PH12014000281A1 - Injectable coco cellulose microparticle for tissue augmentation dermal and subdermal filler - Google Patents
Injectable coco cellulose microparticle for tissue augmentation dermal and subdermal filler Download PDFInfo
- Publication number
- PH12014000281A1 PH12014000281A1 PH12014000281A PH12014000281A PH12014000281A1 PH 12014000281 A1 PH12014000281 A1 PH 12014000281A1 PH 12014000281 A PH12014000281 A PH 12014000281A PH 12014000281 A PH12014000281 A PH 12014000281A PH 12014000281 A1 PH12014000281 A1 PH 12014000281A1
- Authority
- PH
- Philippines
- Prior art keywords
- cellulose
- lip
- glucose
- lib
- coco
- Prior art date
Links
- 229920002678 cellulose Polymers 0.000 title claims abstract description 99
- 239000001913 cellulose Substances 0.000 title claims abstract description 98
- 239000000945 filler Substances 0.000 title abstract description 5
- 230000002500 effect on skin Effects 0.000 title abstract description 3
- 230000003416 augmentation Effects 0.000 title abstract 2
- 239000011859 microparticle Substances 0.000 title abstract 2
- 229920002749 Bacterial cellulose Polymers 0.000 claims description 110
- 239000005016 bacterial cellulose Substances 0.000 claims description 110
- 230000015572 biosynthetic process Effects 0.000 claims description 30
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 18
- 108010040093 cellulose synthase Proteins 0.000 claims description 17
- 239000008103 glucose Substances 0.000 claims description 16
- 239000012528 membrane Substances 0.000 claims description 13
- 241000589220 Acetobacter Species 0.000 claims description 9
- 210000001724 microfibril Anatomy 0.000 claims description 8
- 229930091371 Fructose Natural products 0.000 claims description 7
- 239000005715 Fructose Substances 0.000 claims description 7
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims description 7
- 230000007246 mechanism Effects 0.000 claims description 7
- 235000002837 Acetobacter xylinum Nutrition 0.000 claims description 6
- 230000037361 pathway Effects 0.000 claims description 6
- 229910019142 PO4 Inorganic materials 0.000 claims description 4
- 230000008166 cellulose biosynthesis Effects 0.000 claims description 4
- 239000010452 phosphate Substances 0.000 claims description 4
- IZSRJDGCGRAUAR-MROZADKFSA-M 5-dehydro-D-gluconate Chemical compound OCC(=O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O IZSRJDGCGRAUAR-MROZADKFSA-M 0.000 claims description 3
- NBSCHQHZLSJFNQ-GASJEMHNSA-N D-Glucose 6-phosphate Chemical compound OC1O[C@H](COP(O)(O)=O)[C@@H](O)[C@H](O)[C@H]1O NBSCHQHZLSJFNQ-GASJEMHNSA-N 0.000 claims description 3
- VFRROHXSMXFLSN-UHFFFAOYSA-N Glc6P Natural products OP(=O)(O)OCC(O)C(O)C(O)C(O)C=O VFRROHXSMXFLSN-UHFFFAOYSA-N 0.000 claims description 3
- 230000006860 carbon metabolism Effects 0.000 claims description 3
- 150000002632 lipids Chemical class 0.000 claims description 3
- 239000011148 porous material Substances 0.000 claims description 3
- 239000002028 Biomass Substances 0.000 claims description 2
- XJLXINKUBYWONI-DQQFMEOOSA-N [[(2r,3r,4r,5r)-5-(6-aminopurin-9-yl)-3-hydroxy-4-phosphonooxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2s,3r,4s,5s)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate Chemical compound 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](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-DQQFMEOOSA-N 0.000 claims 5
- RGHNJXZEOKUKBD-SQOUGZDYSA-M D-gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O RGHNJXZEOKUKBD-SQOUGZDYSA-M 0.000 claims 2
- 229940050410 gluconate Drugs 0.000 claims 2
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 claims 2
- VBUYCZFBVCCYFD-JJYYJPOSSA-N 2-dehydro-D-gluconic acid Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C(=O)C(O)=O VBUYCZFBVCCYFD-JJYYJPOSSA-N 0.000 claims 1
- 101710162174 D-glyceraldehyde dehydrogenase (NADP(+)) Proteins 0.000 claims 1
- 241001136169 Komagataeibacter xylinus Species 0.000 claims 1
