CN104324375A - Nucleic acid nano structure carrier-precious metal photosensitive contrast agent composite for living organism photo-acoustic imaging, preparation method and applications thereof - Google Patents
Nucleic acid nano structure carrier-precious metal photosensitive contrast agent composite for living organism photo-acoustic imaging, preparation method and applications thereof Download PDFInfo
- Publication number
- CN104324375A CN104324375A CN201410426106.8A CN201410426106A CN104324375A CN 104324375 A CN104324375 A CN 104324375A CN 201410426106 A CN201410426106 A CN 201410426106A CN 104324375 A CN104324375 A CN 104324375A
- Authority
- CN
- China
- Prior art keywords
- nucleic acid
- nano structure
- contrast agent
- acid nano
- noble metal
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 106
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 106
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 106
- 239000002086 nanomaterial Substances 0.000 title claims abstract description 93
- 239000002872 contrast media Substances 0.000 title claims abstract description 75
- 238000003384 imaging method Methods 0.000 title claims abstract description 36
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 239000002131 composite material Substances 0.000 title abstract description 13
- 239000010970 precious metal Substances 0.000 title abstract 4
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 55
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 41
- 238000005516 engineering process Methods 0.000 claims abstract description 19
- 239000010931 gold Substances 0.000 claims abstract description 18
- 229910052737 gold Inorganic materials 0.000 claims abstract description 18
- 239000000523 sample Substances 0.000 claims abstract description 15
- 230000008836 DNA modification Effects 0.000 claims abstract description 10
- 239000002078 nanoshell Substances 0.000 claims abstract description 3
- 229910000510 noble metal Inorganic materials 0.000 claims description 64
- 238000000034 method Methods 0.000 claims description 25
- 238000001816 cooling Methods 0.000 claims description 17
- 239000000243 solution Substances 0.000 claims description 16
- 238000000137 annealing Methods 0.000 claims description 14
- 238000001962 electrophoresis Methods 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 13
- 230000008569 process Effects 0.000 claims description 13
- 241001524679 Escherichia virus M13 Species 0.000 claims description 12
- 108091034117 Oligonucleotide Proteins 0.000 claims description 12
- 238000010276 construction Methods 0.000 claims description 12
- 230000004087 circulation Effects 0.000 claims description 11
- 238000001338 self-assembly Methods 0.000 claims description 11
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 claims description 9
- 239000002246 antineoplastic agent Substances 0.000 claims description 7
- 229940041181 antineoplastic drug Drugs 0.000 claims description 7
- 238000001514 detection method Methods 0.000 claims description 7
- 238000000746 purification Methods 0.000 claims description 7
- 201000008275 breast carcinoma Diseases 0.000 claims description 6
- 239000000872 buffer Substances 0.000 claims description 6
- 230000012010 growth Effects 0.000 claims description 6
- 230000015572 biosynthetic process Effects 0.000 claims description 5
- 230000008878 coupling Effects 0.000 claims description 5
- 238000010168 coupling process Methods 0.000 claims description 5
- 238000005859 coupling reaction Methods 0.000 claims description 5
- 238000003786 synthesis reaction Methods 0.000 claims description 5
- 206010006187 Breast cancer Diseases 0.000 claims description 4
- 208000026310 Breast neoplasm Diseases 0.000 claims description 4
- 208000017897 Carcinoma of esophagus Diseases 0.000 claims description 4
- 241000701959 Escherichia virus Lambda Species 0.000 claims description 4
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 4
- 206010025323 Lymphomas Diseases 0.000 claims description 4
- 206010030155 Oesophageal carcinoma Diseases 0.000 claims description 4
- 206010033128 Ovarian cancer Diseases 0.000 claims description 4
- 206010061535 Ovarian neoplasm Diseases 0.000 claims description 4
- 239000011543 agarose gel Substances 0.000 claims description 4
- 239000013078 crystal Substances 0.000 claims description 4
- 201000005619 esophageal carcinoma Diseases 0.000 claims description 4
- 238000013467 fragmentation Methods 0.000 claims description 4
- 238000006062 fragmentation reaction Methods 0.000 claims description 4
- 201000010536 head and neck cancer Diseases 0.000 claims description 4
- 208000014829 head and neck neoplasm Diseases 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 201000005296 lung carcinoma Diseases 0.000 claims description 4
- 230000004048 modification Effects 0.000 claims description 4
- 238000012986 modification Methods 0.000 claims description 4
- 201000001514 prostate carcinoma Diseases 0.000 claims description 4
- 208000000649 small cell carcinoma Diseases 0.000 claims description 4
- 239000012634 fragment Substances 0.000 claims description 3
- 230000003902 lesion Effects 0.000 claims description 3
- 229910052751 metal Inorganic materials 0.000 claims description 3
- 239000002184 metal Substances 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 230000002085 persistent effect Effects 0.000 claims description 3
- 230000001419 dependent effect Effects 0.000 claims description 2
- 238000000338 in vitro Methods 0.000 claims description 2
- 238000011031 large-scale manufacturing process Methods 0.000 claims description 2
- 230000035772 mutation Effects 0.000 claims description 2
- 239000002135 nanosheet Substances 0.000 claims description 2
- 239000013612 plasmid Substances 0.000 claims description 2
- 238000013518 transcription Methods 0.000 claims description 2
- 230000035897 transcription Effects 0.000 claims description 2
- 230000008685 targeting Effects 0.000 abstract description 6
- 230000008901 benefit Effects 0.000 abstract description 4
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 abstract 1
- 238000010521 absorption reaction Methods 0.000 abstract 1
- 230000002708 enhancing effect Effects 0.000 abstract 1
- 239000002091 nanocage Substances 0.000 abstract 1
- 229910052709 silver Inorganic materials 0.000 abstract 1
- 239000004332 silver Substances 0.000 abstract 1
- 108020004414 DNA Proteins 0.000 description 73
- 102000053602 DNA Human genes 0.000 description 72
- 210000004027 cell Anatomy 0.000 description 17
- 239000000463 material Substances 0.000 description 11
- 230000000694 effects Effects 0.000 description 10
- 238000002347 injection Methods 0.000 description 9
- 239000007924 injection Substances 0.000 description 9
- 210000001519 tissue Anatomy 0.000 description 8
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 238000009826 distribution Methods 0.000 description 7
- 239000003814 drug Substances 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 6
- 239000007853 buffer solution Substances 0.000 description 6
- 201000011510 cancer Diseases 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 238000013461 design Methods 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 238000003745 diagnosis Methods 0.000 description 5
- 239000002105 nanoparticle Substances 0.000 description 5
- 239000002773 nucleotide Substances 0.000 description 5
- 125000003729 nucleotide group Chemical group 0.000 description 5
- 241000699670 Mus sp. Species 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 230000000259 anti-tumor effect Effects 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000009396 hybridization Methods 0.000 description 4
- 238000011068 loading method Methods 0.000 description 4
- 239000011777 magnesium Substances 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 238000000482 two photon fluorescence microscopy Methods 0.000 description 4
- INGWEZCOABYORO-UHFFFAOYSA-N 2-(furan-2-yl)-7-methyl-1h-1,8-naphthyridin-4-one Chemical compound N=1C2=NC(C)=CC=C2C(O)=CC=1C1=CC=CO1 INGWEZCOABYORO-UHFFFAOYSA-N 0.000 description 3
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 3
- 238000001069 Raman spectroscopy Methods 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 229910052802 copper Inorganic materials 0.000 description 3
- 239000010949 copper Substances 0.000 description 3
- 108010002255 deoxyhemoglobin Proteins 0.000 description 3
- 239000003937 drug carrier Substances 0.000 description 3
- 239000000975 dye Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 238000012552 review Methods 0.000 description 3
- 101710134784 Agnoprotein Proteins 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N acetic acid Substances CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- 230000002411 adverse Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 229910002091 carbon monoxide Inorganic materials 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000007876 drug discovery Methods 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 239000002122 magnetic nanoparticle Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000002082 metal nanoparticle Substances 0.000 description 2
- 238000010895 photoacoustic effect Methods 0.000 description 2
- 239000003504 photosensitizing agent Substances 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 238000010926 purge Methods 0.000 description 2
- 239000002096 quantum dot Substances 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000009182 swimming Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 2
