CN113896906B - 一种电荷引导的微纳米可黏附水凝胶及其制备方法与应用 - Google Patents
一种电荷引导的微纳米可黏附水凝胶及其制备方法与应用 Download PDFInfo
- Publication number
- CN113896906B CN113896906B CN202111030861.0A CN202111030861A CN113896906B CN 113896906 B CN113896906 B CN 113896906B CN 202111030861 A CN202111030861 A CN 202111030861A CN 113896906 B CN113896906 B CN 113896906B
- Authority
- CN
- China
- Prior art keywords
- hydrogel
- cartilage
- charge
- hama
- nano
- 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.)
- Active
Links
- 239000000017 hydrogel Substances 0.000 title claims abstract description 75
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 239000004005 microsphere Substances 0.000 claims abstract description 103
- 239000002502 liposome Substances 0.000 claims abstract description 63
- 239000003814 drug Substances 0.000 claims abstract description 54
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 claims abstract description 14
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims abstract description 9
- REYJJPSVUYRZGE-UHFFFAOYSA-N Octadecylamine Chemical compound CCCCCCCCCCCCCCCCCCN REYJJPSVUYRZGE-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229920002674 hyaluronan Polymers 0.000 claims abstract description 9
- 229960003160 hyaluronic acid Drugs 0.000 claims abstract description 9
- DCUFMVPCXCSVNP-UHFFFAOYSA-N methacrylic anhydride Chemical compound CC(=C)C(=O)OC(=O)C(C)=C DCUFMVPCXCSVNP-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229920000469 amphiphilic block copolymer Polymers 0.000 claims abstract description 7
- 229960003638 dopamine Drugs 0.000 claims abstract description 7
- 239000004327 boric acid Substances 0.000 claims abstract description 6
- -1 boric acid ester Chemical class 0.000 claims abstract description 6
- 238000004132 cross linking Methods 0.000 claims abstract description 4
- 230000002194 synthesizing effect Effects 0.000 claims abstract description 4
- 238000002156 mixing Methods 0.000 claims abstract description 3
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 claims description 58
- 229940079593 drug Drugs 0.000 claims description 40
- 210000001612 chondrocyte Anatomy 0.000 claims description 27
- 229940074391 gallic acid Drugs 0.000 claims description 25
- 235000004515 gallic acid Nutrition 0.000 claims description 25
- 201000008482 osteoarthritis Diseases 0.000 claims description 25
- 238000000034 method Methods 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 9
- 238000006243 chemical reaction Methods 0.000 claims description 7
- 150000002632 lipids Chemical class 0.000 claims description 6
- 229920001503 Glucan Polymers 0.000 claims description 5
- 239000005662 Paraffin oil Substances 0.000 claims description 4
- 208000020084 Bone disease Diseases 0.000 claims description 3
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 claims description 3
- 230000001105 regulatory effect Effects 0.000 claims description 3
- YIFVGFJRFVNSTD-UHFFFAOYSA-N OC(C)(C)C(C)(C)O.C(O)C1=CC=C(C=C1)OB(O)O Chemical compound OC(C)(C)C(C)(C)O.C(O)C1=CC=C(C=C1)OB(O)O YIFVGFJRFVNSTD-UHFFFAOYSA-N 0.000 claims description 2
- 230000002209 hydrophobic effect Effects 0.000 claims description 2
- 210000000845 cartilage Anatomy 0.000 abstract description 57
- 102000055008 Matrilin Proteins Human genes 0.000 abstract description 26
- 108010072582 Matrilin Proteins Proteins 0.000 abstract description 26
- 229920002307 Dextran Polymers 0.000 abstract description 19
- 238000012377 drug delivery Methods 0.000 abstract description 10
- 230000035515 penetration Effects 0.000 abstract description 9
- 239000000969 carrier Substances 0.000 abstract description 6
- 230000008685 targeting Effects 0.000 abstract description 4
- 239000000463 material Substances 0.000 abstract description 2
- 210000003321 cartilage cell Anatomy 0.000 abstract 1
- 229920001690 polydopamine Polymers 0.000 description 42
- OOIBFPKQHULHSQ-UHFFFAOYSA-N (3-hydroxy-1-adamantyl) 2-methylprop-2-enoate Chemical compound C1C(C2)CC3CC2(O)CC1(OC(=O)C(=C)C)C3 OOIBFPKQHULHSQ-UHFFFAOYSA-N 0.000 description 23
- 210000004027 cell Anatomy 0.000 description 21
