Freeze-Dried Human Platelet-Rich Plasma Retains Activation and Growth Factor Expression after an Eight-Week Preservation Period

Yasuhiro Shiga,Go Kubota,Sumihisa Orita,Kazuhide Inage,Hiroto Kamoda,Masaomi Yamashita,Toru Iseki,Michihiro Ito,Kazuyo Yamauchi,Yawara Eguchi,Takeshi Sainoh,Jun Sato,Kazuki Fujimoto,Koki Abe,Hirohito 대한척추외과학회 2017 Asian Spine Journal Vol.11 No.3

Study Design: Controlled laboratory study. Purpose: This study aimed to evaluate the efficacy of platelet-rich plasma (PRP) stored at room temperature (RT), frozen, or after freeze-drying. Overview of Literature: PRP enriches tissue repair and regeneration, and is a novel treatment option for musculoskeletal pathologies. However, whether biological activity is preserved during PRP storage remains uncertain. Methods: PRP was prepared from blood of 12 healthy human volunteers (200 mL/person) and stored using three methods: PRP was stored at RT with shaking, PRP was frozen and stored at –80°C, or PRP was freeze-dried and stored at RT. Platelet counts and growth factor content were examined immediately after preparation, as well as 2, 4, and 8 weeks after storage. Platelet activation rate was quantified by flow cytometry. Results: Platelet counts were impossible to determine in many RT samples after 2 weeks, but they remained at constant levels in frozen and freeze-dried samples, even after 8 weeks of storage. Flow cytometry showed approximately 80% activation of the platelets regardless of storage conditions. Almost no growth factors were detected in the RT samples after 8 weeks, while low but significant expression was detected in the frozen and freeze-dried PRP. Over time, the mean relative concentrations of various growth factors decreased significantly or disappeared in the RT group. In the frozen group, levels were maintained for 4 weeks, but decreased significantly by 8 weeks (p <0.05). The freeze-dried group maintained baseline levels of growth factors for the entire 8-week duration. Conclusions: Freeze-drying enables PRP storage while maintaining bioactivity and efficacy for extended periods.

Time-Course Changes in Bone Metabolism Markers and Density in Patients with Osteoporosis Treated with Romosozumab: A Multicenter Retrospective Study

Kazuhide Inage,Sumihisa Orita,Yawara Eguchi,Yasuhiro Shiga,Masao Koda,Yasuchika Aoki,Toshiaki Kotani,Tsutomu Akazawa,Takeo Furuya,Junichi Nakamura,Hiroshi Takahashi,Miyako Suzuki-Narita,Satoshi Maki,S 연세대학교의과대학 2021 Yonsei medical journal Vol.62 No.9

Purpose: In this multicenter retrospective observational study, we examined the early effects of romosozumab in patients with severeosteoporosis in terms of time-course changes in bone metabolism marker, improvement in bone density, and adverse effects. Materials and Methods: Patients with severe osteoporosis were included. We investigated the progress of TRACP 5b and P1NPbefore and 1–2 months after the administration of romosozumab. We also investigated the bone density of lumbar spine, femoralneck, and the entire femur, measured by the DXA method, before and 5–7 months after the administration of romosozumab. Results: A total of 70 patients (7 males and 63 females, age 75.0±3.6 years) participated in this study. Significant improvements inTRACP 5b and P1NP levels were observed before and 1–2 months after romosozumab administration. The average bone densityof lumbar spine, femoral neck, and the entire femur were measured before and 5–7 months after romosozumab administration;and a significant increase only observed in the lumbar spine. Conclusion: Consistent with the findings of previous clinical studies, romosozumab has both bone formation-enhancing andbone resorption effects (dual effect). In addition, romosozumab also demonstrated improvement in bone density from the earlyphase after the administration, though the result was only seen in the lumbar spine.

Diffusion Tensor Imaging of the Spinal Canal in Quantitative Assessment of Patients with Lumbar Spinal Canal Stenosis

Norimoto Masaki,Eguchi Yawara,Kanamoto Hirohito,Oikawa Yasuhiro,Matsumoto Koji,Masuda Yoshitada,Furuya Takeo,Orita Sumihisa,Inage Kazuhide,Maki Satoshi,Shiga Yasuhiro,Kinoshita Hideyuki,Abe Koki,Inoue 대한척추외과학회 2021 Asian Spine Journal Vol.15 No.2