- NWXMGUDVXFXRIG-WESIUVDSSA-N (4s,4as,5as,6s,12ar)-4-(dimethylamino)-1,6,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4,4a,5,5a-tetrahydrotetracene-2-carboxamide Chemical compound C1=CC=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]4(O)C(=O)C3=C(O)C2=C1O NWXMGUDVXFXRIG-WESIUVDSSA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 229930195573 Amycin Natural products 0.000 description 1
- 238000011729 BALB/c nude mouse Methods 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- 108020003215 DNA Probes Proteins 0.000 description 1
- 239000003298 DNA probe Substances 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- 108020004682 Single-Stranded DNA Proteins 0.000 description 1
- 108091027568 Single-stranded nucleotide Proteins 0.000 description 1
- FMNDTHNESSYWKB-UHFFFAOYSA-N [Ag+].[Au+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O Chemical compound [Ag+].[Au+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O FMNDTHNESSYWKB-UHFFFAOYSA-N 0.000 description 1
- ABUBSBSOTTXVPV-UHFFFAOYSA-H [U+6].CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O Chemical compound [U+6].CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O ABUBSBSOTTXVPV-UHFFFAOYSA-H 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006065 biodegradation reaction Methods 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000004700 cellular uptake Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000004691 chief cell of stomach Anatomy 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000007334 copolymerization reaction Methods 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229960004679 doxorubicin Drugs 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 239000003596 drug target Substances 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000000635 electron micrograph Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- FDWREHZXQUYJFJ-UHFFFAOYSA-M gold monochloride Chemical compound [Cl-].[Au+] FDWREHZXQUYJFJ-UHFFFAOYSA-M 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 239000012216 imaging agent Substances 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- UEGPKNKPLBYCNK-UHFFFAOYSA-L magnesium acetate Chemical compound [Mg+2].CC([O-])=O.CC([O-])=O UEGPKNKPLBYCNK-UHFFFAOYSA-L 0.000 description 1
- 239000011654 magnesium acetate Substances 0.000 description 1
- 210000005075 mammary gland Anatomy 0.000 description 1
- 102000006240 membrane receptors Human genes 0.000 description 1
- 238000002715 modification method Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 239000002073 nanorod Substances 0.000 description 1
- 238000011580 nude mouse model Methods 0.000 description 1
- 229940002988 pegasys Drugs 0.000 description 1
- 108010092853 peginterferon alfa-2a Proteins 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000005204 segregation Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910000033 sodium borohydride Inorganic materials 0.000 description 1
- 239000012279 sodium borohydride Substances 0.000 description 1
- 238000004544 sputter deposition Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses a nucleic acid nano structure carrier, a nucleic acid nano structure carrier-precious metal photosensitive contrast agent composite comprising the same, and preparation methods and applications of the carrier and the composite. The nucleic acid nano structure is a random two-dimensional and/or three-dimensional nano structure constructed by a DNA paper-folding technology. The precious metal components are used to carry out DNA modification on the surface of the carrier and selected from one or more of gold nano rods, gold nano shells, silver-coated gold nano rods, and gold nano cages. The provided composite is used as a photo-acoustic imaging probe, is capable of guaranteeing the absorption contrast enhancing effect of the photo-acoustic contrast agent which is coupled to the composite, and moreover can prominently improve the targeting performance of the contrast agent on tumors. Furthermore, the precious metal photosensitive components which are coupled to the carrier are enriched in the inner of tumor tissues, and thus the solved is the problems that conventional contrast agent is distributed over the whole human body or the contrast agent can only stay on the tumor surfaces. The preparation method has the advantages of simple technology, low cost, convenience, and easy application.
Description
Technical field
The invention belongs to photoacoustic imaging field, relate to a kind of nucleic acid nano structure carrier, comprise its photosensitive contrast agent complex of nucleic acid nano structure carrier-noble metal and their preparation method and application, specifically, the present invention relates to a kind of nucleic acid nano structure carrier for live body photoacoustic imaging, comprise its photosensitive contrast agent complex of nucleic acid nano structure carrier-noble metal and their preparation method and application.
Background technology
Malignant tumor is a kind of important diseases of serious harm human health.In the face of global Tumor incidence and the continuous of mortality rate are risen, and the limitation of current diagnosis and Therapeutic Method, the diagnosis and treatment integrated programme of development high-efficiency low-toxicity has become extremely urgent key subjects.How effective transport molecules image reagent carries out navigating in the earlier detection of tumor and excision, and how realizing detecting and treat integrated, is the hot issue in cancer research now.
Object, under periodically variable illumination, produces hyperacoustic phenomenon and is referred to as optoacoustic effect (Photoacoustic effect) because of expanded by heating.The supersonic sounding element being positioned at organizer surface is utilized to receive this ultrasonic signal, rebuild two dimension or three-dimensional light in tissue according to photoacoustic signal and absorb distributed image, thus the method for carrying out imaging is called as photoacoustic imaging (Photoacoustic imaging).Photoacoustic imaging well combines the advantage of optics and ultra sonic imaging two kinds of technology, obtains better imaging depth and resolution, simultaneously safer.This technology will bring more early stage and effective clinical tumor detection method.
Noble metal nanometer material (nobel metal nano-materials) is the photosensitive contrast agent of a class being caused extensive concern by its photothermal deformation effect.As gold nanorods (gold nanorods, GNRs) near infrared region has good plasma resonance effect (Surface Plasmon Resonance, SPR), and the plasma resonance peak (LSPR) of its longitudinal direction makes gold nanorods be easy to induced by the near infrared light near with its LSPR spike appearance and produce heat (Adv Mater, 2012,24 (36): 4811-41.); Utilize the near two-photon laser of its LSPR spike appearance to carry out exciting, produce obvious two-photon fluorescence (Proc Natl Acad Sci U S A, 2005,102 (44): 15752-6.) and photoacoustic signal (Optics Express 2008,16 (23): 18605-18615; Journal of applied physics, 2007,102 (6): 064701.), the contrast agent of living imaging can be used as.
Applying nano technical development is the key effectively solved the problem for the detection system of malignant tumor.From last century, researcher utilizes nanotechnology to carry out systematic study for the Diagnosis and Treat of malignant tumor.At present existing a large amount of nano-carrier is used successfully among medicament transport and antitumor research and treatment, as bag amycin (Doxorubicin) and Polyethylene Glycol (PEG) interferon-' alpha ' 2a (Pegasys) (Nature Reviews Drug Discovery in the liposome of U.S. FDA approval, 2010,9,615-627; Nature Reviews Drug Discovery, 2005,4,145 – 160; J.Drug Target.2006,14,301 – 310; Nature Review Cancer 2006,6,688 – 701.).The preclinical study simultaneously utilizing nano-particle loading fluorometric reagent to carry out surgical navigational also has greater advance.Although based on nanotechnology, still there is many key issues not yet solve in multi-modal surgical navigational probe, the research field such as multi-functional antitumor probe that integrate test-and-treat in the precedent of existing a lot of delivery system successful Application.In addition, current research nanoparticle drug delivery system comparatively fully, as liposome and macromolecular material, its shape and particle size distribution range wider, thus affect the efficiency that carrier enters cell; Or as metal nanoparticle, although particle diameter and shape easily control, material exists potential source biomolecule safety risks, hinder the further application of clinical research.In sum, more effectively to carry out the Diagnosis and Treat of malignant tumor, one of the key problem being badly in need of solving be how to build a series of micro nano structure make its meet efficient, targeting is controlled, security needs, and in this, as the carrier of molecular image reagent and medicament transport; This carrier system need meet that form and particle diameter easily accurately control, the chemical constitution of its constituent material is easily modified into thus be easy to practical function, can load different kinds of molecules image reagent simultaneously thus realize multi-modality imaging, can load medicine simultaneously thus realize multi-functional, stability and the essential condition such as biocompatibility is better.