- 239000003642 reactive oxygen metabolite Substances 0.000 description 20
- 230000000694 effects Effects 0.000 description 15
- 230000006907 apoptotic process Effects 0.000 description 11
- 238000010186 staining Methods 0.000 description 11
- 239000012620 biological material Substances 0.000 description 10
- 239000010410 layer Substances 0.000 description 10
- 239000002105 nanoparticle Substances 0.000 description 10
- 239000003963 antioxidant agent Substances 0.000 description 9
- 239000002086 nanomaterial Substances 0.000 description 9
- 230000036542 oxidative stress Effects 0.000 description 9
- 239000012071 phase Substances 0.000 description 9
- 239000000243 solution Substances 0.000 description 8
- 238000011282 treatment Methods 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 230000001965 increasing effect Effects 0.000 description 7
- 239000002245 particle Substances 0.000 description 7
- PFKFTWBEEFSNDU-UHFFFAOYSA-N 1,1'-Carbonyldiimidazole Substances C1=CN=CN1C(=O)N1C=CN=C1 PFKFTWBEEFSNDU-UHFFFAOYSA-N 0.000 description 6
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 239000012466 permeate Substances 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- 102000008186 Collagen Human genes 0.000 description 5
- 108010035532 Collagen Proteins 0.000 description 5
- 241000700159 Rattus Species 0.000 description 5
- 239000000853 adhesive Substances 0.000 description 5
- 230000001070 adhesive effect Effects 0.000 description 5
- 230000003078 antioxidant effect Effects 0.000 description 5
- 229920001436 collagen Polymers 0.000 description 5
- 231100000135 cytotoxicity Toxicity 0.000 description 5
- 230000003013 cytotoxicity Effects 0.000 description 5
- 230000035699 permeability Effects 0.000 description 5
- 230000004044 response Effects 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 102000000503 Collagen Type II Human genes 0.000 description 4
- 108010041390 Collagen Type II Proteins 0.000 description 4
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 4
- 229920002683 Glycosaminoglycan Polymers 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 4
- 241000283973 Oryctolagus cuniculus Species 0.000 description 4
- 230000004888 barrier function Effects 0.000 description 4
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 4
- 238000000684 flow cytometry Methods 0.000 description 4
- 230000004054 inflammatory process Effects 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 210000000629 knee joint Anatomy 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 238000004445 quantitative analysis Methods 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- OARRHUQTFTUEOS-UHFFFAOYSA-N safranin Chemical compound [Cl-].C=12C=C(N)C(C)=CC2=NC2=CC(C)=C(N)C=C2[N+]=1C1=CC=CC=C1 OARRHUQTFTUEOS-UHFFFAOYSA-N 0.000 description 4
- 230000035882 stress Effects 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- 230000001640 apoptogenic effect Effects 0.000 description 3
- 208000015100 cartilage disease Diseases 0.000 description 3
- 238000010609 cell counting kit-8 assay Methods 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 239000006185 dispersion Substances 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 238000011068 loading method Methods 0.000 description 3
- 210000001365 lymphatic vessel Anatomy 0.000 description 3
- 230000000149 penetrating effect Effects 0.000 description 3
- 230000000638 stimulation Effects 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 2
- 108090001008 Avidin Proteins 0.000 description 2
- 206010007710 Cartilage injury Diseases 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 108010067787 Proteoglycans Proteins 0.000 description 2
- 102000016611 Proteoglycans Human genes 0.000 description 2
- 108010087230 Sincalide Proteins 0.000 description 2
- GZZBZWITJNATOD-UHFFFAOYSA-N [4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methanol Chemical compound O1C(C)(C)C(C)(C)OB1C1=CC=C(CO)C=C1 GZZBZWITJNATOD-UHFFFAOYSA-N 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 210000001188 articular cartilage Anatomy 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 2
