Abstract: Aims/Introduction: We previously reported that stem cell transplantation into limb skeletal muscle improved diabetic polyneuropathy (DPN) in diabetic animal models. In this study, we examined whether the secreted factors from stem cells from human exfoliated deciduous teeth (SHED) had beneficial effects on DPN. Materials and Methods: The conditioned medium from SHED(SHED-CM) was collected 48 hours after culturing in serum-free DMEM and was separated into four fractions according to molecular weight. Dorsal root ganglion (DRG) neurons isolated from C57BL/6J mice were cultured with SHED-CM, each fraction of SHED-CM or DMEM. Streptozotocin induced diabetic mice were injected with 100µl of SHED-CM or DMEM into unilateral hindlimb muscles twice a week over 4 weeks. Peripheral nerve functions were evaluated by the plantar test and motor and sensory nerve conduction velocities (MNCV and SNCV). Intraepidermal nerve fiber densities (IENFDs), capillary number-to-muscle fiber ratio (CNMFR) and morphometry of sural nerves were also evaluated. The angiogenic profile of SHED-CM was evaluated by MTT, transwell migration, and wound healing assay using human umbilical vein endothelial cells (HUVECs). Results: SHED-CM significantly promoted neurite outgrowth of DRG neurons. Only less than 6 kDa of SHED-CM promoted the neurite outgrowth. SHED-CM significantly prevented decline in SNCVs compared with DMEM in diabetic mice. SHED-CM did not cause any change in MNCVs or sensory functions. Though SHED-CM did not improve IENFDs or morphometry of sural nerves, CNMFR was ameliorated by the administration of SHED-CM. SHED-CM significantly increased the proliferation and the migration of HUVECs. Conclusions: These results suggested that SHED-CM had the therapeutic effect on DPN by promoting neurite outgrowths and improving microcirculation in peripheral nerves. Disclosure E. Miura-Yura: None. S. Tsunekawa: None. K. Naruse: None. M. Kawai: None. M. Kato: None. H. Shimoda: None. Y. Yamada: None. M. Motegi: None. S. Asano: None. T. Himeno: None. M. Kondo: None. Y. Kato: Speaker's Bureau; Self; Merck & Co., Inc. J. Nakamura: Research Support; Self; Astellas Pharma Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo Company, Limited, Eli Lilly and Company, Japan Tobacco Inc., Kissei Pharmaceutical Co., Ltd., Merck Sharp & Dohme Corp., Novartis Pharmaceuticals Corporation, Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Sanofi K.K., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited. Speaker's Bureau; Self; Astellas Pharma Inc., AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo Company, Limited, Eli Lilly and Company, Kowa Pharmaceu. Co. Ltd., Merck Sharp & Dohme Corp., Mitsubishi Tanabe Pharma Corporation, Novartis Pharmaceuticals Corporation, Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Sanofi K.K., Takeda Pharmaceutical Company Limited, Terumo Medical Corporation. H. Kamiya: Speaker's Bureau; Self; Astellas Pharma Inc., Eli Lilly Japan K.K., MSD K.K., Novartis Pharma K.K., Novo Nordisk Oharma Ltd., Ono Pharmaceutical Co., Ltd., Sanofi K.K.