Biomacromolecule DNA (deoxyribonucleic acid) (deoxyribonucleic acid, DNA) is the carrier of the hereditary information of the various life entity of nature.In structural DNA technology (structural DNA nanotechnology), DNA is not re-used as the carrier of hereditary information, but by carrying out sequential design in advance, as the primitive forming self-assembled nano structures further.DNA paper folding technology (DNA origami) is the DNA self-assembled nanometer technology of a kind of novelty and uniqueness, namely long single stranded nucleotide sequence and some short oligonucleotide sequences are utilized to be undertaken by base pair complementarity hybridizing the structure being formed and design in advance, be widely used for now making the two dimension of various nanoscale and/or the DNA pattern of three-dimensional and shape (Nature from bottom to top from DNA (Bottom-up), 2006,440,297-302.).Utilize the nanostructured of DNA paper folding technical construction to have structure-controllable, be easy to the feature of modification, utilize DNA paper folding technique construction small molecule, anti-tumor drug carrier to can be used as the new development direction in drug carrier field, have boundless application prospect.
Characteristic based on nucleotide sequence is analyzed, can find compared with traditional anti-tumor medicine transport agent, nucleic acid nano structure has fairly obvious advantage: 1) self assembly nucleic acid nano material is made up of biomacromolecule itself, can biodegradation, have the good biocompatibility that other carriers are incomparable, research shows that self assembly nucleic acid nano material does not have obvious cytotoxicity and immunogenicity; 2) DNA structure itself constructs according to base pair complementarity principle, the package assembly formed has very predictable, along with DNA paper folding art and the development of other self-assembling techniques, folded by the hybridization of DNA chain, required graphic structure can be constructed, the complicated controlled nano-scale patterns of further structure and shape, by DNA probe size reasonable in design, meet in integral level and transport tumor passive target; 3) short chain on DNA nanostructure surface can be used for assembling and has molecular image material, as water-soluble quantum dot, upper conversion fluorescent nano particle, magnetic nanoparticle, Raman dyestuff, gold nanorods etc., and can the accurately kind of control group package material, quantity and locus, realize effective transport and the multi-modality imaging of imaging agents; 4) DNA paper folding probe by designing reasonable size and shape, can extend its half-life of being detained in blood with its supporting molecular image reagent, better realizes the transport of tumor passive target; 5) outer surface of DNA nanostructure can use short chain DNA to hybridize, thus the modification of targeting functional group is carried out at the position selected, as modified ligand groups or the tumor microenvironment targeting group of tumor cell surface receptor-specific, strengthen the active targeting of DNA nano-probe, better realize tumour-specific imaging; 6) quantum dot, upper conversion fluorescent nano particle, magnetic nanoparticle, Raman dyestuff, gold nanorods etc. of DNA vector transport can carry out the fluorescence imaging of living cells/live body, NMR (Nuclear Magnetic Resonance)-imaging, Raman image or photoacoustic imaging, can in the distribution of cellular level and the delivery of live body level observation probe and convergence, and navigate in the earlier detection being expected to carry out tumor clinical and excision; 7) functional modification can be carried out in DNA nanostructure inside, can the activated multiple anti-tumor function composition of efficient loading tool, thus realize the medicine carrying function of DNA nano-probe, and then realize the multi-functional diagnosis and treatment integration nano platform of collection cancer target, test-and-treat.
Nucleic acid nano structure has above-mentioned advantage, have simultaneously and nucleic acid nano structure is used for antineoplastic drug carrier carries out oncotherapy report in cellular level and animal level, but there is no report nucleic acid nano structure being applied to human body or the treatment of animal tumor test and monitoring as photosensitizer carrier at present.Mainly because the bio distribution of molecular image probe in animal level using nucleic acid nano structure as photosensitizer vector construction also lacks understanding.In other words, also do not study the enrichment that can prove whether nucleic acid nano structure can promote photosensitive composition in tumor tissues as the carrier of noble metal photosensitive composition at present thus more effectively carry out live body photoacoustic imaging.
Therefore, nucleic acid nano structure can overcome the technology prejudice of this area as the correlational study of photosensitive composition carrier, and discloses the development trend of the photosensitive contrast agent complex of nucleic acid nano structure-noble metal in diagnosing tumor field.
Summary of the invention
The object of the present invention is to provide a kind of nucleic acid nano structure carrier, comprise its photosensitive contrast agent complex of nucleic acid nano structure carrier-noble metal and their preparation method and application, it can promote the enrichment of photosensitive composition in tumor tissues thus more effectively carry out live body photoacoustic imaging.
For realizing object of the present invention, by the following technical solutions:
A first aspect of the present invention is to provide a kind of nucleic acid nano structure carrier, it has the two dimension and/or 3-D nano, structure that are built by DNA paper folding technology (DNA origami), the nucleic acid nano structure that self assembly is formed particular by scaffold chain (scaffold chains) and folding assisted staple chain (staple strands) carry out hybridizing, described scaffold chain and described staple chain are hybridized by base pair complementarity principle;
Wherein said scaffold chain can be selected from M13 phage genome DNA, the various single stranded circle DNA transformed are carried out based on M13 phage genome DNA, Lambda phage genome DNA, utilize point mutation technology and site-extension non-dependent clone (SLIC) technology from given plasmid large-scale production length-adjustable single stranded circle DNA chain, the M13 phage genome DNA fragmentation obtained by PCR, Lambda phage genome DNA fragmentation and utilize in vitro transcription to obtain RNA fragment in a kind of or mixture of at least two kinds.In addition, also can use other scaffold chain, as long as it can meet the base pair complementarity on specific site, enable staple chain auxiliary foot hand cradle chain carry out folding self assembly and form nucleic acid nano structure.Preferably, described scaffold is M13 phage genome DNA.
Described folding assisted staple chain is the oligonucleotide sequence of synthetic.It is chosen 5-20 site and extends acquisition sequence, the complementary that its acquisition sequence and follow-up DNA modification noble metal nano photosensitive composition adopt.
As can be seen here, preferred nucleic acid nano structure carrier is that the oligonucleotide sequence of M13 phage genome DNA and synthetic carries out hybridization self assembly by base pair complementarity principle and formed.
Preferably, described nucleic acid nano structure carrier is built into triangle, rectangle by use DNA paper folding technology, nanotube-shaped, tetrahedron, Buckie are spherical, nano-sheet, band shape or caged, preferred triangle.
A second aspect of the present invention is the preparation method of the nucleic acid nano structure carrier provided described in first aspect present invention, it comprises the following steps: joined by described staple chain mixed solution in described scaffold chain solution, the mol ratio of described scaffold chain and described staple chain is 1:2-50, mixing; Carry out cooling with 90-100 DEG C for initial temperature and be annealed to 15-25 DEG C, whole process lasts 8-28h, obtained described nucleic acid nano structure carrier.
The mol ratio of wherein said scaffold chain and described staple chain can be such as 1:2,1:3,1:4,1:5,1:8,1:10,1:12,1:15,1:18,1:20,1:22,1:28,1:32,1:35,1:38,1:40,1:42,1:45,1:48 or 1:49, preferred 1:5-30, more preferably 1:10-25, particularly preferably 1:20;
The initial temperature of wherein said cooling annealing can be such as 90 DEG C, 91 DEG C, 92 DEG C, 93 DEG C, 94 DEG C, 95 DEG C, 96 DEG C, 97 DEG C, 98 DEG C, 99 DEG C or 100 DEG C, preferred 92-98 DEG C, more preferably 95 DEG C;
The outlet temperature of wherein said cooling annealing can be such as 15 DEG C, 16 DEG C, 17 DEG C, 18 DEG C, 19 DEG C, 20 DEG C, 21 DEG C, 22 DEG C, 23 DEG C, 24 DEG C or 25 DEG C, preferred 18-22 DEG C, more preferably 20 DEG C;
The persistent period of wherein said cooling annealing process can be 9h, 11h, 13h, 14h, 15h, 18h, 20h, 21h, 22h, 23h, 25h or 27h, preferred 10-26h, more preferably 12-24h.
A third aspect of the present invention is to provide a kind of nucleic acid nano structure carrier-noble metal photosensitive contrast agent complex, and described nucleic acid nano structure carrier-noble metal photosensitive contrast agent complex comprises the photosensitive contrast agent of noble metal and nucleic acid nano structure carrier as above; The photosensitive contrast agent of described noble metal be surface carry out oligonucleotide sequence and modify gold nanorods, gold nanoshell, surperficial contracted payment gold nanorods, the mixture of a kind of in gold nanometer cage or at least two kinds; Described nucleic acid nano structure carrier and the photosensitive contrast agent of described noble metal are by oligonucleotide sequence hybridized coupling.