Study Design: Retrospective observational study. Purpose: Lumbar spinal stenosis (LSS) has traditionally been evaluated morphologically, there is a paucity of literature on quantitative assessment of LSS. The purpose of this study was to investigate whether intraspinal diffusion tensor imaging (DTI) parameters such as apparent diffusion coefficient (ADC) and fractional anisotropy (FA) are useful for assessing LSS. Overview of Literature: Quantitative assessment of LSS is challenging. Methods: Study participants comprised five healthy volunteers (mean age, 27.2 years) and 27 patients with LSS (mean age, 58.4 years) who were individually assessed using 3.0 Tesla magnetic resonance imaging. Intraspinal ADC and FA values of 10 intervertebral discs from healthy volunteers and 52 intervertebral discs from LSS patients were measured. Also, intraspinal canal area, Schizas classification (A: normal, B: mild stenosis, C: severe stenosis) and correlations with symptoms were investigated. Clinical symptoms were checked for the presence of low back pain (LBP), intermittent claudication (IMC), and bladder and bowel dysfunction (BBD). Results: Compared to healthy individuals, LSS patients had significantly lower ADC (p <0.05) and significantly higher FA values (p <0.01). In Schizas classification, stenosis worsened from A to C. ADC values decreased significantly while FA values increased significantly in that order (p <0.05). A positive correlation was found between intraspinal canal area and ADC values (r =0.63, p <0.01) and a negative correlation between intraspinal canal area and FA values (p =−0.61, p <0.01). No correlations were noted between LBP and ADC or FA values. On the other hand, ADC values were significantly lower (p <0.05) and FA values were significantly higher (p <0.05) in patients with IMC or BBD. Conclusions: Intraspinal DTI parameters such as ADC and FA values were associated with the Schizas classification, intraspinal canal area, and clinical symptoms, suggesting that ADC and FA may be useful for quantitative assessment of LSS.

Association between Osteoporosis and Skeletal Muscle Mass in Men

Mizutani Masaya,Eguchi Yawara,Toyoguchi Toru,Orita Sumihisa,Inage Kazuhide,Shiga Yasuhiro,Maki Satoshi,Nakamura Junichi,Hagiwara Shigeo,Aoki Yasuchika,Inoue Masahiro,Koda Masao,Takahashi Hiroshi,Akaza 대한척추외과학회 2024 Asian Spine Journal Vol.18 No.1

Study Design: Cross-sectional study.Purpose: This cross-sectional study aimed to investigate the risk factors for osteoporosis in men by assessing bone mineral density (BMD), skeletal muscle mass, body fat mass, grip strength, and advanced glycation end products (AGEs).Overview of Literature: Fewer studies have reported the correlation between BMD and skeletal muscle mass in women. Moreover, a few studies have examined the relationship between osteoporosis and skeletal muscle mass.Methods: This study included 99 men (mean age, 74.9 years; range, 28–93 years) who visited Qiball Clinic for BMD and body composition examinations. The osteoporosis group consisted of 24 patients (mean age, 72.5 years; range, 44–92 years), and the control group consisted of 75 individuals (mean age, 74.9 years; range, 28–93 years). Whole-body skeletal muscle mass was measured using a bioelectrical impedance analyzer. BMD was measured by dual X-ray absorptiometry. Skin autofluorescence (SAF), a marker of dermal AGE accumulation, was measured using a spectroscope. Osteoporosis was defined as a bone density T score of –2.5 or less. Physical findings, skeletal muscle mass, BMD, grip strength, and SAF were compared between the osteoporosis and control groups.Results: The osteoporosis group had significantly lower trunk muscle mass (23.1 kg vs. 24.9 kg), lower leg muscle mass (14.4 kg vs. 13.0 kg), and skeletal mass index (7.1 kg/m² vs. 6.7 kg/m²) than the control group (all <i>p</i><0.05). Lower limb muscle mass was identified as a risk factor for osteoporosis in men (odds ratio, 0.64; <i>p</i>=0.03).Conclusions: Conservative treatment of osteoporosis in men will require an effective approach that facilitates the maintenance or strengthening of skeletal muscle mass, including exercise therapy with a focus on lower extremities and nutritional supplementation.

