KR102650228B1 - Ion-supported bioactive glass nanoparticle and preparation method thereof - Google Patents
Ion-supported bioactive glass nanoparticle and preparation method thereof Download PDFInfo
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- KR102650228B1 KR102650228B1 KR1020210132700A KR20210132700A KR102650228B1 KR 102650228 B1 KR102650228 B1 KR 102650228B1 KR 1020210132700 A KR1020210132700 A KR 1020210132700A KR 20210132700 A KR20210132700 A KR 20210132700A KR 102650228 B1 KR102650228 B1 KR 102650228B1
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- 239000002105 nanoparticle Substances 0.000 title claims abstract description 46
- 239000005313 bioactive glass Substances 0.000 title claims abstract description 41
- 238000002360 preparation method Methods 0.000 title description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims abstract description 84
- 229910052742 iron Inorganic materials 0.000 claims abstract description 32
- -1 iron ion Chemical class 0.000 claims abstract description 32
- 229910052588 hydroxylapatite Inorganic materials 0.000 claims abstract description 13
- 238000004519 manufacturing process Methods 0.000 claims abstract description 13
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 8
- 230000003592 biomimetic effect Effects 0.000 claims abstract description 5
- 239000011575 calcium Substances 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 17
- 239000002245 particle Substances 0.000 claims description 13
- 229910052791 calcium Inorganic materials 0.000 claims description 10
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims description 9
- 239000011148 porous material Substances 0.000 claims description 8
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 claims description 6
- 239000007795 chemical reaction product Substances 0.000 claims description 6
- SZQUEWJRBJDHSM-UHFFFAOYSA-N iron(3+);trinitrate;nonahydrate Chemical compound O.O.O.O.O.O.O.O.O.[Fe+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O SZQUEWJRBJDHSM-UHFFFAOYSA-N 0.000 claims description 6
- 239000011259 mixed solution Substances 0.000 claims description 6
- 238000001354 calcination Methods 0.000 claims description 4
- ZHJGWYRLJUCMRT-UHFFFAOYSA-N 5-[6-[(4-methylpiperazin-1-yl)methyl]benzimidazol-1-yl]-3-[1-[2-(trifluoromethyl)phenyl]ethoxy]thiophene-2-carboxamide Chemical compound C=1C=CC=C(C(F)(F)F)C=1C(C)OC(=C(S1)C(N)=O)C=C1N(C1=C2)C=NC1=CC=C2CN1CCN(C)CC1 ZHJGWYRLJUCMRT-UHFFFAOYSA-N 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 150000002500 ions Chemical class 0.000 abstract description 28
- 230000004806 ferroptosis Effects 0.000 abstract description 13
- 206010005949 Bone cancer Diseases 0.000 abstract description 9
- 208000018084 Bone neoplasm Diseases 0.000 abstract description 9
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 11
- ODINCKMPIJJUCX-UHFFFAOYSA-N calcium oxide Inorganic materials [Ca]=O ODINCKMPIJJUCX-UHFFFAOYSA-N 0.000 description 8
- 150000001768 cations Chemical class 0.000 description 8
- 239000002243 precursor Substances 0.000 description 8
- BRPQOXSCLDDYGP-UHFFFAOYSA-N calcium oxide Chemical compound [O-2].[Ca+2] BRPQOXSCLDDYGP-UHFFFAOYSA-N 0.000 description 7
- 239000000292 calcium oxide Substances 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 206010028980 Neoplasm Diseases 0.000 description 6
- 239000003642 reactive oxygen metabolite Substances 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- 229910004283 SiO 4 Inorganic materials 0.000 description 5
- 238000003917 TEM image Methods 0.000 description 5
- 201000011510 cancer Diseases 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 239000000377 silicon dioxide Substances 0.000 description 5
- 210000004881 tumor cell Anatomy 0.000 description 5
- 239000011521 glass Substances 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 229910052710 silicon Inorganic materials 0.000 description 4
- 238000013268 sustained release Methods 0.000 description 4
- 239000012730 sustained-release form Substances 0.000 description 4
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 229910001424 calcium ion Inorganic materials 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000000354 decomposition reaction Methods 0.000 description 3
- 239000003607 modifier Substances 0.000 description 3
- 239000011858 nanopowder Substances 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 239000010703 silicon Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- UUEWCQRISZBELL-UHFFFAOYSA-N 3-trimethoxysilylpropane-1-thiol Chemical compound CO[Si](OC)(OC)CCCS UUEWCQRISZBELL-UHFFFAOYSA-N 0.000 description 2
- CNODSORTHKVDEM-UHFFFAOYSA-N 4-trimethoxysilylaniline Chemical compound CO[Si](OC)(OC)C1=CC=C(N)C=C1 CNODSORTHKVDEM-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 239000012692 Fe precursor Substances 0.000 description 2
- 208000006644 Malignant Fibrous Histiocytoma Diseases 0.000 description 2
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 208000015778 Undifferentiated pleomorphic sarcoma Diseases 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 230000030833 cell death Effects 0.000 description 2
- 230000006721 cell death pathway Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000002977 hyperthermial effect Effects 0.000 description 2
- 238000007654 immersion Methods 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 229910014472 Ca—O Inorganic materials 0.000 description 1
- 208000005243 Chondrosarcoma Diseases 0.000 description 1
- 201000009047 Chordoma Diseases 0.000 description 1
- 230000005778 DNA damage Effects 0.000 description 1
- 231100000277 DNA damage Toxicity 0.000 description 1
- 208000006168 Ewing Sarcoma Diseases 0.000 description 1
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 description 1
- 229910017135 Fe—O Inorganic materials 0.000 description 1
- 201000008808 Fibrosarcoma Diseases 0.000 description 1
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 1
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 229910004298 SiO 2 Inorganic materials 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 238000002441 X-ray diffraction Methods 0.000 description 1
- 238000000026 X-ray photoelectron spectrum Methods 0.000 description 1