- 229960003957 dexamethasone Drugs 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 230000003628 erosive effect Effects 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 230000036732 histological change Effects 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 238000003125 immunofluorescent labeling Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000002757 inflammatory effect Effects 0.000 description 2
- 239000000787 lecithin Substances 0.000 description 2
- 229940067606 lecithin Drugs 0.000 description 2
- 235000010445 lecithin Nutrition 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 230000008506 pathogenesis Effects 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 2
- 210000001179 synovial fluid Anatomy 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 238000013042 tunel staining Methods 0.000 description 2
- 239000003981 vehicle Substances 0.000 description 2
- 238000005406 washing 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
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 description 1
- 244000061520 Angelica archangelica Species 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- 229920001287 Chondroitin sulfate Polymers 0.000 description 1
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 235000001287 Guettarda speciosa Nutrition 0.000 description 1
- 238000012404 In vitro experiment Methods 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 241000702777 Lachesilla anna Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- CERQOIWHTDAKMF-UHFFFAOYSA-N Methacrylic acid Chemical compound CC(=C)C(O)=O CERQOIWHTDAKMF-UHFFFAOYSA-N 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- NWGKJDSIEKMTRX-AAZCQSIUSA-N Sorbitan monooleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O NWGKJDSIEKMTRX-AAZCQSIUSA-N 0.000 description 1
- 208000031737 Tissue Adhesions Diseases 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000002424 anti-apoptotic effect Effects 0.000 description 1
- 229940124599 anti-inflammatory drug Drugs 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 229920001222 biopolymer Polymers 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- YCIMNLLNPGFGHC-UHFFFAOYSA-N catechol Chemical group OC1=CC=CC=C1O YCIMNLLNPGFGHC-UHFFFAOYSA-N 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000008809 cell oxidative stress Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000002648 chondrogenic effect Effects 0.000 description 1
- 229940059329 chondroitin sulfate Drugs 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 229940109262 curcumin Drugs 0.000 description 1
- 235000012754 curcumin Nutrition 0.000 description 1
- 239000004148 curcumin Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- DCOPUUMXTXDBNB-UHFFFAOYSA-N diclofenac Chemical compound OC(=O)CC1=CC=CC=C1NC1=C(Cl)C=CC=C1Cl DCOPUUMXTXDBNB-UHFFFAOYSA-N 0.000 description 1
- 229960001259 diclofenac Drugs 0.000 description 1
- VFLDPWHFBUODDF-UHFFFAOYSA-N diferuloylmethane Natural products C1=C(O)C(OC)=CC(C=CC(=O)CC(=O)C=CC=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-UHFFFAOYSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 235000013345 egg yolk Nutrition 0.000 description 1
- 210000002969 egg yolk Anatomy 0.000 description 1
- 238000001523 electrospinning Methods 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000005713 exacerbation Effects 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 238000001125 extrusion Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000010408 film Substances 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 238000007306 functionalization reaction Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000034659 glycolysis Effects 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000008798 inflammatory stress Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- JDNTWHVOXJZDSN-UHFFFAOYSA-N iodoacetic acid Chemical compound OC(=O)CI JDNTWHVOXJZDSN-UHFFFAOYSA-N 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000005461 lubrication Methods 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000003349 osteoarthritic effect Effects 0.000 description 1