In a specific embodiment of the present invention, confirm that the nucleic acid nano structure carrier after loading gold nanorods has electrophoretic band clearly by electrophoretogram.The DNA paper folding structure detecting this carrier again with electromicroscopic photograph with the photosensitive contrast agent coupling of noble metal before and after structure change, the photosensitive contrast agent complex of visible nucleic acid nanostructured carrier-noble metal is formed well, and assembling and purge process do not have adverse effect to DNA paper folding-gold nanorods complex.
The photosensitive contrast agent of described noble metal can use any known process preparation in field.In a preferred embodiment, the photosensitive contrast agent of described noble metal is the gold nanorods that sulfhydrylation DNA modification is carried out on surface.
One skilled in the art will appreciate that the preparation method of the photosensitive contrast agent of noble metal of the present invention must not depend on above-mentioned detailed process and parameter, what provide is only preferably technical scheme herein.The photosensitive contrast agent of noble metal needed for those skilled in the art can prepare according to its experience.
A specific embodiments of the present invention using the gold nanorods of sulfhydrylation DNA modification as the photosensitive contrast agent of noble metal, by nucleic acid nano structure carrier of the present invention delivered enter zooblast and animal body after measure its imaging effect.Those skilled in the art can know, and the photosensitive contrast agent of existing noble metal and the photosensitive contrast agent later developed all can carry out coupling with nucleic acid nano structure carrier of the present invention and be prepared into complex, all fall into scope of the present invention.
A fourth aspect of the present invention is the preparation method providing the photosensitive contrast agent complex of the structure carrier-noble metal of the nucleic acid nano described in third aspect present invention, it is characterized in that, comprises the following steps:
(1) nucleic acid nano structure carrier is prepared;
(2) the photosensitive contrast agent of noble metal is prepared;
(3) the noble metal contrast agent that nucleic acid nano structure carrier step (1) obtained and step (2) obtain is that 1:1.5-5 is mixed in solution according to mol ratio, to lower the temperature annealing in the mode of gradient circulation;
(4) solution that step (3) obtains is carried out electrophoresis and remove the photosensitive contrast agent of excessive noble metal, purification obtains nucleic acid nano structure-photosensitive contrast agent complex.
This preparation process is mixed with the photosensitive contrast agent of described noble metal by described nucleic acid nano structure carrier, and DNA hybridization coupling forms the photosensitive contrast agent complex of described nucleic acid nano structure carrier-noble metal.Described nucleic acid nano structure carrier solution mixes with the photosensitive contrast agent solution of described noble metal by it, to lower the temperature gradually annealing in the mode of gradient circulation; The photosensitive contrast agent of the noble metal of excessive separation under certain deposition condition again, purification obtains the photosensitive contrast agent complex of described nucleic acid nano structure carrier-noble metal.
In step (1), the preparation method of nucleic acid nano structure carrier is preparation method according to a second aspect of the present invention, such as by the oligonucleotide sequence of M13 phage genome DNA and described synthetic in molar ratio 1:2-50, preferably 1:5-30, more preferably 1:10-25, particularly preferably 1:20 are mixed in solution, with 90-100 DEG C, preferred 92-98 DEG C, more preferably 95 DEG C is initial temperature, and cooling is annealed to 15-25 DEG C gradually, preferred 18-22 DEG C, more preferably 20 DEG C.The cooling annealing process persistent period is 8-28h, and preferred 10-26h, more preferably 12-24h, obtain nucleic acid nano structure carrier;
Described in step (2), the photosensitive contrast agent of noble metal is prepared by following methods: utilize the gold nanorods that crystal seed induced growth legal system is wrapped up for surface C TAB; Use the sulfhydrylation oligonucleotide sequence of synthesis to replace the CTAB on gold rod surface, complete the DNA finishing of gold nanorods thus; Or particularly, be prepared by the synthesis of the gold nanorods of sulfhydrylation DNA modification mentioned above and surface modification method;
In step (3), the mol ratio of described nucleic acid nano structure carrier and described noble metal contrast agent is 1:1.5,1:2,1:3,1:4,1:5, preferred 1:2; Described cooling is annealed into from temperature 40-45 DEG C, and cooling is annealed to temperature 20-25 DEG C gradually, and this cooling annealing process is a circulation, 30-60 circulation altogether, preferred 30-45 circulation, and every DEG C is a gradient, and every gradient keeps 3-5min;
In step (4), the condition of described electrophoresis is: 0.5-1% agarose gel, electrophoresis ambient temperature 4-10 DEG C, electrophoretic buffer is 0.5-1 × tbe buffer liquid and containing 5-11mM Mg
2+, electrophoretic voltage is 10-15V/cm and electrophoresis time 30-50min.
A fifth aspect of the present invention is to provide the nucleic acid nano structure carrier described in first aspect present invention preparing the application in antitumor drug or lesion detection metal contrast agent.Described nucleic acid nano structure carrier loads antitumor drug or the photosensitive contrast agent of noble metal, and described tumor is the combination of a kind of in ovarian cancer, breast carcinoma, non-small cell carcinoma, head and neck cancer, the esophageal carcinoma, hepatocarcinoma, lung carcinoma glanular cell, carcinoma of prostate and Fei Hejin lymphomas or at least two kinds.
A sixth aspect of the present invention is to provide the photosensitive contrast agent complex of the structure carrier-noble metal of the nucleic acid nano described in third aspect present invention preparing the application in live body photoacoustic imaging contrast agent or tumor operation navigation probe, and described tumor is the combination of a kind of in ovarian cancer, breast carcinoma, non-small cell carcinoma, head and neck cancer, the esophageal carcinoma, hepatocarcinoma, lung carcinoma glanular cell, carcinoma of prostate and Fei Hejin lymphomas or at least two kinds.
In the present invention, " staple chain ", " oligonucleotide sequence " and " short nucleotide sequence " etc. represent that the term of nucleotide fragments or phrase all mean same object and have identical function, namely refer to play the short nucleotide sequence of folding assisted work in order to make its self assembly form specific two dimension and/or three dimensional structure by base pair complementarity principle to scaffold chain.Described " staple chain " is a kind of expression of image, means it and different loci of scaffold chain is nailed together as staple." acquisition sequence " then refers to choosing 5-20 staple chain site and extend the short dna sequence of acquisition sequence, the complementary that its acquisition sequence and DNA modification noble metal nano photosensitive composition adopt.
In the present invention, term " scaffold chain " refers to long DNA or RNA sequence, refers in particular to single-stranded DNA sequence, and it is the body material forming nucleic acid nano structure, and under the folding assisted effect of staple chain, self assembly forms specific two dimension and/or three dimensional structure.
In the present invention, term " nucleic acid nano structure " refers to the scaffold chain specific two dimension that self assembly is formed under the folding assisted effect of staple chain and/or three dimensional structure.It should be noted that, although this title is containing " nanometer " word, but the object of its actual indication might not be defined as Nano grade, those skilled in the art will appreciate that, although the granule of Nano grade is common as pharmaceutical carrier, also some pharmaceutical carrier can reach the rank of micron, therefore, nanometer of the present invention is a general designation, and in fact nucleic acid nano structure represents the structure possessing the technology of the present invention feature of nanometer or micron level.
In the present invention, term " the photosensitive contrast agent complex of nucleic acid nano structure carrier-noble metal " refers to nucleic acid nano structural load corresponding light sensitive ingredient, the structural complexity that (refering in particular to noble metal nano particles in the present invention) or its active component are formed.
Beneficial effect of the present invention is:
(1) the present invention is by DNA paper folding technology, by scaffold chain and folding assisted staple chain with catch chain and carry out hybridization by base pair complementarity principle and complete self assembly and formed with acquisition sequence nucleic acid nano structure carrier, this nucleic acid nano structure carrier can the photosensitive contrast agent of carried noble metal, realizes the combination of nucleic acid nano structure carrier and the photosensitive contrast agent of noble metal.
(2) shown in, nucleic acid nano structure carrier-noble metal photosensitive contrast agent complex has highly sensitive photoacoustic imaging ability, can as image probe, prove in the experiment of cellular level and animal level, this complex not only can ensure the two-photon fluorescence imaging of living cells level, and can realize the photoacoustic imaging ability of Noninvasive and high sensitivity in live body level.
