Our research (Under construction) 研究の紹介(更新中)      © Atomi&Shimizu Lab.  2013

2019

Cell Tissue Res. 2019 Apr;376(1):123-135. doi: 10.1007/s00441-018-2954-3. Epub 2018 Nov 17.

Solubilized eggshell membrane supplies a type III collagen-rich elastic dermal papilla

加水分解卵殻膜はIII型コラーゲンに富む弾性のある(ハリのある)真皮乳頭層を与える


Eri Ohto-Fujita1, Miho Shimizu1*, Shoei Sano1, Masashi Kurimoto1, Kai Yamazawa1, Tomoaki Atomi2, Takashi Sakurai3,4, Yoshihiko Murakami5, Taku Takami5, Tomoaki Murakami6, Kotaro Yoshimura7, Yukio Hasebe8, Yoriko Atomi1*#

大戸ー藤田恵理1、清水美穂1*、佐野将英1、栗本大嗣1、山澤開1、跡見友章2、桜井隆史3,4、村上義彦5、高見拓5 、村上智亮6

吉村浩太郎7、長谷部由紀夫8、跡見順子1*#

1Material Health Science Laboratory, Tokyo University of Agriculture and Technology, 2-24-16 Nakacho, Koganei-shi, Tokyo, 184-8588, Japan       2Faculty of Health Sciences, Department of Physical Therapy Kyorin University, 5-4-1 Shimorenjaku, Mitaka-shi, Tokyo 181-8612 JAPAN, Japan         3Department of Life Sciences, The Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1, Meguro-ku, Tokyo, 153-8902, Japan     4Current adress: Astellas Institute for Regenerative Medicine, 33 Locke Drive Marlborough, MA 01752, USA                                                                5Department of Organic and Polymer Materials Chemistry, Faculty of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Nakacho, Koganei-shi, Tokyo, 184-8588, Japan                                                                                                                                                                  6Laboratory of Veterinary Toxicology, Cooperative Department of Veterinary Medicine, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu-shi, Tokyo 183-8509, Japan.                                                                                                                                            7Department of Plastic Surgery, Jichi Medical University, 3311-1 Yakushiji, Shimotsuke-shi, Tochigi-ken, 329-0498, Japan                              8Almado inc., 3-6-18 Kyobashi, Chuo-ku, Tokyo, 104-0031, Japan                                                                                                                               *Equal conteribution, # Corresponding author

1東京農工大学工学府材料健康科学、2杏林大学保健学部理学療法学科、3東京大学大学院総合文化研究科生命環境科学系、4現所属:アステラス再生医療研究所、マサチューセッツ州(米国)、5農工大・工学部・有機材料化学科、6東京農工大学農学研究院 動物生命科学部門 獣医毒性学研究室、7自治医科大学形成外科、8株式会社アルマード


Abstract

Signs of aging in facial skin correlate with lifespan and chronic disease; however, the health of aging skin has not been extensively studied. In healthy young skin, the dermis forms a type III collagen-rich dermal papilla, where capillary vessels supply oxygen and nutrients to basal epidermal cells. Chicken eggshell membranes (ESMs) have been used as traditional medicines to promote skin wound healing in Asian countries for many years. Previously, we designed an experimental system in which human dermal fibroblasts (HDFs) were cultured on a dish with a solubilized ESM (S-ESM) bound to an artificial phosphorylcholine polymer; we found that genes that promoted the health of the papillary dermis, such as those encoding type III collagen were induced in the S-ESM environment. The present study found that a gel with a ratio of 20% type III/80% type I collagen, similar to that of the baby skin resulted in a higher elasticity than 100% type I collagen (p < 0.05) and that HDFs on the gel showed high mitochondrial activity. Thus, we decided to perform further evaluations to identify the effects of S-ESM on gene expression in the skin of hairless mice and found significant increase of type III collagen in S-ESM. Picrosirius Red staining showed that type III collagen significantly increased in the papillary dermis after S-ESM treatment. Moreover, S-ESM application significantly improved human arm elasticity and reduced facial wrinkles. Thus, ESMs may have applications in extending lifespan by reducing the loss of tissue elasticity through the increase of type III collagen.