- 208000005368 osteomalacia Diseases 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 150000008105 phosphatidylcholines Chemical class 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 229920003213 poly(N-isopropyl acrylamide) Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002028 premature Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 229940002612 prodrug Drugs 0.000 description 1
- 239000000651 prodrug Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000005316 response function Methods 0.000 description 1
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000001878 scanning electron micrograph Methods 0.000 description 1
- 238000004626 scanning electron microscopy Methods 0.000 description 1
- 238000007493 shaping process Methods 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000002344 surface layer Substances 0.000 description 1
- 238000013269 sustained drug release Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 238000011269 treatment regimen Methods 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
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1629—Organic macromolecular compounds
- A61K9/1652—Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid
-
- 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
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
- A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
-
- 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
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F251/00—Macromolecular compounds obtained by polymerising monomers on to polysaccharides or derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
- C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
- C08J3/075—Macromolecular gels
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/28—Treatment by wave energy or particle radiation
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J7/00—Chemical treatment or coating of shaped articles made of macromolecular substances
- C08J7/12—Chemical modification
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
- C08J2305/02—Dextran; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2351/00—Characterised by the use of graft polymers in which the grafted component is obtained by reactions only involving carbon-to-carbon unsaturated bonds; Derivatives of such polymers
- C08J2351/02—Characterised by the use of graft polymers in which the grafted component is obtained by reactions only involving carbon-to-carbon unsaturated bonds; Derivatives of such polymers grafted on to polysaccharides
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/02—Dextran; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2451/00—Characterised by the use of graft polymers in which the grafted component is obtained by reactions only involving carbon-to-carbon unsaturated bonds; Derivatives of such polymers
- C08J2451/02—Characterised by the use of graft polymers in which the grafted component is obtained by reactions only involving carbon-to-carbon unsaturated bonds; Derivatives of such polymers grafted on to polysaccharides
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Polymers & Plastics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nanotechnology (AREA)
- Epidemiology (AREA)
- Physical Education & Sports Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Rheumatology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Dispersion Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Crystallography & Structural Chemistry (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- General Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Biophysics (AREA)
- Dermatology (AREA)
- Neurosurgery (AREA)
- Biotechnology (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- General Physics & Mathematics (AREA)
- Manufacturing & Machinery (AREA)
- Immunology (AREA)
- Biomedical Technology (AREA)
- Medicinal Preparation (AREA)
Abstract
本发明属于药用载体技术领域,提供了一种电荷引导的微/纳米可黏附水凝胶微球及其制备方法与应用,该水凝胶微球制备方法如下:(1)以硼酸酯和葡聚糖反应形成两亲性嵌段共聚物,加入硬脂胺,得到以两亲性嵌段共聚物为核心的带正电荷的脂质体;(2)以透明质酸和甲基丙烯酸酐合成水凝胶,利用微流控装置,将水凝胶和脂质体混合,制备水凝胶微球,紫外光交联,形成交联水凝胶微球;(3)在交联水凝胶微球表面接枝多巴胺,获得电荷引导的微/纳米可黏附水凝胶微球。本发明建立了一种高效的软骨粘附、软骨基质穿透、软骨细胞靶向的给药系统,解决了药物难以渗透软骨基质的难题,是一种极具潜力的软骨药物高效递送材料。