(3) the photoacoustic imaging experiment of animal level proves, nucleic acid nano structure carrier effectively improves the passive targeting of the photosensitive contrast agent of noble metal, make antitumor drug in the inner obviously enrichment of tumor tissues, significantly improve the character that independent noble metal photosensitive composition only rests on tumor tissues surface.There is a large amount of distribution in deoxyhemoglobin 3h after the photosensitive contrast agent of the described nucleic acid nano structure carrier-noble metal of injection, and distributed areas and nucleic acid nano structure-gold nanorods complex signal coincide preferably, after 3-24 hour, nucleic acid nano structure-gold nanorods complex signal distributes at tumor tissues inner homogeneous simultaneously.
(4) the present invention prepares the method for described nucleic acid nano structure carrier, only need according to suitable ratio, folding assisted staple chain to be joined in scaffold chain solution, after mixing, lower the temperature to realize self assembly in suitable temperature range, do not need too many human intervention, therefore technique is simple and convenient and easy.
Nucleic acid nano structure carrier prepared by the present invention loads the photosensitive contrast agent of noble metal can make Complex Probes, as image-forming contrast medium and surgical navigational probe, for detecting and/or treatment tumor in tumor screening and operation.
Accompanying drawing explanation
Fig. 1 is the electrophoretogram of gold nanorods and nucleic acid nano structure carrier-gold nanorods complex.
Fig. 2 is the electron micrograph of leg-of-mutton nucleic acid nano structure carrier and nucleic acid nano structure carrier-gold nanorods complex, and scale is 20nm.
Fig. 3 is the comparison diagram that gold nanorods group and nucleic acid nano structure-gold nanorods complex group carry out two-photon fluorescence imaging effect in living cells, and 20 times of oily mirror observations, scale is 20 μm.
Fig. 4 be the photosensitive contrast agent of noble metal after live body horizontal injection, after different time carries out photoacoustic imaging, material is convergence effect figure in transplanted tumor.
Fig. 5 be the photosensitive contrast agent complex of nucleic acid nano structure carrier-noble metal after live body horizontal injection, after different time carries out photoacoustic imaging, material is convergence effect figure in transplanted tumor.
Detailed description of the invention
Technical scheme of the present invention is further illustrated by detailed description of the invention below in conjunction with accompanying drawing.
The equipment that the present invention is used and material:
Equipment: Mastercycler Pro grads PCR instrument (Eppendorf, Germany), 5810R small-sized high speed centrifugal machine (Eppendorf, Germany), UV-2450 ultraviolet-visible spectrophotometer (Shimadzu, Japan), transmission electron microscope (Tecnei, Japan), two-photon fluorescence microscope (Olympus, Japan), toy photoacoustic imaging system (iThera, Germany).
Raw material: short nucleotide sequence (staple chain, Nature, 2006,440,297-302) is purchased from Shanghai Ying Weijie base Bioisystech Co., Ltd, and M13 phage genome DNA is purchased from New England Biolabs company.CTAB, gold chloride, sodium borohydride, the raw material of the synthesis such as silver nitrate gold nanorods is purchased from Sigma-Aldrich company.
Reagent: buffer solution used in experiment has TAE/Mg
2+buffer solution (pH 8.0) and PBS buffer solution (pH 7.4).Wherein, 1 × TAE/Mg
2+the component of buffer solution (pH 8.0) is: 4 × 10
-2mol L
-1tris, 2 × 10
-2mol L
-1acetic acid, 2.0 × 10
-3mol L
-1eDTA and 1.25 × 10
-2mol L
-1magnesium acetate; 1 × PBS buffer solution (pH 7.4) consists of: 136.9 × 10
-3mol L
-1(8.00g L
-1) NaCl, 2.68 × 10
-3mol L
-1(0.20g L
-1) KCl, 9.75 × 10
-3mol L
-1(1.56g L
-1) Na
2hPO
4h
2o and 1.47 × 10
-3mol L
-1(0.20g L
-1) KH
2pO
4; These buffer solution reagent used is analytical pure, purchased from Sigma-Aldrich company.
Cell: 4T1 mouse mammary carcinoma cell line, purchased from Institute of Basic Medical Sciences of China Concord Medical Science University
Culture medium: RPMI1640 culture medium, add 10% hyclone, cell is inoculated in 100mm
2in culture dish, be placed in 5%CO
2incubator, 37 DEG C of cultivations, when Growth of Cells to degrees of fusion about 80% time go down to posterity; Used medium and hyclone are purchased from Life Technology company.
BALB/c nude mice: purchased from Department Of Medicine, Peking University.
Embodiment 1 prepares nucleic acid nano structure carrier, the photosensitive contrast agent of noble metal and the photosensitive contrast agent complex of nucleic acid nano structure carrier-noble metal
The staple chain of to be the M13 phage genome DNA of 5nM and 100 μ L final concentrations by 50 μ L final concentrations be 50nM and catch chain mixed solution and fully mix at 1mL 1 × TAE/Mg
2+(pH=8.0) in solution, from 95 DEG C, cooling is annealed to 20 DEG C gradually, and annealing incubation time is 12-24h, prepares the nucleic acid nano structure having and design geometric shape in advance.Triangle DNA paper folding used in this experiment to be modified (Nature based on the Rothemund design of 2006,2006,440,297-302.), extend in 7 sites and catch chain (the original staple chain-ordering-3 of 5 '-AAAAAAAAAAAAAAA-'), the complementary that its acquisition sequence and follow-up DNA modification gold rod adopt.
Utilize crystal seed induced growth method with cetyl trimethyl ammonium bromide (CTAB) for surfactant, regulation and control AgNO
3concentration prepares gold nanorods (Chem Mater, 2003,15 (10): 1957-62.).Concrete grammar is as follows:
A) synthesis of crystal seed.Be that the CTAB of 100mM adds in 25mL round-bottomed flask by the concentration of 7.5mL, add 50 μ L concentration 2% (w/v) HAuCl
4.Under stirring, add rapidly 600 μ L concentration is 10mM pre-cooling NaBH
4, stir.Treat that solution colour becomes khaki from colourless, stop stirring, 28-30 DEG C of standing 2h is for subsequent use.
B) growth of gold rod.CTAB and 80 μ L 2% (w/v) HAuCl that 10mL concentration is 100mM is added in 25mL round-bottomed flask
4.Under agitation, adding 75 μ L concentration is 10mM AgNO
3, 50 μ L concentration are ascorbic acid and the 20 μ L seed-solution of 100mM.After stirring 1min, solution is placed in 28 DEG C of tepidarium growth 5h.
C) purification of gold nanorods.Above-mentioned growth solution is added test tube centrifugal (8000rpm, 30min), abandon supernatant, the nano-particle ultra-pure water be sunken at the bottom of pipe is disperseed again and recentrifuge (3000rpm, 25min).Discard precipitation, get supernatant recentrifuge (8000rpm, 30min), the AuNR be sunken at the bottom of pipe 100 μ L ultra-pure waters are disperseed again, utilize ultraviolet-visible spectrophotometer to detect gold nanorods concentration.
Utilize sulfhydrylation DNA to carry out finishing for gold nanorods, concrete grammar is as follows:
Sulfhydrylation DNA (ACGCTTTTTTTTTTTTTTT-SH, 100 μMs) TCEP (20mM, 200 times are excessive) reduction 6h, excessive TCEP G-25 volume-exclusion post centrifugal segregation.Sulfhydrylation DNA (100 μMs) after purification is added and modifies in buffer (containing 0.01% (w/v) SDS, 89mM Tris, 89mM boric acid, 2mM EDTA, 500mM NaCl, pH 5-6), add AuNR (100nM, AuNR:DNA=1:1000) during earthquake and be placed in 28 DEG C of tepidariums and modify.Modify after spending the night and gold rod (AuNR-DNA) centrifugal (8000rpm, the 30min) that modified DNA is removed excessive DNA.
The assembling of DNA paper folding structure and gold nanorods:
DNA paper folding structure, after the assembling of PCR temperature programmed control, utilizes molecular cut off remove excessive DNA staple chain for 100kDa centrifugal column purification and catch chain.The gold rod (AuNR-DNA) of DNA paper folding structure and coated DNA is mixed with the ratio of 1:1.2-1:5, is placed in PCR anneal, prepares gold nanorods-DNA paper folding complex.Annealing conditions is: be a circulation from 45 DEG C-25 DEG C, totally 30 circulations, and every DEG C is a gradient, and every gradient keeps 3min.