顔の皮膚の老化の徴候は、寿命および慢性疾患と相関する。しかし、老化した皮膚の健康は広く研究されていない。健康な若い皮膚では、真皮はIII型コラーゲンが豊富な真皮乳頭を形成し、毛細血管が酸素と栄養素を基底表皮細胞に供給する。鶏卵殻膜(ESM)は、アジア諸国の皮膚治療のための民間療法として長年使われてきた。我々は以前に、人工のホスホリルコリンポリマーに結合した可溶化卵殻膜(S-ESM)を付けた培養皿上でヒト皮膚線維芽細胞(HDF)を培養する実験系を設計し、S-ESM環境で誘導されるIII型コラーゲンなどの若い乳頭真皮を促進する遺伝子を見出した。本研究では、赤ちゃんの皮膚と同様のタイプIII / 80% I型コラーゲン20%の比を有するゲルが100% I型コラーゲンより高い弾性をもたらし(p <0.05)、そのゲル上のHDFは高いミトコンドリア活性を示した。このように、我々は、ヘアレスマウスの皮膚における遺伝子発現に対するS-ESMの効果を同定するためのさらなる評価を行うことに決め、S-ESMではIII型コラーゲンの有意な増加を見出した。 ピクロシリウスレッド染色により、S-ESM処置後の乳頭真皮においてIII型コラーゲンが有意に増加することが示された。さらに、S-ESM塗布は、ヒトの腕の弾力性を有意に改善し、顔のしわを低減した。したがって、ESMは、III型コラーゲンの増加により、組織弾性の喪失を減少させることによって、寿命を延ばすために使用することができるだろう。    

Keywords: Extracelllular matrix, eggshell membrane, papillary dermis, geroscience, type III collagen                                                                       細胞外マトリクス、卵殻膜、真皮乳頭層、老化科学、III型コラーゲン 

19CTR introduction for HP


Result 1 Type III collagen synthesized by HDFs and the high modulus of elasticity in gels with collagen ratios similar to those of young skin

結果1 HDFによって合成されたIII型コラーゲンと、若い皮膚と同様のコラーゲン比を有するゲル中の高い弾性率

19CTR fig1 for HP


Result 2 The mitochondrial activity of fibroblasts was significantly higher when grown on 20% type III collagen

結果2 線維芽細胞のミトコンドリア活性は、20%のIII型コラーゲンで培養させた場合に有意に高い

アセット 2


Result 3 S-ESM application for 10 days upregulated type III collagen, MMP2, and decorin in the skin of hairless mice by altering the type III/I collagen ratio

結果3 III型 / I型コラーゲン比を変更することによりヘアレスマウスの皮膚でのIII型コラーゲン、MMP2、デコリンを上方制御する10日間のS-ESM塗布

19CTR fig3 for HP


Result 4 S-ESM application increased type III collagen in the papillary dermis

結果4 S-ESM塗布は真皮乳頭層におけるIII型コラーゲンを増加させた

19CTR fig4 for HP


Result 5 Effects of S-ESM on the elasticity of arm skin in women

結果5 女性の肌の弾力性に及ぼすS-ESMの影響


Result 6 Topical application of S-ESM improved facial wrinkles

結果6 卵殻膜溶液の局所塗布により目尻のしわが改善された

19CTR fig6 for HP


In summary, in this study, we found that ESM had anti-aging effects by improving the ECM environment in the papillary layer. ESM is available worldwide and is a biomaterial with application in medical, food, and cosmetic fields. Thus, natural products, such as ESM, from traditional Asian medicine may provide new approaches to improving health.

