WO2023041275A1 - Besiedelung von oberflaechen mit biologischen zellen - Google Patents
Besiedelung von oberflaechen mit biologischen zellen Download PDFInfo
- Publication number
- WO2023041275A1 WO2023041275A1 PCT/EP2022/072853 EP2022072853W WO2023041275A1 WO 2023041275 A1 WO2023041275 A1 WO 2023041275A1 EP 2022072853 W EP2022072853 W EP 2022072853W WO 2023041275 A1 WO2023041275 A1 WO 2023041275A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cells
- biological cells
- hfm
- azide
- dbco
- Prior art date
Links
- 238000000034 method Methods 0.000 claims abstract description 44
- 238000004519 manufacturing process Methods 0.000 claims abstract description 8
- 210000004027 cell Anatomy 0.000 claims description 90
- 239000012528 membrane Substances 0.000 claims description 53
- ZUHQCDZJPTXVCU-UHFFFAOYSA-N C1#CCCC2=CC=CC=C2C2=CC=CC=C21 Chemical compound C1#CCCC2=CC=CC=C2C2=CC=CC=C21 ZUHQCDZJPTXVCU-UHFFFAOYSA-N 0.000 claims description 37
- 210000002889 endothelial cell Anatomy 0.000 claims description 31
- 238000011534 incubation Methods 0.000 claims description 28
- HGMISDAXLUIXKM-LIADDWGISA-N [(2r,3s,4r,5s)-3,4,6-triacetyloxy-5-[(2-azidoacetyl)amino]oxan-2-yl]methyl acetate Chemical compound CC(=O)OC[C@H]1OC(OC(C)=O)[C@@H](NC(=O)CN=[N+]=[N-])[C@@H](OC(C)=O)[C@@H]1OC(C)=O HGMISDAXLUIXKM-LIADDWGISA-N 0.000 claims description 22
- 238000012650 click reaction Methods 0.000 claims description 21
- 239000000376 reactant Substances 0.000 claims description 21
- IVRMZWNICZWHMI-UHFFFAOYSA-N azide group Chemical group [N-]=[N+]=[N-] IVRMZWNICZWHMI-UHFFFAOYSA-N 0.000 claims description 20
- 210000004072 lung Anatomy 0.000 claims description 20
- -1 phosphine ester Chemical class 0.000 claims description 20
- 150000001540 azides Chemical class 0.000 claims description 17
- 239000011116 polymethylpentene Substances 0.000 claims description 17
- 229920000306 polymethylpentene Polymers 0.000 claims description 16
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 15
- RRCXYKNJTKJNTD-UHFFFAOYSA-N dbco-peg4-nhs ester Chemical compound C1C2=CC=CC=C2C#CC2=CC=CC=C2N1C(=O)CCC(=O)NCCOCCOCCOCCOCCC(=O)ON1C(=O)CCC1=O RRCXYKNJTKJNTD-UHFFFAOYSA-N 0.000 claims description 14
- 239000012510 hollow fiber Substances 0.000 claims description 14
- 230000021615 conjugation Effects 0.000 claims description 12
- 238000002444 silanisation Methods 0.000 claims description 10
- XYFCBTPGUUZFHI-UHFFFAOYSA-N Phosphine Chemical compound P XYFCBTPGUUZFHI-UHFFFAOYSA-N 0.000 claims description 7
- 239000004743 Polypropylene Substances 0.000 claims description 7
- 229920001155 polypropylene Polymers 0.000 claims description 7
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 6
- 150000002148 esters Chemical class 0.000 claims description 6
- 239000001301 oxygen Substances 0.000 claims description 6
- 229910052760 oxygen Inorganic materials 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 5
- 210000003734 kidney Anatomy 0.000 claims description 4
- 229910000073 phosphorus hydride Inorganic materials 0.000 claims description 4
- 235000000346 sugar Nutrition 0.000 claims description 4
- 230000002792 vascular Effects 0.000 claims description 4
- 238000009832 plasma treatment Methods 0.000 claims description 3
- AFNOHTDETQTADW-YLRIPHBZSA-N 2-azido-n-[(3s,4r,5s,6r)-2,4,5-trihydroxy-6-(hydroxymethyl)oxan-3-yl]acetamide Chemical compound OC[C@H]1OC(O)[C@@H](NC(=O)CN=[N+]=[N-])[C@@H](O)[C@@H]1O AFNOHTDETQTADW-YLRIPHBZSA-N 0.000 claims description 2
- HGMISDAXLUIXKM-ALTVCHKUSA-N [(2r,3s,4r,5r)-3,4,6-triacetyloxy-5-[(2-azidoacetyl)amino]oxan-2-yl]methyl acetate Chemical compound CC(=O)OC[C@H]1OC(OC(C)=O)[C@H](NC(=O)CN=[N+]=[N-])[C@@H](OC(C)=O)[C@@H]1OC(C)=O HGMISDAXLUIXKM-ALTVCHKUSA-N 0.000 claims 1
- 210000004369 blood Anatomy 0.000 description 41
- 239000008280 blood Substances 0.000 description 41
- 239000012591 Dulbecco’s Phosphate Buffered Saline Substances 0.000 description 32
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 description 28
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 24
- 238000004458 analytical method Methods 0.000 description 18
- 210000001772 blood platelet Anatomy 0.000 description 18
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- 210000002381 plasma Anatomy 0.000 description 14
- 238000001514 detection method Methods 0.000 description 13
- 238000010186 staining Methods 0.000 description 11
- 230000004913 activation Effects 0.000 description 10
- 230000003511 endothelial effect Effects 0.000 description 10
- 108020004999 messenger RNA Proteins 0.000 description 10
- 238000011529 RT qPCR Methods 0.000 description 9
- 108010000134 Vascular Cell Adhesion Molecule-1 Proteins 0.000 description 9
- 102100023543 Vascular cell adhesion protein 1 Human genes 0.000 description 9
- 108010072035 antithrombin III-protease complex Proteins 0.000 description 9
- 229920000669 heparin Polymers 0.000 description 9
- 108010024212 E-Selectin Proteins 0.000 description 8
- 102100023471 E-selectin Human genes 0.000 description 8
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 8
- 108010064593 Intercellular Adhesion Molecule-1 Proteins 0.000 description 8
- 102000015271 Intercellular Adhesion Molecule-1 Human genes 0.000 description 8
- 230000015572 biosynthetic process Effects 0.000 description 8
- 238000000799 fluorescence microscopy Methods 0.000 description 8
- 102000006602 glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 8
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 8
- 229960002897 heparin Drugs 0.000 description 8
- 238000005406 washing Methods 0.000 description 8
- 102100036154 Platelet basic protein Human genes 0.000 description 7
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 7
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 7
- 230000008901 benefit Effects 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 7
- 238000000576 coating method Methods 0.000 description 7
- 239000010410 layer Substances 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 238000001466 metabolic labeling Methods 0.000 description 7
- STZCRXQWRGQSJD-GEEYTBSJSA-M methyl orange Chemical compound [Na+].C1=CC(N(C)C)=CC=C1\N=N\C1=CC=C(S([O-])(=O)=O)C=C1 STZCRXQWRGQSJD-GEEYTBSJSA-M 0.000 description 7
- 229940012189 methyl orange Drugs 0.000 description 7
- 239000013642 negative control Substances 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 238000007619 statistical method Methods 0.000 description 7
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 6
- 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 6
- JWFYJJIXFMMPNH-UHFFFAOYSA-N N-(3-azidopropyl)-6-[(2E)-3,3-dimethyl-2-[(E)-3-(1,3,3-trimethylindol-1-ium-2-yl)prop-2-enylidene]indol-1-yl]hexanamide chloride Chemical compound [Cl-].CN1\C(=C\C=C\C2=[N+](CCCCCC(=O)NCCCN=[N+]=[N-])C3=C(C=CC=C3)C2(C)C)C(C)(C)C2=CC=CC=C12 JWFYJJIXFMMPNH-UHFFFAOYSA-N 0.000 description 6
- 125000003277 amino group Chemical group 0.000 description 6
- 239000006143 cell culture medium Substances 0.000 description 6
- 239000011248 coating agent Substances 0.000 description 6
- 238000007306 functionalization reaction Methods 0.000 description 6
- 230000002757 inflammatory effect Effects 0.000 description 6
- 238000001543 one-way ANOVA Methods 0.000 description 6
- 230000010118 platelet activation Effects 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- 230000027455 binding Effects 0.000 description 5
- BQRGNLJZBFXNCZ-UHFFFAOYSA-N calcein am Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(=O)OCOC(C)=O)CC(=O)OCOC(C)=O)=C(OC(C)=O)C=C1OC1=C2C=C(CN(CC(=O)OCOC(C)=O)CC(=O)OCOC(=O)C)C(OC(C)=O)=C1 BQRGNLJZBFXNCZ-UHFFFAOYSA-N 0.000 description 5
- 230000004154 complement system Effects 0.000 description 5
