マイオスタチン ニ タイスル RNA カンショウホウ ニヨル コッカクキン ケイセイ ノ チョウセツ

Skeletal muscle is one of the most important morpho-functional organs, and its atrophy causes severe conditions such as muscular dystrophies. In the craniofacial region, skeletal deformities and malocclusion are often caused by hyper-and hypotrophy of masticatory muscles or the tongue. Recently, RNA interference (RNAi)-based gene therapy has been expected as a safe remedy without side effects ; therefore, we used RNAi to control the expression of myostatin, a negative regulator of skeletal muscle formation, for the purpose of developing a new method to regulate skeletal muscle volume in this study. We constructed a plasmid vector containing the full-length of myostatin cDNA (pcDNA3.1-Mst-myc) and a vector for silencing mRNA for either myostatin (pSi-Mst) or green fluorescence protein (GFP) sequence as a negative control (pSi-N). When these vectors were transiently co-transfected in COS-1 cells, the myostatin expression level was notedly reduced. For better understanding of the effect of gene silencing by pSi-Mst on mouse myoblastic cells, we generated stable transfectants overexpressing pcDNA3.1-Mst-myc in mouse myoblast cell line C2C12. The proliferation ability in C2C12 cells overexpressing myostatin was accelerated by transfection with pSi-Mst, but not with pSi-N. Furtheremore, the myogenic differentiation of C2C12 cells overexpressing myostatin, transfected with pSi-Mst, showed a higher expression of MyoD and myogenin than control cells transfected with pSi-N. In addition, they exhibited slight acceleration of myotube formation and a slight increase in myosin heavy chain (MHC) expression. Finally, to investigate the effect of synthetic 19 nucleotide-long double-stranded RNA (Mst-siRNA) in vivo, we injected Mst-siRNA/atelocollagen complex into the masseter and biceps femoris muscles in four mice. After two weeks, myostatin protein levels in those muscles were significantly decreased, but not in the contralateral muscles. Moreover, dramatically increased muscle mass and myofibril size were observed following myostatin down-regulation. These results suggest that it may be possible to regulate skeletal muscle volume and to develop a new treatment for various muscle disorders by RNAi technique

RNA カンショウホウ オ モチイタ キン イシュクセイ シッカン ニ タイスル カクサン イヤク ノ カイハツ ケンキュウ

Skeletal muscle is one of the most important morpho-functional organs, and its atrophy causes severe conditions such as muscular dystrophies. RNA interference (RNAi)-based gene therapy has been expected as a safe remedy without side effects. However, the rapid degradation of small interfering RNAs (siRNAs) and their limited duration of activity require efficient delivery methods. Atelocollagen (ATCOL)- or cationic liposome-mediated administration of siRNAs is a promising approach to disease treatment, including muscular atrophy. In thie study, we used RNAi to control the expression of myostatin, a negative regulator of skeletal muscle formation, for the purpose of developing a good as new method to regulate skeletal muscle volume, and report herein that ATCOL- or cationic liposome-mediated administration of a myostatin-targeting siRNA into skeletal muscles of normal and diseased mice induced a marked increase in muscle mass and a recovery of myofunction. We are now trying to examine the effectiveness of systemic administration of cationic liposome-delivered myostatin-targeting siRNA. Since cationic liposomes delivered siRNA to muscles effectively and are safe and cost-effective, they may represent a powerful therapeutic tool for disease treatment, including muscular atrophy. In this review, I would like to summarize about possibility to regulate skeletal muscle volume and to develop a new treatment for various muscle disorders by RNAi technique

Treatment of a patient with skeletal open bite and temporomandibular joint disorders

The treatment of severe skeletal anterior open bite is extremely difficult in adults, and orthognathic surgery is generally selected for its treatment. We report the case of an 18-year-old adult patient with skeletal anterior open bite and temporomandibular disorders who was successfully treated using temporary anchorage devices. She had an open bite of -2.0 mm and an increased facial height. Miniplates were implanted in both the maxilla and mandible, and molar intrusion resulted in counterclockwise rotation of the mandible over a period of 12 months. After active treatment, her upper and lower first molars were intruded by approximately 2 mm and her overbite became +2.5 mm. Her retrognathic profile improved with counterclockwise rotation of the mandible. Orthodontic treatment aided with skeletal anchorage is beneficial for intrusion of bimaxillary molars in patients with anterior open bite