Purified in electrophoresis: prepare 1% agarose gel, adds the gold of coated DNA rod, DNA paper folding and DNA paper folding-gold nanorods in glue hole and carries out electrophoresis, and electrophoresis environment is 0.5 × tbe buffer liquid and containing 11mMMg
2+.Under white light, gel is taken pictures after electrophoresis terminates.Under white light, cut out target stripe, utilize gel-purified post 4 DEG C of centrifugal concentratings.
Electronic Speculum: the DNA paper folding-gold nanorods composite construction after 7 μ L purification, through plasma processor glow discharge sputtering, is deposited on copper mesh by plating carbon supporting film (copper mesh), inhales after 10min and abandons, with 0.7% uranium acetate dyeing 30s.Dyeing liquor is abandoned in suction, after copper mesh bone dry, carry out transmission electron microscope observation, and voltage is 80kV, and high contrast pattern is taken pictures.
As shown in Figure 1, the left side first swimming lane is the gold nanorods of sulfhydrylation DNA modification, and four, right side swimming lane is DNA paper folding-gold nanorods composite construction; The band in agarose gel forward position is the gold nanorods band of DNA modification, is the band of target dna paper folding-gold nanorods composite construction in white dashed line frame, is the polymer of composite construction on target stripe.Nucleic acid nano structure after loading gold nanorods as seen from Figure 1 has electrophoretic band clearly.As shown in Figure 2, left figure is DNA paper folding nanostructured Electronic Speculum figure, right figure is DNA paper folding-gold nanorods composite construction Electronic Speculum figure.Assemble the triangle DNA paper folding structure before and after gold nanorods as seen from Figure 2, visible DNA paper folding-gold nanorods complex is formed well, and assembling and purge process do not have adverse effect to DNA paper folding-gold nanorods complex.
The living cells of embodiment 2 nucleic acid nano structure-gold nanorods complex is taken in
Mammary carcinoma 4T1 cell is inoculated in 100mm
2in culture dish, utilize RPMI1640 culture medium (10% hyclone and dual anti-) in 5%CO
2incubator, 37 DEG C of cultivations.When Growth of Cells is to degrees of fusion about 80%, peptic cell, is inoculated in 35mm
2overnight incubation in the burnt capsule of copolymerization, adds gold rod, DNA paper folding-gold nanorods composite construction (0.1nM, complete medium dilutes) and matched group and hatches 24h.Each group of medicinal liquid is abandoned in suction, and utilize two-photon fluorescence microscope to observe after cleaning cell with PBS, excitation wavelength is 800nm.
Observed result as shown in Figure 3, picture is respectively matched group, gold nanorods processed group and DNA paper folding-gold nanorods composite construction processed group from top to bottom, is respectively the superposed signal of light field imaging, two-photon fluorescence imaging and two-photon passage and cell membrane dyestuff from left to right.Can be learnt by Fig. 3, it is good that nucleic acid nano structure effectively can deliver gold nanorods two-photon fluorescence imaging in living cells; Under identical gold nanorods concentration conditions, the cellular uptake successful of nucleic acid nano structure-gold nanorods complex is better than independent gold nanorods.This result illustrates nucleic acid nano structure-gold nanorods complex carries out imaging ability in living cells level.
The live body photoacoustic imaging of embodiment 3 nucleic acid nano structure-gold nanorods complex
Cultivate 4T1 cell to exponential phase, trypsinization, collecting cell, adjustment concentration of cell suspension is 1 × 10
7individual/mL, gets 100 μ L in-situ inoculatings in 5-6 week female BAl BIc/c nude mice dorsal sc, carries out the modeling of subcutaneous mammary gland transplantation tumor.Modeling is after 1 week, when tumor size is to 100mm
3, be divided into 2 groups to carry out administration process, be respectively gold nanorods group (3nM × 150 μ L), nucleic acid nano structure-gold nanorods complex group (3nM × 150 μ L), intravenous drug.Injection after 0,3,24h time utilize toy photoacoustic imaging system to carry out live body photoacoustic imaging for mice with tumor.
Figure 4 shows that the mice with tumor photoacoustic imaging result of injection gold rod rear 0h, 3h and 24h.As seen from Figure 4, after utilizing independent gold nanorods process, after 1-3 hour, contrast agent has been gathered in mice with tumor tumor region, and a large amount of distribution appears in deoxyhemoglobin after injection 3h, and distributed areas and gold rod signal coincide preferably; After injection, 24h only has the photoacoustic signal of gold rod in tumor top layer.Figure 5 shows that the mice with tumor photoacoustic imaging result of 0h, 3h and 24h after injection DNA paper folding-gold nanorods composite construction.And as seen from Figure 5, with gold rod is similar separately, after nucleic acid nano structure-gold nanorods complex process, there is a large amount of distribution in deoxyhemoglobin after injection 3h, and distributed areas and nucleic acid nano structure-gold nanorods complex signal coincide preferably, after 3-24h, nucleic acid nano structure-gold nanorods complex signal distributes at tumor tissues inner homogeneous simultaneously.This result proves that nucleic acid nano structure-gold nanorods complex is a kind of good nano-probe material further, in living cells, with vivo tumor inside, there is good distribution, Noninvasive, real-time photoacoustic imaging can be carried out for tumor tissues in live body level, be expected to provide basis for developing nucleic acid nano structure further as lesion detection contrast agent and excision navigation probe, there is great using value.
Applicant states, the present invention illustrates method detailed of the present invention by above-described embodiment, but the present invention is not limited to above-mentioned method detailed, does not namely mean that the present invention must rely on above-mentioned method detailed and could implement.Person of ordinary skill in the field should understand, any improvement in the present invention, to equivalence replacement and the interpolation of auxiliary element, the concrete way choice etc. of each raw material of product of the present invention, all drops within protection scope of the present invention and open scope.
Claims (10)
1. a nucleic acid nano structure carrier, it is characterized in that, described nucleic acid nano structure carrier has two dimension by DNA paper folding technique construction and/or 3-D nano, structure, the nucleic acid nano structure that self assembly is formed particular by scaffold chain and folding assisted staple chain carry out hybridizing, wherein said scaffold chain is M13 phage genome DNA, the various single stranded circle DNA transformed are carried out based on M13 phage genome DNA, Lambda phage genome DNA, utilize point mutation technology and site-extension non-dependent clone technology from given plasmid large-scale production length-adjustable single stranded circle DNA chain, the M13 phage genome DNA fragmentation obtained by PCR, Lambda phage genome DNA fragmentation or utilize in vitro transcription to obtain RNA fragment in a kind of or mixture of at least two kinds.
2. nucleic acid nano structure carrier according to claim 1, is characterized in that, described scaffold chain and described staple chain are hybridized by base pair complementarity principle;
Preferably, described scaffold is M13 phage genome DNA;
Preferably, described staple chain is the oligonucleotide sequence of synthetic;
Preferably, described nucleic acid nano structure carrier is built into triangle, rectangle by use DNA paper folding technology, nanotube-shaped, tetrahedron, Buckie are spherical, nano-sheet, band shape or caged, preferred triangle.
3. the preparation method of nucleic acid nano structure carrier according to claim 1 and 2, it is characterized in that, comprise the following steps: joined by described staple chain mixed solution in described scaffold chain solution, the mol ratio of described scaffold chain and described staple chain is 1:2-50, mixing; Carry out cooling with 90-100 DEG C for initial temperature and be annealed to 15-25 DEG C, whole process lasts 8-28h, obtained described nucleic acid nano structure carrier.
4. preparation method according to claim 3, is characterized in that, the mol ratio 1:5-30 of described scaffold chain and described staple chain, more preferably 1:10-25, particularly preferably 1:20;
Preferably, the initial temperature of described cooling annealing is 92-98 DEG C, more preferably 95 DEG C;
Preferably, the outlet temperature of described cooling annealing is 18-22 DEG C, more preferably 20 DEG C;
Preferably, the persistent period of described cooling annealing process is 10-26h, more preferably 12-24h.