- 238000002618 extracorporeal membrane oxygenation Methods 0.000 description 5
- 238000000684 flow cytometry Methods 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 4
- 102000009123 Fibrin Human genes 0.000 description 4
- 108010073385 Fibrin Proteins 0.000 description 4
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 4
- 101000947178 Homo sapiens Platelet basic protein Proteins 0.000 description 4
- 102100023472 P-selectin Human genes 0.000 description 4
- 102000008790 VE-cadherin Human genes 0.000 description 4
- 238000004820 blood count Methods 0.000 description 4
- 108010018828 cadherin 5 Proteins 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 230000015271 coagulation Effects 0.000 description 4
- 238000005345 coagulation Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 210000003743 erythrocyte Anatomy 0.000 description 4
- 229950003499 fibrin Drugs 0.000 description 4
- 238000002372 labelling Methods 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 230000000638 stimulation Effects 0.000 description 4
- 101800003265 Beta-thromboglobulin Proteins 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 102000016359 Fibronectins Human genes 0.000 description 3
- 108010067306 Fibronectins Proteins 0.000 description 3
- 102000003886 Glycoproteins Human genes 0.000 description 3
- 108090000288 Glycoproteins Proteins 0.000 description 3
- 206010062506 Heparin-induced thrombocytopenia Diseases 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 3
- 241001421711 Mithras Species 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 108090000631 Trypsin Proteins 0.000 description 3
- 102000004142 Trypsin Human genes 0.000 description 3
- 229940098773 bovine serum albumin Drugs 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 210000003855 cell nucleus Anatomy 0.000 description 3
- 230000003833 cell viability Effects 0.000 description 3
- 229910052802 copper Inorganic materials 0.000 description 3
- 239000010949 copper Substances 0.000 description 3
- 210000004292 cytoskeleton Anatomy 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 210000003038 endothelium Anatomy 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- 125000000524 functional group Chemical group 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- 210000000265 leukocyte Anatomy 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000003068 static effect Effects 0.000 description 3
- 210000000130 stem cell Anatomy 0.000 description 3
- 238000002054 transplantation Methods 0.000 description 3
- 239000012588 trypsin Substances 0.000 description 3
- HFCUBKYHMMPGBY-UHFFFAOYSA-N 2-methoxyethyl prop-2-enoate Chemical compound COCCOC(=O)C=C HFCUBKYHMMPGBY-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 208000006545 Chronic Obstructive Pulmonary Disease Diseases 0.000 description 2
- 101000622137 Homo sapiens P-selectin Proteins 0.000 description 2
- 108010028275 Leukocyte Elastase Proteins 0.000 description 2
- 102000016799 Leukocyte elastase Human genes 0.000 description 2
- 108010035766 P-Selectin Proteins 0.000 description 2
- 108010067372 Pancreatic elastase Proteins 0.000 description 2
- 102000016387 Pancreatic elastase Human genes 0.000 description 2
- 229930040373 Paraformaldehyde Natural products 0.000 description 2
- 102000004211 Platelet factor 4 Human genes 0.000 description 2
- 108090000778 Platelet factor 4 Proteins 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 238000010461 azide-alkyne cycloaddition reaction Methods 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- ZPWOOKQUDFIEIX-UHFFFAOYSA-N cyclooctyne Chemical group C1CCCC#CCC1 ZPWOOKQUDFIEIX-UHFFFAOYSA-N 0.000 description 2
- 238000003795 desorption Methods 0.000 description 2
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- 210000004263 induced pluripotent stem cell Anatomy 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- 238000007427 paired t-test Methods 0.000 description 2
- 229920002866 paraformaldehyde Polymers 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 230000010412 perfusion Effects 0.000 description 2
- YHHSONZFOIEMCP-UHFFFAOYSA-O phosphocholine Chemical compound C[N+](C)(C)CCOP(O)(O)=O YHHSONZFOIEMCP-UHFFFAOYSA-O 0.000 description 2
- 229950004354 phosphorylcholine Drugs 0.000 description 2
- 230000004962 physiological condition Effects 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 230000031915 positive regulation of coagulation Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 230000009257 reactivity Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000004626 scanning electron microscopy Methods 0.000 description 2
- ZFXYFBGIUFBOJW-UHFFFAOYSA-N theophylline Chemical compound O=C1N(C)C(=O)N(C)C2=C1NC=N2 ZFXYFBGIUFBOJW-UHFFFAOYSA-N 0.000 description 2
- 229950003937 tolonium Drugs 0.000 description 2
- HNONEKILPDHFOL-UHFFFAOYSA-M tolonium chloride Chemical compound [Cl-].C1=C(C)C(N)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 HNONEKILPDHFOL-UHFFFAOYSA-M 0.000 description 2
- 210000003606 umbilical vein Anatomy 0.000 description 2
- 230000035899 viability Effects 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 238000012800 visualization Methods 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 239000012103 Alexa Fluor 488 Substances 0.000 description 1
- APKFDSVGJQXUKY-KKGHZKTASA-N Amphotericin-B Natural products O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1C=CC=CC=CC=CC=CC=CC=C[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 APKFDSVGJQXUKY-KKGHZKTASA-N 0.000 description 1
- 108091093088 Amplicon Proteins 0.000 description 1
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 201000003883 Cystic fibrosis Diseases 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 241000252212 Danio rerio Species 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 108700039887 Essential Genes Proteins 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- 208000029523 Interstitial Lung disease Diseases 0.000 description 1
- 208000019693 Lung disease Diseases 0.000 description 1
- BAWFJGJZGIEFAR-NNYOXOHSSA-M NAD(1-) Chemical class NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP([O-])(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-M 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- SNIOPGDIGTZGOP-UHFFFAOYSA-N Nitroglycerin Chemical compound [O-][N+](=O)OCC(O[N+]([O-])=O)CO[N+]([O-])=O SNIOPGDIGTZGOP-UHFFFAOYSA-N 0.000 description 1
- 241000233805 Phoenix Species 0.000 description 1
- 108091026813 Poly(ADPribose) Proteins 0.000 description 1
- 206010037423 Pulmonary oedema Diseases 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 208000004756 Respiratory Insufficiency Diseases 0.000 description 1
- 238000003800 Staudinger reaction Methods 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 238000010162 Tukey test Methods 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 150000001345 alkine derivatives Chemical class 0.000 description 1
- 125000002355 alkine group Chemical group 0.000 description 1
- APKFDSVGJQXUKY-INPOYWNPSA-N amphotericin B Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/C=C/C=C/C=C/[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 APKFDSVGJQXUKY-INPOYWNPSA-N 0.000 description 1
- 229960003942 amphotericin b Drugs 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 230000001174 ascending effect Effects 0.000 description 1
- 125000000852 azido group Chemical group *N=[N+]=[N-] 0.000 description 1
- OIRCOABEOLEUMC-GEJPAHFPSA-N bivalirudin Chemical compound C([C@@H](C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)CNC(=O)CNC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 OIRCOABEOLEUMC-GEJPAHFPSA-N 0.000 description 1