Description
技术领域
本发明属于药用载体技术领域,具体涉及一种电荷引导的微/纳米可黏附水凝胶微球及其制备方法与应用。
背景技术
众所周知,利用生物材料加载药物,配合生物材料的功能化,是提高药物疗效的主要途径。其中,提高药物利用率是生物材料最重要的功能之一,它可以减少药物的用量,减少药物的毒副作用。提高药物穿透组织的能力是目前提高药物利用率最重要的方法之一。常规生物材料(如水凝胶和静电纺丝)可有效地将药物渗透到病变部位,如具有良好渗透性的组织(如皮肤、粘膜、肌肉等)。然而,一些组织或器官(如肿瘤和大脑)由于其特殊的生物结构或组成,往往导致药物渗透困难。
软骨也同样面临着药物渗透困难的问题。软骨是一种没有血管、神经和淋巴的组织,其基质由胶原蛋白、纤维网和蛋白多糖组成。胶原纤维网络由大量的II型胶原和少量的Ⅸ型、Ⅺ型胶原组成,孔径为60~200nm,形成高密度的细胞外基质。蛋白多糖中存在大量的带负电荷的硫酸软骨素粘多糖链,使软骨基质具有很强的负静电屏障作用。因此,这两个巨大的屏障阻碍了软骨的药物渗透。目前,由于尚缺乏有效的给药系统来突破这些障碍,难以满足临床对软骨疾病药物疗效的需求。
设计合适的药物递送载体,是克服软骨给药两大障碍和实现有效给药的重点。目前,药物传递载体通常设计在纳米尺度上,以克服软骨基质的致密结构带来的给药障碍。Lin et al.设计了可降解的聚(n-异丙基丙烯酰胺)纳米颗粒,用于负载抗炎肽,使得软骨细胞摄入的纳米颗粒可以抑制软骨炎症。鉴于软骨基质的负电荷性质,人们在药物上修饰阳离子基团以增加其渗透性。Bajpayee et al.以带正电的亲和素作为模型药物,研究了电荷驱动的软骨运输和药物运输,证明了带正电的亲和素能更好地被软骨吸收,并且在软骨中停留的时间更长。此外,Bajpayee et al.通过将地塞米松前药与带正电的壳聚糖共价结合,使药物更好地渗透到软骨基质中。因此,使用带正电荷的纳米粒子修饰药物是克服两大软骨障碍的可能解决方案之一。然而,实验证明,纳米粒子可能引起的炎症反应和带正电荷的药物修饰导致的疗效降低或改变都是该方案的严重缺陷。
脂质体由于其高生物相容性、不易引起炎症反应、物理化学性质(如大小、导电性、渗透性)具有可塑性,目前已被认为是软骨药物的理想载体之一。Ji et al.使用饱和磷脂酰胆碱改性脂质体,负载抗炎药物,以达到持续药物释放和改善骨关节炎环境的润滑性的作用。此外,Elron-Gross et al.应用载药地塞米松和双氯芬酸脂质体治疗软骨病表现出显著提高疗效,降低了药物初始剂量。Elsaid et al.使用罗丹明标记的脂质体来证明脂质体穿透软骨基质的能力,结果表明,细胞极化肽修饰的脂质体具有较强的软骨细胞结合能力。因此,带正电的脂质体载体可以克服软骨致密结构的阻碍和带负电的软骨,穿透软骨基质,实现药物的控释,是一种理想的软骨疾病药物纳米载体。
骨关节炎(OA)的发病机制之一是软骨细胞的氧化应激反应,导致软骨细胞凋亡,软骨基质降解,关节软骨损伤加重。目前,有效的OA治疗方法之一,是向关节腔注射抗氧化剂,清除软骨细胞中的ROS(生物体内活性氧)。研究表明,当归多糖、姜黄素、没食子酸(GA)等抗氧化剂能显著抑制氧化应激诱导的软骨细胞凋亡,改善OA疾病的进展。然而,这些抗氧化剂具有化学活性,容易与滑液中所含成分发生反应,从而降低了药物对ROS的去除效率。此外,它们不能积极穿透软骨基质,这进一步降低了药物的效果。
使用含抗氧化剂的带正电荷脂质体可有效地实现药物渗透软骨基质。然而,关节腔内的纳米颗粒由于直径过小很容易被毛细血管和淋巴管清除,只有当净流入足够大时,才能确保在被清除前有足够的治疗药物浓度,而大剂量注射药物会增加药物的毒性副作用。可注射的水凝胶微球载体目前已被引入,以克服毛细血管和淋巴管的药物去除。如Yanget al.利用水凝胶的微球特性作为微米大小的颗粒,可以长时间停留在关节腔内,通过表面改性,水凝胶微球可在关节腔内起到轴承润滑作用。Zhang et al.将药物装入水凝胶微球中,既能防止药物被移除,又能控制药物的释放。适当大小的水凝胶微球可以最大限度地避免被血管或淋巴管清除,从而延长半衰期;然而,微米级粒子载体既不能穿透软骨,也不能与软骨结合,它悬浮在滑膜液中,不断将药物释放到关节腔内,不利于药物的利用。
因此,如何利用生物材料载体,延长抗氧化剂在关节腔内的滞留时间,同时使药物能穿透软骨基质并靶向软骨,是目前亟待突破的技术瓶颈。
发明内容
本发明的目的就是为了解决上述技术问题,从而提供一种电荷引导的微/纳米可黏附水凝胶微球及其制备方法与应用。本发明创造性地构建了具有电荷导向的二级纳米结构的可注射黏附水凝胶微球。在电荷引导下,带正电荷的二级纳米结构从水凝胶微球中释放并渗透软骨,从而建立了一种高效的软骨粘附、软骨基质渗透、软骨细胞靶向的给药系统,解决了药物难以渗透软骨基质的难题。
本发明的目的之一是提供一种电荷引导的微/纳米可黏附水凝胶微球的制备方法,其包括以下步骤:
(1)以疏水性硼酸酯和亲水性葡聚糖进行接枝反应,形成两亲性嵌段共聚物,然后以硬脂胺作为脂质层调节电荷的阳性成分,得到以两亲性嵌段共聚物为核心的带正电荷的脂质体;
(2)采用透明质酸和甲基丙烯酸酐合成水凝胶,利用微流控装置,将所述水凝胶和步骤(1)所得脂质体混合,制备水凝胶微球,进行紫外光交联,形成交联水凝胶微球;
(3)在交联水凝胶微球表面接枝多巴胺,获得电荷引导的微/纳米可黏附水凝胶微球。
由于软骨基质致密的结构和所携带的高密度负电荷的特点,均严重阻碍药物渗透进入软骨组织。目前,难以有效突破这些阻碍,进行软骨细胞靶向给药。本发明首次构建了一种具有电荷导向的二级纳米结构的可注射可黏附水凝胶微球,其是一种微/纳米组合水凝胶微球,本发明建立了一种高效的软骨粘附、软骨基质渗透、软骨细胞靶向的给药系统,很好解决了药物难以渗透软骨基质的难题。
本发明的水凝胶微球作为给药系统,在电荷引导下,带正电荷的二级纳米结构从水凝胶微球中释放出来并穿透软骨,而微球表面独特的多巴胺修饰结构可以将微球附着在软骨表面。带正电的脂质体可携带药物穿透软骨基质,在ROS刺激下释放药物,作用于软骨细胞。在实施例中,本发明通过大鼠OA模型进一步验证了电荷引导微/纳米水凝胶微球对OA的作用。本发明的电荷引导的微/纳米可黏附水凝胶微球可显著渗透软骨基质,提高GA疗效,抑制氧化应激诱导的软骨细胞凋亡,缓解OA进展。总的来说,电荷引导的微/纳米可黏附水凝胶微球可以提供一种潜在的生物材料,用于软骨药物的高效输送。
本发明的上述合成方法中,首先,成功合成了带有硼酸酯(如4-羟甲基苯硼酸频哪醇酯)修饰的葡聚糖(PHB-dextran)作为ROS响应的生物聚合物;随后,使用硬脂胺作为脂质层调节电荷的阳性成分,制备以PHB-dextran为核心的带正电荷的脂质体,从而可以被带负电的软骨基质吸引,渗透深入软骨基质,释放GA靶向作用于软骨细胞;最后,利用透明质酸(HA)和甲基丙烯酸酐(MA)合成HAMA,将脂质体搭载于多巴胺修饰的HAMA微球,既能提高脂质体在关节腔内抗清除能力,又利用微球黏附于关节表面,实现药物在软骨附近富集。
进一步的是,步骤(1)中所述硼酸酯为4-羟甲基苯硼酸频哪醇酯,所述硼酸酯与葡聚糖的质量比为2:1~4。
进一步的是,步骤(2)中所述透明质酸与甲基丙烯酸酐的质量比为10:1~20。
进一步的是,步骤(2)中在采用微流控装置进行制备时,是以水凝胶和脂质体的混合物为分散相,以石蜡油为连续相。
进一步的是,步骤(2)中所述水凝胶和脂质体的质量比为10:1~5。
本发明的目的之二是提供一种电荷引导的微/纳米可黏附水凝胶微球,其是由如上所述方法制备得到。
在本发明的电荷引导的微/纳米可黏附水凝胶微球的脂质体内部,可以负载没食子酸(GA)。
本发明的目的之三是提供上述电荷引导的微/纳米可黏附水凝胶微球的应用,所述应用是将其作为软骨细胞靶向给药的载体,用于递送骨疾病如骨关节炎方面的药物。
本发明的有益效果如下:
本发明构建了具有带正电荷的二级纳米结构的可黏附水凝胶微球,该二级纳米结构可以在电荷的引导下渗透软骨基质,从而作为黏附软骨、渗透软骨基、靶向软骨细胞的高效药物递送体系,实现了药物的高效递送。体内外实验表明,在模拟骨性关节炎的环境中,该水凝胶微球成功的将抗氧化剂递送至软骨细胞,将软骨细胞在氧化应激环境下的凋亡率由38.36±5.48%降至12.86±4.27%,并显著优于直接药物治疗组(28.43±5.87%)。该可渗透软骨基的可注射粘附性微/纳水凝胶微球通过使负载药物高效渗透软骨基质并ROS响应释放,实现了药物利用率和疗效的提升,是一种极具潜力的软骨药物高效递送材料。
附图说明
图1为本发明实施例中水凝胶微球的合成路线图;A)GA负载正电荷脂质体的合成;B)微流控装置制备Lipo@HAMA微球和PDA@Lipo@HAMA微球;C)基于渗透软骨、ROS响应性药物释放和抑制软骨细胞凋亡的治疗OA的电荷引导微/纳米水凝胶微球的设计。
图2为带正电脂质体的特性;A)PHB-dextran单体的核磁共振氢谱;B)带正电的脂质体的透射电镜图;C)脂质体在水溶液中的粒径分布;D)脂质体的Zeta电位;E)图片显示了在PBS和含1mM H2O2的PBS中,ROS触发了PHB-dextran纳米颗粒的水解;F)脂质体从HAMA水凝胶到上层溶液的释放行为;G)用FITC标记脂质体后,将兔软骨盘浸泡其中,用PBS清洗兔软骨盘,去除未渗透的脂质体,并在成像前对半切割;H)荧光显微镜图像显示,带负电荷的脂质体(i,ii)和带正电荷的脂质体(iii,iv)的渗透性。