Myostatin-siRNAおよびActivinIIB型受容体融合タンパク質の共導入による骨格筋形成制御効果の検討

Background: Myostatin (Mstn) is a secreted TGF-β family member that controls skeletal muscle growth, and binds with high affinity to the activin type IIB receptor (ActRIIB). The soluble ligand-binding domain of ActRIIB fused to the Fc domain of IgG (ActRIIB-Fc) potently binds and inhibits TGF-β family members in muscle, leading to rapid and marked muscle growth. The present study was designed to assess the combinative effects of myostatin-targeting siRNA (Mstn-siRNA) and ActRIIB-Fc on murine myoblast in vitro and in vivo. Materials and Methods: C2C12 cells were treated by Mstn-siRNA with or without ActRIIB-Fc at 0 and 48 h after differentiation. Myotube size was measured, and gene expression of Mstn, MuRF-1, MyoD and myogenin were analyzed. Furthermore, 11-week-old, male C57BL/6 mice were injected with atelocollagen (ATCOL)-mediated Mstn-siRNA and Mstn-siRNA/ActRIIB-Fc locally into the masseter muscle twice a week. Histological and biochemical analyses were performed using the dissected muscles. Results: Transfection of Mstn-siRNA and Mstn-siRNA/ActRIIB-Fc resulted in significant increases in the myotube diameter of the C2C12 cells compared with untreated control. Also, treatment with Mstn-siRNA and Mstn-siRNA/ActRIIB-Fc could lead to an upregulation of MyoD and myogenin gene expression and downregulation of Mstn and MuRF-1. In vivo, muscle fibril hypertrophy was observed in both Mstn-siRNA and Mstn-siRNA/ActRIIB-Fc treated groups. Moreover, western blotting analysis showed that the p-Smad2/3 expression level was decreased by treatment of Mstn-siRNA/ActRIIB-Fc. In contrast, MyoD and myogenin protein levels were increased by combined treatment, compared with the other groups. Conclusions: These suggest that double inhibition of myostain is potentially useful for myogenesis and muscle growth promotion. This may be a good as new treatment remedy for patients with various muscle atrophies, including muscular dystrophy

Effectiveness of PNAM on nasal form

Objectives: To evaluate the effectiveness of pre-surgical nasoalveolar molding (PNAM) in patients with unilateral cleft lip nasal deformities. Methods: This was a retrospective study involving 29 patients with unilateral cleft lip and palate defects, of whom 13 were treated with palatal devices with nasal stents (PNAM group) and 16 were treated with palatal devices without nasal stents or surgical tapes (control group). Submental oblique photographs and orthodontic models were longitudinally obtained at the initial visit (T1) and immediately before (T2) and after cheiloplasty (T3). Asymmetry of the external nose, degree of columellar shifting, nasal tip/ala nose ratio, nasal base angle, interalveolar gap, and the sagittal difference in the alveolar gap were measured. The study was conducted in the Orthodontic Clinic at Tokushima University Hospital, Tokushima, Japan between 1997 and 2012. Results: At T1, there were no significant intergroup differences in the first 4 asymmetry parameters. At T2, the PNAM group showed a significant improvement in all values compared to the control group. At T3, the PNAM group showed significant improvement in nasal asymmetry and columellar shifting. Model analysis showed significantly greater changes in the inter-alveolar gap and the sagittal difference of the alveolar cleft gap from T1 to T2 in the PNAM group. Conclusion: The use of PNAM is indispensable for pre-surgical orthodontic treatment at the early postnatal age

サンジゲン デジタル モデル オ モチイタ ヘンソクセイ コウシンレツ コウガイレツ カンジャ ノ コウゴウ ヒョウカ ニ カンスル ケントウ

【目的】Goslon Yardstickは片側性口唇裂口蓋裂患者の咬合状態から治療の難易度を評価する方法で，簡便かつ再現性が高いことから広く用いられている。しかし，評価に際し相当数の歯列模型を用いるため，その管理がしばしば困難となる。一方，三次元デジタルモデル（以下，3DDモデル）は石膏模型をデジタルデータに変換するため保存が容易であるが，デジタルデータより構築した画像は石膏模型とは異なった印象を受けることもある。今回我々は，石膏模型より作成した3DDモデルを用いてGoslon Yardstickによる評価を行い，口腔模型を用いた場合との差異について検討を行った。 【資料および方法】資料として，徳島大学病院矯正歯科を受診した片側性口唇裂口蓋裂患者37症例の石膏模型を用いた。それぞれの石膏模型をスキャニングし，3DDモデル（OrthoCAD，CADENT）を作成した。8年以上の臨床経験をもつ矯正歯科医4名が，石膏模型と3DDモデルをそれぞれ2回ずつ，5日以上の間隔をあけて評価を行った。評価はMarsらの方法に従った。また，評価の一致度を求めるために重み付きkappa値を算出した。kappa値は0.81～1.0をgood agreement，0.61～0.80をsubstantial，0.41～0.60をmoderate，0.21～0.40をfair，0.20以下をpoorとした。 【結果】石膏模型と3DDモデルについて評価者内でのkappa値はそれぞれ0.82～0.91，0.77～0.85となり，ともに評価は一致していた。このことから，石膏模型，3DDモデルのいずれを用いた場合でも評価の再現性が高いことが示された。さらに同一評価者内の石膏模型と3DDモデルの評価の一致度については，kappa値は0.75～0.86となり，石膏模型と3DDモデルにおける評価は比較的一致していた。一致していなかったものに着目すると，3DDモデルの方がGoslon Yardstickの値をやや高く評価する傾向にあった。 【考察】Goslon Yardstickの評価にあたって，3DDモデルの使用は石膏模型と比較しても遜色がないことが示された。三次元デジタルモデルを用いた片側性口唇裂口蓋裂患者の咬合評価に関する検