- KHMFYFVXTICBEL-UHFFFAOYSA-N [4-(4-fluorophenyl)phenyl]boronic acid Chemical compound C1=CC(B(O)O)=CC=C1C1=CC=C(F)C=C1 KHMFYFVXTICBEL-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- MCDLETWIOVSGJT-UHFFFAOYSA-N acetic acid;iron Chemical compound [Fe].CC(O)=O.CC(O)=O MCDLETWIOVSGJT-UHFFFAOYSA-N 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- VSGNNIFQASZAOI-UHFFFAOYSA-L calcium acetate Chemical compound [Ca+2].CC([O-])=O.CC([O-])=O VSGNNIFQASZAOI-UHFFFAOYSA-L 0.000 description 1
- 239000001639 calcium acetate Substances 0.000 description 1
- 235000011092 calcium acetate Nutrition 0.000 description 1
- 229960005147 calcium acetate Drugs 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000012830 cancer therapeutic Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- OTARVPUIYXHRRB-UHFFFAOYSA-N diethoxy-methyl-[3-(oxiran-2-ylmethoxy)propyl]silane Chemical compound CCO[Si](C)(OCC)CCCOCC1CO1 OTARVPUIYXHRRB-UHFFFAOYSA-N 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000005684 electric field Effects 0.000 description 1
- 238000002149 energy-dispersive X-ray emission spectroscopy Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229960002089 ferrous chloride Drugs 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 238000007496 glass forming Methods 0.000 description 1
- JELBFNWRSIMKML-UHFFFAOYSA-N hydroxy-tris(methylperoxy)silane Chemical compound COO[Si](O)(OOC)OOC JELBFNWRSIMKML-UHFFFAOYSA-N 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000002354 inductively-coupled plasma atomic emission spectroscopy Methods 0.000 description 1
- NMCUIPGRVMDVDB-UHFFFAOYSA-L iron dichloride Chemical compound Cl[Fe]Cl NMCUIPGRVMDVDB-UHFFFAOYSA-L 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- QZRHHEURPZONJU-UHFFFAOYSA-N iron(2+) dinitrate nonahydrate Chemical compound O.O.O.O.O.O.O.O.O.[Fe+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O QZRHHEURPZONJU-UHFFFAOYSA-N 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- RUTXIHLAWFEWGM-UHFFFAOYSA-H iron(3+) sulfate Chemical compound [Fe+3].[Fe+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O RUTXIHLAWFEWGM-UHFFFAOYSA-H 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 229910000360 iron(III) sulfate Inorganic materials 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000013335 mesoporous material Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002114 nanocomposite Substances 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 238000000696 nitrogen adsorption--desorption isotherm Methods 0.000 description 1
- 238000002429 nitrogen sorption measurement Methods 0.000 description 1
- 230000009818 osteogenic differentiation Effects 0.000 description 1
- 201000008968 osteosarcoma Diseases 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 208000016691 refractory malignant neoplasm Diseases 0.000 description 1
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- 238000003980 solgel method Methods 0.000 description 1
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- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/5115—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/26—Iron; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5192—Processes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
본 발명은 철 이온 생활성 유리 나노입자(Fe-BGn, Fe ions-bioactive glass nanoparticle) 및 이의 제조방법에 관한 것으로, 상기 생활성 유리 나노입자에 철(Fe) 이온을 담지하여 초소형 크기(< 20 nm), 고도의 메조다공성, 생체모방용액(SBF) 내에서 하이드록시아파타이트(hydroxyapatite)를 형성시킬 수 있는 우수한 생활성을 나타내고, 거의 0차 방출 역학으로 Fe3+ 이온이 지속적인 방출되어 페롭토시스(ferroptosis)-기반 골암 치료에 적용될 수 있는 철 이온 생활성 유리 나노입자 및 이의 제조 방법을 제공한다. The present invention relates to iron ion bioactive glass nanoparticles (Fe-BGn, Fe ions-bioactive glass nanoparticles) and a method for manufacturing the same. The bioactive glass nanoparticles are loaded with iron (Fe) ions to produce ultra-small size (< 20 nm), exhibits excellent bioactivity capable of forming hydroxyapatite in a highly mesoporous, biomimetic solution (SBF), and Fe 3+ ions are continuously released with almost zero-order release dynamics, leading to ferroptosis. Provided are iron ion bioactive glass nanoparticles that can be applied to (ferroptosis)-based bone cancer treatment and a method for manufacturing the same.
Description
본 발명은 철 이온이 담지된 생활성 유리 나노입자 및 이의 제조방법에 관한 것으로, 보다 구체적으로는, 생활성 유리 나노입자에 Fe 이온을 담지하여 초소형 크기(< 20 nm), 고도의 메조다공성, 우수한 생활성을 나타내고, 거의 0차 방출 역학으로 Fe3+ 이온의 지속적인 방출을 나타내는 철 이온 생활성 유리 나노입자 및 이의 제조 방법에 관한 것이다. The present invention relates to bioactive glass nanoparticles loaded with iron ions and a method for producing the same. More specifically, the present invention relates to bioactive glass nanoparticles loaded with Fe ions, which have ultra-small size (< 20 nm), high mesoporosity, and bioactive glass nanoparticles. The present invention relates to iron ion bioactive glass nanoparticles that exhibit excellent bioactivity and sustained release of Fe 3+ ions with nearly zero-order release kinetics, and a method for producing the same.
최근에 발견된 페롭토시스(ferroptosis)는 철 이온이 매개하는 펜톤(fenton)의 반응을 통해 활성산소(ROS)를 발생시켜 암세포를 죽이는 유망한 접근법으로 암 치료의 연구분야에서 대단한 관심을 받고 있다. 페롭토시스- 기반 암 치료제 중 페롭토시스의 암 치료를 방해하는 금속 함유 나노 물질은 복잡한 세포 신호 전달없이 종양 세포의 페롭토시스를 빠르고 강력하게 유도하는 데 매우 효과적이다. Fe는 Fe-함유 생체 재료가 온열 암 치료를 위해 적용됨에 따라 핵심 TE 중 하나이다. Fe-함유 생체 재료의 온열 효과 외에도, Fe 이온의 방출은 종양 세포 내부에서 촉매적 H2O2 분해 및 ROS 생성이 가능하다. 실제로, Fe 촉매에 의한 H2O2 분해는 펜톤(fenton)의 반응(Fe3+ + H2O2 = Fe2++ ·OOH + H+ & Fe2+ + H2O2 = Fe3+ + ·OH + HO-)으로 알려져 있으며, 이때 히드록실 라디칼(·OH)과 같은 높은 ROS가 종양 세포에서 막 지질, 단백질 및 DNA 손상의 신속한 산화를 위한 강력한 제제로 생성된다. The recently discovered ferroptosis is a promising approach to kill cancer cells by generating reactive oxygen species (ROS) through the Fenton reaction mediated by iron ions, and is receiving great attention in the field of cancer treatment research. Among ferroptosis-based cancer therapeutics, metal-containing nanomaterials that interfere with ferroptosis in cancer treatment are highly effective in rapidly and robustly inducing ferroptosis of tumor cells without complex cell signaling. Fe is one of the key TEs as Fe-containing biomaterials are applied for hyperthermic cancer treatment. In addition to the hyperthermic effect of Fe-containing biomaterials, the release of Fe ions is capable of catalytic H 2 O 2 decomposition and ROS generation inside tumor cells. In fact, the decomposition of H 2 O 2 by Fe catalyst is Fenton's reaction (Fe 3+ + H 2 O 2 = Fe 2+ + ·OOH + H + & Fe 2+ + H 2 O 2 = Fe 3+ + ·OH + HO - ), where high ROS such as hydroxyl radicals (·OH) are generated as powerful agents for rapid oxidation of membrane lipids, proteins and DNA damage in tumor cells.
