1.Regulation of the Wnt Signaling Pathways during Cell Culture of Human Mesenchymal Stem Cells for Efficient Bone Regeneration
Wataru Katagiri ; Yoichi Yamada ; Sayaka Nakamura ; Kenji Ito ; Kenji Hara ; Hideharu Hibi ; Minoru Ueda
Oral Science International 2010;7(2):37-46
Tissue engineering and bone regeneration techniques using mesenchymal stem cells (MSCs) have started to be applied to the field of oral and maxillofacial surgery. Clinically, a shortened treatment time and improved efficiency are necessary because of the patients' needs and the running cost of cell culture. In the present study, the cultivation process for human MSCs (hMSCs) was examined by regulating the Wnt signaling pathway. We activated Wnt signaling with LiCl and inhibited Wnt signaling with sFRP-3 (secreted Frizzled-Related Protein-3). The proliferation of LiCl-treated hMSCs was examined by studying the cell growth rate and performing BrdU assays. Osteogenic differentiation of sFRP-3-treated hMSCs was examined by alizarin red staining, and osteogenic gene expression on days 7 and 14 after induction was examined by reverse-transcription polymerase chain reaction (RT-PCR) analysis and quantitative real-time RT-PCR analysis. LiCl-treated hMSCs showed increased cell numbers and BrdU-positive cells as compared to the untreated cells. Alizarin red staining showed early mineralization of hMSCs on day 7 of the sFRP-3 treatment. A high expression level of the alkaline phosphatase gene on days 7 and 14 of sFRP-3 treatment was also demonstrated. These results suggest that the regulation of the Wnt signaling pathway contributes to the increased cell numbers and the early osteogenic differentiation of hMSCs. This study supports the possibility that the regulation of the Wnt signaling pathway contributes to the development of effective and efficient bone regeneration techniques.
2.CURRENT TREND IN SKI INJURIES AND BOOT TOP FRACTURE
SETSURO KURIYAMA ; WATARU KAWASHIMA ; SHOKICHI UEMURA ; YOSHIKATSU KUROKI ; ETSUO FUJIMAKI ; YOSHIAKI MORI ; TOMOO KATAGIRI ; KEIZO SAKAMOTO ; HIDEMASA SEKI
Japanese Journal of Physical Fitness and Sports Medicine 1980;29(3):177-187
1) The objectives of author's present study were 42, 245 patients with ratio of males to females of 7 to 3 in 44, 327 affected locations, covering the period of 23 years from the December of 1956 to the end of March of 1979.
2) All the injuries thus treated can be classified into 18, 815 (42.5%) of sprains, 11, 838 (26.7 %) of fractures, 9, 650 (21.8%) of lacerations, 2, 812 (6.4%) of contusions, 752 (1.7%) of dislocations and 460 (1.0%) of others.
3) Looking from auther's statistical study of the ski injuries in the past 23 years, the development of the ski equipments have largely effected on the ski injuries.
4) The lacerations on the head, face and arm are increased when the safety bindings have come into wide use. So, we expect the effect of the ski-stopper replacing the strap, in order to decrease the lacerations.
5) Injuries of the ankle have been decreased because of the plastic ski boots. Especially, “the ski fractures”, or, the abducted and external rotated fractures of the lateral malleolus are decreased remarkably.
6) The knee sprains, or, the ligamentous strains of medial colateral ligament of knee joint have been increased, because the stiff and high backed plastic ski boots have come into wide use.
7) Fractures of the lower legs and boot top fractures have been increased, according the development of the ski boots, such as from the leather boots to buckled boots to plastic boots.
3.Angiogenesis in newly regenerated bone by secretomes of human mesenchymal stem cells.