本研究では、真皮乳頭層のECM環境を改善することにより、卵殻膜が老化防止効果を有することを見出した。 卵殻膜は世界中で利用可能で、医療、食品、および美容分野での応用可能な生体材料である。したがって、伝統的なアジアの医学からの卵殻膜などの天然産物は、健康を改善するための新しいアプローチを提供する可能性がある。    


2018

Geroscience From Cell-body Dynamics and Proteostasis Cooperation Supported by αB-crystallin and Human will ~ A Proposal of “Body-Mind Integrative Science”

Yoriko Atomi*, Miho Shimizu, Eri Ohto-Fujita, Aya Atomi, Saaya Hayasaki, Yoshikazu Higashi, Tomoaki Atomi 

In: Asea A., Kaur P. (eds) Regulation of Heat Shock Protein Responses. Heat Shock Proteins, vol 13. pp307-360, Springer, Cham  [Invited book chapter]


The importance of small heat shock protein HSP in geroscience is increasing. We believe that research progress from life science should contribute to well-being. Molecular chaperone studies are considered superior when performed on model substrates and model animals. However, by itself, concrete measures for extending the healthy life of human beings are not provided. It is important to analyze the cell-animal-human results, interpret the relationship, and promote comprehensively studied HSP research. αB-crystallin (CRYAB) was identified for key molecule to explain the mechanism of exercise adaptation of slow-twitch muscle for a long ago. It is only human beings that stand up against gravity and move all day in their standing position. With CRYAB and tubulin/microtubule as a key word, we will introduce the principle of clarifying not only cells but also how to control the human body. Mechanistically fiber structure produced after protein assembly has not only multifunction but also is available as materials to make “body” and can sustain body weight by tension development at both micro- and macro-levels. This links cell’s autonomous regulating ability and human will to keep standing. This manuscript may contribute to develop a new direction of HSP Geroscience research proposing real program to healthy aging and mature human world.

Keywords:αB-Crystallin (CRYAB) Cytoskeleton Dynamic Instability Heat Shock Protein Mechanical Stress Bipedal standing 


2017

The free moment is associated with torsion between the pelvis and the foot during gait

Gait Posture. 2017 Oct;58:415-420. doi: 10.1016/j.gaitpost.2017.09.002. Epub 2017 Sep 5.

Ohkawa T, Atomi T, Hasegawa K, Atomi Y.

Abstract

BACKGROUND: During walking, the friction between the foot and the ground surface causes a free moment (FM), which influences the torsional stress on the lower extremity. However, few studies have investigated the FM during natural walking. The main aim of this study was to examine the relationship between the FM and the absolute and relative rotation angles of the foot and pelvis.

METHODS: The rotation angles of foot and pelvic were measured in 18 healthy men using a motion capture system. Rotation angles were measured in absolute and relative coordinates as well as in reference to the line connecting the center of pressure (CoP) line under the right and left feet to evaluate the effects of the opposite lower limb on the FM. The absolute and relative rotation angles of the foot and pelvis were entered into forced-entry linear regression models to evaluate the influence on the FM.

FINDINGS: Only the relative angle of rotation between the foot and pelvis could explain the prediction equations significantly. In the Pearson's product-moment correlation coefficient, the rotation angles of the foot and pelvis defined using the bilateral CoP points had not significantly correlated with FM. No joint rotation movement was correlated with FM.

INTERPRETATION: The torsion of the entire lower extremity should be performed principally through hip internal rotation. When evaluating the FM as a torsional stress, focusing on the rotation of the entire lower extremity, rather than on one segment, is beneficial.

Keywords Free moment; Friction; Gait; Rotation; Torsional stress

PMID: 28898801 DOI: 10.1016/j.gaitpost.2017.09.002


下肢に生じる捻じれストレスが高まってしまう歩き方を解明

・歩行中の下肢に生じる捻じれストレスが、足部と骨盤間の相対的な捻れ量(角度)に相関することを世界で初めて明らかにした。

・フリーモーメントというパラメータに着目し歩行を解析したところ、足が地面を蹴りだす際の股関節の柔らかさによって下肢に生じる捻じれストレスが制御されていることが示唆された。

・変形性膝関節症などのロコモティブ症候群へのリハビリや予防に活用することが期待できる。

東京農工大プレスリリース


2016

Small Heat Shock Protein αB-Crystallin Controls Shape and Adhesion of Glioma and Myoblast Cells in the Absence of Stress

PLoS One. 2016 Dec 15;11(12):e0168136. doi: 10.1371/journal.pone.0168136. eCollection 2016.