따라서, 본 발명자들은 새로운 Fe-BGn을 제조하고, 입자 크기, 조직 특성, 이온 방출 및 시험관 내 뼈와 같은 HA 형성능에 대한 Fe2O3 첨가 효과를 조사하였고, Fe-BGn이 Fe3+ 이온의 지속적인 방출을 보임으로서, 페롭토시스-기반 골암 나노 치료제로 활용될 수 있음을 확인하였다. Therefore, we prepared a new Fe-BGn, investigated the effect of Fe 2 O 3 addition on the particle size, texture properties, ion release and ability to form bone-like HA in vitro, and found that Fe-BGn has a high concentration of Fe 3+ ions. By showing sustained release, it was confirmed that it could be used as a ferroptosis-based bone cancer nanotherapeutic agent.
본 명세서에서 언급된 특허문헌 및 참고문헌은 각각의 문헌이 참조에 의해 개별적이고 명확하게 특정된 것과 동일한 정도로 본 명세서에 참조로 삽입된다.Patent documents and references mentioned herein are herein incorporated by reference to the same extent as if each individual document was individually and specifically identified by reference.
본 발명자들은 철 이온 생활성 유리 나노입자의 장점, 즉, 제조된 졸-겔 유래 Fe-BGn은 초소형 크기(< 20 nm), 고도의 메조다공성, 우수한 생활성을 나타내고, 거의 0차 방출 역학으로 Fe3+ 이온의 지속적인 방출을 실험적으로 증명함으로써 본 발명을 완성하게 되었다. The present inventors have demonstrated the advantages of iron ion bioactive glass nanoparticles, namely, the prepared sol-gel derived Fe-BGn exhibits ultrasmall size (<20 nm), high mesoporosity, excellent bioactivity, and nearly zero-order emission kinetics. The present invention was completed by experimentally proving the continuous release of Fe 3+ ions.
따라서, 본 발명의 목적은 다음의 (a) PEG를 용해시켜 템플레이트 용액을 제조하는 단계; (b) 상기 템플레이트 용액에 산화칼슘 전구체 및 FeNN(Iron nitrate nonahydrate)을 첨가하여 산화칼슘-철-템플레이트 혼합용액을 제조하는 단계; (c) 상기 산화칼슘-철-템플레이트 혼합용액에 실리카 전구체 용액을 첨가하면서 초음파 처리하여 반응 생성물을 제조하는 단계; 및 (d) 상기 반응 생성물을 원심분리한 후 세척하고 건조 및 소성하여 철 이온 생활성 유리 나노입자를 제조하는 단계를 포함하는 철 이온 생활성 유리 나노입자(Fe-BGn, Fe ions-bioactive glass nanoparticle)의 제조 방법을 제공하는 것에 있다. Therefore, the object of the present invention is to prepare a template solution by dissolving PEG (a); (b) adding a calcium oxide precursor and iron nitrate nonahydrate (FeNN) to the template solution to prepare a calcium oxide-iron-template mixed solution; (c) preparing a reaction product by sonicating the calcium oxide-iron-template mixed solution while adding a silica precursor solution; and (d) centrifuging the reaction product, then washing, drying, and calcining to prepare iron ion bioactive glass nanoparticles (Fe-BGn, Fe ions-bioactive glass nanoparticle). ) is to provide a manufacturing method.
본 발명의 다른 목적은 상기 방법으로 제조된 철 이온 생활성 유리 나노입자를 제공하는 것에 있다.Another object of the present invention is to provide iron ion bioactive glass nanoparticles prepared by the above method.
본 발명의 또 다른 목적은 상기 방법으로 제조된 철 이온 생활성 유리 나노입자를 유효성분으로 포함하는 골암 치료용 조성물을 제공하는 것에 있다. Another object of the present invention is to provide a composition for treating bone cancer containing the iron ion bioactive glass nanoparticles prepared by the above method as an active ingredient.
본 발명의 다른 목적 및 기술적 특징은 이하의 발명의 상세한 설명, 청구의 범위 및 도면에 의해 보다 구체적으로 제시된다.Other objects and technical features of the present invention are presented in more detail by the following detailed description, claims, and drawings.
본 발명의 일 양태에 따르면, 다음의 단계를 포함하는 철 이온 생활성 유리 나노입자(Fe-BGn, Fe ions-bioactive glass nanoparticle)의 제조 방법을 제공한다: (a) PEG를 용해시켜 템플레이트 용액을 제조하는 단계; (b) 상기 템플레이트 용액에 산화칼슘 전구체 및 철 전구체를 첨가하여 산화칼슘-철-템플레이트 혼합용액을 제조하는 단계; (c) 상기 산화칼슘-철-템플레이트 혼합용액에 실리카 전구체 용액을 첨가하면서 초음파 처리하여 반응 생성물을 제조하는 단계; 및 (d) 상기 반응 생성물을 원심분리한 후 세척하고 건조 및 소성하여 철 이온 생활성 유리 나노입자를 제조하는 단계. According to one aspect of the present invention, a method for producing iron ion bioactive glass nanoparticles (Fe-BGn, Fe ions-bioactive glass nanoparticle) is provided, comprising the following steps: (a) dissolving PEG to create a template solution; manufacturing step; (b) preparing a calcium oxide-iron-template mixed solution by adding a calcium oxide precursor and an iron precursor to the template solution; (c) preparing a reaction product by sonicating the calcium oxide-iron-template mixed solution while adding a silica precursor solution; and (d) preparing iron ion bioactive glass nanoparticles by centrifuging the reaction product, washing it, drying it, and calcining it.
본 발명의 일 구현예에 따르면, 상기 단계 (b)의 산화칼슘 전구체는 질산칼슘(calcium nitrate tetrahydrate), 염화칼슘(calcium chloride), 아세트산칼슘(calcium acetate), 칼슘메톡시에톡사이드(calcium methoxyethoxide) 또는 이의 혼합물일 수 있다.According to one embodiment of the present invention, the calcium oxide precursor in step (b) is calcium nitrate tetrahydrate, calcium chloride, calcium acetate, and calcium methoxyethoxide. Or it may be a mixture thereof.