Abstract: Background/Aim: We previously reported that conditioned medium (CM) of stem cells from human exfoliated deciduous teeth (SHED) improved diabetic polyneuropathy in diabetic mice due to an increase of capillary blood flow in the vasa nervorum, and in in vitro study only secreted factors of less than 6 kDa fraction of SHED-CM promoted neurite outgrowth of mouse dorsal root ganglion neurons. In this study, we examined whether SHED-CM had beneficial effects on angiogenesis. Methods: SHED-CM was collected 48 hours after culturing in serum-free DMEM, and was separated into two fractions according to molecular weight (less than 6 kDa and more than 6 kDa). Exosomes were isolated from SHED-CM by ultracentrifugation. Human umbilical-vein endothelial cells (HUVECs) were cultured with six different media (DMEM, DMEM with VEGF, SHED-CM, less than 6 kDa, more than 6 kDa or DMEM with exosomes) for 12-48 hours, then MTT assay, wound healing assay, Boyden chamber assay and tube formation assay were performed to evaluate the cell viability, migration ability and tube formation ability in HUVECs. Rat aortic ring assay and mouse Matrigel plug assay were performed to assess neovascularization and endothelial cell migration. Results: SHED-CM, especially more than 6 kDa fraction, significantly promoted cell viability, migration and tube formation in HUVECs, compared with DMEM. Meanwhile, less than 6 kDa fraction promoted only tube formation, and the effects of exosomes were negligible. In aortic ring assay and Matrigel plug assay, SHED-CM accelerated neovascularization and increased endothelial cell migration in HUVECs, and the effects of more than 6 kDa fraction were stronger than that of less than 6 kDa fraction. These data suggested that the secreted factors in more than 6 kDa fraction played an essential role in promoting angiogenesis. Conclusion: Soluble factors from SHED might have angiogenesis-promoting effects and hold promise for comprehensive clinical applications in vascular diseases and diabetic complications. Disclosure M. Kato: None. S. Tsunekawa: None. N. Nakamura: None. E. Miura-Yura: None. Y. Yamada: None. Y. Hayashi: None. R. Inoue: None. M. Mohiuddin: Other Relationship; Self; Abbott Japan Co. Ltd, ARKRAY, Astellas Pharma Inc., Astellas Pharma Inc., AstraZeneca K.K., Boehlinger Ingelheim Japan Co. Ltd., Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo, Daiichi Sankyo, Eli Lilly Japan K.K., Eli Lilly Japan K.K., Fukuda Denshi, Japan Tobacco Inc., Kaken Pharmaceutical Co., Ltd., Kissei Pharmaceutical Co., Ltd., Kowa pharmaceu. co. Ltd., Kowa Pharmaceu. Co., Ltd., Kyowa Kirin Co., Ltd., Mitsubishi Tanabe Pharma Corporation, Mitsubishi Tanabe Pharma Corporation, MSD K.K., MSD K.K., Mylan N. V., Novartis Pharma K.K., Novartis Pharma K.K., Novo Nordisk Pharma Ltd., Novo Nordisk Pharma Ltd., Ono Pharmaceu. Co., Ltd., Ono Pharmaceutical Co., Ltd., Otsuka Pharmaceutical Co., Ltd., Pfizer Japan Inc., Sanofi K.K., Sanofi K.K., Sanwa Kagaku Kenkyusho, Sanwa Kagaku Kenkyusho, Shionogi & Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited, Takeda Pharmaceutical Company Limited, Terumo Co. Ltd. Y. Morishita: None. T. Himeno: None. M. Kondo: None. Y. Kato: None. H. Kamiya: Speaker’s Bureau; Self; Astellas Pharma Inc., AstraZeneca K.K., Boehringer Ingelheim K.K., Daiichi Sankyo, Eli Lilly Japan K.K., Fukuda Denshi, Kissei Pharmaceutical Co., Ltd., Kowa Company, Ltd., Kyowa Hakko Kirin Co., Ltd., Mitsubishi Tanabe Pharma Corporation, MSD K.K., Novartis Pharma K.K., Novo Nordisk Pharma Ltd., Ono Pharmaceutical Co., Ltd., Sanofi K.K., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited. K. Naruse: None. J. Nakamura: Research Support; Self; Astellas Pharma Inc., Boehlinger Ingelheim Japan Co., Ltd., Daiichi Sankyo, Eli Lilly Japan K.K., Japan Tobacco Inc., Kaken Pharmaceutical Co., Ltd., Kowa Company, Ltd., Kyowa Hakko Kirin Co., Ltd., Mitsubishi Tanabe Pharma Corporation, MSD K.K., Novartis Pharma K.K., Novo Nordisk Pharma Ltd., Ono Pharmaceutical Co., Ltd., Otsuka Pharmaceutical Co., Ltd., Pfizer Japan Inc., Sanofi K.K., Sanwa Kagaku Kenkyusho, Shionogi & Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited. Speaker’s Bureau; Self; Abbott Japan Co., Ltd., ARKRAY, Astellas Pharma Inc., AstraZeneca K.K., Boehlinger Ingelheim Japan Co., Ltd.,, Daiichi Sankyo, Eli Lilly Japan K.K., Fukuda Denshi, Kissei Pharmaceutical Co., Ltd., Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation, MSD K.K., Mylan, Novartis Pharma K.K., Novo Nordisk Pharma Ltd, Ono Pharmaceutical Co., Ltd., Sanofi, Sanwa Kagaku Kenkyusho, Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited, Terumo Medical Corporation.