- HSCJRCZFDFQWRP-JZMIEXBBSA-N UDP-alpha-D-glucose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OP(O)(=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-JZMIEXBBSA-N 0.000 claims 1
- HSCJRCZFDFQWRP-UHFFFAOYSA-N Uridindiphosphoglukose Natural products OC1C(O)C(O)C(CO)OC1OP(O)(=O)OP(O)(=O)OCC1C(O)C(O)C(N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-UHFFFAOYSA-N 0.000 claims 1
- 230000034659 glycolysis Effects 0.000 claims 1
- GSXOAOHZAIYLCY-HSUXUTPPSA-N keto-D-fructose 6-phosphate Chemical compound OCC(=O)[C@@H](O)[C@H](O)[C@H](O)COP(O)(O)=O GSXOAOHZAIYLCY-HSUXUTPPSA-N 0.000 claims 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims 1
- 230000029058 respiratory gaseous exchange Effects 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 20
- 230000008569 process Effects 0.000 abstract description 12
- 239000001963 growth medium Substances 0.000 abstract description 10
- 235000013305 food Nutrition 0.000 abstract description 7
- 230000002829 reductive effect Effects 0.000 abstract description 6
- 229920001340 Microbial cellulose Polymers 0.000 abstract description 5
- 231100000252 nontoxic Toxicity 0.000 abstract description 5
- 230000003000 nontoxic effect Effects 0.000 abstract description 5
- 241001602876 Nata Species 0.000 abstract description 3
- 235000020197 coconut milk Nutrition 0.000 abstract description 2
- 230000001815 facial effect Effects 0.000 abstract description 2
- 230000000774 hypoallergenic effect Effects 0.000 abstract description 2
- 238000012360 testing method Methods 0.000 abstract description 2
- 101100243022 Mus musculus Pcnt gene Proteins 0.000 abstract 2
- 206010070834 Sensitisation Diseases 0.000 abstract 2
- 230000008313 sensitization Effects 0.000 abstract 2
- 206010024604 Lipoatrophy Diseases 0.000 abstract 1
- 239000004480 active ingredient Substances 0.000 abstract 1
- 239000012467 final product Substances 0.000 abstract 1
- 230000002163 immunogen Effects 0.000 abstract 1
- 230000007794 irritation Effects 0.000 abstract 1
- 231100000344 non-irritating Toxicity 0.000 abstract 1
- ARIWANIATODDMH-UHFFFAOYSA-N rac-1-monolauroylglycerol Chemical compound CCCCCCCCCCCC(=O)OCC(O)CO ARIWANIATODDMH-UHFFFAOYSA-N 0.000 abstract 1
- 238000010008 shearing Methods 0.000 abstract 1
- 230000037303 wrinkles Effects 0.000 abstract 1
- 235000010980 cellulose Nutrition 0.000 description 93
- 238000004519 manufacturing process Methods 0.000 description 35
- 244000235858 Acetobacter xylinum Species 0.000 description 28
- 241000894006 Bacteria Species 0.000 description 26
- 210000004027 cell Anatomy 0.000 description 23
- 238000003786 synthesis reaction Methods 0.000 description 22
- 229920000642 polymer Polymers 0.000 description 21
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 18
- 230000000694 effects Effects 0.000 description 18
- 238000011282 treatment Methods 0.000 description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 18
- 208000027418 Wounds and injury Diseases 0.000 description 17
- 206010052428 Wound Diseases 0.000 description 16
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 15
- 239000000243 solution Substances 0.000 description 14
- 241000196324 Embryophyta Species 0.000 description 13
- 210000001519 tissue Anatomy 0.000 description 13
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 12
- 210000004379 membrane Anatomy 0.000 description 12
- RARSHUDCJQSEFJ-UHFFFAOYSA-N p-Hydroxypropiophenone Chemical compound CCC(=O)C1=CC=C(O)C=C1 RARSHUDCJQSEFJ-UHFFFAOYSA-N 0.000 description 12
- HSCJRCZFDFQWRP-RDKQLNKOSA-N UDP-D-glucose Chemical group O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)OC1OP(O)(=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-RDKQLNKOSA-N 0.000 description 11
- 238000006243 chemical reaction Methods 0.000 description 11
- 239000000123 paper Substances 0.000 description 11
- 229920001503 Glucan Polymers 0.000 description 10
- 230000001580 bacterial effect Effects 0.000 description 10