본 발명의 일 구현예에 따르면, 상기 단계 (b)의 철 전구체는 질산제2철(FeNN, Ferric nitrate nonahydrate), 염화제1철(Ferrous chloride), 염화제2철(Ferric chloride), 황산제1철(Ferrous sulfate), 황산제2철(Ferric sulfate) 및 초산제1철(Ferrous acetate) 또는 이의 혼합물일 수 있다. According to one embodiment of the present invention, the iron precursor in step (b) is ferric nitrate nonahydrate (FeNN), ferrous chloride, ferric chloride, and sulfuric acid. It may be ferrous sulfate, ferric sulfate, ferrous acetate, or a mixture thereof.
본 발명의 일 구현예에 따르면, 상기 단계 (c)의 실리카 전구체는 TEOS(Tetraethyl orthosilicate), TMOS(trimethoxy orthosilicate), GPTMS((3-glycidoxypropyl)methyldiethoxysilane), MPS(3-mercaptopropyl trimethoxysilane), GOTMS(γtrimethoxysilane), APTMOS(aminophenyl trimethoxysilane) 또는 이의 혼합물일 수 있다.According to one embodiment of the present invention, the silica precursor in step (c) is tetraethyl orthosilicate (TEOS), trimethoxy orthosilicate (TMOS), (3-glycidoxypropyl)methyldiethoxysilane (GPTMS), 3-mercaptopropyl trimethoxysilane (MPS), and GOTMS ( It may be γtrimethoxysilane), aminophenyl trimethoxysilane (APTMOS), or a mixture thereof.
본 발명에서, 상기 생활성 유리 나노입자는 주형(template)으로 서 PEG를 사용하여 초음파 졸-겔법을 이용하여 알칼리 조건 하에서 제조된 것일 수 있다. 상기 주형(template)으로서 PEG를 C1-4 알코올 중에 용해시킨 다음 pH를 9 내지 13으로 조절하고, 상기 PEG 용액에 산화칼슘 전구체를 첨가하여 혼합한 후 실리카 전구체를 첨가하면서 초음파 처리하고 교반하여 반응 생성물을 제조한 뒤, 이를 원심분리한 후 세척하고 건조 및 소성함으로써 메조다공성 생활성 유리 나노입자를 수득할 수 있다. 이때 상기 pH 조절은 NH4OH, NaOH, KOH 또는 Tris로 수행할 수 있다. 또한, 초음파 처리는 출력 전력 200 W 내지 240 W, 10 s on/ 10 s off 사이클로 15분 내지 25분 동안 수행할 수 있고, 소성은 대기하에서 550℃ 내지 650℃ 온도범위에서 5시간 내지 7시간 동안 수행할 수 있다. 상기 메조다공성 생활성 유리 나노입자는 실리카(SiO2)와 산화칼슘(CaO)을 60:40 내지 95:5의 몰 비율로 포함하는 것일 수 있다. In the present invention, the bioactive glass nanoparticles may be prepared under alkaline conditions using an ultrasonic sol-gel method using PEG as a template. As the template, PEG was dissolved in C 1-4 alcohol, the pH was adjusted to 9 to 13, a calcium oxide precursor was added to the PEG solution, mixed, and a silica precursor was added while sonicating and stirring to react. After preparing the product, it can be centrifuged, washed, dried, and calcined to obtain mesoporous bioactive glass nanoparticles. At this time, the pH adjustment can be performed with NH 4 OH, NaOH, KOH, or Tris. In addition, ultrasonic treatment can be performed for 15 to 25 minutes with an output power of 200 W to 240 W and a 10 s on/10 s off cycle, and calcination can be performed for 5 to 7 hours at a temperature range of 550°C to 650°C in air. It can be done. The mesoporous bioactive glass nanoparticles may contain silica (SiO 2 ) and calcium oxide (CaO) at a molar ratio of 60:40 to 95:5.
본 발명의 다른 양태에 따르면, 본 발명은 상기 방법으로 제조된 철 이온 생활성 유리 나노입자를 제공한다.According to another aspect of the present invention, the present invention provides iron ion bioactive glass nanoparticles prepared by the above method.
본 발명에서 “생활성 유리 나노입자(bioactive glass nanoparticle, BGn)”는 실리콘, 칼슘, 인 등의 무기성분으로 구성된 나노입자를 의미하는데, 상기 BGn는 PEG 용액에 칼슘을 포함하는 화합물과 실리콘을 포함하는 화합물을 가하여 반응시킴으로써, PEG, 칼슘 및 실리콘이 응결된 응집체를 수득하고, 상기 응집체를 소결시켜서, PEG를 제거함으로써 제조할 수 있다. 생활성 유리 나노입자(BGn)는 나노필러(nanofiller)로서 나노복합체를 제조할 수 있으며 생분자를 전달할 수 있는 등 많은 장점을 가지고 있다. 또한, 줄기세포의 골분화 촉진, 약물의 로딩과 전달 및 치아의 재미네랄화(dentinremineralization)와 같은 우수한 생물학적 특성을 나타낼 수 있다. 더 나아가, BGn은 졸-겔 제조과정에서 이의 구조 내부에 특정 이온을 결합시킬 수 있고, 고도의 다공 구조가 생성된다.In the present invention, “bioactive glass nanoparticle (BGn)” refers to nanoparticles composed of inorganic components such as silicon, calcium, and phosphorus. BGn contains a compound containing calcium and silicon in a PEG solution. It can be prepared by adding and reacting a compound to obtain an aggregate in which PEG, calcium, and silicon have solidified, and sintering the aggregate to remove PEG. Bioactive glass nanoparticles (BGn) have many advantages, such as being able to manufacture nanocomposites as nanofillers and delivering biomolecules. In addition, it can exhibit excellent biological properties such as promoting osteogenic differentiation of stem cells, drug loading and delivery, and dentinremineralization of teeth. Furthermore, BGn can bind specific ions inside its structure during the sol-gel manufacturing process, and a highly porous structure is created.
생활성 유리 나노입자(BGn)는 종래의 마이크로입자 형태의 BG((bioactive glass)와 비교하여 나노스케일의 크기를 가지면서 더욱 큰 표면적을 가져 이와 관련된 물리화학적 특성 및 생활성 특성이 더욱 우수하다. 이에 더하여, 본 발명의 메조다공성을 가지는 BGn은 세포내 흡수는 가능하면서 치료 분자의 효과적인 담지 및 전달이 가능하다. 결과적으로, 나노스케일 크기 및 실리카-기초의 조성을 갖는 BGn은 우수한 골-생활성 및 세포 및 조직 적합성을 갖는다.Bioactive glass nanoparticles (BGn) have a nanoscale size and a larger surface area compared to conventional microparticle-type BG (bioactive glass), and thus have superior physicochemical and bioactivity properties. In addition, the mesoporous BGn of the present invention is capable of effective loading and delivery of therapeutic molecules while allowing intracellular absorption. As a result, BGn with nanoscale size and silica-based composition has excellent bone bioactivity and Has cell and tissue compatibility.