Abstract: In this study, we propose a novel DPN severity classification based on coefficient of variance of R-R interval on ECG (CVRR) and nerve conduction study (NCS). The subjects consisted of 182 patients (mean age 58.9 years-old; duration of diabetes 10.2 years, HbA1c 9.72% and BMI 26.0) with type 2 diabetes admitted to our hospital from 2015 to 2017. Based on NCS and CVRR during rest, the subjects were classified into six categories as follows; 1) N0C0: sural nerve sensory action potential (SP) & gt;5μV and tibial nerve compound motor action potential (CP) & gt;5mV + CVRR & gt;2%, 2) N1C0: SP & lt;5 and CP & gt;5 + CVRR & gt;2, 3) N2C0: SP & lt;5 and CP & lt;5 + CVRR & gt;2, 4) N0C1: SP & gt;5 and CP & gt;5 + CCRR & lt;2, 5) N1C1: SP & lt;5 and CP & gt;5 + CVRR & lt;2 and 6) N2C1: SP & lt;5 and CP & lt;5 + CR & lt;2. We examined the relationship between each category with urinary albumin excretion rate (logarithmic transformation: LogACR), eGFR, presence/absence of nephropathy (ACR & gt;300mg/gCre and/or eGFR & lt;30) or retinopathy, and PWV as an index of arteriosclerosis. For statistical analysis, one-way analysis of variance (ANOVA), multiple comparison (Tukey), covariance analysis and logistic regression analysis were used. The one-way analysis revealed significant differences between each category in terms of age, the duration of diabetes, LogACR, eGFR and PWV. LogACR and PWV were significantly higher in the N2C1 than in the N0C0 (p & lt;0.05). Even after correction with age, duration and gender, a significant difference (p=0.008) between groups was observed in LogACR and a trend in PWV (p=0.068). In the logistic regression analysis corrected by age, duration and gender, the odds ratio (OR) for retinopathy was 5.36 in N0C1, 11.10 in N1C0, 8.23 in N1C1 and 11.78 in N2C1 compared with N0C0 (p & lt;0.01). The OR for nephropathy in N0C1 was 6.56, 7.91 in N1C1, 15.54 in N2C0 and 23.55 in N2C1 (p & lt;0.05). Severity classification of DPN based on NCS and CVRR during rest was well co-related to other diabetic complications, suggesting the usefulness of this novel classification to evaluate the severity of DPN. Disclosure H. Shimoda: None. H. Kamiya: Other Relationship; Self; MSD K.K., Ono Pharmaceutical Co., Ltd., Sanofi K.K., AstraZeneca, Astellas Pharma KK, Eli Lilly and Company, Novartis Pharma K.K., Dainippon Sumitomo Pharma Co., Ltd, Boehringer Ingelheim Pharmaceuticals, Inc., Takeda Pharmaceutical Co., Ltd, Ono Pharmaceutical Co., Ltd.. A. Watarai: None. T. Himeno: None. Y. Shibata: None. M. Motegi: None. M. Kato: None. Y. Yamada: None. E. Miura-Yura: None. M. Kondo: None. S. Tsunekawa: None. Y. Kato: Speaker's Bureau; Self; Merck Sharp & Dohme Corp., Sanofi, Takeda Pharmaceutical Company, Eli Lilly Japan. J. Nakamura: Other Relationship; Self; Astellas Pharma US, Inc., AstraZeneca, Ono Pharmaceutical Co., Ltd., MSD K.K., Kyowa Hakko Kirin Co., Ltd., Kowa Pharmaceuticals America, Inc., Sanofi K.K., Taisho Pharmaceutical Co., Ltd., Takeda Development Center Asia, Pte. Ltd., Mitsubishi Tanabe Pharma Corporation, Eli Lilly and Company, Novartis Pharma K.K., Pfizer Inc..