Shimizu M, Tanaka M, Atomi Y.

Abstract

Cell shape and adhesion and their proper controls are fundamental for all biological systems. Mesenchymal cells migrate at an average rate of 6 to 60 μm/hr, depending on the extracellular matrix environment and cell signaling. Myotubes, fully differentiated muscle cells, are specialized for power-generation and therefore lose motility. Cell spreading and stabilities of focal adhesion are regulated by the critical protein vinculin from immature myoblast to mature costamere of differentiated myotubes where myofibril Z-band linked to sarcolemma. The Z-band is constituted from microtubules, intermediate filaments, cell adhesion molecules and other adapter proteins that communicate with the outer environment. Mesenchymal cells, including myoblast cells, convert actomyosin contraction forces to tension through mechano-responsive adhesion assembly complexes as Z-band equivalents. There is growing evidence that microtubule dynamics are involved in the generation of contractile forces; however, the roles of microtubules in cell adhesion dynamics are not well determined. Here, we show for the first time that αB-crystallin, a molecular chaperon for tubulin/microtubules, is involved in cell shape determination. Moreover, knockdown of this molecule caused myoblasts and glioma cells to lose their ability for adhesion as they tended to behave like migratory cells. Surprisingly, αB-crystallin knockdown in both C6 glial cells and L6 myoblast permitted cells to migrate more rapidly (2.7 times faster for C6 and 1.3 times faster for L6 cells) than dermal fibroblast. On the other hand, overexpression of αB-crystallin in cells led to an immortal phenotype because of persistent adhesion. Position of matured focal adhesion as visualized by vinculin immuno-staining, stress fiber direction, length, and density were clearly αB-crystallin dependent. These results indicate that the small HSP αB-crystallin has important roles for cell adhesion, and thus microtubule dynamics are necessary for persistent adhesion.

PMID: 27977738 PMCID: PMC5158045 DOI: 10.1371/journal.pone.0168136


人間が立つことを支えるストレスタンパク質・αB-クリスタリンが、

一細胞レベルでもふんばりを支え、かたちを制御していることを発見

東京農工大学大学院工学研究院応用化学専攻有機材料科学・材料健康科学寄付講座 跡見順子客員教授、清水美穂客員准教授をはじめとする研究チームは、ストレスタンパク質・αB-クリスタリン(αB)の多寡が、かたちの保持や接着といった細胞の基本的な機能を制御していることを、遺伝子工学的な手法を用いて世界で初めてあきらかにしました。αBは、不安定な二足立位姿勢を選んだ動物としての人間の筋活動を支えるタンパク質を追求するなかで我々が見出したタンパク質で、ヒラメ筋や脊柱起立筋といった抗重力筋で多く発現し、宇宙の微小重力環境実験動物モデルで顕著に減少します。細胞が重力に対抗してかたちを保持するためには、細胞外マトリクスとの接着に加えて、細胞骨格タンパク質との接着点における安定性が必要で、αBがその機能を担っていることが本研究から明らかになりました。この研究成果は寝たきりなど超高齢社会において重要となる疾病を予防するためのメカニズム解明につながることが期待されます。

東京農工大プレスリリース


2015

Gravitational Effects on Human Physiology.

Subcell Biochem. 2015;72:627-59. doi: 10.1007/978-94-017-9918-8_29.

Atomi Y.