Improvements in Intractable Lumbar and LowerExtremity Symptoms after Systemic Administration of Tocilizumab, an Anti-interleukin-6 Receptor Antibody

Sainoh Takeshi,Orita Sumihisa,Miyagi Masayuki,Suzuki-Narita Miyako,Sakuma Yoshihiro,Oikawa Yasuhiro,Kubota Go,Sato Jun,Shiga Yasuhiro,Fujimoto Kazuki,Eguchi Yawara,Koda Masao,Aoki Yasuchika,Akazawa Ts 대한척추외과학회 2022 Asian Spine Journal Vol.16 No.1

Study Design: Prospective cohort study (open-label, single-arm, and non-blinded).Purpose: This study aims to determine the effects of systemic administration of tocilizumab, an anti-interleukin-6 (IL-6) receptor antibody on refractory low back pain and leg symptoms.Overview of Literature: IL-6 overexpression is associated with neuropathic pain pathogenesis, which is potentially followed by chronic low back pain, including leg pain and numbness. This finding suggest that inhibition of IL-6 at the site of pain or in the transmission pathway could provide novel therapeutic targets for chronic low back pain.Methods: This prospective, single-arm study included 11 patients (eight men; mean age, 62.7 years) with ≥3-months’ chronic pain history due to lumbar disease. Subcutaneous TCZ injections were administered twice, at a 2-week interval. We evaluated low back pain, leg pain, and leg numbness using numeric rating scales and the Oswestry Disability Index (ODI; baseline and 6 months postinjection); serum IL-6 and tumor necrosis factor-α levels (baseline and 1 month postinjection); and clinical adverse events.Results: Intractable symptoms reduced after TCZ administration. Low back pain improved for 6 months. Improvements in leg pain and numbness peaked at 4 and 1 month, respectively. Improvements in ODI were significant at 1 month and peaked at 4 months. Serum IL-6 was increased at 1 month. IL-6 responders (i.e., patients with IL-6 increases >10 pg/mL) showed particularly significant improvements in leg pain at 2 weeks, 1 month, and 2 months compared with nonresponders. We observed no apparent adverse events.Conclusions: Systemic TCZ administration improved symptoms effectively for 6 months, with peak improvements at 1–4 months and no adverse events. Changing serum IL-6 levels correlated with leg pain improvements; further studies are warranted to elucidate the mechanistic connections between lumbar disorders and inflammatory cytokines.

The Time Course Changes in Bone Metabolic Markers after Administering the Anti-Receptor Activator of Nuclear Factor-Kappa B Ligand Antibody and Drug Compliance among Patients with Osteoporosis

Kazuhide Inage,Sumihisa Orita,Kazuyo Yamauchi,Yoshihiro Sakuma,Go Kubota,Yasuhiro Oikawa,Takeshi Sainoh,Jun Sato,Kazuki Fujimoto,Yasuhiro Shiga,Kazuhisa Takahashi,Seiji Ohtori 대한척추외과학회 2015 Asian Spine Journal Vol.9 No.3

Study Design: Retrospective study. Purpose: We conducted a study to investigate the time course changes in bone metabolic markers after the administration of the anti-receptor activator of nuclear factor-kappa B ligand (RANKL) antibody and to assess drug compliance among osteoporotic patients. Overview of Literature: The anti-RANKL antibody is expected to provide an improvement in those with a bone metabolism disorder. However there are only a few clinical reports available on the effect of treatment. Methods: We included 40 post-menopausal osteoporotic patients who received the anti-RANKL antibody. To determine the time course changes in the bone metabolic markers, we measured the serum tartrate-resistant acid phosphatase 5b (TRACP 5b; a bone resorption marker) and the serum N-terminal propeptide of type 1 collagen (P1NP; a bone formation marker) levels prior to and 1 month after administrating the anti-RANKL antibody. To evaluable drug compliance, we assessed the dropout rate during treatment and at 6 months after treatment. Results: The average TRACP 5b level significantly decreased from 574.8 mU/dL before treatment to 153.2 mU/dL 1 month after treatment (p <0.05). There was no significant difference in the average P1NP level, which was 56.9 μG/L and 35.1 μG/L before and 1 month after treatment, respectively (p >0.05). As for drug compliance, we did not have any dropouts during the treatment or after 6 months (dropout rate: 0%). Conclusions: Our study suggests that anti-RANKL antibody treatment suppresses bone resorption and maintains bone formation.