Abstract: Introduction: It is indicated that sodium-glucose cotransporter 2 inhibitors (SGLT2i), glucose lowering agents, may increase a level of glucagon, and simultaneously, compete with their antiglycemic effect. Aim: To elucidate the alteration of glucagon concentration and blood glucose level during administration of SGLT2i, Empagliflozin (EMPA) was administered to diabetic mice with homozygous (gcg−/−) or heterozygous (gcg+/−) deficiency of glucagon gene. Methods: After induction of diabetes using 150 mg/kg of streptozotocin, gcg−/− and control gcg+/− mice of 12-14 weeks old were administered 30 mg/kg/day of EMPA for 3-5 weeks. Casual blood glucose, fasting blood glucose, oral glucose tolerance test (OGTT), insulin tolerance test (ITT), blood glucagon, blood insulin, blood beta-hydroxybutyric acid were evaluated. Additionally, metabolic parameters including respiratory exchange ratio (RER), activity, feeding, and drinking, were monitored utilizing a home-cage chronic laboratory animal monitoring system (CLAMS). Results: Glucagon concentration in control gcg+/− mice were not altered by induction of diabetes or administration of EMPA. In gcg−/−, blood glucagon was not detected. The OGTT of diabetic gcg+/− and gcg−/− revealed that the area under the curve (AUC) of blood glucose in mice treated with EMPA were decreased compared with that without EMPA (p & lt;0.05). However, reduction ratios of the AUC showed no significant difference between gcg+/− and gcg−/−. In ITT, there was no significant difference in the reduction rate of glucose levels between mice treated with or without EMPA. CLAMS revealed no significant difference induced by EMPA in metabolic parameters: RER, activity, food intake, and drinking. Conclusion: Administration of EMPA for almost one month in mice did not induce noticeable change in glucagon level or metabolic parameters. The gcg deficiency did not have any impact on glucose lowering effect of EMPA. Disclosure H. Shimoda: None. Y. Seino: None. T. Himeno: None. R. Inoue: None. M. Motegi: None. T. Hayami: None. E. Asano: None. S. Asano: None. M. Kato: None. Y. Yamada: None. E. Miura-Yura: None. M. Kondo: None. S. Tsunekawa: None. Y. Kato: Speaker's Bureau; Self; Merck & Co., Inc. K. Kato: None. Y. Hayashi: Speaker's Bureau; Self; Astellas Pharma Inc., Merck & Co., Inc., Novartis Pharmaceuticals Corporation, Novo Nordisk Inc., Sumitomo Dainippon Pharma Co., Ltd. J. Nakamura: Research Support; Self; Astellas Pharma Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo Company, Limited, Eli Lilly and Company, Japan Tobacco Inc., Kissei Pharmaceutical Co., Ltd., Merck Sharp & Dohme Corp., Novartis Pharmaceuticals Corporation, Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Sanofi K.K., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited. Speaker's Bureau; Self; Astellas Pharma Inc., AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo Company, Limited, Eli Lilly and Company, Kowa Pharmaceu. Co. Ltd., Merck Sharp & Dohme Corp., Mitsubishi Tanabe Pharma Corporation, Novartis Pharmaceuticals Corporation, Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Sanofi K.K., Takeda Pharmaceutical Company Limited, Terumo Medical Corporation. H. Kamiya: Speaker's Bureau; Self; Astellas Pharma Inc., Eli Lilly Japan K.K., MSD K.K., Novartis Pharma K.K., Novo Nordisk Oharma Ltd., Ono Pharmaceutical Co., Ltd., Sanofi K.K.

Abstract: Nerve conduction study (NCS) is a reliable examination to diagnose and to evaluate diabetic symmetric sensorimotor polyneuropathy (DSPN). However, NCS is not feasible in daily medical practice, because it requires skilled laboratory technicians and an expensive equipment. A point-of-care device, DPNCheck™can measure sural nerve (SN) action potential amplitude (Amp) and conduction velocity (CV) in less than 5 minutes without special techniques. Here, we evaluated the accuracy and the usefulness of DPNCheck™ for diagnosis of DSPN in Japanese diabetic patients. We selected 207 diabetic patients. 1. We examined bilateral SN both by electromyography (EM) and DPNCheck™ and compared the differences. 2. To evaluate the reproducibility and inter-rater reliability of DPNCheck™, Amp and CV of SN were measured twice by an examiner and once by another examiner. 3. Based on the results of EM, we classified the severity of DSPN into 3 grades as follows; grade 0: Amp of SN (SNAP) 5μV or more and Amp of tibial nerve (CMAP) 5mV or more, grade 1: SNAP & lt;5μV and CMAP 5mV or more, and grade 2: SNAP & lt;5μV and CMAP & lt;5mV. Then, a formula to predict the grades of DSPN severity by using the Amp and CV from DPNCheck™ was created. For statisticalanalyses SPSS was used. 1. The correlation coefficient (R) for CV and Amp between DPNCheck™ and EM were R=0.7663 and 0.6178, respectively. 