- 229960001500 bivalirudin Drugs 0.000 description 1
- 108010055460 bivalirudin Proteins 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 230000002612 cardiopulmonary effect Effects 0.000 description 1
- 239000013553 cell monolayer Substances 0.000 description 1
- 238000003570 cell viability assay Methods 0.000 description 1
- 239000012094 cell viability reagent Substances 0.000 description 1
- 125000003636 chemical group Chemical group 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000000038 chest Anatomy 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 229940001468 citrate Drugs 0.000 description 1
- 230000024203 complement activation Effects 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000009109 curative therapy Methods 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- IZEKFCXSFNUWAM-UHFFFAOYSA-N dipyridamole Chemical compound C=12N=C(N(CCO)CCO)N=C(N3CCCCC3)C2=NC(N(CCO)CCO)=NC=1N1CCCCC1 IZEKFCXSFNUWAM-UHFFFAOYSA-N 0.000 description 1
- 229960002768 dipyridamole Drugs 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000003822 epoxy resin Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 238000002073 fluorescence micrograph Methods 0.000 description 1
- 238000012757 fluorescence staining Methods 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- BBKFSSMUWOMYPI-UHFFFAOYSA-N gold palladium Chemical compound [Pd].[Au] BBKFSSMUWOMYPI-UHFFFAOYSA-N 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- 230000002439 hemostatic effect Effects 0.000 description 1
- ZFGMDIBRIDKWMY-PASTXAENSA-N heparin Chemical compound CC(O)=N[C@@H]1[C@@H](O)[C@H](O)[C@@H](COS(O)(=O)=O)O[C@@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O[C@H]2[C@@H]([C@@H](OS(O)(=O)=O)[C@@H](O[C@@H]3[C@@H](OC(O)[C@H](OS(O)(=O)=O)[C@H]3O)C(O)=O)O[C@@H]2O)CS(O)(=O)=O)[C@H](O)[C@H]1O ZFGMDIBRIDKWMY-PASTXAENSA-N 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 238000011503 in vivo imaging Methods 0.000 description 1
- 102000006495 integrins Human genes 0.000 description 1
- 108010044426 integrins Proteins 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 229940127554 medical product Drugs 0.000 description 1
- 238000011880 melting curve analysis Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000007431 microscopic evaluation Methods 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000002953 phosphate buffered saline Substances 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920001690 polydopamine Polymers 0.000 description 1
- 229920000647 polyepoxide Polymers 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 208000005333 pulmonary edema Diseases 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 201000004193 respiratory failure Diseases 0.000 description 1
- 230000004043 responsiveness Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 229910000077 silane Inorganic materials 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 229960001790 sodium citrate Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000006557 surface reaction Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000012353 t test Methods 0.000 description 1
- 229960000278 theophylline Drugs 0.000 description 1
- 238000002411 thermogravimetry Methods 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 230000009424 thromboembolic effect Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 239000003656 tris buffered saline Substances 0.000 description 1
- 238000007492 two-way ANOVA Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
- A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
- A61L27/3808—Endothelial cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/005—Ingredients of undetermined constitution or reaction products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/18—Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment
Definitions
- the present invention relates to a method for producing a surface that can be colonized with biological cells, a device with a surface that can be colonized with biological cells, and a method for colonizing a surface with biological cells.
- COPD chronic obstructive pulmonary disease
- extracorporeal membrane oxygenation is performed by continuously pumping the patient's blood through a gas-permeable membrane oxygenator (artificial lung) that mimics the gas exchange process of the lungs.
- Standard blood oxygenators consist of bundles of microporous hollow fiber membranes (HFM), most commonly made of polypropylene (PP) or polymethylpentene (PMP), with a large surface area ( ⁇ 2 m 2 ) to achieve clinically required O2 and CO2 transport rates.
- HFM microporous hollow fiber membranes
- PP polypropylene
- PMP polymethylpentene
- heparin [Pappalardo, F., et al., Bioline heparin-coated ECMO with bivalirudin anticoagulation in a patient with acute heparin-induced thrombocytopenia: the immune reaction appeared to continue unabated. Perfusion, 2009. 24(2): pp. 135-137] or poly-2-methoxyethyl acrylate (PMEA) [Eisses, MJ, et al., Effect of polymer coating (poly 2-methoxyethyl acrylate) of the oxygenator on hemostatic markers during cardiopulmonary bypass in children. J Cardiothorac Vase Anesth, 2007. 21(1): pp. 28-34.].
- the coating of surfaces with polydopamine is known, for example, from CN 110545754, and the coating with polymers from US Pat. No. 6,309,660. However, both approaches have not proven themselves in practice.
- heparin The most common approach is to coat the surfaces with heparin, as described for example in WO 2020/190214.
- the heparin can detach from the surface and lead to the formation of thrombi.
- long-term exposure to heparin carries the risk that the patient will develop heparin-induced thrombocytopenia (HIT).
- HIT heparin-induced thrombocytopenia
- PF4 platelet factor 4
- cRGD Moller, L, et al., Towards a biocompatible artificial lung: Covalent functionalization of poly(4-methylpent-1-ene) (TPX) with cRGD pentapeptide. Beilstein Journal of Organic Chemistry, 2013. 9: pp. 270-277] or fibronectin [Cornelissen, CG, et al., Fibronectin coating of oxygenator membranes enhances endothelial cell attachment. Biomed Eng Online, 2013. 12:p.7] to aid in endothelialization.
- a method for producing a surface that can be colonized with biological cells in which the disadvantages of the prior art are avoided or at least reduced.
- a surface should be provided that can be populated with biological cells in such a way that a high degree of biocompatibility is achieved.
- This object is achieved by a method for producing a surface that can be colonized with biological cells, which has the following steps:
- a “surface” includes any area that is to be populated with biological cells, for example endothelial cells. This includes, but is not limited to, surfaces of medical products or medical devices, such as artificial lungs of oxygenators, e.g. the walls of membranes where gas exchange takes place. Plastic or metal surfaces are also included according to the invention.
- biological cells means any type of cell, such as endothelial cells, but also stem cells, including mesenchymal stem cells, and progenitor cells thereof or cells derived therefrom, such as endothelial progenitor cells (EPCs) or endothelial cells derived from induced pluripotent stem cells (“iPSC-derived endothelial cells”).
- EPCs endothelial progenitor cells
- iPSC-derived endothelial cells induced pluripotent stem cells
- “Silanization” in step 2 of the method according to the invention is understood as meaning the chemical attachment of a silane compound to the surface. This can be done, for example, but not exclusively, by reacting the free hydroxyl groups with 3-aminopropyltriethoxysilane (APTES).