口唇裂・口蓋裂患者の歯数異常に関する調査

口唇裂・口蓋裂患者は，軟組織や骨の癒合不全だけでなく，永久歯の先天欠如や過剰などの歯数異常が認められ，歯科矯正学的対応に苦慮することも多い．そこで，口唇裂・口蓋裂患者における永久歯の歯数異常の実態を明らかにすることを目的として，1995年1月から2011年12月までの17年間に出生し，徳島大学病院矯正歯科を受診した口唇裂・口蓋裂患者（症候群を含まない）を対象とした調査を行い，以下の結果を得た． 1．調査資料が揃っている患者101名の男女比は，1：1.02であった． 2．顎裂保有者82名の顎裂部位は，1 ▼ 3の型が最も多く，次いで1 ▼23の型であった．（▼は顎裂部位を示す） 3．永久歯における歯数異常の発現率は63.4％であり，歯の欠如のみを有するものが50.5％，過剰歯のみを有するものは8.9％，歯の欠如と過剰歯をともに有するものは4.0％であった． 4．歯の欠如の歯種別頻度は，側切歯が最も多く，次いで第二小臼歯の順であった． 5．顎裂保有者のみを対象とすると，歯の欠如の発現頻度は披裂側で59.8％，非披裂側で24.3％であった． 以上のことから，口唇裂・口蓋裂患者での先天欠如歯の発現率は高いため，治療計画立案時に補綴治療を含めた包括的歯科治療の必要性が示唆された

Myostatin-Targeting siRNA and ActRIIB-Fc Fusion Protein

Myostatin, a member of the TGF-β superfamily, is a negative regulator of skeletal muscle cell growth and differentiation, and binds with high affinity to the activin type IIB receptor (ActRIIB). The soluble ligand-binding domain of ActRIIB fused to the Fc domain of IgG (ActRIIB-Fc) potently binds and inhibits TGF-β family members in muscle, leading to rapid and marked muscle growth. The present study was designed to assess the effectiveness of the co-delivery of myostatin-targeting siRNA (Mstn-siRNA) and ActRIIB-Fc into skeletal muscle as a potential treatment of atrophic myopathies. Eleven-week-old, male C57BL/6 mice were injected with atelocollagen (ATCOL)-mediated Mstn-siRNA with/without ActRIIB-Fc locally into the masseter muscle twice a week. Inhibition of myostatin function by the combination of Mstn-siRNA and ActRIIB-Fc increased muscle weight and myofibril size in murine masseter muscle. Real-time RT-PCR analysis revealed significant downregulation of myostatin mRNA expression in both the Mstn-siRNA-treated and the combination treatment group. Furthermore, myogenin mRNA expression was upregulated in the combination treatment group, while MuRF-1 and Atrogin-1 mRNA expression was downregulated compared to administration of each compound alone. These findings suggest that double inhibition of myostatin is a potentially useful treatment strategy to increase muscle mass and fiber size and could be a useful treatment of patients with various muscle atrophies, including muscular dystrophy

Skeletal anchorage for intrusion of bimaxillary molars in a patient with skeletal open bite and temporomandibular disorders

The treatment of severe skeletal anterior open bite is extremely difficult in adults, and orthognathic surgery is generally selected for its treatment. We report the case of an 18-year-old adult patient with skeletal anterior open bite and temporomandibular disorders who was successfully treated using temporary anchorage devices. She had an open bite of −2.0 mm and an increased facial height. Miniplates were implanted in both the maxilla and mandible, and molar intrusion resulted in counterclockwise rotation of the mandible over a period of 12 months. After active treatment, her upper and lower first molars were intruded by approximately 2 mm and her overbite became +2.5 mm. Her retrognathic profile improved with counterclockwise rotation of the mandible. Orthodontic treatment aided with skeletal anchorage is beneficial for intrusion of bimaxillary molars in patients with anterior open bite