2. The reproducibility and inter-rater reliability of DPNCheckTM were “superior” and “acceptable”, respectively. 3. The formula to predict the severity of DSPN by DPNCheck™ was as follows; “grade = 2.235+0.009xAge-0.015xCV-0.041xAmp.” The grade calculated by this formula was relatively “acceptable” compared with the real grade determined by EM (correlation coefficient: R=0.645). These results indicate that DPNCheck™ is a useful device to measure SN functions in Japanese diabetic patients and should be used more frequently in the bedside to diagnose and evaluate DSPN. Disclosure Y. Shibata: None. H. Kamiya: Other Relationship; Self; MSD K.K., Ono Pharmaceutical Co., Ltd., Sanofi K.K., AstraZeneca, Astellas Pharma KK, Eli Lilly and Company, Novartis Pharma K.K., Dainippon Sumitomo Pharma Co., Ltd, Boehringer Ingelheim Pharmaceuticals, Inc., Takeda Pharmaceutical Co., Ltd, Ono Pharmaceutical Co., Ltd.. T. Himeno: None. M. Motegi: None. H. Shimoda: None. M. Kato: None. Y. Yamada: None. E. Miura-Yura: None. M. Kondo: None. S. Tsunekawa: None. Y. Kato: Speaker's Bureau; Self; Merck Sharp & Dohme Corp., Sanofi, Takeda Pharmaceutical Company, Eli Lilly Japan. J. Nakamura: Other Relationship; Self; Astellas Pharma US, Inc., AstraZeneca, Ono Pharmaceutical Co., Ltd., MSD K.K., Kyowa Hakko Kirin Co., Ltd., Kowa Pharmaceuticals America, Inc., Sanofi K.K., Taisho Pharmaceutical Co., Ltd., Takeda Development Center Asia, Pte. Ltd., Mitsubishi Tanabe Pharma Corporation, Eli Lilly and Company, Novartis Pharma K.K., Pfizer Inc..

Abstract: Background: Glucagon gene-derived peptides are produced as glucagon-like peptide 1 (GLP-1) in intestinal L cells and glucagon in pancreatic α cells, and play an important physiological role. However, the physiological roles of those peptides in various tissues are not fully elucidated. In this study, we examined the physiological roles of these peptides in the peripheral nervous system (PNS). Methods: Expression of glucagon receptor in the PNS was examined in 18-21 week-old C57BL6/J (WT) mice utilizing immunohistochemical staining. Neurite outgrowth was assessed after 24-48 hours of primary culture of dorsal root ganglion (DRG) neurons supplemented with or without glucagon (10⁻⁷ to 10⁻⁵ mol/l). Furthermore, the sensory function of the plantar pedis was evaluated by thermal plantar test in glucagon gene deficient mice (gcg+/⁻, gcg⁻/⁻) of 8, 12, 18, 24, and 30 week-old. Results: The immunohistological staining validated the expression of glucagon receptor in most DRG neurons and satellite glial cells. In the primary culture of DRG neurons, the neurite length was significantly increased in the group cultured with glucagon. (control: 761.4±294.9 μm/cell, glucagon 10⁻⁵ mol/l: 2276.5±835.6 μm/cell, p & lt;0.01). The thermal plantar test revealed a hypersensitivity in 12 week-old gcg-/- mice (WT: 8.8 second, gcg ⁺/⁻: 6.9, gcg⁻/⁻: 5.5, p & lt; 0.01: WT vs. gcg⁻/⁻) and in 18 week-old gcg-/- and gcg+/- mice (WT: 8.8, gcg⁺/⁻: 7.1, gcg⁻/⁻: 6.6, p & lt; 0.01: WT vs. gcg⁺/⁻ or gcg⁻/⁻). However, the hypersensitivity disappeared in 24 week-old mice (WT: 8.4, gcg+/-: 8.9, gcg⁻/⁻: 8.5). Discussion: These data suggested that glucagon may have a protective role for DRG neurons. Further investigation should be performed to elucidate the role of glucagon gene-derived peptides in the PNS in the future. Disclosure M. Motegi: None. T. Himeno: None. H. Kamiya: Other Relationship; Self; MSD K.K., Ono Pharmaceutical Co., Ltd., Sanofi K.K., AstraZeneca, Astellas Pharma KK, Eli Lilly and Company, Novartis Pharma K.K., Dainippon Sumitomo Pharma Co., Ltd, Boehringer Ingelheim Pharmaceuticals, Inc., Takeda Pharmaceutical Co., Ltd, Ono Pharmaceutical Co., Ltd.. H. Shimoda: None. M. Kato: None. Y. Yamada: None. E. Miura-Yura: None. M. Kondo: None. S. Tsunekawa: None. Y. Kato: Speaker's Bureau; Self; Merck Sharp & Dohme Corp., Sanofi, Takeda Pharmaceutical Company, Eli Lilly Japan. Y. Hayashi: Other Relationship; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Kissei Pharmaceutical Co., Ltd., Takeda Pharmaceuticals Co., Ltd., Novartis Pharma K.K., Novo Nordisk A/S, Astellas Pharma US, Inc. J. Nakamura: Other Relationship; Self; Astellas Pharma US, Inc., AstraZeneca, Ono Pharmaceutical Co., Ltd., MSD K.K., Kyowa Hakko Kirin Co., Ltd., Kowa Pharmaceuticals America, Inc., Sanofi K.K., Taisho Pharmaceutical Co., Ltd., Takeda Development Center Asia, Pte. Ltd., Mitsubishi Tanabe Pharma Corporation, Eli Lilly and Company, Novartis Pharma K.K., Pfizer Inc..