- APTES 3-aminopropyltriethoxysilane
- step 3 of the method of the invention the reactant capable of reacting in a copper-free click reaction with azides, such as, but not limited to, dibenzocyclooctyne (DBCO), is conjugated to the silanized surface using methods known to those skilled in the art.
- azides such as, but not limited to, dibenzocyclooctyne (DBCO)
- a “click reaction”, also synonymously referred to as “click chemistry”, means a reaction between azide and alkyne to provide a 1,5-disubstituted 1,2,3-triazole. According to the invention, this takes place without the use of copper or other cytotoxic catalysts under physiological conditions, e.g. in the cell culture medium. This represents a departure from the "click responses" used in the prior art.
- Cu-AAC copper-catalyzed azide-alkyne cycloadditions
- the surfaces produced according to the invention are now capable of reacting with azides in a copper-free click reaction and thereby covalently binding structures which expose azide groups (N3) to the surface.
- azide groups N3
- endothelial cells can be functionalized in such a way that they have glycoproteins with azide groups on their surfaces. These groups can be covalently conjugated to the reactant such as DBCO in a copper-free click reaction.
- the method according to the invention thus provides surfaces which can be colonized efficiently and in a targeted manner with azide-modified biological cells. Because this reaction is very specific, only those biological cells that expose azide groups are bound to the surface. This prevents the non-specific binding of undesired cell populations.
- the method according to the invention can also be used to bind the patient's own endothelial progenitor cells (EPCs) or endothelial cells which are derived from induced pluripotent stem cells (“iPSC-derived endothelial cells”) to desired surfaces of blood-contacting materials.
- EPCs endothelial progenitor cells
- iPSC-derived endothelial cells endothelial cells which are derived from induced pluripotent stem cells
- the reactant of step 3 is selected from the group consisting of: cycloalkyne, cycloalkyne ester, phosphine and phosphine ester.
- This measure has the advantage that those reactants are used which are suitable for being able to react with azides in a copper-free click reaction and are therefore particularly functional according to the invention.
- phosphine groups react with the azide groups in a so-called Staudinger reaction.
- This reaction shows slow reaction kinetics of about 10' 3 M' 1 s' 1 .
- Cycloalkynes react with the azide groups in a so-called 'strain-promoted azide-alkyne cycloaddition reaction' (SPAAC).
- SPAAC 'strain-promoted azide-alkyne cycloaddition reaction'
- the second-order reaction has a moderate rate constant of 10' 2 — 1 M' 1 s' 1 .
- the cycloalkyne dibenzocyclooctyne (DBCO) and/or the cycloalkyne ester DBCO-PEG4-NHS ester is used as the reactant.
- DBCO cycloalkyne dibenzocyclooctyne
- DBCO-PEG4-NHS ester is used as the reactant.
- step 1 the surface containing hydroxyl groups is provided by oxygen plasma treatment of a surface.
- This measure has the advantage that a method is used that is particularly suitable for generating hydroxyl groups on the surface.
- the surface is one of a medical device, preferably selected from the group consisting of: artificial lung, oxygenator, artificial kidney, prosthesis, vascular prosthesis, stent, artificial heart.
- This measure has the advantage that the method is used where the colonization of surfaces with biological cells for the functionality and Biocompatibility of the devices is particularly important. When used in patients, it can thus be prevented that the device is recognized as "foreign".
- the surface is a surface of a hollow fiber membrane (HFM), preferably a polymethylpentene or polypropylene HFM.
- HFM hollow fiber membrane
- This measure has the advantage that the method is used for blood-contacting surfaces that are particularly relevant in practice, which are used, for example, in oxygenators.
- the biological cells are endothelial cells.
- the biological cells have azide groups (-N3) on their surface.
- Another object of the invention relates to a device with a surface that can be colonized with biological cells, preferably an artificial lung, an oxygenator, an artificial kidney, a prosthesis, a vascular prosthesis, a stent, an artificial heart or a hollow fiber membrane (HFM) such as a polymethylpentene or polypropylene HFM, wherein the surface has a reactant that reacts with azides in a copper-free click reaction.
- biological cells preferably an artificial lung, an oxygenator, an artificial kidney, a prosthesis, a vascular prosthesis, a stent, an artificial heart or a hollow fiber membrane (HFM) such as a polymethylpentene or polypropylene HFM, wherein the surface has a reactant that reacts with azides in a copper-free click reaction.
- HFM hollow fiber membrane
- the surface which can be colonized with biological cells was preferably obtained using the method according to the invention.
- the features, properties, advantages and embodiments of the manufacturing method according to the invention also apply in
- Another object of the present invention relates to a method for colonizing a surface with biological cells, which has the following steps:
- the surface having a reactant reacting with azides in a copper-free click reaction was obtained using the production method according to the invention.
- the biological cells which have azide groups (-N3) on their surface were obtained by incubating biological cells with an azide sugar which is preferably selected from the group consisting of: Ac4ManNAz (N- azidoacetylmannosamine-tetraacylated), N-azidoacetylglucosamine-tetraacylated (Ac4GlcNAz), N-azidoacetylgalactosamine-tetraacylated (Ac4GalNAz), and other azide-functionalized glycoconjugates.
- an azide sugar which is preferably selected from the group consisting of: Ac4ManNAz (N- azidoacetylmannosamine-tetraacylated), N-azidoacetylglucosamine-tetraacylated (Ac4GlcNAz), N-azidoacetylgalactosamine-tetraacylated (Ac4GalNAz), and other azide-functionalized
- nucleic acid By this measure, biological cells are obtained in an advantageous manner, which expose azide groups (-N3) on their surface.
- the azide sugars specified are particularly suitable for the functionalization of the cells. Like their natural analogues, the azide sugars are taken up into the cells and incorporated into glycoproteins. This causes the cells to have glycoproteins on the cell surface that are provided with azide groups (-N3). These groups can be covalently conjugated to the surface using click chemistry.
- Incubating the modified cells with the modified HFMs results in a bio-orthogonal reaction, the alkyne group of the reactant or DBCO reacts with the azide on the surface of the cells. This happens very specifically, so only modified cells with azide molecules can bind to the reactants on the membrane. Binding of the cells to the membrane and further incubation in cell culture medium leads to proliferation and ultimately colonization of the membrane with a confluent monolayer of cells.
- the biological cells are endothelial cells.
- the surfaces are populated with cells that are of particular importance for the functionality of important medical surfaces or devices.
- Particularly suitable are primary endothelial cells isolated from umbilical cord blood, so-called Human Umbilical Vein Endothelial Celis (HUVECs), which are cultivated in VascuLife® EnGS Endothelial Medium Complete Kit until they are endothelialized.
- HUVECs Human Umbilical Vein Endothelial Celis
- colonization with the endothelial cells can take place in EGM-2 Endothelial Cell Growth Medium-2 BulletKit.
- Polymethylpentene HFMs (PMP, OXYPLUS, 3M Membrana, Wuppertal, Germany), APTES, methanol, toluene (Sigma-Aldrich, Darmstadt), DBCO-PEG4-NHS ester (Jena Bioscience, Jena, Germany), Dulbecco's phosphate-buffered saline (DPBS), (Invitrogen), N-azidoacetylmannosamine-tetraacylated (Ac4ManNAz, Sigma Aldrich, Germany), DMSO (Sigma-Aldrich).
