

A number of studies have recently demonstrated the effectiveness of CO2 laser irradiation for the repair and regeneration of scar tissue from injuries or surgical wounds. However, such studies of the oral mucosa are highly limited. Previous studies using CO2 laser irradiation have indicated that two factors contribute to esthetic healing, namely, artificial scabs, which are a coagulated and carbonized blood layer formed on the wound surface, and photobiomodulation therapy (PBMT) for suppressing wound scarring and promoting wound healing. This review outlines basic research and clinical studies of esthetic healing with the use of a CO2 laser for both artificial scab formation by high-intensity laser therapy and PBMT in the treatment of injuries and surgical wounds with small parenchymal defects in oral soft tissues. The results showed that the wound surface was covered by an artificial scab, enabling the accumulation of blood and the perfusion necessary for tissue regeneration and repair. Subsequent PBMT also downregulated the expression of transformation growth factor-b1, which is involved in tissue scarring, and decreased the appearance of myofibroblasts. Taken together, artificial scabs and PBMT using CO2 lasers contribute to the suppression of scarring in the tissue repair process, leading to favorable esthetic and functional outcomes of wound healing.

Space-making effect for new bone formation by suppressing scar contraction of mucosal epithelium of rat tooth extraction wound using diode laser and CO2 laser treatment.

Background/purpose: As an extraction wound closes, the mucosal epithelium of the tooth extraction wound impedes the space for new bone formation by invading into the extraction socket. Thus, the height of the alveolar crest decreases, causing significant depression of the alveolar mucosa. In this study, we created a rat tooth extraction model and examined the effects of laser irradiation by CO2 and diode on the dynamics of myofibroblast expression through α-SMA, and TGF-ß1. Materials and methods: After tooth extraction of five-week-old male Wistar rats, they were divided into two laser treatment groups (CO2 laser or diode laser was irradiated into tooth extraction socket) and non-laser treatment group (control group). Surrounding tissues, including the extraction socket, were removed at 3, 5, 7, and 21 days after tooth extraction and the expression of α-SMA and TGF-ß1 was verified using immunohistological techniques (6 animals in each group and each period, 72 animals in total). Results: α-SMA-positive cells and TGF-ß1-positive areas were significantly lower in the two laser treatment groups than in the control group. Particularly, the diode group almost had no TGF-ß1-positive areas on the 21st day when healing after tooth extraction was deemed to be completed. Conclusion: Both CO2 and diode laser irradiation of tooth extraction wounds decreases α-SMA-positive cells and TGF-ß1-positive areas. Further, it causes a decrease in myofibroblast expression and suppresses the invasion of mucosal epithelium into the extraction socket. Therefore, laser irradiation may exert a space-making effect for new bone formation and also contribute to socket preservation.

Healing of Open Upper Lip Vermillion Wounds Irradiated with CO2 Laser Immediately After Injury.

Objective: To evaluate the healing of open upper lip vermillion wounds irradiated with CO2 laser immediately after injury. Background: There are reports of using CO2 laser therapy on healed wounds for scar tissue reduction. However, limited data exist regarding its use immediately after an injury. Thus, the role of CO2 laser in wound healing remains unclear. Methods: Two patients with open upper lip vermillion wounds were treated using CO2 laser irradiation to the area postsuturing. Results: Good functional and aesthetic results were obtained from the therapy, with no postoperative pain complaints or infection. Conclusions: CO2 laser irradiation, performed immediately after an injury, could be an effective treatment option for open vermillion wounds.

Wound Healing and Cell Dynamics Including Mesenchymal and Dental Pulp Stem Cells Induced by Photobiomodulation Therapy: An Example of Socket-Preserving Effects after Tooth Extraction in Rats and a Literature Review.