Abstract: Objective and Methods: Eating behavior questionnaire issued by the Japan Society for the Study of Obesity was administered to 41 patients who received semaglutide injection (SI) and 52 patients who received oral semaglutide (OS) in our outpatient clinic. Changes in HbA1c levels, body weight, eating behavior, and their correlations after 3 months of treatment were examined. Results: The mean dose after 3 months of treatment was 0.50 ± 0.17 mg/week in the SI group and 6.35 ± 2.29 mg/day in the OS group. Mean HbA1c significantly decreased from 7.34 ± 0.97% to 6.85 ± 0.72% in the SI group and from 7.65 ± 0.94% to 7.± 0.91% in the OS group (p & lt;0.001) . Weight significantly decreased from 79.1 ± 16.3 kg to 76.8 ± 16.8 kg in the SI group and from 74.8 ± 15.2 kg to 73.2 ± 15.4 kg in the OS group (p & lt;0.001) . In the SI group, “Recognition for weight and constitution,” “External eating behavior,” “Sense of hunger,” “Eating style,” “Food preference,” “Regularity of eating habit” and the total scores of the questionnaire significantly improved after 3 months, while in the OS group, all factors improved significantly. In the SI group, there was a significant positive correlation between the changes in HbA1c level over 3 months and the changes in scores for “Regularity of eating habit” (r=0.401, p=0.013) and between the changes in weight and the changes in total scores (r=0.330, p=0.043) . In the OS group, the changes in weight was significantly and positively correlated with the changes in scores for “External eating behavior,” “Emotional eating behavior,” “Sense of hunger,” and “Regularity of eating habit,” and the changes in total scores. Adverse events were nausea (22.6%) , diarrhea (14.0%) , constipation and stomach pain (5.4% each) , fatigue (3.2%) , abdominal distension (2.2%) , soft stool, abdominal pain, and headache (1.1% each) in the total. Conclusion: These results suggest that semaglutide improves glycemic control and eating behavior in Japanese patients with type 2 diabetes. Disclosure S.T.Sato: None. H.Kamiya: Research Support; Abbott Japan Co., Ltd., Eli Lilly and Company, Japan Tobacco Inc., Kissei Pharmaceutical Co., Ltd., Kowa Company, Ltd., Merck & Co., Inc., Mitsubishi Tanabe Pharma Corporation, Novo Nordisk, Ono Pharmaceutical Co., Ltd., Sanwa Kagaku Kenkyusho, Sumitomo Dainippon Pharma Co., Ltd., Taiho Pharmaceutical Co. Ltd., Takeda Pharmaceutical Company Limited, Terumo Corporation, Speaker's Bureau; Astellas Pharma Inc., AstraZeneca, Boehringer Ingelheim International GmbH, Daiichi Sankyo, Eli Lilly and Company, Kissei Pharmaceutical Co., Ltd., Kowa Company, Ltd., Merck & Co., Inc., Mitsubishi Tanabe Pharma Corporation, Novartis Pharma K.K., Novo Nordisk, Ono Pharmaceutical Co., Ltd., Sanofi K.K., Sanwa Kagaku Kenkyusho, Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Holdings Co., Ltd. T.Tosaki: Other Relationship; Eli Lilly and Company, Novo Nordisk, Sumitomo Dainippon Pharma Co., Ltd. A.Kotorii: None. M.Kondo: n/a. E.Miura-yura: None. S.Tsunekawa: n/a. T.Himeno: None. Y.Kato: None. J.Nakamura: None.