Freeze-Dried Platelet-Rich Plasma Induces Osteoblast Proliferation via Platelet-Derived Growth Factor Receptor-Mediated Signal Transduction

Hideyuki Kinoshita,Sumihisa Orita,Kazuhide Inage,Kazuki Fujimoto,Yasuhiro Shiga,Koki Abe,Masahiro Inoue,Masaki Norimoto,Tomotaka Umimura,Takeshi Ishii,Tsukasa Yonemoto,Hiroto Kamoda,Toshinori Tsukanis 대한척추외과학회 2020 Asian Spine Journal Vol.14 No.1

Study Design: Controlled laboratory study. Purpose: This study aimed to evaluate the in vitro pharmacological activity of growth factors (GFs) in freeze-dried platelet-rich plasma (FD-PRP) after storage for 4 weeks. Overview of Literature: Freshly prepared PRP is a rich source of many GFs. We reported that FD-PRP stored for 8 weeks accelerated bone union in a rat posterolateral fusion model equally well as fresh-PRP. However, the pharmacological activity of FD-PRP after long-term storage has not been shown in vitro. Methods: Immediately after preparation, as well as 4 weeks after freeze-dried storage, the platelet count was measured. Human osteoblasts were treated with fresh-PRP and FD-PRP, respectively. Western blotting was used to assess the phosphorylation of the platelet-derived growth factor (PDGF) receptor (PDGFR) and its downstream target, extracellular signal-regulated kinase (ERK). The proliferation rates of osteoblasts were investigated by immunocytochemistry and MTT cell viability assays. Furthermore, we used western blotting to evaluate the effect of PDGFR knockdown on the phosphorylation of ERK stimulated with fresh-PRP and FD-PRP. Results: Platelet counts in both the fresh-PRP and FD-PRP samples were approximately 10-fold higher than in peripheral blood samples. The phosphorylation and activation of the PDGFR and ERK were evenly induced by fresh-PRP and FD-PRP stimulation. Both fresh-PRP and FD-PRP significantly induced osteoblast proliferation in MTT cell viability assays. Furthermore, osteoblast PDGFR knockdown attenuated the downstream ERK activation by fresh PRP and FD-PRP. Conclusions: We demonstrated the pharmacological activity of PDGF in FD-PRP in vitro after 4 weeks of storage.

Long-Term Outcomes of In Situ Fusion for Treating Dysplastic Spondylolisthesis

Kazuhide Inage,Sumihisa Orita,Kazuyo Yamauchi,Miyako Suzuki,Yoshihiro Sakuma,Go Kubota,Yasuhiro Oikawa,Takeshi Sainoh,Jun Sato,Kazuki Fujimoto,Yasuhiro Shiga,Koki Abe,Hirohito Kanamoto,Masahiro Inoue 대한척추외과학회 2017 Asian Spine Journal Vol.11 No.2

Study Design: Retrospective, observational, single-center study. Purpose: To investigate the long-term outcomes of in situ fusion procedures for treating dysplastic spondylolisthesis. Overview of Literature: In situ fusion performed in patients with dysplastic spondylolisthesis avoids the development of nerve complications. Methods: In total, 12 of 28 patients who underwent in situ fusion for treating dysplastic spondylolisthesis at Chiba University Hospital from 1974 to 2004 were followed up in August 2013. Surgical complications were evaluated. Low back pain and leg pain were assessed using a visual analog scale (VAS). Vertebral alignment, including the lumbosacral angle and lumbar lordosis angle measurement on radiographic images (profile view in the neutral standing position), was evaluated during preoperative, postoperative, and final examinations. Results: The mean follow-up duration, patient age at the final examination, and patient age at operation were 20.0±7.2, 42.3±13.3, and 22.3±11.4 years, respectively. No complications were reported. Mean VAS scores for low back pain and leg pain were significantly lower at the final examination than at the preoperative examination (p <0.05). At the preoperative, postoperative, and final examinations, the mean lumbosacral angle was 32.3°±14.2°, 33.7°±11.8°, and 36.5°±16.4°, while the mean lumbar lordosis angle was 51.0°±14.8°, 48.6°±18.8°, and 49.6°±15.5°, respectively. No significant differences were noted among these values across the different time periods (p <0.05). Conclusions: In situ fusion performed in patients with dysplastic spondylolisthesis avoids the development of nerve complications such as nerve paralysis that may occur after repositioning operation and maintains appropriate long-term sagittal alignment, even 20 years after operation.