High-intensity laser therapy (HILT) and photobiomodulation therapy (PBMT) are two types of laser treatment. According to recent clinical reports, PBMT promotes wound healing after trauma or surgery. In addition, basic research has revealed that cell differentiation, proliferation, and activity and subsequent tissue activation and wound healing can be promoted. However, many points remain unclear regarding the mechanisms for wound healing induced by PBMT. Therefore, in this review, we present an example from our study of HILT and PBMT irradiation of tooth extraction wounds using two types of lasers with different characteristics (diode laser and carbon dioxide laser). Then, the effects of PBMT on the wound healing of bone tissues are reviewed from histological, biochemical, and cytological perspectives on the basis of our own study of the extraction socket as well as studies by other researchers. Furthermore, we consider the feasibility of treatment in which PBMT irradiation is applied to stem cells including dental pulp stem cells, the theme of this Special Issue, and we discuss research that has been reported on its effect.

Utility of High-Intensity Laser Therapy Combined with Photobiomodulation Therapy for Socket Preservation After Tooth Extraction.

Objective: High-intensity laser therapy (HILT) combined with photobiomodulation therapy (PBMT) using a diode or CO2 laser was administered after extraction of the left first molar in rats. Effects on socket preservation (preservation of the alveolar bone and healing time after extraction) were evaluated histopathologically. Background: Irradiation using a diode or CO2 laser has been shown to hasten wound healing, but the effects remain controversial. Methods: Five-week-old male Wistar rats that underwent extraction of the left maxillary first molar were divided into three groups: diode laser irradiation (diode group), CO2 laser irradiation (CO2 group), and no laser irradiation (control group). HILT (27 J) was performed immediately after tooth extraction to enhance blood coagulation, followed by PBMT (0.7 J) 1 day later to enhance healing. Tissues, including the extraction socket, were removed en bloc 3, 5, 7, 10, and 21 days postextraction to determine the morphological characteristics of wound healing and the distribution of myofibroblasts involved in scar formation. Results: In the diode and CO2 groups, new bone formation and cancellous bone maturation were observed at an early stage of wound healing. The number of myofibroblasts was significantly lower in the laser treatment groups than the control (p < 0.001), and both treatment groups had a significantly higher alveolar crest height (p < 0.01), with almost no concavity in the mucosa of the extraction wound. Conclusions: Combined HILT and PBMT following tooth extraction hastened wound healing and preserved alveolar crest height, suggesting a role in socket preservation.

Placental extract suppresses differentiation of 3T3-L1 preadipocytes to mature adipocytes via accelerated activation of p38 MAPK during the early phase of adipogenesis.

BACKGROUND: Adipogenesis, the process of preadipocyte differentiation to mature adipocytes accompanied by accumulation of intracytoplasmic lipid droplets, is regulated by various genetic and environmental factors, and closely associated with the development of obesity. Numerous recent studies suggest that some bioactive peptides and proteins derived from animals, and chemical compounds isolated from plants may be useful for prevention and treatment of obesity and obesity-related chronic diseases. In the present study, we examined the broad spectrum of effects of placental extract, with a focus on the influence of placental extract on adipogenesis. METHOD: We cultured 3T3-L1 cells, which are widely used as a model of white preadipocytes, under differentiation conditions in the presence of porcine placental extract (PPE) for 8 days, and then stained the lipid droplets accumulated in the cytoplasm with Oil Red O. We also analyzed the effects of PPE on the mitogen-activated protein kinases (MAPKs) signaling, mitotic clonal expansion (MCE) and gene expressions associated with 3T3-L1 differentiation. RESULTS: When we cultured 3T3-L1 cells with PPE under differentiation conditions, the accumulation of lipid droplets and expression of adipocyte differentiation marker genes (Cebpa, Pparg, Slc2a4, Fasn and Adipoq) were dramatically attenuated. The suppressive activity of PPE against adipogenesis was heat-stable and recovered in a low-molecular-weight fraction after ultrafiltration (< 3 kDa) and gel-filtration chromatography (fraction No. 9). We also found that the suppressive activity of PPE affected the early phase of adipocyte differentiation (Days 0-2) without influencing the expression levels of C/EBPß and C/EBPδ. The PPE and fraction No. 9 obtained from gel-filtration chromatography both promoted mitotic clonal expansion of 3T3-L1 while accelerating p38 MAPK phosphorylation. In addition, SB203580, a p38 MAPK inhibitor, partially restored the accumulation of lipid droplets and the expression of adipocyte differentiation marker genes that were suppressed by fraction No. 9. CONCLUSION: These results indicate that PPE suppresses the differentiation of preadipocytes via accelerated activation of p38 MAPK during the early phase of adipogenesis, suggesting PPE or its functional component could be a potential therapy for treating obesity.