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 9
- 229910052799 carbon Inorganic materials 0.000 description 9
- 230000012010 growth Effects 0.000 description 9
- 239000000463 material Substances 0.000 description 9
- GHXZTYHSJHQHIJ-UHFFFAOYSA-N Chlorhexidine Chemical compound C=1C=C(Cl)C=CC=1NC(N)=NC(N)=NCCCCCCN=C(N)N=C(N)NC1=CC=C(Cl)C=C1 GHXZTYHSJHQHIJ-UHFFFAOYSA-N 0.000 description 8
- 229960003260 chlorhexidine Drugs 0.000 description 8
- 230000001419 dependent effect Effects 0.000 description 8
- 150000004676 glycans Chemical class 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- -1 pentose phosphate Chemical class 0.000 description 8
- 229920001282 polysaccharide Polymers 0.000 description 8
- 239000005017 polysaccharide Substances 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- UEJSSZHHYBHCEL-UHFFFAOYSA-N silver(1+) sulfadiazinate Chemical compound [Ag+].C1=CC(N)=CC=C1S(=O)(=O)[N-]C1=NC=CC=N1 UEJSSZHHYBHCEL-UHFFFAOYSA-N 0.000 description 8
- 210000003491 skin Anatomy 0.000 description 8
- 230000003068 static effect Effects 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 7
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 7
- 230000007423 decrease Effects 0.000 description 7
- 239000000835 fiber Substances 0.000 description 7
- 239000001301 oxygen Substances 0.000 description 7
- 229910052760 oxygen Inorganic materials 0.000 description 7
- 238000006116 polymerization reaction Methods 0.000 description 7
- 210000000845 cartilage Anatomy 0.000 description 6
- 239000000306 component Substances 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 238000002955 isolation Methods 0.000 description 6
- 238000000746 purification Methods 0.000 description 6
- 239000000758 substrate Substances 0.000 description 6
- 230000000699 topical effect Effects 0.000 description 6
- 206010058041 Wound sepsis Diseases 0.000 description 5
- 238000005273 aeration Methods 0.000 description 5
- 238000013019 agitation Methods 0.000 description 5
- 210000000170 cell membrane Anatomy 0.000 description 5
- 239000006071 cream Substances 0.000 description 5
- 239000000645 desinfectant Substances 0.000 description 5
- 238000000855 fermentation Methods 0.000 description 5
- 230000004151 fermentation Effects 0.000 description 5
- 230000002269 spontaneous effect Effects 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- WJLVQTJZDCGNJN-UHFFFAOYSA-N Chlorhexidine hydrochloride Chemical compound Cl.Cl.C=1C=C(Cl)C=CC=1NC(N)=NC(N)=NCCCCCCN=C(N)N=C(N)NC1=CC=C(Cl)C=C1 WJLVQTJZDCGNJN-UHFFFAOYSA-N 0.000 description 4
- 208000025865 Ulcer Diseases 0.000 description 4
- 239000004599 antimicrobial Substances 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 229960004504 chlorhexidine hydrochloride Drugs 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 238000012136 culture method Methods 0.000 description 4
- RXKJFZQQPQGTFL-UHFFFAOYSA-N dihydroxyacetone Chemical compound OCC(=O)CO RXKJFZQQPQGTFL-UHFFFAOYSA-N 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 230000001976 improved effect Effects 0.000 description 4
- 230000001965 increasing effect Effects 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 230000014759 maintenance of location Effects 0.000 description 4
- FJQXCDYVZAHXNS-UHFFFAOYSA-N methadone hydrochloride Chemical compound Cl.C=1C=CC=CC=1C(CC(C)N(C)C)(C(=O)CC)C1=CC=CC=C1 FJQXCDYVZAHXNS-UHFFFAOYSA-N 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 241000894007 species Species 0.000 description 4
- 238000012546 transfer Methods 0.000 description 4
- 230000029663 wound healing Effects 0.000 description 4
- RGHNJXZEOKUKBD-SQOUGZDYSA-N D-gluconic acid Chemical class OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 3
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 3
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 3
- 229920002488 Hemicellulose Polymers 0.000 description 3