图3为水凝胶微球的表征;A)PDA@Lipo@HAMA微凝胶的显微图像:(i)分散的微球,(ii)单微球,(iii)冻干Lipo@HAMA,(iv)冻干PDA@Lipo@HAMA;B)PDA@Lipo@HAMA的直径;C)微球的SEM图像:(i)HAMA微球,(ii)Lipo@HAMA微球,(iii)PDA@Lipo@HAMA微球;D)PDA@HAMA-GA和PDA@Lipo-GA@HAMA的GA释放曲线(n=3);E)PDA@Lipo-GA@HAMA分别在PBS和含1mm H2O2的PBS里的GA释放曲线(n=3);F)PDA@Lipo@HAMA微球与软骨粘附示意图;G)数码照片展示,将HAMA微球或PDA@HAMA微球置于软骨表面,用水清洗,红色箭头表示漂移的微球。
图4为微球的细胞毒性和抗氧化应激作用;A)将Lipo-GA和PDA@Lipo-GA@HAMA与软骨细胞共培养的活/死染色结果,显示微球的细胞毒性;B)活/死染色的定量分析(n=3);C)CCK-8检测结果显示Lipo-GA和PDA@Lipo-GA@HAMA对软骨细胞的细胞毒性(n=5);D)流式细胞术显示含ROS细胞的数量;E)流式细胞术显示凋亡细胞数量;F)与IL-1β、IL-1β+PDA@HAMA-GA、IL-1β+PDA@Lipo-GA@HAMA共培养软骨细胞的活/死染色结果;G)活/死染色的定量分析(n=3);H)流式细胞术的定量分析(n=3),(NS:没有意义,***P<0.05,P<0.01,***P<0.001)。
图5为PDA@Lipo@HAMA减轻OA的病情进展(每个分组6只);A)动物实验概述;B)各组有代表性的H&E染色图像;C)各组关节软骨OARSI评分;D)Safranin O-fast greenstaining(S-F staining)显示五组软骨的组织学变化;E)各组相对糖胺聚糖(GAG)含量;F)凋亡细胞TUNEL染色的代表性切片;G)TUNEL阳性细胞定量;H)II型胶原免疫荧光染色的代表性图像;I)II型胶原阳性细胞的定量分析,(黑色箭头表示软骨糜烂),(*P<0.05,**P<0.01,***P<0.001)。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白,以下结合实施例对本发明进行具体描述,有必要指出的是,以下实施例仅仅用于对本发明进行解释和说明,并不用于限定本发明。本领域技术人员根据上述发明内容所做出的一些非本质的改进和调整,仍属于本发明的保护范围。
实施例
本实施例中,带正电荷脂质体的合成以及PDA@Lipo@HAMA微球的制备整体流程如图1所示。
原料:4-羟甲基苯硼酸频哪醇酯(PBAP)、甲酰胺、葡聚糖和4-二甲氨基吡啶(DMAP)来自上海麦克林生化科技有限公司;1,1’-羰基二咪唑(CDI)、GA(没食子酸)、胆固醇、卵磷脂(来自蛋黄)来自生工生物工程(上海)有限公司。硬脂胺(SA)来自梯希爱(上海)化成工业发展有限公司。
1.1脂质体的制备及物理性质
本实施例中采用改进的薄膜分散法制备脂质体,方法如下:
两亲性嵌段共聚物(PHB-dextran)的合成:将PBAP(5.85g,25mmol)和CDI(8.11g,50mmol)溶于无水二氯甲烷(30mL)中。反应30min后,硫酸镁干燥过夜,浓缩后真空干燥,生成PBAP-CDI。用甲酰胺溶解葡聚糖,加入PBAP-CDI和4-二甲胺基吡啶,在25℃下搅拌过夜,然后用透析法从反应溶液中去除溶剂和未反应的PBAP-CDI。经1H NMR证实,冷冻干燥得到的白色粉末(PHB-dextran)即为硼酸酯与葡聚糖的共价接枝产物。
脂质体的合成:将60mg卵磷脂、20mg胆固醇和8.0mg硬脂胺溶于30mL氯仿中。然后,将有机溶剂在35℃(约1小时)完全蒸发,得到附着在圆底烧瓶底部的脂质膜。然后在烧瓶中加入3mL重蒸馏水与PHB-dextran的混合物,在25℃下超声分解20min,使膜完全溶解于水中,得到微米大小的脂质体双分子层。为了制备粒径更小的脂质体,使用5min强探头超声溶液(60单脉冲/min,130W)。
HAMA水凝胶的合成:将浓度为10%(w/v)的透明质酸(HA)于60℃溶解于PBS缓冲溶液中,加入甲基丙烯酸(MA)溶液,在50℃下混合反应1h。使用5倍稀释的PBS缓冲液停止反应,所得化合物在40℃下透析1周,过滤杂质(14kDa截止分子量)。HAMA水溶液进行冷冻干燥,得到乳白色,多孔结构泡沫。
PDA@Lipo@HAMA微球的合成:PDA@Lipo@HAMA采用改进的微流控装置。按水相(5wt%HAMA,2.5wt%脂质体均匀混合在PBS中,加入0.5wt%光稳定剂)和油相(5wt%Span80,石蜡油)加入到微流控装置,通过连接注射器泵的注射器控制水相和油相的水流速度。所得的单分散乳液滴在紫外光下进行光学交联。将微球转移至离心管,然后加入1mL异丙醇,振荡洗涤,4000rpm离心收集微球。随后,用多巴胺处理微球,10mM的Tris-HCl缓冲液(pH=8.5),37℃,60rpm摇晃0.5小时。然后,对这些微球进行清洗,以去除多余的多巴胺。
本发明方法在复水过程中反复超声得到单分散脂质体。其中,SA(硬脂胺)作为脂质层调节电荷的阳性成分,使得磷双分子层携带正电荷。此外,为了提高抗氧化剂的有效性,确保GA(没食子酸)在接触ROS之前尽可能长时间地被包封在生物材料中,合成了PHB-dextran,使脂质体核心具有ROS响应特性。PHB-dextran的质子核磁共振谱证实了共聚物的成功合成。通过与标准葡聚糖信号进行比较,在2.51ppm时的附加信号由-CH3提供,7.33和7.64ppm的信号来源于苯环上的氢。说明PHB与dextran的反应是有效的,PHB在化学上与dextran的羟基结合(如图2A)。
脂质体的透射电镜如图2B所示。颗粒形状为完美的球形,具有明显的壳核结构。外部浅色物质是脂质层,内部深色物质是具有ROS响应的内相。脂质体分散体的粒径集中在91.2nm左右,Zeta电位约为28.40mV,说明脂质体分散体粒径分布均匀,表面带强正电荷(如图2C-D)。
随后,将2mL制备的PHB-dextran纳米颗粒放入样品瓶中,1mM的H2O2注射提供ROS,观察60min,拍照验证PHB-dextran对ROS的响应功能(如图2E)。样品瓶中的液体在ROS刺激下逐渐由浑浊变为澄清,说明纳米颗粒正在逐渐降解。PHB-dextran可被证实具有良好的ROS反应性。
1.2脂质体的释放和软骨渗透性
本发明使用带正电荷的脂质体作为具有电荷引导特性的二级纳米结构渗透软骨基质。因此,该脂质体穿透软骨的能力有待进一步验证。实验设置如图2F所示,用于研究HAMA释放脂质体到外液体中的能力。
首先,将5wt%的HAMA水凝胶和2.5wt%的脂质体混合,置于瓶子底部,并进行紫外光交联。同时,在水凝胶上层注入蒸馏水。随着时间的变化,可以观察到水凝胶和蒸馏水的交界面随着时间的推移会逐渐向上移动。此外,在t=48h时,HAMA水凝胶上部的液体由全透明变为乳白色。随后,取HAMA水凝胶上层的液体并混合至均匀,对该液体样品进行透射电镜观察,发现其中含有脂质体(如图2F)。因此,有强有力的证据表明,HAMA水凝胶可以将脂质体释放到外部液体中。同时,在膝关节腔的高压环境中,水凝胶孔隙会因挤压而扩大,更有利于脂质体的释放。
另外,利用异硫氰酸荧光素标记脂质体,验证带正电荷脂质体的软骨渗透性。取兔的膝关节软骨圆片,将软骨面的一侧暴露于脂质体溶液中。将渗透(24h)和洗涤(24h)后的软骨对半切开,以排除软骨原片侧面与脂质体溶液接触造成的误差,并用荧光显微镜观察软骨剖面(如图2G)。带负电荷脂质体在此期间的渗透范围局限于软骨表层,不能进入软骨深层。而带正电荷的脂质体仍能在深层软骨层检测到强荧光。由此证明,带正电的脂质体可以深入渗透软骨(如图2H)。
综上所述,带正电荷的二级纳米结构可以从HAMA水凝胶中释放出来,并在电荷引导下有效穿透软骨基质。
1.3利用微流控装置制备单分散水凝胶微球
为了更好地发挥带正电荷的纳米结构穿透软骨基质的功能,又能不被人体过早清除,我们将其与可注射水凝胶微球相结合。通过微流控装置(可参考文献“N.B.H.A.S.L.Anna,Applied Physics Letters 2003,364.”),制备了聚多巴胺@脂质体@HAMA水凝胶微球(PDA@Lipo@HAMA)。将HAMA水凝胶和脂质体的混合物做为中间通道的分散相,石蜡油为连续相(外通道)。随着连续相的挤压,分散相的混合物受到的剪切力增大,使混合物分裂,形成乳状液液滴。随后,液滴在紫外线下交联成Lipo@HAMA微球(如图3A.i-iii)。进一步,通过碱性环境下的交联反应将Lipo@HAMA微球表面接枝多巴胺,获得具有粘附功能的PDA@Lipo@HAMA微球(如图3A.iv)。最后,使用明场显微镜测量了复合微球的直径(如图3B)。
通过扫描电镜观察冻干后的HAMA微球、Lipo@HAMA微球和PDA@Lipo@HAMA微球。可以观察到HAMA微球的直径约为200μm,其内部网状多孔的结构可以持有足够的纳米颗粒形成新型复合材料,并具有可扩展功能(如图3C.i)。此外,Lipo@HAMA微球的网孔内可观察到完整的单分散的脂质体(如图3C.ii)。进一步,在Lipo@HAMA微球表面进行聚多巴胺接枝,利用聚多巴胺的邻苯二酚结构与组织粘附性能,制备具有黏附性能的PDA@Lipo@HAMA微球。最后,扫描电镜显示聚多巴胺被修饰在微球上(如图3C.iii)。