- Fig. 1 Schematic representation of the functionalization of the PMP hollow fiber membranes (HFMs).
- HFMs PMP hollow fiber membranes
- Fig.2 Detection of the generated functional groups on PMP hollow fiber membranes (HFMs).
- Fig. 3 Result of the SEM analysis. There are no visual differences on the
- FIG. 6 Metabolic labeling of HUVECs with Ac4ManNAz and cell viability analysis. 2x10 5 HUVECs were incubated for 48 h with and without 50 pM Ac4ManNAz.
- B) Cells were incubated for 1 hour with 5 pM DBCO-Sulfo-Cy3 and, after washing with DPBS, flow cytometric analyzes were performed. Results are presented as mean ⁇ SD (n 3).
- Fig. 8 Analysis of the response of the endothelialized surface to an inflammatory stimulus using qRT-PCR.
- Endothelialized HFM were treated with 50 ng/ml TNF- ⁇ and the expression of adhesion molecules (E-selectin, VCAM-1 and ICAM-1) was determined by qRT-PCR.
- TAT thrombin-antithrombin III complex
- sC5b-9 complement system
- PMN- Elastase inflammation
- HIVECs Human Umbilical Vein Endothelial Cells
- HUVECs (ATCC®, Manassas, USA) were seeded in T75 cell culture flasks coated with 0.1% gelatin and cultured at 37°C and 5% CO2 in Vasculife® EnGS EC culture medium (CellSystems, Troisdorf, Germany) with VascuLife EnGS LifeFactors Kit, 50 mg/ml gentamicin and 0.05 mg/ml amphotericin B (Thermo Fisher Scientific, Waltham, USA). The medium was changed every 3 days. After reaching 80% confluency, the cells were detached with trypsin/EDTA (0.04%/0.03%, PromoCell, Heidelberg, Germany).
- the surface of polymethylpentene HFMs (PMP, OXYPLUS, 3M Membrana, Wuppertal, Germany) was coated in a three-stage process ( Figure 1). First the surface was treated with 02 plasma and in the second step the silanization was carried out. In the third step, dibenzylcyclooctyne-PEG4-NHS-ester (DBCO-PEG4-NHS-ester) was conjugated to the silanized surface.
- DBCO-PEG4-NHS-ester dibenzylcyclooctyne-PEG4-NHS-ester
- PMP membranes were cut to a size of 3 x 3 cm and treated with 02 plasma (Denta Pias®, Diener electronic, Ebhausen, Germany) for 30 min at a pressure of 0.3 mbar ( ⁇ 0.20 mbar ) and an output of 80% ( ⁇ 5%) hydroxyl groups.
- the HFMs were then incubated for 30 minutes in toluene (Sigma-Aldrich, Darmstadt, Germany) with 2% (3-aminopropyl)triethoxysilane (APTES, Sigma-Aldrich) on a shaker at 20 rpm.
- the membranes were then sonicated in 100% toluene, 50% toluene/50% methanol, and 100% methanol for 2 minutes each to remove unbound or weakly bound APTES. After drying, the membranes were soaked in 400 M DBCO-PEG4-NHS-ester (Jena Bioscience, Jena, Germany) with Dulbecco's phosphate-buffered saline for 30 minutes (DPBS) (Invitrogen, Carlsbad, USA) to introduce cyclooctyne groups and washed three times with fresh DPBS.
- DPBS Dulbecco's phosphate-buffered saline for 30 minutes
- the membranes were incubated for 5 hours in 0.5 mM methyl orange (Sigma-Aldrich, Darmstadt, Germany) at 37°C on a shaker, followed by washing three times with 1 mM HCl -Solution. Desorption of bound methyl orange from the HFM surface was performed with 0.4 mL of a 1 mM NaOH solution overnight at 37°C on a shaker. For detection of methyl orange, 100 ⁇ l of the desorption solution was placed in triplicate in a 96-well plate. The absorbance was measured at 465 nm with a microplate reader (Eon Synergy 2, Bio Tek Instruments Inc., Winooski USA).
- the HFM in DPBS without Ca 2+ /Mg 2+ were incubated for 1 hour with 80 pg/ml Cy3-azide (Sigma-Aldrich) to convert the Cy3-azide bind to the DBCO-functionalized HFM surface.
- the HFMs were then washed five times with 100% ethanol and the staining of the membranes was detected using fluorescence microscopy (Axiovert 135, Carl Zeiss AG, Oberkochen, Germany).
- the fluorescence intensity on the membranes was quantified using a microplate reader (Mithras 940, Berthold Technologies, Bad Wildbad, Germany) at an excitation wavelength of 500 nm and an emission wavelength of 600 nm. HFM with no treatment and with APTES treatment only were used as controls.
- HUVECs 1 .4 Metabolic labeling of HUVECs with azide groups and detection of the azide label
- 2 x 10 5 HUVECs were seeded per well of a 6-well plate and tetraacylated with or without 50 pM N-azidoacetylmannosamine (Ac4ManNAz, Sigma Aldrich) in 2 ml medium for Treated with 5% CO2 at 37°C for 48 h.
- the cells were washed with DPBS without Ca 2+ /Mg 2+ and with 5 pM DBCO-Sulfo-Cy3 (Jena Bioscience, Germany) in DPBS with Ca 2+ /Mg 2+ for 1 h at 37° C and 5% CO2.
- the cells were washed three times with 10 ml DPBS, detached with trypsin/EDTA (0.04%/0.03%) and centrifuged at 300 ⁇ g for 5 minutes.
- the cell pellet was resuspended in 0.5 ml DPBS and the Cy3 labeling of 10,000 cells was analyzed by flow cytometry (FACS SCAN, BD, Heidelberg, Germany).
- HUVECs The influence of metabolic labeling on the viability of HUVECs was examined 48 hours after labeling with 50 pM Ac4ManNAz using the PrestoBlue assay (Invitrogen, Carlsbad, USA). For this purpose, 500 pL 1 ⁇ Presto Blue cell viability reagent were added per well of a 6-well plate and incubated at 37° C. for 1.5 h. The fluorescence intensity of 100 pL of supernatant was measured in triplicate at an excitation wavelength of 530 nm and an emission wavelength of 600 nm using a multimode microplate reader (Mithras LB 940, Berthold Technologies, Bad Wildbad, Germany).
- DBCO-coated and uncoated PMP membranes (3 x 3 cm) were incubated with 5.2 x 10 6 Ac4ManNAz-labeled HUVECs in 13 ml of EGMTM-2 endothelial cell growth medium BulletKitTM (Lonza, Basel, Switzerland). Incubation was carried out for 24 hours at 10 rpm with a rotator (Phoenix Instrument, Garbsen, Germany) in one humidified incubator with 5% CO2 at 37°C. After the rotary incubation, the membranes were placed in each well of a 6-well plate and further incubated for 1 or 5 days under static conditions.
- the HLIVECs attached to the membranes were stained with 250 ng/mL calcein acetoxymethyl ester (Calcein-AM; Invitrogen) for 10 minutes at 37°C.
- the stained cells were detected using fluorescence microscopy (Axio 135, Carl Zeiss AG, Oberkochen, Germany).
- fluorescence signal on the surface of the HFMs before and after calcein-AM staining was detected at an excitation wavelength of 494 nm and an emission wavelength of 517 nm using a multimode microplate reader (Mithras LB 940, Berthold Technologies).
- the formation of a confluent endothelial cell monolayer and the presence of cell-cell contacts were detected by staining the cells with an anti-human VE-cadherin antibody.