- 241000589516 Pseudomonas Species 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- HXXFSFRBOHSIMQ-VFUOTHLCSA-N alpha-D-glucose 1-phosphate Chemical compound OC[C@H]1O[C@H](OP(O)(O)=O)[C@H](O)[C@@H](O)[C@@H]1O HXXFSFRBOHSIMQ-VFUOTHLCSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 3
- 229920001222 biopolymer Polymers 0.000 description 3
- 238000002425 crystallisation Methods 0.000 description 3
- 230000008025 crystallization Effects 0.000 description 3
- 230000007547 defect Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 210000002744 extracellular matrix Anatomy 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 229950010772 glucose-1-phosphate Drugs 0.000 description 3
- ACGUYXCXAPNIKK-UHFFFAOYSA-N hexachlorophene Chemical compound OC1=C(Cl)C=C(Cl)C(Cl)=C1CC1=C(O)C(Cl)=CC(Cl)=C1Cl ACGUYXCXAPNIKK-UHFFFAOYSA-N 0.000 description 3
- 229960004068 hexachlorophene Drugs 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 239000002243 precursor Substances 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- 230000001698 pyrogenic effect Effects 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 239000008399 tap water Substances 0.000 description 3
- 235000020679 tap water Nutrition 0.000 description 3
- 231100000397 ulcer Toxicity 0.000 description 3
- 239000002699 waste material Substances 0.000 description 3
- CPKVUHPKYQGHMW-UHFFFAOYSA-N 1-ethenylpyrrolidin-2-one;molecular iodine Chemical compound II.C=CN1CCCC1=O CPKVUHPKYQGHMW-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 229920003043 Cellulose fiber Polymers 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 102000030595 Glucokinase Human genes 0.000 description 2
- 108010021582 Glucokinase Proteins 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 102000001105 Phosphofructokinases Human genes 0.000 description 2
- 108010069341 Phosphofructokinases Proteins 0.000 description 2
- 102000009569 Phosphoglucomutase Human genes 0.000 description 2
- 108091000080 Phosphotransferase Proteins 0.000 description 2
- 229920000954 Polyglycolide Polymers 0.000 description 2
- 239000004372 Polyvinyl alcohol Substances 0.000 description 2
- 229920000153 Povidone-iodine Polymers 0.000 description 2
- LCTONWCANYUPML-UHFFFAOYSA-M Pyruvate Chemical compound CC(=O)C([O-])=O LCTONWCANYUPML-UHFFFAOYSA-M 0.000 description 2
- 241000589180 Rhizobium Species 0.000 description 2
- 241000192023 Sarcina Species 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 230000000274 adsorptive effect Effects 0.000 description 2
- 239000012670 alkaline solution Substances 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 230000000845 anti-microbial effect Effects 0.000 description 2
- 230000002421 anti-septic effect Effects 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 230000004888 barrier function Effects 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 229960002152 chlorhexidine acetate Drugs 0.000 description 2
- YZIYKJHYYHPJIB-UUPCJSQJSA-N chlorhexidine gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O.C1=CC(Cl)=CC=C1NC(=N)NC(=N)NCCCCCCNC(=N)NC(=N)NC1=CC=C(Cl)C=C1 YZIYKJHYYHPJIB-UUPCJSQJSA-N 0.000 description 2
- 229960003333 chlorhexidine gluconate Drugs 0.000 description 2
- 235000020415 coconut juice Nutrition 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 235000013325 dietary fiber Nutrition 0.000 description 2
- 229940120503 dihydroxyacetone Drugs 0.000 description 2
- 235000011180 diphosphates Nutrition 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 230000004110 gluconeogenesis Effects 0.000 description 2
- 235000012208 gluconic acid Nutrition 0.000 description 2
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 2
- KWIUHFFTVRNATP-UHFFFAOYSA-N glycine betaine Chemical compound C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 description 2
- 230000035876 healing Effects 0.000 description 2
- 150000002402 hexoses Chemical class 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 239000000543 intermediate Substances 0.000 description 2