Wataru KATAGIRI ; Takamasa KAWAI ; Masashi OSUGI ; Yukiko SUGIMURA-WAKAYAMA ; Kohei SAKAGUCHI ; Taku KOJIMA ; Tadaharu KOBAYASHI
Maxillofacial Plastic and Reconstructive Surgery 2017;39(3):8-
BACKGROUND: For an effective bone graft for reconstruction of the maxillofacial region, an adequate vascular network will be required to supply blood, osteoprogenitor cells, and growth factors. We previously reported that the secretomes of bone marrow-derived mesenchymal stem cells (MSC-CM) contain numerous growth factors such as insulin-like growth factor (IGF)-1, transforming growth factor (TGF)-β1, and vascular endothelial growth factor (VEGF), which can affect the cellular characteristics and behavior of regenerating bone cells. We hypothesized that angiogenesis is an important step for bone regeneration, and VEGF is one of the crucial factors in MSC-CM that would enhance its osteogenic potential. In the present study, we focused on VEGF in MSC-CM and evaluated the angiogenic and osteogenic potentials of MSC-CM for bone regeneration. METHODS: Cytokines in MSC-CM were measured by enzyme-linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells (HUVECs) were cultured with MSC-CM or MSC-CM with anti-VEGF antibody (MSC-CM + anti-VEGF) for neutralization, and tube formation was evaluated. For the evaluation of bone and blood vessel formation with micro-computed tomography (micro-CT) and for the histological and immunohistochemical analyses, a rat calvarial bone defect model was used. RESULTS: The concentrations of IGF-1, VEGF, and TGF-β1 in MSC-CM were 1515.6 ± 211.8 pg/mL, 465.8 ± 108.8 pg/mL, and 339.8 ± 14.4 pg/mL, respectively. Tube formation of HUVECs, bone formation, and blood vessel formation were increased in the MSC-CM group but decreased in the MSC-CM + anti-VEGF group. Histological findings suggested that new bone formation in the entire defect was observed in the MSC-CM group although it was decreased in the MSC-CM + anti-VEGF group. Immunohistochemistry indicated that angiogenesis and migration of endogenous stem cells were much more abundant in the MSC-CM group than in the MSC-CM + anti-VEGF group. CONCLUSIONS: VEGF is considered a crucial factor in MSC-CM, and MSC-CM is proposed to be an adequate therapeutic agent for bone regeneration with angiogenesis.
Animals
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Blood Vessels
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Bone Regeneration
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Culture Media, Conditioned
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Cytokines
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Enzyme-Linked Immunosorbent Assay
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Human Umbilical Vein Endothelial Cells
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Humans*
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Immunohistochemistry
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Insulin-Like Growth Factor I
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Intercellular Signaling Peptides and Proteins
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Mesenchymal Stromal Cells*
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Osteogenesis
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Rats
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Stem Cells
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Transforming Growth Factors
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Transplants
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Vascular Endothelial Growth Factor A
4.Masticatory muscle tendon‑aponeurosis hyperplasia that was initially misdiagnosed for polymyositis: a case report and review of the literature
Wataru KATAGIRI ; Daisuke SAITO ; Satoshi MARUYAMA ; Makiko IKE ; Hideyoshi NISIYAMA ; Takafumi HAYASHI ; Jun‑ichi TANUMA ; Tadaharu KOBAYASHI
Maxillofacial Plastic and Reconstructive Surgery 2023;45(1):18-
Background:
Masticatory muscle tendon-aponeurosis hyperplasia (MMTAH) is a relatively newly identified clinical condition that manifests as trismus with a square-shaped mandible. Herein, we report a case of MMATH that was initially misdiagnosed for polymyositis due to trismus and simultaneous lower limb pain, with literature review.Case presentation A 30-year-old woman had a history of lower limb pain after exertion for 2 years. Initial physical examination had been performed at the Department of General Medicine in our hospital. There was also redness in the hands and fingers. Although polymyositis was suspected, it was denied. The patient visited our department for right maxillary wisdom tooth extraction.Clinical examination revealed that the patient had a square-shaped mandible. The maximal mouth opening was 22 mm. There was no temporomandibular joint pain at the time of opening. Furthermore, there was awareness of clenching while working. Panoramic radiography revealed developed square mandibular angles with flattened con‑ dyles. Computed tomography showed enlarged masseter muscles with high-density areas around the anterior and lateral fascia. Magnetic resonance imaging also showed thickened tendons and aponeuroses on the anterior surface and inside bilateral masseter muscles. Finally, the patient was diagnosed with MMTAH. Bilateral aponeurectomy of the masseter muscles with coronoidectomy and masseter muscle myotomy was performed under general anesthesia.The maximum opening during surgery was 48 mm. Mouth opening training was started on day 3 after surgery. Histo‑ pathological examination of the surgical specimen showed that the muscle fibers were enlarged to 60 μm. Immuno‑ histochemistry testing for calcineurin, which was associated with muscle hypertrophy due to overload in some case reports, showed positive results. Twelve months after surgery, the mouth self-opening and forced opening were over 35 mm and 44 mm, respectively.
Conclusions
Herein, we report a case of MMATH. Lower limb pain due to prolonged standing at work and overload due to clenching were considered risk factors for symptoms onset of MMATH.