Abstract: We previously reported that stem cell transplantation into limb skeletal muscle improved diabetic polyneuropathy (DPN) in diabetic animal models without the differentiation of transplanted cells into neuron, suggesting that the improvement was due to a paracrine effect. The aim of our study is to examine whether the secreted factors from dental pulp stem cells from human exfoliated deciduous teeth (SHED) has a beneficial effect on DPN. Conditioned medium from SHED (SHED-CM) was collected 48 hours after culturing in serum-free DMEM. Incubation of dorsal root ganglion (DRG) neurons excised from C57BL/6J mice with SHED-CM significantly promoted neurite outgrowth compared that with DMEM. (SHED-CM: 3793.2±907.9 um/neuron, DMEM: 370.9±321.7 um/neuron) Among 4 fractions of SHED-CM according to molecular weight (less than 6kDa, 6-20kDa, 20-100kDa, and more than 100kDa), only fraction of less than 6kDa significantly increased neurite outgrowth. Incubation of DRG neurons with exosome isolated from SHED-CM didn’t have any effect on neurite outgrowth, indicating that only soluble factors from SHED-CM contributed to neurite outgrowth of DRG neurons. In in vivo study, 12 weeks after streptozotosin (STZ) administration, 100ul of SHED-CM or DMEM was injected into unilateral lower limb muscle of STZ-induced diabetic mice twice a week over a 4 weeks period. SHED-CM or DMEM injections didn’t change serum glucose levels and body weight in diabetic mice. SHED-CM significantly prevented decline in sensory nerve conduction velocity (SNCV), compared with DMEM. In diabetic mice, capillary number-to-muscle fiber ratio (CNMFR) and intraepidermal nerve fiber densities (IENFDs) decreased less than those in nondiabetic mice. SHED-CM significantly improved the decline of CNMFR, but not IENFDs, suggesting that SHED-CM in in vivo alleviated SNCV by improving of nerve blood flow. These data suggested that SHED-CM might have potential as a novel strategy for treatment of DPN. Disclosure E. Miura-Yura: None. S. Tsunekawa: None. T. Himeno: None. K. Naruse: None. M. Motegi: None. H. Shimoda: None. M. Kato: None. Y. Yamada: None. M. Kondo: None. Y. Kato: Speaker's Bureau; Self; Merck Sharp & Dohme Corp., Sanofi, Takeda Pharmaceutical Company, Eli Lilly Japan. H. Kamiya: Other Relationship; Self; MSD K.K., Ono Pharmaceutical Co., Ltd., Sanofi K.K., AstraZeneca, Astellas Pharma KK, Eli Lilly and Company, Novartis Pharma K.K., Dainippon Sumitomo Pharma Co., Ltd, Boehringer Ingelheim Pharmaceuticals, Inc., Takeda Pharmaceutical Co., Ltd, Ono Pharmaceutical Co., Ltd. J. Nakamura: Other Relationship; Self; Astellas Pharma US, Inc., AstraZeneca, Ono Pharmaceutical Co., Ltd., MSD K.K., Kyowa Hakko Kirin Co., Ltd., Kowa Pharmaceuticals America, Inc., Sanofi K.K., Taisho Pharmaceutical Co., Ltd., Takeda Development Center Asia, Pte. Ltd., Mitsubishi Tanabe Pharma Corporation, Eli Lilly and Company, Novartis Pharma K.K., Pfizer Inc..