5. the photosensitive contrast agent complex of nucleic acid nano structure carrier-noble metal, it is characterized in that, described nucleic acid nano structure carrier-noble metal photosensitive contrast agent complex comprises the photosensitive contrast agent of noble metal and nucleic acid nano structure carrier according to claim 1 and 2; The photosensitive contrast agent of described noble metal is the mixture that a kind of in the gold nanorods of oligonucleotide sequence modification, gold nanoshell, the gold nanorods of surperficial contracted payment, gold nanometer cage or at least two kinds are carried out in surface; Described nucleic acid nano structure carrier and the photosensitive contrast agent of described noble metal are by the coupling of oligonucleotide sequence phase.
6. the photosensitive contrast agent complex of nucleic acid nano structure carrier-noble metal according to claim 5, it is characterized in that, the photosensitive contrast agent of described noble metal is the gold nanorods that sulfhydrylation DNA modification is carried out on surface.
7. the preparation method of the photosensitive contrast agent complex of nucleic acid nano structure carrier-noble metal according to claim 5 or 6, is characterized in that, comprise the following steps:
(1) nucleic acid nano structure carrier is prepared;
(2) the photosensitive contrast agent of noble metal is prepared;
(3) the noble metal contrast agent that nucleic acid nano structure carrier step (1) obtained and step (2) obtain is that 1:1.5-5 is mixed in solution according to mol ratio, to lower the temperature annealing in the mode of gradient circulation;
(4) solution that step (3) obtains is carried out electrophoresis and remove the photosensitive contrast agent of excessive noble metal, purification obtains the photosensitive contrast agent complex of nucleic acid nano structure-noble metal.
8. preparation method according to claim 7, is characterized in that, in step (1), the preparation method of nucleic acid nano structure carrier is the preparation method according to claim 3 or 4;
Preferably, described in step (2), the photosensitive contrast agent of noble metal is prepared by the following method: utilize crystal seed induced growth legal system for gold nanorods; Cetyl trimethyl ammonium bromide CTAB is used to modify; Use the sulfhydrylation oligonucleotide sequence of synthesis to substitute CTAB, complete the DNA finishing of gold nanorods thus;
Preferably, in step (3), the mol ratio of described nucleic acid nano structure carrier and described noble metal contrast agent is 1:2; Described cooling is annealed into from temperature 40-45 DEG C, and cooling is annealed to temperature 20-25 DEG C gradually, and this annealing temperature-fall period is a circulation, 30-60 circulation altogether, preferred 30-45 circulation, and every degree Celsius DEG C is a gradient, and every gradient keeps 3-5min;
Preferably, in step (4), the condition of described electrophoresis is: 0.5-1% agarose gel, electrophoresis ambient temperature 4-10 DEG C, electrophoretic buffer is 0.5-1 × tbe buffer liquid and containing 5-11mM Mg
2+, electrophoretic voltage is 10-15V/cm and electrophoresis time 30-50min.
9. nucleic acid nano structure carrier according to claim 1 and 2 is preparing the application in antitumor drug or lesion detection metal contrast agent, it is characterized in that, described nucleic acid nano structure carrier loads antitumor drug or the photosensitive contrast agent of noble metal, and described tumor is the combination of a kind of in ovarian cancer, breast carcinoma, non-small cell carcinoma, head and neck cancer, the esophageal carcinoma, hepatocarcinoma, lung carcinoma glanular cell, carcinoma of prostate and Fei Hejin lymphomas or at least two kinds.
10. the photosensitive contrast agent complex of nucleic acid nano structure carrier-noble metal according to claim 5 or 6 is preparing the application in live body photoacoustic imaging contrast agent or tumor operation navigation probe, and described tumor is the combination of a kind of in ovarian cancer, breast carcinoma, non-small cell carcinoma, head and neck cancer, the esophageal carcinoma, hepatocarcinoma, lung carcinoma glanular cell, carcinoma of prostate and Fei Hejin lymphomas or at least two kinds.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410426106.8A CN104324375A (en) | 2014-08-26 | 2014-08-26 | Nucleic acid nano structure carrier-precious metal photosensitive contrast agent composite for living organism photo-acoustic imaging, preparation method and applications thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410426106.8A CN104324375A (en) | 2014-08-26 | 2014-08-26 | Nucleic acid nano structure carrier-precious metal photosensitive contrast agent composite for living organism photo-acoustic imaging, preparation method and applications thereof |
Publications (1)
Publication Number | Publication Date |
---|---|
CN104324375A true CN104324375A (en) | 2015-02-04 |
Family
ID=52399261
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201410426106.8A Pending CN104324375A (en) | 2014-08-26 | 2014-08-26 | Nucleic acid nano structure carrier-precious metal photosensitive contrast agent composite for living organism photo-acoustic imaging, preparation method and applications thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN104324375A (en) |
Cited By (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104840966A (en) * | 2015-05-05 | 2015-08-19 | 国家纳米科学中心 | Nucleic acid nanometer structure anticancer compound medicine and preparation method and application thereof |
CN104873989A (en) * | 2015-06-11 | 2015-09-02 | 李剑波 | Preparation method of technetium-99m labeled DNA trigonal bipyramid nanoparticles |
CN104962615A (en) * | 2015-06-04 | 2015-10-07 | 西南大学 | DNA flexagon nano structure-nanogold biosensor based on adapter modification and preparing method and application of DNA flexagon nano structure-nanogold biosensor |
CN106141201A (en) * | 2016-08-26 | 2016-11-23 | 首都师范大学 | A kind of method improving gold nanorods light thermal property and photo and thermal stability |
CN106620725A (en) * | 2017-02-16 | 2017-05-10 | 国家纳米科学中心 | Optical and opto-acoustic integration dual-mode molecular image probe as well as preparation method and application thereof |
CN106893722A (en) * | 2017-02-20 | 2017-06-27 | 国家纳米科学中心 | A kind of stimuli responsive type nucleic acid nano structure carrier chirality noble metal nano compound and its preparation method and application |
CN107469088A (en) * | 2017-06-27 | 2017-12-15 | 郑州大学 | A kind of construction method of accurate identification targeted nano carrier based on DNA paper folding arts and its application |
CN107488661A (en) * | 2017-09-21 | 2017-12-19 | 国家纳米科学中心 | A kind of nucleic acid nano structure and its preparation method and application |
CN107760759A (en) * | 2017-11-07 | 2018-03-06 | 上海纳米技术及应用国家工程研究中心有限公司 | The method for detecting prostate cancer target methyl amimoacetic acid |
CN108273056A (en) * | 2018-02-01 | 2018-07-13 | 中国科学院长春应用化学研究所 | A kind of modified gold nano-material/nucleic acid probe nanometer system and preparation method thereof, application |
CN108593558A (en) * | 2018-04-04 | 2018-09-28 | 深圳大学 | One kind being used for Hg2+Photo-acoustic detection probe of detection and preparation method thereof |
CN109477096A (en) * | 2017-06-02 | 2019-03-15 | 清华大学 | DNA paper folding unit substep construction from part |
CN110279713A (en) * | 2019-07-22 | 2019-09-27 | 广西医科大学 | A kind of pharmaceutical combination preparations and preparation method thereof for targeting therapy on tumor |
CN114177312A (en) * | 2021-12-14 | 2022-03-15 | 国家纳米科学中心 | Nucleic acid nano-drug carrier and preparation method and application thereof |
CN114392354A (en) * | 2022-01-14 | 2022-04-26 | 国家纳米科学中心 | DNA nano system for tumor targeting and preparation method and application thereof |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007127020A2 (en) * | 2006-04-21 | 2007-11-08 | Dana-Farber Cancer Institute | Nucleic acid nanotube liquid crystals and use for nmr structure determination of membrane proteins |