- the membranes coated with DBCO-PEG4-NHS ester and endothelialized were washed with DPBS and fixed in 4% paraformaldehyde (v/v) for 10 minutes at RT. After washing with DPBS, blocking was performed with Tris-buffered saline (50 mM Tris base, 150 mM sodium chloride, pH 7.5) containing 5% goat serum (Thermo Fisher Scientific, Waltham, USA) for 30 minutes at RT.
- the membranes were washed with DPBS and with the primary antibody mouse anti-human VE-cadherin (CD144 (16B1), eBioscience, Invitrogen, USA) diluted 1:100 in DPBS with 1% bovine serum albumin (BSA) for 1 h at RT incubated. The membranes were then washed three times with DPBS and incubated with 1:400 diluted goat anti-mouse IgG (H+L) cross-adsorbed secondary antibody (ThermoFisher, USA) labeled with Alexa Fluor 488 (ThermoFisher, USA) for 1 h in PBS with 1% BSA .
- the cell nuclei were stained for 5 minutes with DAPI (Sigma Aldrich, Darmstadt, Germany) diluted 1:2000 in DPBS. As a negative control, the membranes were also incubated with an isotype control antibody. The stained cells were detected using fluorescence microscopy (Axio 135, Carl Zeiss AG, Aoberkochen, Germany). 1 .8 Reactivity of the endothelium to HFMs to the inflammatory stimulus
- Endothelialized HFMs were incubated for 4 hours without or with 50 ng/mL tumor necrosis factor ⁇ (TNF- ⁇ , Sigma-Aldrich) in EGMTM-2 endothelial cell growth medium without hydrocortisone. After incubation, the HFMs were washed twice with DPBS and total RNA isolated. Expression of the activation markers, endothelial leukocyte adhesion molecule 1 (E-selectin), vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1), was detected by qRT-PCR. The mRNA levels were normalized to the GAPDH mRNA levels and the results were presented relative to the expression levels in endothelialized HFM without TNF- ⁇ stimulation.
- TNF- ⁇ tumor necrosis factor ⁇
- RNA was isolated with the Aurum Total RNA Mini Kit (Bio-Rad, Kunststoff, Germany) according to the manufacturer's instructions. 300 ng of RNA was reverse transcribed into DNA copies (cDNA) using the iScript Kit (Bio-Rad). The transcripts were amplified using the primers listed in Table 1 in a final concentration of 300 nM under the following conditions: one cycle of 3 min at 95°C, followed by 40 cycles of 95°C for 15 s and 72°C for 10 s After 40 cycles, a melting curve analysis was performed to detect the specific amplicons.
- ICAM-1 CTTGAGGGCACCTACCTCTGTC CGGCTGTCTACCACAGTGATG
- VCAM-1 ACACTTTATGTCAATGTTGCCCC GAGGCTGTAGCTCCCCGTTAG
- Tubes containing (0.3 mL citrate solution/3 mL blood, 0.106 M CeHsNasO? x 2H2O, Sartstedt) were used to detect PMN elastase and TAT.
- blood was collected in 2.7 mL CTAD tubes containing 270 pL of 0.109 M CTAD solution containing buffered sodium citrate, theophylline, adenosine and dipyridamole (BD Vacutainer CTAD, Becton-Dickinson GmbH, Heidelberg, Germany ), transferred and placed on ice for 15 minutes.
- the EDTA and CTAD preparations were centrifuged at 2500 xg for 20 minutes at 4°C.
- the citrated blood preparations were centrifuged at 1800 ⁇ g for 18 min at RT.
- the blood plasma each Sample was snap frozen in liquid nitrogen and stored at -80°C until further investigation.
- the numbers of erythrocytes, leukocytes and thrombocytes were measured in the collected blood samples with an automatic cell counting system (ABX Micros 60, HORIBA Medical, Minami-ku, Kyoto, Japan).
- thrombin-antithrombin III complex TAT
- sC5b-9 complement system
- ß-thromboglobulin ß-thromboglobulin
- neutrophils commercially available ELISA Kits (enzyme-linked immunosorbent assays) used according to manufacturer's instructions.
- sC5b-9 (MicroVueTM Complement, Quidel Germany GmbH & AnDiaTec Division, Kornwestheim, Germany), TAT (Enzygnost® TAT micro, Siemens Healthcare, Er Weg, Germany), polymorphonuclear (PMN) elastase (Demeditec Diagnostics, Kiel, Germany) and ⁇ -TG (Asserachrom® ⁇ -TG, Diagnostica Stago, Parsippany, NJ, USA) ELISAs.
- TAT Enzygnost® TAT micro, Siemens Healthcare, Er Weg, Germany
- PMN polymorphonuclear
- ⁇ -TG Assserachrom® ⁇ -TG, Diagnostica Stago, Parsippany, NJ, USA
- Uncoated, DBCO-coated, or endothelialized HFMs were incubated with blood as described above. After incubating the HFMs with human blood for 90 minutes, the blood was collected and diluted 1:5 with DPBS. The samples were treated for 30 minutes at RT with 10 ⁇ l mouse anti-human CD41-FITC antibody (company) for the detection of platelets and 10 ⁇ l mouse anti-human CD62P-PE antibody (BD Biosciences, Heidelberg, Germany ) to detect platelets expressing P-selectin (CD62P), indicating platelet activation. Samples were fixed with 0.5% paraformaldehyde and platelet activation analyzed by flow cytometry. Scanning electron microscopy (SEM) analyzes of the HFM surfaces
- HFMs were rinsed with 0.9% saline (Fresenius Kabi, Bad Homburg, Germany) and immersed in 2.5% glutaraldehyde (Sigma Aldrich, Darmstadt, Germany) fixed in DPBS. After the fixation step, the samples were washed with DPBS for 15 min at RT and then dehydrated in 15 min steps at RT with an ascending ethanol series (40%-100% ethanol; Merck-Millipore, Darmstadt, Germany).
- 0.9% saline Fresenius Kabi, Bad Homburg, Germany
- glutaraldehyde Sigma Aldrich, Darmstadt, Germany
- Hydroxyl groups on the surface of materials can react with chemical groups on other molecules to form a covalent bond.
- the free hydroxyl groups react with APTES.
- Diluting APTES in anhydrous toluene is a widely used method in silanization.
- the membranes were incubated for 30 min in 2% APTES (APTES, Sigma-Aldrich, Darmstadt) diluted in toluene (Sigma-Aldrich, Darmstadt, Germany) at 20 rpm and RT.
- Three washing steps were then carried out in an ultrasonic bath for 2 min to remove unbound or weakly bound APTES.
- DBCO Conjugation (Step 3 in Figure 1)
- the silanized membranes were then diluted with 400 pM DBC0-PEG4-NHS ester (Jena Bioscience, Jena, Germany) in Dulbecco's phosphate-buffered saline (DPBS, Invitrogen) and incubated for 30 min at RT. After the incubation, three washing steps were carried out in DPBS for 5 min each.
- the DBCO used contains a PEG4 chain to extend the DBCO to enable easier binding to the metabolically modified cells. Furthermore, there is an activated carboxylic acid (-NHS ester) on the DBCO used, which can react with amino groups from APTES and thus form covalent bonds.
- the pressure was monitored with a pressure gauge (RS 1113, RS Components, Corby, UK) connected to the system.