본 발명의 일 구현예에 따르면, 상기 철 이온 생활성 유리 나노입자는 Si, Ca 및 Fe을 포함한다.According to one embodiment of the present invention, the iron ion bioactive glass nanoparticles include Si, Ca, and Fe.
본 발명의 일 구현예에 따르면, 철 이온 생활성 유리 나노입자에서 Si : Ca : Fe의 중량비는 80 ~ 90 : 5 ~ 15 : 1 ~ 10인 것이 바람직하다. Fe 이온은 종양 세포 내부에서 촉매적 H2O2 분해 및 ROS 생성이 가능하여 암세포를 죽이는 항암효과를 가지며, 또한, BGn 제조시 Ca2+와 Fe3+ 이온 사이의 이온 장(ionic field) 강도에 영향을 주어, 더 단단하고 초소형 크기(< 20 nm)의 나노입자를 형성케 하고, 상기의 중량비율 범위에서 가장 안정한 3차원의 나노기공구조가 형성된다. According to one embodiment of the present invention, the weight ratio of Si:Ca:Fe in iron ion bioactive glass nanoparticles is preferably 80 to 90:5 to 15:1 to 10. Fe ions are capable of catalytic H 2 O 2 decomposition and ROS generation inside tumor cells, which has an anticancer effect that kills cancer cells. Additionally, when manufacturing BGn, the ionic field strength between Ca 2+ and Fe 3+ ions affects the formation of harder and ultra-small size (<20 nm) nanoparticles, and the most stable three-dimensional nanopore structure is formed in the above weight ratio range.
본 발명의 일 구현예에 따르면, 상기 철 이온 생활성 유리 나노입자는 SiO4 4-, Ca2+ 및 Fe3+ 이온을 방출한다. 상기 SiO4 4- (260-340 ppm) 및 Ca2+(175-450 ppm) 이온과 함께, 21-66 ppm(0.38-1.18 mM) 범위의 Fe3+ 이온이 4주 동안 방출될 수 있다. According to one embodiment of the present invention, the iron ion bioactive glass nanoparticles emit SiO 4 4- , Ca 2+ and Fe 3+ ions. Along with the SiO 4 4- (260-340 ppm) and Ca 2+ (175-450 ppm) ions, Fe 3+ ions in the range of 21-66 ppm (0.38-1.18 mM) can be released over a period of 4 weeks.
본 발명의 일 구현예에 따르면, 상기 철 이온 생활성 유리 나노입자는 입자 크기가 < 20 nm이다.According to one embodiment of the present invention, the iron ion bioactive glass nanoparticles have a particle size of <20 nm.
본 발명의 일 구현예에 따르면, 상기 철 이온 생활성 유리 나노입자는 비표면적이 100~300 m2/g이다. According to one embodiment of the present invention, the iron ion bioactive glass nanoparticles have a specific surface area of 100 to 300 m 2 /g.
본 발명에서 “비표면적(specific surface area, SSA)”은 분체, 입자체의 단위 중량 또는 단위 부피당 겉넓이를 의미한다. 비표면적은 또 분체의 입도를 나타내는 특성값으로서 이용된다. 1g의 분체가 갖는 표면적 Sw(cm2/g)로 나타내는 것이 가장 많지만, 단위 부피에 대한 표면적 Sv(cm2/cm3)로 나타내는 것도 있다. 입자가 작아지는데에 따라 이 값은 커지며 계면 현상에 있어 중요한 값이 된다. 비표면적이 넓을수록 실제 나노입자의 표면적이 증가된다. In the present invention, “specific surface area (SSA)” refers to the surface area per unit weight or unit volume of powder or particles. The specific surface area is also used as a characteristic value indicating the particle size of the powder. The surface area of 1 g of powder is most often expressed as Sw (cm 2 /g), but it can also be expressed as the surface area per unit volume, Sv (cm 2 /cm 3 ). As the particle gets smaller, this value increases and becomes an important value in the interface phenomenon. The larger the specific surface area, the greater the surface area of the actual nanoparticle.
본 발명의 일 구현예에 따르면, 상기 철 이온 생활성 유리 나노입자는 총 기공 부피가 0.2~0.5 cm3/g이다. According to one embodiment of the present invention, the iron ion bioactive glass nanoparticles have a total pore volume of 0.2 to 0.5 cm 3 /g.
본 발명의 일 구현예에 따르면, 상기 철 이온 생활성 유리 나노입자는 생체모방용액(SBF) 내에서 하이드록시아파타이트(hydroxyapatite, HA)를 형성시킬 수 있음을 확인하였다.According to one embodiment of the present invention, it was confirmed that the iron ion bioactive glass nanoparticles can form hydroxyapatite (HA) in a biomimetic solution (SBF).
본 발명의 또 다른 양태에 따르면, 철 이온 생활성 유리 나노입자를 유효성분으로 포함하는 골암 치료용 조성물을 제공한다.According to another aspect of the present invention, a composition for treating bone cancer is provided comprising iron ion bioactive glass nanoparticles as an active ingredient.
상기 골암은 연골육종, 골육종, 유잉 육종, 척삭종, 섬유육종 및 악성 섬유성 조직구종(MFH)으로부터 선택될 수 있다.The bone cancer may be selected from chondrosarcoma, osteosarcoma, Ewing's sarcoma, chordoma, fibrosarcoma, and malignant fibrous histiocytoma (MFH).
본 발명의 일 구현예에 따르면, 상기 골암 치료용 조성물은 페롭토시스(ferroptosis)-기반일 수 있다.According to one embodiment of the present invention, the composition for treating bone cancer may be ferroptosis-based.
본 발명에서 “페롭토시스(ferroptosis)”이란 세포막의 지질과산화에 의해 발생하는 철(Ferrous)-의존적 세포사멸 경로이며, 최근 항암제 내성암을 비롯한 다양한 난치암의 효과적인 세포사멸 경로로서 주목받고 있다.In the present invention, “ferroptosis” is an iron-dependent cell death pathway caused by lipid peroxidation of the cell membrane, and has recently been attracting attention as an effective cell death pathway for various incurable cancers, including anticancer drug-resistant cancer.