Anti-inflammatory Effect of the Water-Soluble Portion of Porcine Placental Extract in Lipopolysaccharide-Stimulated RAW264.7 Murine Macrophage Cells.

Porcine placental extract (PPE) is used as a nonprescription drug for analeptics and in health foods and cosmetics in Japan, Korea and China. It was reported that PPE has anti-oxidative and anti-inflammatory activities; however, the mechanisms and the responsible molecules involved in these activities are still unclear. Here, we investigated how enzymatically prepared PPE affects proinflammatory factors such as interleukin (IL)-1ß, IL-6, and tumor necrosis factor (TNF)-α in a cultured macrophage cell line, RAW264.7, when co-stimulated with lipopolysaccharide (LPS). Enhanced production of IL-1ß, IL-6 and TNF-α by LPS was significantly reduced by the addition of PPE and these effects were dose dependent. Nitric oxide (NO) production induced in cultured macrophages by LPS was also inhibited by PPE. Real-time PCR after the reverse transcription of total RNAs isolated from cells treated with PPE revealed that the mRNA expressions of IL-1ß, IL-6, TNFα, and NO synthase (NOS)-2 were reduced. The necessary concentration of PPE prepared by enzymatic digestion to mediate anti-inflammatory effects compared with the reported value of that extracted by phosphate buffered saline without digestion was proportional to the amount of extracted materials from the same amount of placenta (about 10-fold). This suggests that the molecules responsible for the anti-inflammatory activity exists in the placenta and can be extracted by phosphate buffered saline, and thus might survive enzymatic digestion.

Influence of carbon dioxide laser irradiation on the healing process of extraction sockets.

OBJECTIVE: To clarify the healing-promoting effects of carbon dioxide laser irradiation in high and low reactive-level laser therapies (HLLT and LLLT, respectively) on extraction sockets after tooth extraction. MATERIAL AND METHODS: Forty-two 5-week-old male Wistar rats were divided into laser irradiation and non-irradiation (control) groups and compared. The laser-irradiation group underwent HLLT immediately after tooth extraction and then LLLT 1 day post-extraction. Tissue was excised 6 h and 3, 7, or 21 days after extraction and histopathologically investigated. The alveolar crest height was measured osteomorphometrically 21 days post-extraction, and granulation tissue in the extraction socket surface layer was immunohistologically investigated using anti-α-smooth muscle actin (anti-α-SMA) antibody 3 and 7 days post-extraction. RESULTS: Many osteoclasts appeared and active bone resorption was noted in the irradiation group 3 days after extraction compared to the controls. On Day 7, new bone formation started around the extraction socket in the control group, but from the superficial to over the middle layer of the socket in the irradiation group. On Day 21, a concavity existed in the alveolar crest region in the controls, whereas this region was flat, with no concavity, in the irradiation group. On osteomorphometry, the alveolar crest height was significantly higher in the irradiation (0.7791 ± 0.0122) than the control (0.6516 ± 0.0181) group (P < 0.05). On immunostaining, many α-SMA-positive cells were noted in the control group, but very few in the irradiation group. CONCLUSION: Laser-irradiated extraction wound healing showed characteristics different from those of the normal healing process, suggesting a favorable healing-promoting effect.