- the cells were washed with DPBS without Ca 2+ /Mg 2+ and with 5 pM DBCO-Sulfo-Cy3 (Jena Bioscience, Germany) in DPBS with Ca 2+ /Mg 2+ for 1 h at 37°C and 5% CO2 incubated.
- the cells were then washed three times with 1 ml DPBS, detached with trypsin/EDTA (0.04%/0.03%) and centrifuged at 300 ⁇ g for 5 min.
- the cell pellet was resuspended in 0.5 ml DPBS and the Cy3 labeling of 10,000 cells was analyzed by flow cytometry (FACS SCAN, BD, Heidelberg, Germany).
- the HLIVECs were treated with 50 pM Ac4ManNAz for 48 hours. To determine if Na groups are present on the cell surface, cells were then incubated with 5 pM DBCO-Sulfo-Cy3 for 1 hour. The presence of N3 on the cell surface led to conjugation of DBCO and labeling of cells with Cy3 ( Figure 6A). The cells without Ac4ManNAz treatment were not stained. In addition, flow cytometric analyzes showed that about 98% of the cells analyzed were Cy3-positive (FIG. 6B). Treatment of the cells with Ac4ManNAz had no effect on the viability of the HUVECs ( Figure 6C).
- DBCO-coated and uncoated HFMs were incubated with 5.2 x 10 6 Ac4ManNAz-labeled HUVECs for 24 hours under rotation and then for 24 hours under static conditions.
- the attached HUVECs were detected by staining with calcein AM ( Figure 7A). Fluorescence microscopy analysis showed that Ac4ManNAz-treated HUVECs efficiently bound to the DBCO-functionalized HFM surface and resulted in almost complete endothelialization.
- the detection of the fluorescence intensity of the HFMs also showed that the DBCO functionalization led to a significantly increased adhesion of the HUVECs compared to uncoated or APTES-coated HFMs (Figure 7B).
- the tight cell-cell contacts of HUVECs on the HFM membranes were detected by staining with an anti-VE-cadherin antibody ( Figure 7C).
- E-selectin The expression of the adhesion molecules E-selectin, VCAM-1 and ICAM-1 was analyzed after stimulation of the endothelial layer on HFMs with TNF- ⁇ and compared with the non-stimulated endothelial layer using qRT-PCR (Figure 8).
- the stimulation of Endothelial layer with TNF- ⁇ resulted in significantly higher expression of E-selectin (199-fold), VCAM-1 (328-fold) and ICAM-1 (162-fold) compared to the unstimulated endothelial layer. This demonstrated the responsiveness of the generated endothelial layer to HFMs.
- the P-selectin expression on the platelets was also analyzed as a marker for platelet activation. As shown in FIG. 9, a significantly higher number of activated platelets was detected on uncoated and DBCO-coated HFM compared to the blood samples without HFM. However, the endothelialized membranes prevented platelet activation.
- thrombin-antithrombin III complex TAT
- ⁇ -thromboglobulin a marker of platelet activation
- the inventors provide a method by which biological cells can be covalently attached to surfaces in a very specific and selective manner. This enables efficient colonization of the surfaces, for example with endothelial cells, in order to prevent the surfaces from being recognized as "foreign".
- the method can also be used to attach patient EPCs or "iPSC-derived" endothelial cells to desired surfaces of blood-contacting materials.
- Off-the-shelf products can be manufactured that can be populated with the patient's own endothelial cells if required. The conjugation of the cells to the surface takes place without a catalyst under physiological conditions.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Engineering & Computer Science (AREA)
- Transplantation (AREA)
- Dermatology (AREA)
- Medicinal Chemistry (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Cell Biology (AREA)
- Surgery (AREA)
- Vascular Medicine (AREA)
- Heart & Thoracic Surgery (AREA)
- Zoology (AREA)
- Botany (AREA)
- Urology & Nephrology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA3231506A CA3231506A1 (en) | 2021-09-14 | 2022-08-16 | Colonization of surfaces with biological cells |
EP22768269.7A EP4401803A1 (de) | 2021-09-14 | 2022-08-16 | Besiedelung von oberflaechen mit biologischen zellen |
KR1020247012314A KR20240055861A (ko) | 2021-09-14 | 2022-08-16 | 생물학적 세포를 사용한 표면의 집락화 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE102021123779.3 | 2021-09-14 | ||
DE102021123779.3A DE102021123779A1 (de) | 2021-09-14 | 2021-09-14 | Besiedelung von Oberflächen mit biologischen Zellen |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2023041275A1 true WO2023041275A1 (de) | 2023-03-23 |
Family
ID=83271222
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP2022/072853 WO2023041275A1 (de) | 2021-09-14 | 2022-08-16 | Besiedelung von oberflaechen mit biologischen zellen |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP4401803A1 (de) |
KR (1) | KR20240055861A (de) |
CA (1) | CA3231506A1 (de) |
DE (1) | DE102021123779A1 (de) |
WO (1) | WO2023041275A1 (de) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6309660B1 (en) | 1999-07-28 | 2001-10-30 | Edwards Lifesciences Corp. | Universal biocompatible coating platform for medical devices |
CN110545754A (zh) | 2017-04-13 | 2019-12-06 | 祥丰医疗私人有限公司 | 用聚多巴胺和抗体涂覆的医疗设备 |
WO2020190214A1 (en) | 2019-03-19 | 2020-09-24 | Jmedtech Coating Technologies Pte Ltd | Medical device comprising covalently bonded heparin coating |
-
2021
- 2021-09-14 DE DE102021123779.3A patent/DE102021123779A1/de active Pending
-
2022
- 2022-08-16 KR KR1020247012314A patent/KR20240055861A/ko unknown
- 2022-08-16 CA CA3231506A patent/CA3231506A1/en active Pending
- 2022-08-16 EP EP22768269.7A patent/EP4401803A1/de active Pending
- 2022-08-16 WO PCT/EP2022/072853 patent/WO2023041275A1/de active Application Filing
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6309660B1 (en) | 1999-07-28 | 2001-10-30 | Edwards Lifesciences Corp. | Universal biocompatible coating platform for medical devices |
CN110545754A (zh) | 2017-04-13 | 2019-12-06 | 祥丰医疗私人有限公司 | 用聚多巴胺和抗体涂覆的医疗设备 |
WO2020190214A1 (en) | 2019-03-19 | 2020-09-24 | Jmedtech Coating Technologies Pte Ltd | Medical device comprising covalently bonded heparin coating |
Non-Patent Citations (16)
Title |
---|
AVCI-ADALI, M. ET AL.: "Current concepts and new developments for autologous in vivo endothelialisation of biomaterials for intravascular applications", EUROPEAN CELLS & MATERIALS, vol. 21, 2011, pages 157 - 176 |
CORNELISSEN, C.G. ET AL.: "Fibronectin coating of oxygenator membranes enhances endothelial cell attachment", BIOMED ENG ONLINE, vol. 12, 2013, pages 7, XP021146904, DOI: 10.1186/1475-925X-12-7 |
DE MEL, A.B.G. COUSINSA.M. SEIFALIAN: "Surface modification of biomaterials: a quest for blood compatibility", INT J BIOMATER, vol. 2012, 2012, pages 707863 |