상기 철(Ferrous)-의존적 세포사멸은 철(Fe)을 조효소로 사용하여 활성산소 생성을 유도하는 세포사멸을 의미한다.The iron (Ferrous)-dependent cell death refers to cell death that induces the production of active oxygen by using iron (Fe) as a coenzyme.
본 발명은 생활성 유리 나노입자에 Fe 이온을 담지하여 초소형 크기(< 20 nm), 고도의 메조다공성, 생체모방용액(SBF) 내에서 하이드록시아파타이트(hydroxyapatite)를 형성시킬 수 있는 우수한 생활성을 나타내고, 거의 0차 방출 역학으로 Fe3+ 이온의 지속적인 방출을 나타내는 철 이온 생활성 유리 나노입자 및 이의 제조 방법을 제공한다. The present invention supports Fe ions in bioactive glass nanoparticles to achieve ultra-small size (< 20 nm), high mesoporosity, and excellent bioactivity that can form hydroxyapatite in a biomimetic solution (SBF). The present invention provides iron ion bioactive glass nanoparticles that exhibit sustained release of Fe 3+ ions with nearly zero-order release kinetics, and a method for producing the same.
도 1은 0Fe-BGn(a), 5Fe-BGn(b) 및 10Fe-BGn(c)의 TEM 이미지, 10Fe-BGn의 광폭 XPS 조사(d)와 Fe2p 코어 수준의 협폭 XPS 스캔(d에 삽입됨) 결과, 및 10Fe-BG 나노분말의 광학 사진 이미지(d에 삽입됨)를 나타낸 것이다.
도 2는 N2 흡착-탈착 등온선(a), NLDFT 기공 크기 분포(a에 삽입됨) 및 BET-plot(b)를 나타낸 것이다.
도 3은 실리케이트와 칼슘 이온(a), 철 이온(b)의 방출 프로파일 및 철 이온 방출의 선형 피팅(b에 삽입됨)을 나타낸 것이다.
도 4a 및 도 4b는 10Fe-BGn의 뼈와 같은 HA 형성 능력을 보여주는 것으로, SBF 침지 21일 후의 XRD 분석 결과(a)와 TEM 이미지(b)(원은 Fe-BGn 위치를 나타냄)를 나타낸 것이다,
도 4c는 BGn 및 Fe-BGn 나노분말의 광학 사진 이미지를 나타낸 것이다.
도 4d는 Fe 이온을 방출하는 Fe-BGn을 페롭토시스(ferroptosis)-기반 골암 나노치료제로서 제안되는 새로운 용도를 설명하는 개략도이다. Figure 1 shows TEM images of 0Fe-BGn (a), 5Fe-BGn (b) and 10Fe-BGn (c), a wide ) results, and an optical photographic image of the 10Fe-BG nanopowder (inserted in d).
Figure 2 shows the N2 adsorption-desorption isotherm (a), NLDFT pore size distribution (inset in a), and BET-plot (b).
Figure 3 shows the emission profiles of silicate and calcium ions (a) and iron ions (b) and a linear fit of the iron ion emission (inset in b).
Figures 4a and 4b show the bone-like HA forming ability of 10Fe-BGn, showing XRD analysis results (a) and TEM images (b) after 21 days of SBF immersion (circles indicate the positions of Fe-BGn). ,
Figure 4c shows optical photographic images of BGn and Fe-BGn nanopowders.
Figure 4d is a schematic diagram illustrating the proposed new use of Fe-BGn, which releases Fe ions, as a ferroptosis-based bone cancer nanotherapeutic.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다, 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명한 것이다, Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples according to the gist of the present invention. This is self-evident to those skilled in the art.
실험 재료 및 방법Experimental materials and methods
1. Fe-BGn의 제조1. Preparation of Fe-BGn
Fe-BGn은 초음파-결합 졸-겔 합성에 의해 제조되었다. Fe2O3는 CaO 대신에 0, 5 및 10wt%로 바이너리(binary) 85% SiO2 - 15% CaO에 첨가되었다. 간단히 말해서, CaNT 및 FeNN을 PEG 용액(5g PEG, 120 ml 에탄올/30 ml 암모니아)에 용해시켰다. 이 후, 20분 동안 격렬한 교반 및 고출력 초음파 조사(10초 켜기/10초 끄기) 하에 TEOS 용액(0.95 ml TEOS/30 ml 에탄올)을 적가하였다. 24시간 동안 격렬하게 교반한 후 수집된 침전물을 세척하고 70℃의 오븐에서 밤새 건조시켰다. 마지막으로, 건조된 분말을 5시간 동안 기류하에 600℃에서 열처리하였다. Fe-BGn was prepared by ultrasound-coupled sol-gel synthesis. Fe 2 O 3 was added to binary 85% SiO 2 - 15% CaO at 0, 5 and 10 wt% instead of CaO. Briefly, CaNT and FeNN were dissolved in PEG solution (5 g PEG, 120 ml ethanol/30 ml ammonia). Afterwards, TEOS solution (0.95 ml TEOS/30 ml ethanol) was added dropwise under vigorous stirring and high-power ultrasonic irradiation (10 seconds on/10 seconds off) for 20 minutes. After vigorous stirring for 24 hours, the collected precipitate was washed and dried in an oven at 70°C overnight. Finally, the dried powder was heat treated at 600°C under airflow for 5 hours.
2. Fe-BGn의 특성 2. Characteristics of Fe-BGn
Fe-BGn은 TEM/EDS 분석으로 조사하였다. Fe-BGn의 평균 입자 크기는 입자 직경의 TEM 측정에 의해 결정되었다(n = 150). XRD는 비정질 구조를 확인하는데 사용되었으며, XPS는 표면 요소를 감지하는 데 사용되었다. BET-SSA, NLDFT 기공 부피/크기는 N2-흡착으로부터 평가하였다. Fe-BGn의 제타 전위는 pH 7.4에서 측정되었다. SiO4 4-, Ca2+ 및 Fe3+ 이온의 방출은 pH 7.4 및 37℃에서 10 ml Tris-HCl 완충액에 50 mg Fe-BGn을 담그는 동안 ICP-AES에 의해 측정되었다. 하이드록시아파타이트(hydroxyapatite, HA) 형성 능력은 Kokubo의 생체모방용액(SBF) 담그기 테스트에 의해 평가하였다. Fe-BGn은 최대 21일 동안 37℃에서 SBF(1 mg/ml)에 담가졌다. Fe-BGn was investigated by TEM/EDS analysis. The average particle size of Fe-BGn was determined by TEM measurements of particle diameter (n = 150). XRD was used to confirm the amorphous structure, and XPS was used to detect surface elements. BET-SSA, NLDFT pore volume/size was assessed from N2-sorption. The zeta potential of Fe-BGn was measured at pH 7.4. The release of SiO 4 4− , Ca 2+ and Fe 3+ ions was measured by ICP-AES during immersion of 50 mg Fe-BGn in 10 ml Tris-HCl buffer at pH 7.4 and 37°C. The ability to form hydroxyapatite (HA) was evaluated by Kokubo's biomimetic solution (SBF) soak test. Fe-BGn was soaked in SBF (1 mg/ml) at 37°C for up to 21 days.