Abstract

Physical working capacity decreases with age and also in microgravity. Regardless of age, increased physical activity can always improve the physical adaptability of the body, although the mechanisms of this adaptability are unknown. Physical exercise produces various mechanical stimuli in the body, and these stimuli may be essential for cell survival in organisms. The cytoskeleton plays an important role in maintaining cell shape and tension development, and in various molecular and/or cellular organelles involved in cellular trafficking. Both intra and extracellular stimuli send signals through the cytoskeleton to the nucleus and modulate gene expression via an intrinsic property, namely the "dynamic instability" of cytoskeletal proteins. αB-crystallin is an important chaperone for cytoskeletal proteins in muscle cells. Decreases in the levels of αB-crystallin are specifically associated with a marked decrease in muscle mass (atrophy) in a rat hindlimb suspension model that mimics muscle and bone atrophy that occurs in space and increases with passive stretch. Moreover, immunofluorescence data show complete co-localization of αB-crystallin and the tubulin/microtubule system in myoblast cells. This association was further confirmed in biochemical experiments carried out in vitro showing that αB-crystallin acts as a chaperone for heat-denatured tubulin and prevents microtubule disassembly induced by calcium. Physical activity induces the constitutive expression of αB-crystallin, which helps to maintain the homeostasis of cytoskeleton dynamics in response to gravitational forces. This relationship between chaperone expression levels and regulation of cytoskeletal dynamics observed in slow anti-gravitational muscles as well as in mammalian striated muscles, such as those in the heart, diaphragm and tongue, may have been especially essential for human evolution in particular. Elucidation of the intrinsic properties of the tubulin/microtubule and chaperone αB-crystallin protein complex systems is expected to provide valuable information for high-pressure bioscience and gravity health science.

PMID: 26174402 DOI: 10.1007/978-94-017-9918-8_29


2014

2011

Hydrolyzed eggshell membrane immobilized on phosphorylcholine polymer supplies extracellular matrix environment for human dermal fibroblasts

Cell Tissue Res. 2011 Jul;345(1):177-90. doi: 10.1007/s00441-011-1172-z. Epub 2011 May 20.

Ohto-Fujita E1, Konno T, Shimizu M, Ishihara K, Sugitate T, Miyake J, Yoshimura K, Taniwaki K, Sakurai T, Hasebe Y, Atomi Y.

Abstract

We have found that a water-soluble alkaline-digested form of eggshell membrane (ASESM) can provide an extracellular matrix (ECM) environment for human dermal fibroblast cells (HDF) in vitro. Avian eggshell membrane (ESM) has a fibrous-meshwork structure and has long been utilized as a Chinese medicine for recovery from burn injuries and wounds in Asian countries. Therefore, ESM is expected to provide an excellent natural material for biomedical use. However, such applications have been hampered by the insolubility of ESM proteins. We have used a recently developed artificial cell membrane biointerface, 2-methacryloyloxyethyl phosphorylcholine polymer (PMBN) to immobilize ASESM proteins. The surface shows a fibrous structure under the atomic force microscope, and adhesion of HDF to ASESM is ASESM-dose-dependent. Quantitative mRNA analysis has revealed that the expression of type III collagen, matrix metalloproteinase-2, and decorin mRNAs is more than two-fold higher when HDF come into contact with a lower dose ASESM proteins immobilized on PMBN surface. A particle-exclusion assay with fixed erythrocytes has visualized secreted water-binding molecules around the cells. Thus, HDF seems to possess an ECM environment on the newly designed PMBN-ASESM surface, and future applications of the ASESM-PMBN system for biomedical use should be of great interest.

PMID: 21597915 PMCID: PMC3132421 DOI: 10.1007/s00441-011-1172-z


「生活の知恵を先端科学で解明

―卵殻膜の働きにより細胞が自分で作る健康な環境―」

細胞は30%程度の割合で常に様々なエラーを起こしていますが、健康状態が保たれている間は、修復機能が働いて問題をキャンセルしてくれていますが、加齢に伴いこの修復機能が低下します。皮膚の細胞で考えると、紫外線、活性酸素、などによる様々なダメージの蓄積が加齢とともに増加し、傷の治りが遅く、傷痕も残り、悪い例としては癌化といった細胞の異常状態に至ります。

今回、東京大学-アルマード共同研究プロジェクトでは、中国や日本では人々の間では効果が認められ、”生活の智恵”として古くから使われていながらも、その実態が明らかにされていなかった卵殻膜に着目し、先端科学を駆使してメカニズムの解明に取り組みました。その結果、適度な量の卵殻膜には、細胞自らが正常な状態へ修復するように働きかける効果があるということが証明されました。

東京大学・株式会社アルマード共同プレスリリース



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