- 230000001788 irregular Effects 0.000 description 2
- 239000002085 irritant Substances 0.000 description 2
- 231100000021 irritant Toxicity 0.000 description 2
- 235000019226 kombucha tea Nutrition 0.000 description 2
- 229920005610 lignin Polymers 0.000 description 2
- 239000012533 medium component Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 239000003471 mutagenic agent Substances 0.000 description 2
- 231100000707 mutagenic chemical Toxicity 0.000 description 2
- 239000007783 nanoporous material Substances 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000002085 persistent effect Effects 0.000 description 2
- 108091000115 phosphomannomutase Proteins 0.000 description 2
- 102000020233 phosphotransferase Human genes 0.000 description 2
- 239000008104 plant cellulose Substances 0.000 description 2
- 239000004633 polyglycolic acid Substances 0.000 description 2
- 229920002451 polyvinyl alcohol Polymers 0.000 description 2
- 229960001621 povidone-iodine Drugs 0.000 description 2
- 238000003825 pressing Methods 0.000 description 2
- MCSINKKTEDDPNK-UHFFFAOYSA-N propyl propionate Chemical compound CCCOC(=O)CC MCSINKKTEDDPNK-UHFFFAOYSA-N 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 2
- 230000004936 stimulating effect Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 230000008093 supporting effect Effects 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 230000002110 toxicologic effect Effects 0.000 description 2
- 231100000027 toxicology Toxicity 0.000 description 2
- 230000004102 tricarboxylic acid cycle Effects 0.000 description 2
- 239000000052 vinegar Substances 0.000 description 2
- 235000021419 vinegar Nutrition 0.000 description 2
- 239000002023 wood Substances 0.000 description 2
- 229920001285 xanthan gum Polymers 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 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 description 1
- JNYAEWCLZODPBN-JGWLITMVSA-N (2r,3r,4s)-2-[(1r)-1,2-dihydroxyethyl]oxolane-3,4-diol Chemical compound OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O JNYAEWCLZODPBN-JGWLITMVSA-N 0.000 description 1
- BZSXEZOLBIJVQK-UHFFFAOYSA-N 2-methylsulfonylbenzoic acid Chemical compound CS(=O)(=O)C1=CC=CC=C1C(O)=O BZSXEZOLBIJVQK-UHFFFAOYSA-N 0.000 description 1
- WTLKTXIHIHFSGU-UHFFFAOYSA-N 2-nitrosoguanidine Chemical compound NC(N)=NN=O WTLKTXIHIHFSGU-UHFFFAOYSA-N 0.000 description 1
- MWMOPIVLTLEUJO-UHFFFAOYSA-N 2-oxopropanoic acid;phosphoric acid Chemical compound OP(O)(O)=O.CC(=O)C(O)=O MWMOPIVLTLEUJO-UHFFFAOYSA-N 0.000 description 1
- 108010017192 4-hydroxy-4-methyl-2-oxoglutarate aldolase Proteins 0.000 description 1
- YGUMVDWOQQJBGA-VAWYXSNFSA-N 5-[(4-anilino-6-morpholin-4-yl-1,3,5-triazin-2-yl)amino]-2-[(e)-2-[4-[(4-anilino-6-morpholin-4-yl-1,3,5-triazin-2-yl)amino]-2-sulfophenyl]ethenyl]benzenesulfonic acid Chemical compound C=1C=C(\C=C\C=2C(=CC(NC=3N=C(N=C(NC=4C=CC=CC=4)N=3)N3CCOCC3)=CC=2)S(O)(=O)=O)C(S(=O)(=O)O)=CC=1NC(N=C(N=1)N2CCOCC2)=NC=1NC1=CC=CC=C1 YGUMVDWOQQJBGA-VAWYXSNFSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- GJCOSYZMQJWQCA-UHFFFAOYSA-N 9H-xanthene Chemical compound C1=CC=C2CC3=CC=CC=C3OC2=C1 GJCOSYZMQJWQCA-UHFFFAOYSA-N 0.000 description 1
- 206010058040 Abdominal sepsis Diseases 0.000 description 1
- 241000589234 Acetobacter sp. Species 0.000 description 1
- 241000590020 Achromobacter Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000589158 Agrobacterium Species 0.000 description 1
- 241000588986 Alcaligenes Species 0.000 description 1
- 241000105989 Aleurochiton aceris Species 0.000 description 1
- 108010023063 Bacto-peptone Proteins 0.000 description 1
- 206010004146 Basal cell carcinoma Diseases 0.000 description 1
- 241000219310 Beta vulgaris subsp. vulgaris Species 0.000 description 1
- 235000018185 Betula X alpestris Nutrition 0.000 description 1
- 235000018212 Betula X uliginosa Nutrition 0.000 description 1
- 229940123208 Biguanide Drugs 0.000 description 1