실험 결과Experiment result
1. Fe-BGn의 특성 1. Characteristics of Fe-BGn
0Fe-BGn의 TEM 이미지(도 1a)는 평균 입자 크기가 약 65 nm인 구형 나노입자를 나타내고 있는 반면에, 5Fe-BGn(도 1b) 및 10Fe-BGn(도 1c)의 TEM 이미지는 크기가 20 nm 미만인 초소형 나노입자를 나타내고 있다(표 1). The TEM image of 0Fe-BGn (Figure 1a) shows spherical nanoparticles with an average particle size of approximately 65 nm, while the TEM images of 5Fe-BGn (Figure 1b) and 10Fe-BGn (Figure 1c) show a size of 20 nm. It represents ultra-small nanoparticles less than nm (Table 1).
CaO를 대체하여 첨가된 Fe2O3가 BGn의 입자 크기에 현저한 영향을 미치는 것은 명확하다. 이러한 점은 Ca2+와 Fe3+ 이온 사이의 이온 장(ionic field) 강도의 차이로 설명할 수 있다. 이온 장 강도가 높은 양이온은 더 단단한 유리 네트워크와 산소 패킹 밀도를 증가시켜 개질제(modifiers)를 둘러싼 더 작은 틈새 공간이 만들어진다는 것은 알려져 있다. 양이온의 전계 강도(F)는 F = Zc/(rc + ro)2 = Zc/a2 로 표시되고, 여기서 Zc는 양이온의 전하이고, rc 및 ro는 양이온 및 산소 이온 O2-의 이온 반경(Å)이고, a는 MnOm 산화물에서 양이온-산소(M-O) 결합 거리(Å)이다. r(Ca2+) = 1.06 Å, r(Fe3+) = 0.67 Å 및 r(O2-) = 1.40 Å이 주어지면, F(Ca2+) = 0.33 Å (8배 배위의 Ca과 함께 Ca-O = 2.46 Å 기초하여 계산됨) 및 F (Fe3+) = 0.76 Å (6배 배위의 Fe과 함께 Fe-O = 1.99 Å에 기초하여 계산됨)이다. Dietzel에 따르면, 유리 개질제 양이온의 F는 ≤ 0.4 Å이고, 유리 형성 양이온의 F는 1.3~2Å이고, 중간 양이온의 F는 0.5~1.1 Å이다. Fe3+에 대해 계산된 전계 강도는 Fe3+를 중간 양이온으로 분류한다. 즉, 네트워크 개질자 외에도 유리 형성제로도 작용할 수 있음을 의미한다. 도 1d는 10Fe-BGn의 넓은 XPS 스펙트럼과 Fe 2p 코어 레벨의 좁은 XPS 스캔을 보여준다(삽입). 715 eV 및 ~725 eV에서의 XPS 피크는 Fe3+ 산화 상태에 속하며 각각 Fe 2p3/2 및 Fe 2p1/2에 기인한다. It is clear that Fe 2 O 3 added in place of CaO has a significant effect on the particle size of BGn. This point can be explained by the difference in ionic field strength between Ca 2+ and Fe 3+ ions. It is known that cations with high ionic field strength produce a stiffer glass network and increased oxygen packing density, resulting in a smaller interstitial space surrounding the modifiers. The electric field strength (F) of a cation is given by F = Z c /(r c + r o ) 2 = Z c /a 2 , where Z c is the charge of the cation, r c and r o are the cation and oxygen ion. O 2- is the ionic radius (Å), and a is the cation-oxygen (MO) bond distance (Å) in M n O m oxide. Given r (Ca2+) = 1.06 Å, r (Fe3+) = 0.67 Å and r (O2-) = 1.40 Å, F (Ca2+) = 0.33 Å (based on Ca-O = 2.46 Å with Ca in 8-fold coordination ) and F (Fe3+) = 0.76 Å (calculated based on Fe-O = 1.99 Å with Fe in 6-fold coordination). According to Dietzel, the F of glass modifier cations is ≤ 0.4 Å, the F of glass-forming cations is 1.3 to 2 Å, and the F of intermediate cations is 0.5 to 1.1 Å. The calculated field strength for Fe 3+ classifies Fe 3+ as an intermediate cation. This means that in addition to being a network modifier, it can also act as a glass former. Figure 1d shows the broad XPS spectrum of 10Fe-BGn and the narrow XPS scan of the Fe 2p core level (inset). The XPS peaks at 715 eV and ~725 eV belong to the Fe 3+ oxidation state and are attributed to Fe 2p3/2 and Fe 2p1/2, respectively.
N2-흡착 등온선(도 2a)은 메조다공성 재료의 유형 IV 등온선을 나타내었다. 기공 크기 분포(도 2a에 삽입됨)는 2~10 nm 범위의 주요 메조포어를 나타낸다. BET-플롯(도 2b)은 0Fe-BGn에 비해 5Fe-BGn 및 10Fe-BGn의 비표면적(specific surface area, SSA)이 상당히 더 높음을 보여준다. 또한, Fe-BGn의 표면은 음전하를 띠는 것으로 나타났다(표 1). The N 2 -adsorption isotherm (Figure 2a) represents the type IV isotherm of the mesoporous material. The pore size distribution (inset in Figure 2a) shows major mesopores ranging from 2 to 10 nm. The BET-plot (Figure 2b) shows that the specific surface area (SSA) of 5Fe-BGn and 10Fe-BGn is significantly higher compared to 0Fe-BGn. Additionally, the surface of Fe-BGn was found to be negatively charged (Table 1).