Differences in Risk Factors for Decreased Cervical Lordosis after Multiple-Segment Laminoplasty for Cervical Spondylotic Myelopathy and Ossification of the Posterior Longitudinal Ligament: A Pilot Study

Inoue Takaki,Maki Satoshi,Furuya Takeo,Okimatsu Sho,Yunde Atsushi,Miura Masataka,Shiratani Yuki,Nagashima Yuki,Maruyama Juntaro,Shiga Yasuhiro,Inage Kazuhide,Orita Sumihisa,Eguchi Yawara,Ohtori Seiji 대한척추외과학회 2023 Asian Spine Journal Vol.17 No.4

Study Design: Retrospective study.Purpose: To compare the radiographic risk factors for decreased cervical lordosis (CL) after laminoplasty, focusing on the difference between cervical spondylotic myelopathy (CSM) and cervical ossification of the posterior longitudinal ligament (C-OPLL).Overview of Literature: A few reports compared the risk factors for decreased CL between CSM and C-OPLL although these two pathologies have their characteristics.Methods: This study included 50 patients with CSM and 39 with C-OPLL who underwent multi-segment laminoplasty. Decreased CL was defined as the difference between preoperative and 2-year postoperative neutral C2–7 Cobb angles. Radiographic parameters included preoperative neutral C2–7 Cobb angles, C2–7 sagittal vertical axis (SVA), T1 slope (T1S), dynamic extension reserve (DER), and range of motion. The radiographic risk factors were investigated for decreased CL in CSM and C-OPLL. Additionally, the Japanese Orthopedic Association (JOA) score was assessed preoperatively and 2 years postoperatively.Results: C2–7 SVA (<i>p</i> =0.018) and DER (<i>p</i> =0.002) were significantly correlated with decreased CL in CSM, while C2–7 Cobb angle (<i>p</i> =0.012) and C2–7 SVA (<i>p</i> =0.028) were correlated with decreased CL in C-OPLL. Multiple linear regression analysis revealed that greater C2–7 SVA (<i>B</i> =0.22, <i>p</i> =0.026) and small DER (<i>B</i> =−0.53, <i>p</i> =0.002) were significantly associated with decreased CL in CSM. By contrast, greater C2–7 SVA (<i>B</i> =0.36, <i>p</i> =0.031) was significantly associated with decreased CL in C-OPLL. The JOA score significantly improved in both CSM and C-OPLL (<i>p</i> <0.001).Conclusions: C2–7 SVA was associated with a postoperative decreased CL in both CSM and C-OPLL, but DER was only associated with decreased CL in CSM. Risk factors for decreased CL slightly differed depending on the etiology of the condition.

Use of Bioelectrical Impedance Analysis for the Measurement of Appendicular Skeletal Muscle Mass/Whole Fat Mass and Its Relevance in Assessing Osteoporosis among Patients with Low Back Pain: A Comparative Analysis Using Dual X-ray Absorptiometry

Kazuki Fujimoto,Kazuhide Inage,Yawara Eguchi,Sumihisa Orita,Miyako Suzuki,Go Kubota,Takeshi Sainoh,Jun Sato,Yasuhiro Shiga,Koki Abe,Hirohito Kanamoto,Masahiro Inoue,Hideyuki Kinoshita,Masaki Norimoto 대한척추외과학회 2018 Asian Spine Journal Vol.12 No.5

Study Design: Cross-sectional observational study. Purpose: To compare measurements of appendicular skeletal muscle mass (ASMM) and whole fat mass (WFM) obtained using dualenergy X-ray absorptiometry (DXA) and bioelectrical impedance analysis (BIA) among patients with low back pain (LBP). Moreover, the study investigated the correlation between BIA-based ASMM and DXA-based bone mineral density (BMD). Overview of the Literature: If reliable, BIA may be a useful alternative to DXA as a screening tool for sarcopenia and osteoporosis among patients with LBP. Methods: Measurements were performed in 130 patients, including BMD of the lumbar spine and femoral neck. The correlation between DXA and BIA as well as between BIA-ASMM and BMD were evaluated. Results: BIA and DXA were highly correlated in both male and female patients (r =0.73–0.90, p <0.0001). However, BIA consistently overestimated ASMM by 1.5–2.5 kg on an average (p <0.0001) and underestimated WFM (−4.0 to −2.7 kg) on an average (p <0.0001). BIA-based ASMM correlated with BMD of the lumbar spine in both male and female patients (r =0.28–0.37, p ≤0.02) and that of the femoral neck (r =0.34–0.51, p ≤0.005). Regarding the calculated skeletal muscle index (SMI: ASMM/height [m2]) used as a criterion for sarcopenia, BIA-based SMI correlated with BMD of the lumbar spine in male patients (r =0.44, p =0.0004) and that of the femoral neck in female patients (r =0.33, p =0.009). Conclusions: BIA may be a favorable alternative to DXA as a screening tool for sarcopenia and osteoporosis among patients with LBP. Considering the overestimation of BIA-based ASMM and SMI, we recommend using the cutoff values for sarcopenia of 7.9 kg/m2 for males and 6.1 kg/m2 for females.