1.4软骨粘附与药物释放行为
研究了水凝胶微球的药物释放动力学,以评估PDA@Lipo@HAMA作为药物传递载体的适用性(如图3D)。
将GA分别装入HAMA微球(PDA@HAMA-GA)和微/纳米水凝胶微球(PDA@Lipo-GA@HAMA)中,结果发现PDA@HAMA-GA微球释放迅速,GA在12h后几乎被完全释放。相比之下,PDA@Lipo-GA@HAMA具有更好的封装效果,可将药物封装在载体内很长一段时间。
随后,验证了PDA@Lipo-GA@HAMA载体在ROS响应时释放GA的能力(如图3E)。在ROS的刺激下,载体释放的GA量显著增加。经过1mM H2O2处理12h后,PDA@Lipo-GA@HAMA的GA释放率为65.20%,显著高于对照组(11.66%)。因此,PDA@Lipo@HAMA微球可以实现药物的缓释和ROS响应释放。
进一步,设计了体外软骨实验验证微球黏附(如图3F)。取直径4.5mm的兔膝关节软骨盘用胶水固定在培养皿上,将普通的HAMA微球和多巴胺修饰的PDA@HAMA微球置于软骨表面静置20min,然后用移液管用水冲洗软骨盘上的微球。放大后的图像中可见,由于缺乏粘附软骨的能力,HAMA微球大部分被电流冲走,因此分散在培养皿的水中(红色箭头所示)。PDA@HAMA微球具有良好的粘附能力。虽然由于电流的强冲刷力,部分微球从原来的位置被冲刷掉,但它们仍然能够粘附在软骨样本上,且几乎没有微球被冲刷到培养皿的水中(如图3G)。因此,关节腔内的HAMA微球容易脱离软骨,漂浮在关节液中。
综上所述,PDA@HAMA能有效地粘附在软骨上,将所载药物集中在软骨表面。
1.5体外细胞毒性和抗氧化应激的疗效
由于该生物材料将直接注射到关节腔内,因此研究了Lipo-GA和PDA@Lipo-GA@HAMA对小鼠软骨细胞的体外细胞毒性。
将不同生物材料和细胞共培养1、3、5天后,对细胞进行活/死检测和CCK-8检测。活/死实验显示,在实验期间,三组细胞基本存活,死亡细胞数量较少(如图4A)。细胞计数显示,细胞相对密度随塑形时间的增加而增加(如图4B)。经CCK-8检测,三组细胞在各时间点生长无显著差异(如图4C)。综上所述,该生物材料与软骨细胞具有良好的生物相容性。
OA的发病机制与多种因素有关,其中炎症和氧化应激被认为与OA的病理过程密切相关。局部炎症反应伴随着年龄的增加和机械应力的长期作用导致氧化应激反应的增加。发明人使用IL-1β(20ng)模拟OA发病过程中的炎症微环境,诱导小鼠软骨细胞氧化应激。随后,使用流式细胞仪检测PDA@Lipo-GA@HAMA抗ROS能力和抗凋亡能力(如图4D、E、H)。结果发现,细胞被IL-1β诱导后24h后,和正常细胞相比,其产生活性氧的比例显著增加,细胞凋亡晚期阶段的数量也显著增加。说明利用IL-1β模拟炎症微环境可成功诱导细胞氧化应激反应。
进一步建立了PDA@HAMA-GA和PDA@Lipo-GA@HAMA两个分组。根据上文中两种载体的释放曲线,通过控制载药量,使两种载体在24h内释放相同剂量的GA。结果表明,两种药物载体均能显著抑制氧化应激诱导的ROS生成,减少细胞凋亡。同时PDA@Lipo-GA@HAMA组疗效优于PDA@HAMA-GA组。这是因为载GA的脂质体是纳米级的,可以进入细胞释放GA,增加GA的利用率。
最后,用活/死实验(如图4F-G)再次验证了药物载体的有效性,结果与流式细胞术相似,PDA@Lipo-GA@HAMA由于其独特的载体设计,可以提高GA的抗氧化功效,而且该组的死亡细胞数量也最低。
1.6 OA动物模型的治疗效果
为了验证PDA@Lipo@HAMA微球在体内渗透软骨基质的作用,建立了大鼠OA模型,并将微球注射到关节腔内治疗OA。如果该微球能有效地渗透深入软骨,则可以观察到该微球的疗效将优于其他治疗组。
采用关节内注射碘乙酸(MIA)建立大鼠OA模型。MIA是一种代谢抑制剂,能破坏机体有氧运动的糖酵解途径,进而抑制软骨组织细胞中甘油醛-3-硫酸铵脱氢酶的活性,诱导细胞死亡。关节内注射MIA导致软骨组织细胞总数减少,引起与人OA相似的病理组织学改变。既往报道表明,大鼠关节腔注射MIA可建立与人类OA病理进展相似的有效OA模型。该模型具有重现性好、精度高、入侵少、易于实现、进展快等优点。
造模成功后,各组大鼠分别给予PBS、GA、PDA@HAMA-GA(P@H-G)、PDA@Lipo-GA@HAMA(P@L-G@H)治疗5周。取材后,进行了组织学和免疫荧光评估,研究PDA@Lipo-GA@HAMA对软骨损伤的保护作用(如图5A)。苏木精-伊红(H&E)染色(Hematoxylin and eosin(H&E)staining)(如图5B)和藏红-O染色(Safranin O-fast green staining)(如图5D)都表明,骨关节炎的典型症状(如:不规则表面腐蚀层和裂缝)PBS组更显著,其次是GA组和PDA@HAMA-GA组,而PDA@Lipo-GA@HAMA组是最不显著的。OARSI分数的结果(如图5C)表明,与PBS组相比,其他治疗的OARSI评分普遍降低,PDA@Lipo-GA@HAMA组显示了最好的结果,减少了53.50%。其次是PDA@HAMA-GA组(下降36.05%)和GA组(减少27.95%)。根据各治疗组Safranin O-fast green staining的结果,检测了糖胺聚糖(红色染色)的含量,以PDA@Lipo-GA@HAMA组为最多,PDA@HAMA-GA组次之,GA组最后。可见PDA@Lipo-GA@HAMA组在维持软骨基质方面效果最好(如图5E)。
随后,采用TUNEL染色检测软骨细胞凋亡(如图5F)。可见PDA@Lipo-GA@HAMA组与对照组相比凋亡细胞最少(绿色染色),其次是GA组和PDA@HAMA-GA组。
进一步,对阳性染色的细胞进行了定量分析(如图5G)。与PBS组比较,PDA@Lipo-GA@HAMA组凋亡率最低(66.48%),其次是PDA@HAMA-GA组(40.31%)和GA组(25.88%)。经过PDA@Lipo-GA@HAMA微球处理后,氧化应激下软骨细胞凋亡率由38.36±5.48%降至12.86±4.27%,明显优于GA组(28.43±5.87%)。
此外,用免疫荧光染色检测软骨的主要生物标志物胶原II的表达(如图5H)。与对照组比较,各治疗组II型胶原(红色染色)表达水平均有或多或少的下降,其中PDA@Lipo-GA@HAMA组下降最少,GA组和PDA@HAMA-GA组次之。
对阳性细胞进行定量分析(如图5I)。与PBS组比较,PDA@Lipo-GA@HAMA组II型胶原表达水平最高,增加282.14%,PDA@HAMA-GA组和GA组次之,分别增加105.93%和63.07%。
总体而言,上述结果表明:PDA@Lipo-GA@HAMA可以显著渗透软骨基质,增加GA疗效,抑制氧化应激诱导的骨关节炎软骨细胞凋亡。因此,该电荷引导的微/纳米水凝胶微球提供了一种有吸引力的OA治疗策略。
Claims (9)
1.一种电荷引导的微/纳米可黏附水凝胶微球的制备方法,其特征在于,包括以下步骤:
(1)以疏水性硼酸酯和亲水性葡聚糖进行接枝反应,形成两亲性嵌段共聚物,然后以硬脂胺作为脂质层调节电荷的阳性成分,得到以所述两亲性嵌段共聚物为核心的带正电荷的脂质体;
(2)采用透明质酸和甲基丙烯酸酐合成水凝胶,利用微流控装置,将所述水凝胶和步骤(1)所得脂质体混合,制备水凝胶微球,进行紫外光交联,形成交联水凝胶微球;
(3)在所述交联水凝胶微球表面接枝多巴胺,获得所述电荷引导的微/纳米可黏附水凝胶微球。
2.根据权利要求1所述的制备方法,其特征在于,步骤(1)中所述硼酸酯为4-羟甲基苯硼酸频哪醇酯,所述硼酸酯与葡聚糖的质量比为2:1~4。
3.根据权利要求1所述的制备方法,其特征在于,步骤(2)中所述透明质酸与甲基丙烯酸酐的质量比为10:1~20。
4.根据权利要求1所述的制备方法,其特征在于,步骤(2)中在采用微流控装置进行制备时,是以水凝胶和脂质体的混合物为分散相,以石蜡油为连续相。
5.根据权利要求1所述的制备方法,其特征在于,步骤(2)中所述水凝胶和脂质体的质量比为10:1~5。
6.一种电荷引导的微/纳米可黏附水凝胶微球,其特征在于,是由权利要求1-5任一项所述方法制备得到。
7.根据权利要求6所述的电荷引导的微/纳米可黏附水凝胶微球,其特征在于,所述脂质体内部负载没食子酸。
8.如权利要求6或7所述的电荷引导的微/纳米可黏附水凝胶微球的应用,其特征在于,是将其作为软骨细胞靶向给药的载体,用于递送骨疾病方面的药物。
9.根据权利要求8所述的应用,其特征在于,所述骨疾病包括骨关节炎。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111030861.0A CN113896906B (zh) | 2021-09-03 | 2021-09-03 | 一种电荷引导的微纳米可黏附水凝胶及其制备方法与应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111030861.0A CN113896906B (zh) | 2021-09-03 | 2021-09-03 | 一种电荷引导的微纳米可黏附水凝胶及其制备方法与应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113896906A CN113896906A (zh) | 2022-01-07 |
CN113896906B true CN113896906B (zh) | 2024-02-27 |
Family