4주 동안의 SiO4 4-, Ca2+ 및 Fe3+ 이온의 방출 프로파일을 도 3(a-b)에 나타내었다. SiO4 4- (260-340 ppm) 및 Ca2+(175-450 ppm) 이온과 함께 21-66 ppm(0.38-1.18 mM) 범위의 Fe3+ 이온이 4주 동안 방출되었다. 참고로, 선형성(y5Fe = 0.69 x + 0.43, R2 = 0.973 & y10Fe = 2.2 x + 2.3, R2 = 0.989)에 대한 Fe3+ 이온의 피팅(도 3b에 삽입됨)은 Fe3+ 이온 방출에 대해 거의 0차 방출 동역학을 나타낸다. The release profiles of SiO 4 4- , Ca 2+ and Fe 3+ ions for 4 weeks are shown in Figure 3(ab). Fe 3+ ions in the range of 21-66 ppm (0.38-1.18 mM) along with SiO 4 4- (260-340 ppm) and Ca 2+ (175-450 ppm) ions were released over a period of 4 weeks. For reference, the fit of Fe 3+ ions (inset in Figure 3b) to linearity (y 5Fe = 0.69 x + 0.43, R 2 = 0.973 & y 10Fe = 2.2 x + 2.3, R 2 = 0.989 ) shows that It exhibits nearly zero-order emission kinetics for ion emission.
도 4(a-b)에 도시된 바와 같이, 10Fe-BGn의 HA 형성능은 XRD 및 TEM에 의해 확인되었다. HA 형성은 도 4a에서 HA 인덱싱된 XRD 피크의 진화에서 입증된 바와 같이 2주 이내에 확인되었다. 10Fe-BGn를 21일 동안 SBF에 담근 후의 TEM(도 4b) 이미지는 초소형 나노입자로부터 바늘 모양의 HA 결정의 성장을 명확하게 보여주었다. 5Fe-BGn의 생활성도 XRD에 의해 확인되었다. 도 4c는 BG(백색)와 Fe-BG(노란색)의 나노분말의 광학 사진을 나타낸 것이다. Fe-BGn의 노란색은 유리 매트릭스에 Fe3+ 이온이 존재하여 나타나는 것이다. 마지막으로, 도 4d는 Fe3+ 이온을 방출하는 Fe-BGn을 페롭토시스(ferroptosis)-기반 골암 나노치료제로서 제안되는 새로운 용도를 설명하는 것이다. As shown in Figure 4(ab), the HA forming ability of 10Fe-BGn was confirmed by XRD and TEM. HA formation was confirmed within 2 weeks as evidenced by the evolution of the HA indexed XRD peak in Figure 4a. The TEM image (Figure 4b) after immersing 10Fe-BGn in SBF for 21 days clearly showed the growth of needle-shaped HA crystals from ultrasmall nanoparticles. The bioactivity of 5Fe-BGn was also confirmed by XRD. Figure 4c shows an optical photograph of nanopowders of BG (white) and Fe-BG (yellow). The yellow color of Fe-BGn appears due to the presence of Fe 3+ ions in the glass matrix. Finally, Figure 4d illustrates the proposed new use of Fe-BGn, which releases Fe 3+ ions, as a ferroptosis-based bone cancer nanotherapeutic.
결론conclusion
새로 제조된 졸-겔 유래 Fe-BGn은 초소형 크기(< 20 nm), 고도의 메조다공성, 적절한 생활성을 나타내고, 거의 0차 방출 역학으로 Fe3+ 이온의 지속적인 방출을 나타냈다. 추가적인 세포 및 동물 연구가 필요하지만, Fe-BGn이 종양 세포에 침투하고 Fe3+ 이온을 방출하여 세포 내 펜톤(fenton)의 반응을 일으켜 ROS를 생성하고 궁극적으로 세포의 페롭토시스(ferroptosis)를 유도할 수 있음이 제안된다. 따라서, Fe 이온을 방출하는 Fe-BGn은 페롭토시스-기반 골암 치료에 적용될 수 있다. The newly prepared sol-gel derived Fe-BGn exhibited ultrasmall size (<20 nm), high mesoporosity, appropriate bioactivity, and exhibited sustained release of Fe 3+ ions with nearly zero-order release kinetics. Although additional cell and animal studies are needed, Fe-BGn penetrates tumor cells and releases Fe 3+ ions, causing an intracellular fenton reaction to generate ROS and ultimately cell ferroptosis. It is suggested that it can be derived. Therefore, Fe-BGn, which releases Fe ions, can be applied in ferroptosis-based bone cancer treatment.
Claims (13)
(b) 상기 템플레이트 용액에 칼슘 나이트레이트 테트라하이드레이트(CaNT, calcium nitrate tetrahydrate) 및 질산제2철(FeNN, Ferric nitrate nonahydrate)을 첨가하여 산화칼슘-철-템플레이트 혼합용액을 제조하는 단계;
(c) 상기 산화칼슘-철-템플레이트 혼합용액에 TEOS(Tetraethyl orthosilicate) 용액을 첨가하면서 초음파 처리하여 반응 생성물을 제조하는 단계; 및
(d) 상기 반응 생성물을 원심분리한 후 세척하고 건조 및 소성하는 단계를 포함하는 방법으로 제조된, Si : Ca : Fe의 중량비는 80 ~ 90 : 5 ~ 15 : 1 ~ 10이며, 비표면적이 100 ~ 300 m2/g이고, 총 기공 부피가 0.2 ~ 0.5 cm3/g인 것을 특징으로 하는, Fe3+를 0차 방출 역학으로 지속적으로 방출하는 철 이온 생활성 유리 나노입자의 제조 방법. (a) dissolving PEG to prepare a template solution;
(b) preparing a calcium oxide-iron-template mixed solution by adding calcium nitrate tetrahydrate (CaNT) and ferric nitrate nonahydrate (FeNN) to the template solution;
(c) preparing a reaction product by adding TEOS (tetraethyl orthosilicate) solution to the calcium oxide-iron-template mixed solution and sonicating it; and
(d) prepared by a method comprising the steps of centrifuging the reaction product, then washing, drying, and calcining, the weight ratio of Si:Ca:Fe is 80 to 90:5 to 15:1 to 10, and the specific surface area is A method for producing iron ion bioactive glass nanoparticles that continuously release Fe 3+ with zero-order release kinetics, characterized in that the particle size is 100 to 300 m 2 /g and the total pore volume is 0.2 to 0.5 cm 3 /g.
상기 철 이온 생활성 유리 나노입자는 입자 크기가 < 20 nm인 것을 특징으로 하는 나노입자. According to clause 4,
The iron ion bioactive glass nanoparticles are nanoparticles, characterized in that the particle size is <20 nm.
상기 철 이온 생활성 유리 나노입자는 생체모방용액(SBF) 내에서 하이드록시아파타이트(hydroxyapatite)를 형성시킬 수 있는 것을 특징으로 하는 나노입자. According to clause 4,
The iron ion bioactive glass nanoparticle is a nanoparticle characterized in that it can form hydroxyapatite in a biomimetic solution (SBF).
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