EISSES, M.J. ET AL.: "Effect of polymer coating (poly 2-methoxyethylacrylate) of the oxygenator on hemostatic markers during cardiopulmonary bypass in children", J CARDIOTHORAC VASC ANESTH, vol. 21, no. 1, 2007, pages 28 - 34, XP005874302, DOI: 10.1053/j.jvca.2006.03.023 |
GULACK, B.C.S.A. HIRJIM.G. HARTWIG: "Bridge to lung transplantation and rescue post-transplant: the expanding role of extracorporeal membrane oxygenation", J THORAC DIS, vol. 6, no. 8, 2014, pages 1070 - 1079 |
KOO, H. ET AL.: "Bioorthogonal Copper-Free Click Chemistry In Vivo for Tumor-Targeted Delivery of Nanoparticles", ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, vol. 51, no. 47, 2012, pages 11836 - 11840, XP002758195, DOI: 10.1002/anie.201206703 |
LAUGHLIN, S.T. ET AL.: "In vivo imaging of membrane-associated glycans in developing zebrafish", SCIENCE, vol. 320, no. 5876, 2008, pages 664 - 667, XP055383745, DOI: 10.1126/science.1155106 |
MIYAHARA KOKI ET AL: "A Cu-free clickable surface with controllable surface density", COLLOID & POLYMER SCIENCE, SPRINGER VERLAG, HEIDELBERG, DE, vol. 297, no. 6, 8 May 2019 (2019-05-08), pages 927 - 931, XP036791405, ISSN: 0303-402X, [retrieved on 20190508], DOI: 10.1007/S00396-019-04515-4 * |
MOLLER, L. ET AL.: "Towards a biocompatible artificial lung: Covalent functionalization of poly(4-methylpent-1-ene) (TPX) with cRGD pentapeptide", BEILSTEIN JOURNAL OF ORGANIC CHEMISTRY, vol. 9, 2013, pages 270 - 277 |
PAPPALARDO, F. ET AL.: "Bioline heparin-coated ECMO with bivalirudin anticoagulation in a patient with acute heparin-induced thrombocytopenia: the immune reaction appeared to continue unabated", PERFUSION, vol. 24, no. 2, 2009, pages 135 - 137 |
PIERI, M. ET AL.: "A new phosphorylcholine-coated polymethylpentene oxygenator for extracorporeal membrane oxygenation: a preliminary experience", PERFUSION, vol. 28, no. 2, 2013, pages 132 - 137 |
WALLRODT, S. ET AL.: "Bioorthogonally Functionalized NAD(+) Analogues for In-Cell Visualization of Poly(ADP-Ribose) Formation", ANGEW CHEM INT ED ENGL, vol. 55, no. 27, 2016, pages 7660 - 7664 |
WENDEL, H.P.G. ZIEMER: "Coating-techniques to improve the hemocompatibility of artificial devices used for extracorporeal circulation", EUR J CARDIOTHORAC SURG, vol. 16, no. 3, 1999, pages 342 - 350 |
WIEGMANN BETTINA ET AL: "Developing a biohybrid lung - sufficient endothelialization of poly-4-methly-1-pentene gas exchange hollow-fiber membranes", JOURNAL OF THE MECHANICAL BEHAVIOR OF BIOMEDICAL MATERIALS, ELSEVIER, AMSTERDAM, NL, vol. 60, 11 February 2016 (2016-02-11), pages 301 - 311, XP029556100, ISSN: 1751-6161, DOI: 10.1016/J.JMBBM.2016.01.032 * |
WIEGMANN, B. ET AL.: "Developing a biohybrid lung - sufficient endothelialization of poly-4-methly-1-pentene gas exchange hollow-fiber membranes", J MECH BEHAV BIOMED MATER, vol. 60, 2016, pages 301 - 311, XP029556100, DOI: 10.1016/j.jmbbm.2016.01.032 |
XIE, R. ET AL.: "In vivo metabolic labeling of sialoglycans in the mouse brain by using a liposome-assisted bioorthogonal reporter strategy", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 113, no. 19, 2016, pages 5173 - 5178 |
Also Published As
Publication number | Publication date |
---|---|
EP4401803A1 (de) | 2024-07-24 |
KR20240055861A (ko) | 2024-04-29 |
DE102021123779A1 (de) | 2023-03-16 |
CA3231506A1 (en) | 2023-03-23 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Nakabayashi et al. | Preparation of non-thrombogenic materials using 2-methacryloyloxyethyl phosphorylcholine | |
DE69214466T2 (de) | Polymerbeschichtung | |
DE69932273T2 (de) | Befestigung von Biomolekülen an Oberflächen von medizinischen Geräten | |
EP0748633B1 (de) | Gewebeklebstoff auf der Basis von Fibrinogen und Verwendung desselben zur Herstellung eines Hämostyptikum | |
DE2812174C2 (de) | Die Blutgerinnung nicht fördernde, biologisch verträgliche Gegenstände | |
CN102202771B (zh) | 用于细胞扩增的辐照膜 | |
DE69822872T2 (de) | Verfahren zur oxidativen bindung von glycoproteinen und glycopeptiden an oberflächen von medizinischen artikeln | |
Dimitrievska et al. | Glycocalyx‐Like Hydrogel Coatings for Small Diameter Vascular Grafts | |
DE10004884A1 (de) | Verfahren zum Anbringen von Biomolekülen an Oberflächen mittels funktioneller Amin-Gruppen | |
CN103536958B (zh) | 一种基于溶菌酶和丝蛋白的层层自组装改性纤维素纳米纤维膜及其制备与应用 | |
CN103301506A (zh) | 一种抗凝血丝素膜及其制备方法 | |
Dai et al. | Immobilizing argatroban and mPEG-NH2 on a polyethersulfone membrane surface to prepare an effective nonthrombogenic biointerface | |
Raut et al. | Engineering biomimetic polyurethane using polyethylene glycol and gelatin for blood-contacting applications | |
EP1152778B1 (de) | Hämokompatible oberflächen und verfahren zu deren herstellung | |
Liu et al. | Electrofabrication of flexible and mechanically strong tubular chitosan implants for peripheral nerve regeneration | |
CN102940909B (zh) | 一种修饰组织工程支架的方法 | |
Canjuga et al. | Improving hemocompatibility of artificial lungs by click conjugation of glycoengineered endothelial cells onto blood-contacting surfaces | |
WO2023041275A1 (de) | Besiedelung von oberflaechen mit biologischen zellen | |
CN102477172B (zh) | 壳聚糖-肝素纳米颗粒以及用该纳米颗粒处理的去细胞基质的生物材料 | |
Peng et al. | A robust mixed-charge zwitterionic polyurethane coating integrated with antibacterial and anticoagulant functions for interventional blood-contacting devices | |
DE3026805C2 (de) | ||
EP2452624A1 (de) | Implantierbarer theranostischer Artikel | |
EP0290642B1 (de) | Verfahren zur Besiedlung einer Polymeroberfläche mit menschlichen Gefässinnenhautzellen | |
CN103961750B (zh) | 一种小口径原位组织工程血管及构建方法 | |
US20070059473A1 (en) | Sheet-like covering member used for implant medical device |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 22768269 Country of ref document: EP Kind code of ref document: A1 |
|
WWE | Wipo information: entry into national phase |
Ref document number: 3231506 Country of ref document: CA |
|
REG | Reference to national code |
Ref country code: BR Ref legal event code: B01A Ref document number: 112024004932 Country of ref document: BR |
|
ENP | Entry into the national phase |
Ref document number: 20247012314 Country of ref document: KR Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2022768269 Country of ref document: EP |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 2022768269 Country of ref document: EP Effective date: 20240415 |
|
ENP | Entry into the national phase |
Ref document number: 112024004932 Country of ref document: BR Kind code of ref document: A2 Effective date: 20240313 |