Effects of Irradiation by Carbon Dioxide Laser Equipped With a Water Spray Function on Bone Formation in Rat Tibiae.

BACKGROUND/AIM: Irradiation of tissue with carbon dioxide (CO2) laser shows a characteristic thermal effect that causes vaporization of tissue in the target region. However, the thermal effect in places other than the target region induces tissue damage. Two methods are used: high reactive-level laser therapy (HLLT), aimed at surgical treatment, and low reactive-level laser therapy (LLLT), aimed at cell and tissue activation. In both, vaporization of tissue is induced by thermal damage. A water spray function may ameliorate thermal damage from CO2 laser irradiation. In this study, we irradiated CO2 laser on rat tibiae with or without a water spray function and examined the effects of this technique on bone metabolism. MATERIALS AND METHODS: Bone defects were created in rat tibiae by dental bur in a Bur group and by laser in laser irradiation groups with (Spray group) and without (Air group) water spray function. At 1 week postoperatively, histological analyses of tibiae were performed using hematoxylin and eosin staining, immunohistochemical staining (IHC) with anti-sclerostin antibody, and 3-dimensional (3D) observation using micro-computed tomography. RESULTS: Histological findings and 3D observation confirmed induction of new bone formation following laser irradiation in both the Air and Spray groups. No bone formation was seen in the Bur group. IHC revealed that the activity of osteocytes in the region of irradiated cortical bone was markedly impaired in the Air group, but osteocyte impairment was ameliorated in the Spray group and absent in the Bur group. CONCLUSION: The water spray function appears effective in reducing thermal damage to tissues irradiated by CO2 laser. CO2 lasers with water spray function may be useful in bone regeneration therapy.

Oxytocin Facilitates Dentinogenesis of Rat Dental Pulp Cells.

INTRODUCTION: Oxytocin (OT) is a neurohypophysial hormone that plays a role in lactation and parturition and exerts diverse biological actions via the OT receptor. Recently, several studies have reported that OT stimulates bone formation by osteoblasts in osteoporosis. We focused on OT and hypothesized that OT can stimulate the differentiation of odontoblasts as well as osteoblasts. The aim of this study was to verify whether OT is an essential factor in dentinogenesis; we examined the effects of OT on dentinogenesis using a long-term culture system of rat dental pulp cells. METHODS: Using a culture system of rat dental pulp cells with Otr knocked out by CRISPR-Cas9 genome editing, we examined the effects of OT on odontoblastlike cell differentiation as reflected by dentin formation. RESULTS: We confirmed that OT stimulated mineralized nodule formation and the expression of both dentin sialoprotein and bone Gla protein messenger RNAs (mRNAs) in the culture system. Interestingly, the cultured cells treated with OT also exhibited an increase of both Wnt10a and Lef-1 mRNA. The Otr knockout cells showed inhibition of nodule formation and mRNA expression, and these phenomena remained despite OT treatment. These results indicate the following: OT regulates odontoblastlike cell differentiation via the OT receptor, it stimulates dentin formation, and the Wnt canonical pathway is closely related to these effects. CONCLUSIONS: The present results suggest that OT can promote odontoblastlike cell differentiation, resulting in increased dentin formation, and that OT could be an important factor for dentinogenesis.

Enamel Matrix Derivative in Diffusion Chamber Implanted Subcutaneously in Rat Induces Formation of Fibrous Connective Tissue Containing Abundant Blood Vessels.

BACKGROUND: Enamel matrix derivative (EMD) is widely used for regeneration therapy in dental clinical situations, but the mechanism of EMD bioactivity remains obscure. To clarify this mechanism, we focused on the formation of connective tissue and blood vessels. The aim of this study was to confirm whether EMD induces the formation of connective tissue and blood vessels by using the diffusion chamber (DC) technique. MATERIALS AND METHODS: Individual DCs containing EMD (DC-EMD) or propylene glycol alginate (PGA) were implanted subcutaneously in rat dorsum. At 4 weeks after the implantation, histological analysis of DCs was performed using azan staining. RESULTS: DC-EMD induced the formation of much larger amounts of connective tissue containing abundant blood vessels than did DC-PGA. CONCLUSION: The results indicated that EMD can induce the formation of both connective tissue and blood vessels. This bioactivity may contribute to the mechanism whereby EMD induces tissue regeneration.