Abstract: Background: Glucagon is a peptide hormone produced by the alpha cells of the pancreas. Other than gluconeogenesis, glucagon has many pleiotropic effects on feeding, energy expenditure, and thermogenesis. Our recent studies revealed that glucagon promoted neurite outgrowth in a primary culture of mouse dorsal root ganglion (DRG) neurons. Therefore, glucagon might have a neuroprotective function in the peripheral nervous system. To clarify the protective function, we use an immortalized rat embryonic DRG neuronal cell line 50B11 as an in vitro model to study the effects of glucagon in DRG neurons. Methods: Expression of glucagon receptor in the DRG neurons was examined by RT-PCR. The cells were cultured with or without glucagon (10-8, 10-10 M) for 24 hours. To reproduce the cell stress, cells were cultured with methylglyoxal (MG). Cell cytotoxicity was examined by lactate dehydrogenase assay. Cell viability was evaluated using the MTS assay. The apoptosis was also examined by APOPercentageTM dye. Result: Glucagon receptor was expressed in the 50B11 cell line. The glucagon itself had no effect on cytotoxicity in the cells. The cell viability decreased by MG was significantly increased in the group treated with glucagon. The ratio of apoptotic cells, which was increased by MG, was decreased in the group treated with glucagon. Conclusion: Glucagon may have a protective role in DRG neurons. Further investigation is necessary to determine the detailed action of glucagon in DRG neurons. Disclosure M. Mohiuddin: Other Relationship; Self; Abbott Japan Co. Ltd, ARKRAY, Astellas Pharma Inc., Astellas Pharma Inc., AstraZeneca K.K., Boehlinger Ingelheim Japan Co. Ltd., Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo, Daiichi Sankyo, Eli Lilly Japan K.K., Eli Lilly Japan K.K., Fukuda Denshi, Japan Tobacco Inc., Kaken Pharmaceutical Co., Ltd., Kissei Pharmaceutical Co., Ltd., Kowa pharmaceu. co. Ltd., Kowa Pharmaceu. Co., Ltd., Kyowa Kirin Co., Ltd., Mitsubishi Tanabe Pharma Corporation, Mitsubishi Tanabe Pharma Corporation, MSD K.K., MSD K.K., Mylan N. V., Novartis Pharma K.K., Novartis Pharma K.K., Novo Nordisk Pharma Ltd., Novo Nordisk Pharma Ltd., Ono Pharmaceu. Co., Ltd., Ono Pharmaceutical Co., Ltd., Otsuka Pharmaceutical Co., Ltd., Pfizer Japan Inc., Sanofi K.K., Sanofi K.K., Sanwa Kagaku Kenkyusho, Sanwa Kagaku Kenkyusho, Shionogi & Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited, Takeda Pharmaceutical Company Limited, Terumo Co. Ltd. M. Motegi: None. R. Inoue: None. T. Mizuno: None. K. Suzuki: None. S. Asano: None. T. Hayami: None. Y. Hayashi: None. M. Kato: None. E. Miura-Yura: None. H. Shimoda: None. Y. Morishita: None. T. Himeno: None. M. Kondo: None. S. Tsunekawa: None. Y. Kato: None. H. Kamiya: Speaker’s Bureau; Self; Astellas Pharma Inc., AstraZeneca K.K., Boehringer Ingelheim K.K., Daiichi Sankyo, Eli Lilly Japan K.K., Fukuda Denshi, Kissei Pharmaceutical Co., Ltd., Kowa Company, Ltd., Kyowa Hakko Kirin Co., Ltd., Mitsubishi Tanabe Pharma Corporation, MSD K.K., Novartis Pharma K.K., Novo Nordisk Pharma Ltd., Ono Pharmaceutical Co., Ltd., Sanofi K.K., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited. J. Nakamura: Research Support; Self; Astellas Pharma Inc., Boehlinger Ingelheim Japan Co., Ltd., Daiichi Sankyo, Eli Lilly Japan K.K., Japan Tobacco Inc., Kaken Pharmaceutical Co., Ltd., Kowa Company, Ltd., Kyowa Hakko Kirin Co., Ltd., Mitsubishi Tanabe Pharma Corporation, MSD K.K., Novartis Pharma K.K., Novo Nordisk Pharma Ltd., Ono Pharmaceutical Co., Ltd., Otsuka Pharmaceutical Co., Ltd., Pfizer Japan Inc., Sanofi K.K., Sanwa Kagaku Kenkyusho, Shionogi & Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited. Speaker’s Bureau; Self; Abbott Japan Co., Ltd., ARKRAY, Astellas Pharma Inc., AstraZeneca K.K., Boehlinger Ingelheim Japan Co., Ltd.,, Daiichi Sankyo, Eli Lilly Japan K.K., Fukuda Denshi, Kissei Pharmaceutical Co., Ltd., Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation, MSD K.K., Mylan, Novartis Pharma K.K., Novo Nordisk Pharma Ltd, Ono Pharmaceutical Co., Ltd., Sanofi, Sanwa Kagaku Kenkyusho, Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited, Terumo Medical Corporation.