ID=79188587
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202111030861.0A Active CN113896906B (zh) | 2021-09-03 | 2021-09-03 | 一种电荷引导的微纳米可黏附水凝胶及其制备方法与应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113896906B (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115025049B (zh) * | 2022-05-30 | 2023-04-28 | 浙江大学 | 一种高效负载抗炎药物的水凝胶微球及其制备方法 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20160036523A (ko) * | 2014-09-25 | 2016-04-04 | 서울대학교산학협력단 | 피브로인 미립구를 포함하는 약물 전달체 및 그 제조방법 |
CN108744060A (zh) * | 2018-05-29 | 2018-11-06 | 四川大学 | 一种可注射多重孔隙结构的骨修复材料及其制备方法 |
CN109563199A (zh) * | 2016-06-13 | 2019-04-02 | 麻省理工学院 | 用于减少异物反应和纤维化的生物相容性两性离子聚合物涂层和水凝胶 |
-
2021
- 2021-09-03 CN CN202111030861.0A patent/CN113896906B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20160036523A (ko) * | 2014-09-25 | 2016-04-04 | 서울대학교산학협력단 | 피브로인 미립구를 포함하는 약물 전달체 및 그 제조방법 |
CN109563199A (zh) * | 2016-06-13 | 2019-04-02 | 麻省理工学院 | 用于减少异物反应和纤维化的生物相容性两性离子聚合物涂层和水凝胶 |
CN108744060A (zh) * | 2018-05-29 | 2018-11-06 | 四川大学 | 一种可注射多重孔隙结构的骨修复材料及其制备方法 |
Non-Patent Citations (1)
Title |
---|
双效骨诱导性的有机-无机水凝胶的构建及其骨再生研究;程若昱;燕宇飞;陈皓;齐进;邓廉夫;崔文国;;上海交通大学学报(医学版)(08);第66-75页 * |
Also Published As
Publication number | Publication date |
---|---|
CN113896906A (zh) | 2022-01-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Lin et al. | Charge‐guided micro/nano‐hydrogel microsphere for penetrating cartilage matrix | |
Swanson et al. | Scaffolds with controlled release of pro-mineralization exosomes to promote craniofacial bone healing without cell transplantation | |
Zare et al. | Alginate sulfate-based hydrogel/nanofiber composite scaffold with controlled Kartogenin delivery for tissue engineering | |
Cheng et al. | Mechanically enhanced lipo-hydrogel with controlled release of multi-type drugs for bone regeneration | |
Wang et al. | Microencapsulation using natural polysaccharides for drug delivery and cell implantation | |
Piotrowicz et al. | Nerve guidance channels as drug delivery vehicles | |
CN110123842B (zh) | 一种外泌体缓释体系及其构建方法与应用 | |
US20230020948A1 (en) | Nanofibrous mat containing ceramic particles with releasable dopant | |
Chen et al. | Surface-engineering of glycidyl methacrylated dextran/gelatin microcapsules with thermo-responsive poly (N-isopropylacrylamide) gates for controlled delivery of stromal cell-derived factor-1α | |
CN107496358A (zh) | 一种脂质体增强型水凝胶及其应用 | |
Gupta et al. | Repair and reconstruction of a resected tumor defect using a composite of tissue flap–nanotherapeutic–silk fibroin and chitosan scaffold | |
Nikravesh et al. | Physical structuring of injectable polymeric systems to controllably deliver nanosized extracellular vesicles | |
Bizeau et al. | Design and applications of protein delivery systems in nanomedicine and tissue engineering | |
CN110292652B (zh) | 巯基苯硼酸活化金纳米颗粒、其制备方法及应用 | |
CN112618571B (zh) | 治疗骨科疾病的可注射水凝胶微球及其制备方法与应用 | |
CN107375196A (zh) | 一种含儿茶酚基天然多糖复合水凝胶载体及其制备方法 | |
Xue et al. | Controllable fabrication of alginate/poly-L-ornithine polyelectrolyte complex hydrogel networks as therapeutic drug and cell carriers | |
CN113896906B (zh) | 一种电荷引导的微纳米可黏附水凝胶及其制备方法与应用 | |
WO2021257533A2 (en) | Microparticles for controlled release of extracellular vesicles and methods for fabricating the same | |
Molina-Peña et al. | Nanoparticle-containing electrospun nanofibrous scaffolds for sustained release of SDF-1α | |
WO2015187098A1 (en) | Material and structures for cell delivery | |
Kim et al. | In situ pocket-type microcarrier (PMc) as a therapeutic composite: Regeneration of cartilage with stem cells, genes, and drugs | |
Sethuram et al. | Synthesis, fabrication and biosafety profiles of biobased microemulsions reinforced electrospun nanofibers for wound dressing applications | |
Abbaszadeh et al. | Emerging strategies to bypass transplant rejection via biomaterial-assisted immunoengineering: Insights from islets and beyond | |
Wang et al. | A photopolymerized semi-interpenetrating polymer networks-based hydrogel incorporated with nanoparticle for local chemotherapy of tumors |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |