JP7433685B1 - 真皮線維芽細胞の機能賦活用剤及びそれを含んでなる化粧品 - Google Patents
真皮線維芽細胞の機能賦活用剤及びそれを含んでなる化粧品 Download PDFInfo
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Abstract
Description
(2)DNAの加水分解物が、分子量が12,000以下である画分を10~80%含有するものであり、細胞外小胞が、間葉系幹細胞に由来するものである、前記(1)に記載の機能賦活用剤。
(3)前記(1)又は(2)に記載の機能賦活用剤を含んでなる化粧品。
(1)エクソソーム;50~150nm程度サイズのエンドサイトーシス過程で形成されるエンドソーム膜由来の小胞。
(2)マイクロベシクル;100~1,000nm程度サイズの細胞膜から直接出芽して細胞外へと分泌される小胞。
(3)アポトーシス小胞;アポトーシスを起こした細胞により膜から出芽されるマイクロメートル程度のサイズの小胞(2,000×g程度の低速遠心で分離されるため、他の小胞とは区別がしやすい)。
(2)表面抗原CD105、CD73、CD90が陽性で、CD45、CD34、CD14またはCD11b、CD79aまたはCD19、HLA-DRが陰性であること
(3)試験管内(in vitro)で骨芽細胞・軟骨細胞・脂肪細胞への分化能をもつこと
1 試験の目的及び概要
真皮線維芽細胞が産生するコラーゲンは皮膚のハリ、線維芽細胞が産生するヒアルロン酸は皮膚の柔軟性や潤いに重要な役割を果たしていると考えられている。そこで、真皮線維芽細胞と用いて、被験品(加水分解DNA-Naと間葉系幹細胞由来の細胞外小胞)のコラーゲン及びヒアルロン酸産生促進作用を評価した。
2-1 細胞
ヒト新生児由来の真皮線維芽細胞株NB1RGB細胞(RIKEN BRC,Japan)を、CO2インキュベータ(CO2濃度 5%、37℃)で培養し、本試験を実施した。
10.0%(v/v)Fetal Bovine Serum FBS, Cat No. SH30071.03, Hyclone, UK)および1.0%(v/v)抗真菌剤(Antibiotic-Antimycotic 100X, Cat No. 15240-062, Invitrogen, USA)を含むEagle’s Minimal Essential Medium(EMEM, Cat No.051-07615, Wako, Japan)を用いた。
2-3-1 加水分解DNA-Na(オリゴDNA)
日生バイオ株式会社製の加水分解DNA-Na(製品名)の10%水溶液(オリゴDNA10%水溶液)を被験品として用いた。本品は、前記した方法により、サケ科魚類の精巣(白子)からDNAのNa塩を抽出し、それを加水分解することにより調製したものである。
なお、使用した日生バイオ株式会社製の加水分解DNA-Naのゲルパーミエーションクロマトグラフィー(GPC)による分析結果を図1に、保持時間と分子量の関係は、表1に示す。分子量目安の12,000はCytochrome C(MW 12,400)の保持時間を基とした。
分子量12,000~5,000の画分(溶出時間20分~25分未満):32.0%
分子量5,000以下の画分(溶出時間25分以降):32.4%
以上のことから、日生バイオ社製の加水分解DNA-Naの分子量12,000以下である画分は64.4%である。
セルツーイン社(cell2in Inc.)製のExoVESICLE(製品名)を被験品として用いた。
本品は、ヒト臍帯由来間葉系幹細胞(human umbilical cord-derived mesenchymal stem cell;ソウル大学病院産婦人科から臍帯試料を受けて作成)を、80%コンフルエント程度まで培養し、フェノールレッドや血清を含有しない培地で培養して細胞外小胞を培地に放出(分泌)させ、遠心分離により細胞や細胞の破片を除き、フィルターろ過、限外ろ過により分離したものである。
0.5%FBS含有EMEMによって被験品(オリゴDNA+ExoVESICLE)を希釈して計3濃度(1+0.01%, 1+0.1%, 1+0.2%)を用時調製した。DNA-Naの終濃度は0.1%(1mg/ml)、ExoVESICLE(蛋白質量換算)の終濃度は、それぞれ0.003μg/ml、0.03μg/ml、0.06μg/mlである。対照には0.5%FBS含有EMEMを用いた。
細胞増殖、コラーゲン産生およびヒアルロン酸産生の算出には1つの処理群につき96ウェルプレート(細胞増殖:CatNo. 3860-096、Iwaki、japan。コラーゲン産生およびヒアルロン酸産生:Cat No. 3855, Thermo scientific, USA)の3ウェルを用いた。また、1プレートにつき被験品群、対照群をそれぞれ1群設けて試験を実施した。被験品調製を含め、試験に関わる操作は特に記載のないかぎり室温で実施した。
2-5-1 細胞培養
96ウェルプレートに1.0×104 cells/wellの密度でNB1RGB細胞を播種し、CO2インキュベータ内で24時間培養した。また、培養時の乾燥を防ぐため、試験に用いないウェルにPhosphate buffer saline(PBS(-), Cat No. 198601,Nissui, Japan)を200 μL加えた。
真皮線維芽細胞の増殖に及ぼす被験品の作用を次のとおり評価した。培地を除去した96ウェルプレートを100 μLのPBS(-)で洗浄した。次に、100 μLの0.5 mg/mL 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT, CAS No. 298-93-1,Sigma-Aldrich, USA)溶液を加え、CO2インキュベータ内で2時間インキュベートした。
I型コラーゲンの産生量に対する被験品の効果を直接ELISA法で次のとおり評価した。PBS(-)で4倍希釈した培養上清を高吸着型96ウェルプレートに100 μL添加し、4℃で一晩インキュベートした。標準物質としてI型コラーゲン溶液(Cat No. 009-001-103, RCK, USA)を使用した。
コラーゲンやエラスチンの間で水分を抱え込むと言われているヒアルロン酸の産生量に対する被験品の効果をサンドイッチELISAで次のとおり評価した。
BSA溶液を除去し、200 μLのPBS-Tで洗浄後、PBS(-)で100倍希釈した培養上清を100 μL加え、室温で1時間インキュベートした。標準物質にはヒアルロン酸ナトリウム (Cat No. 087-04511,Wako, Japan) を使用した。
試験ごとに、対照と被験品添加群を対応のないt検定で有意差検定を実施した。検定はいずれも両側で有意水準を5%未満とした。
4-1 オリゴDNA
何れの濃度においても細胞増殖に有意な変化はなかった。一方、細胞当たりのコラーゲン産生率およびヒアルロン酸産生率が、対象と比較して有意に増加した(表2)。この結果から、オリゴDNAは、線維芽細胞のコラーゲン産生およびヒアルロン酸産生を高めることが示唆された。
何れの濃度においても細胞増殖に有意な変化はなかった。一方、細胞あたりのヒアルロン酸産生率が、対照と比較して有意に増加した(表2)。この結果から、ExoVESICLEは、線維芽細胞のヒアルロン酸産生を高めることが示唆された。
何れの濃度においても細胞増殖に有意な変化はなかった。一方、細胞あたりのコラーゲン産生率及びヒアルロン酸産生率が、対照と比較して有意に増加した(表2)。この結果から、オリゴDNAとExoVESICLEは、併用により、線維芽細胞のコラーゲン産生およびヒアルロン酸産生を顕著に高めることが示唆された。
Claims (2)
- サケ科魚類の精巣由来のDNAの加水分解物と細胞外小胞とを有効成分として含む、真皮線維芽細胞のコラーゲン産生促進用剤であって、
前記DNAの加水分解物が、分子量が12,000以下である画分を10~80%含有するものであり、前記DNAの加水分解物の含有量が、0.001質量%以上1質量%以下であり、
前記細胞外小胞が、間葉系幹細胞に由来するものであり、前記細胞外小胞の含有量が、総蛋白質の含有量として0.00001×10 -3 質量%以上0.5×10 -3 質量%以下であり、
前記DNAの加水分解物の含有量と、前記総蛋白質の含有量としての前記細胞外小胞の含有量との質量比が、1:10 -8 ~1:10 -1 である、
前記真皮線維芽細胞のコラーゲン産生促進用剤。 - 請求項1に記載のコラーゲン産生促進用剤を含んでなる化粧品。
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
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WO2006134685A1 (ja) | 2005-06-13 | 2006-12-21 | Nissei Bio Company, Limited | 毛髪手入れ用製剤 |
WO2020022131A1 (ja) | 2018-07-24 | 2020-01-30 | ジェネティックバイオラボ株式会社 | 皮膚の抗老化用剤及び抗老化関連遺伝子発現調節用剤 |
CN113248573A (zh) | 2021-05-18 | 2021-08-13 | 北京戴域生物技术有限公司 | 改善皮肤生理特性的活性肽与间充质干细胞外泌体在药品或化妆品中的应用 |
WO2021200299A1 (ja) | 2020-03-31 | 2021-10-07 | 富士フイルム和光純薬株式会社 | 細胞老化抑制剤、生体組織修復促進剤、遺伝子発現調節剤及び製造方法 |
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WO2006134685A1 (ja) | 2005-06-13 | 2006-12-21 | Nissei Bio Company, Limited | 毛髪手入れ用製剤 |
WO2020022131A1 (ja) | 2018-07-24 | 2020-01-30 | ジェネティックバイオラボ株式会社 | 皮膚の抗老化用剤及び抗老化関連遺伝子発現調節用剤 |
WO2021200299A1 (ja) | 2020-03-31 | 2021-10-07 | 富士フイルム和光純薬株式会社 | 細胞老化抑制剤、生体組織修復促進剤、遺伝子発現調節剤及び製造方法 |
CN113248573A (zh) | 2021-05-18 | 2021-08-13 | 北京戴域生物技术有限公司 | 改善皮肤生理特性的活性肽与间充质干细胞外泌体在药品或化妆品中的应用 |
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