CN102127542A (en) * | 2010-12-27 | 2011-07-20 | 江南大学 | Preparation method of self-assembly material having surface-enhanced Raman activity |
-
2014
- 2014-08-26 CN CN201410426106.8A patent/CN104324375A/en active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007127020A2 (en) * | 2006-04-21 | 2007-11-08 | Dana-Farber Cancer Institute | Nucleic acid nanotube liquid crystals and use for nmr structure determination of membrane proteins |
CN102127542A (en) * | 2010-12-27 | 2011-07-20 | 江南大学 | Preparation method of self-assembly material having surface-enhanced Raman activity |
Non-Patent Citations (5)
Title |
---|
PAI-CHI LI,ET AL: "In vivo Photoacoustic Molecular Imaging with", 《OPTICS EXPRESS》 * |
QIAO JIANG ET AL: "DNA Origami as a Carrier for Circumvention of Drug Resistance", 《JOURNAL OF THE AMERICAN CHEMICAL SOCIETY》 * |
YONGGANG KE ET AL: ""Scaffolded DNA Origami of a DNA"", 《NANO LETTERS》 * |
ZHAO ZHAO,ET AL: ""Encapsulation of Gold Nanoparticles in a DNA Origami Cage"", 《ANGEW. CHEM. INT》 * |
申西波: "DNA折纸结构在手性等离子纳米材料和纳米医药领域的应用研究", 《中国优秀硕士学位论文全文数据库-基础科学辑》 * |
Cited By (26)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104840966A (en) * | 2015-05-05 | 2015-08-19 | 国家纳米科学中心 | Nucleic acid nanometer structure anticancer compound medicine and preparation method and application thereof |
CN104840966B (en) * | 2015-05-05 | 2018-02-02 | 国家纳米科学中心 | A kind of nucleic acid nano structure anticancer combination drug and its preparation method and application |
CN104962615A (en) * | 2015-06-04 | 2015-10-07 | 西南大学 | DNA flexagon nano structure-nanogold biosensor based on adapter modification and preparing method and application of DNA flexagon nano structure-nanogold biosensor |
CN104873989B (en) * | 2015-06-11 | 2017-11-03 | 李剑波 | A kind of preparation method of Tc 99m marker DNA trigonal biyramid nano particle |
CN104873989A (en) * | 2015-06-11 | 2015-09-02 | 李剑波 | Preparation method of technetium-99m labeled DNA trigonal bipyramid nanoparticles |
CN106141201A (en) * | 2016-08-26 | 2016-11-23 | 首都师范大学 | A kind of method improving gold nanorods light thermal property and photo and thermal stability |
CN106620725A (en) * | 2017-02-16 | 2017-05-10 | 国家纳米科学中心 | Optical and opto-acoustic integration dual-mode molecular image probe as well as preparation method and application thereof |
CN106620725B (en) * | 2017-02-16 | 2020-02-14 | 国家纳米科学中心 | Optical and photoacoustic integrated bimodal molecular imaging probe and preparation method and application thereof |
CN106893722A (en) * | 2017-02-20 | 2017-06-27 | 国家纳米科学中心 | A kind of stimuli responsive type nucleic acid nano structure carrier chirality noble metal nano compound and its preparation method and application |
CN106893722B (en) * | 2017-02-20 | 2020-05-08 | 国家纳米科学中心 | Stimulus-responsive nucleic acid nanostructure carrier chiral noble metal nano-composite and preparation method and application thereof |
CN109477096A (en) * | 2017-06-02 | 2019-03-15 | 清华大学 | DNA paper folding unit substep construction from part |
CN109477096B (en) * | 2017-06-02 | 2021-08-31 | 清华大学 | DNA paper folding unit step-by-step assembling method |
CN107469088A (en) * | 2017-06-27 | 2017-12-15 | 郑州大学 | A kind of construction method of accurate identification targeted nano carrier based on DNA paper folding arts and its application |
CN107469088B (en) * | 2017-06-27 | 2020-04-03 | 郑州大学 | Construction method for accurately identifying targeted nano-carrier based on DNA origami and application thereof |
CN107488661A (en) * | 2017-09-21 | 2017-12-19 | 国家纳米科学中心 | A kind of nucleic acid nano structure and its preparation method and application |
CN107488661B (en) * | 2017-09-21 | 2020-12-15 | 国家纳米科学中心 | Nucleic acid nano structure and preparation method and application thereof |
CN107760759A (en) * | 2017-11-07 | 2018-03-06 | 上海纳米技术及应用国家工程研究中心有限公司 | The method for detecting prostate cancer target methyl amimoacetic acid |
CN108273056A (en) * | 2018-02-01 | 2018-07-13 | 中国科学院长春应用化学研究所 | A kind of modified gold nano-material/nucleic acid probe nanometer system and preparation method thereof, application |
CN108593558A (en) * | 2018-04-04 | 2018-09-28 | 深圳大学 | One kind being used for Hg2+Photo-acoustic detection probe of detection and preparation method thereof |
CN108593558B (en) * | 2018-04-04 | 2020-12-01 | 深圳大学 | For Hg2+Detected photoacoustic detection probe and preparation method thereof |
CN110279713B (en) * | 2019-07-22 | 2021-04-16 | 广西医科大学 | Pharmaceutical combination preparation for targeted therapy of tumors and preparation method thereof |
CN110279713A (en) * | 2019-07-22 | 2019-09-27 | 广西医科大学 | A kind of pharmaceutical combination preparations and preparation method thereof for targeting therapy on tumor |
CN114177312A (en) * | 2021-12-14 | 2022-03-15 | 国家纳米科学中心 | Nucleic acid nano-drug carrier and preparation method and application thereof |
CN114177312B (en) * | 2021-12-14 | 2024-02-13 | 国家纳米科学中心 | Nucleic acid nano-drug carrier and preparation method and application thereof |
CN114392354A (en) * | 2022-01-14 | 2022-04-26 | 国家纳米科学中心 | DNA nano system for tumor targeting and preparation method and application thereof |
CN114392354B (en) * | 2022-01-14 | 2023-11-07 | 国家纳米科学中心 | DNA nano system for tumor targeting and preparation method and application thereof |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN104324375A (en) | Nucleic acid nano structure carrier-precious metal photosensitive contrast agent composite for living organism photo-acoustic imaging, preparation method and applications thereof | |
Narayan et al. | Mesoporous silica nanoparticles: A comprehensive review on synthesis and recent advances | |
Zhang et al. | Janus nanoparticles: From fabrication to (bio) applications | |
Jiang et al. | DNA nanomaterials for preclinical imaging and drug delivery | |
He et al. | Rationally programming nanomaterials with DNA for biomedical applications | |
Le et al. | Janus particles: Recent advances in the biomedical applications | |
Sabir et al. | DNA based and stimuli-responsive smart nanocarrier for diagnosis and treatment of cancer: Applications and challenges | |
Locatelli et al. | Surface modifications of gold nanorods for applications in nanomedicine | |
Chen et al. | Gold nanoparticles: from nanomedicine to nanosensing | |
Rao | Shedding light on tumors using nanoparticles | |
Shang et al. | Metal nanoparticles for photodynamic therapy: A potential treatment for breast cancer | |
Zhang et al. | Supramolecular hybrids of AIEgen with carbon dots for noninvasive long-term bioimaging | |
Li et al. | Nanosized Janus AuNR-Pt motor for enhancing NIR-II photoacoustic imaging of deep tumor and Pt2+ ion-based chemotherapy | |
Zeng et al. | The applications of functionalized DNA nanostructures in bioimaging and cancer therapy | |
Song et al. | Near-IR responsive nanostructures for nanobiophotonics: Emerging impacts on nanomedicine | |
Jahanban-Esfahlan et al. | Dynamic DNA nanostructures in biomedicine: Beauty, utility and limits | |
CN106620725B (en) | Optical and photoacoustic integrated bimodal molecular imaging probe and preparation method and application thereof | |
Li et al. | Medical micro-and nanomotors in the body | |
CN102441179B (en) | Silicon dioxide base ultrasonic contrast medium/high intensity focused ultrasound (HIFU) synergist and preparation method thereof | |
Aghajanzadeh et al. | Synergic antitumor effect of photodynamic therapy and chemotherapy mediated by nano drug delivery systems | |
Mishra et al. | Advances in DNA origami–cell interfaces | |
Guan et al. | DNA nanodevice-based drug delivery systems | |
CN108543083A (en) | A kind of multi-modal tumor imaging agent and the preparation method and application thereof of biomembrane package | |
CN104840966A (en) | Nucleic acid nanometer structure anticancer compound medicine and preparation method and application thereof | |
Han et al. | Hybrid mesoporous MnO2-upconversion nanoparticles for image-guided lung cancer spinal metastasis therapy |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150204 |
|
RJ01 | Rejection of invention patent application after publication |