- 241001598984 Bromius obscurus Species 0.000 description 1
- 238000009631 Broth culture Methods 0.000 description 1
- 206010006802 Burns second degree Diseases 0.000 description 1
- 206010006803 Burns third degree Diseases 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 102000005575 Cellulases Human genes 0.000 description 1
- 108010084185 Cellulases Proteins 0.000 description 1
- PTHCMJGKKRQCBF-UHFFFAOYSA-N Cellulose, microcrystalline Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC)C(CO)O1 PTHCMJGKKRQCBF-UHFFFAOYSA-N 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- 241000192020 Clostridium ventriculi Species 0.000 description 1
- GUTLYIVDDKVIGB-OUBTZVSYSA-N Cobalt-60 Chemical compound [60Co] GUTLYIVDDKVIGB-OUBTZVSYSA-N 0.000 description 1
- 208000003322 Coinfection Diseases 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102000000503 Collagen Type II Human genes 0.000 description 1
- 108010041390 Collagen Type II Proteins 0.000 description 1
- 206010011091 Coronary artery thrombosis Diseases 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- HEBKCHPVOIAQTA-NGQZWQHPSA-N D-Arabitol Natural products OC[C@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-NGQZWQHPSA-N 0.000 description 1
- HEBKCHPVOIAQTA-QWWZWVQMSA-N D-arabinitol Chemical compound OC[C@@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-QWWZWVQMSA-N 0.000 description 1
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 1
- HXXFSFRBOHSIMQ-GASJEMHNSA-N D-glucopyranose 1-phosphate Chemical compound OC[C@H]1OC(OP(O)(O)=O)[C@H](O)[C@@H](O)[C@@H]1O HXXFSFRBOHSIMQ-GASJEMHNSA-N 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- HXXFSFRBOHSIMQ-UHFFFAOYSA-N Di-K salt-alpha-D-Pyranose-Galactose 1-dihydrogen phosphate Natural products OCC1OC(OP(O)(O)=O)C(O)C(O)C1O HXXFSFRBOHSIMQ-UHFFFAOYSA-N 0.000 description 1
- QRLVDLBMBULFAL-UHFFFAOYSA-N Digitonin Natural products CC1CCC2(OC1)OC3C(O)C4C5CCC6CC(OC7OC(CO)C(OC8OC(CO)C(O)C(OC9OCC(O)C(O)C9OC%10OC(CO)C(O)C(OC%11OC(CO)C(O)C(O)C%11O)C%10O)C8O)C(O)C7O)C(O)CC6(C)C5CCC4(C)C3C2C QRLVDLBMBULFAL-UHFFFAOYSA-N 0.000 description 1
- 241000588722 Escherichia Species 0.000 description 1
- 108010080379 Fibrin Tissue Adhesive Proteins 0.000 description 1
- 201000003741 Gastrointestinal carcinoma Diseases 0.000 description 1
- 241000032681 Gluconacetobacter Species 0.000 description 1
- 102100031132 Glucose-6-phosphate isomerase Human genes 0.000 description 1
- 108010070600 Glucose-6-phosphate isomerase Proteins 0.000 description 1
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical compound ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102100027050 Inorganic pyrophosphatase Human genes 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 102000011782 Keratins Human genes 0.000 description 1
- 108010076876 Keratins Proteins 0.000 description 1
- 241001468094 Komagataeibacter europaeus Species 0.000 description 1
- 241000589216 Komagataeibacter hansenii Species 0.000 description 1
- 241000933022 Komagataeibacter intermedius Species 0.000 description 1
- 241000923674 Komagataeibacter oboediens Species 0.000 description 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 239000004909 Moisturizer Substances 0.000 description 1
- 206010061308 Neonatal infection Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 description 1
- 206010067482 No adverse event Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 108010069823 Oxaloacetate decarboxylase Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 235000008331 Pinus X rigitaeda Nutrition 0.000 description 1
- 235000011613 Pinus brutia Nutrition 0.000 description 1
- 241000018646 Pinus brutia Species 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 108010009736 Protein Hydrolysates Proteins 0.000 description 1
- 102000016611 Proteoglycans Human genes 0.000 description 1
- 108010067787 Proteoglycans Proteins 0.000 description 1
- 241000588769 Proteus <enterobacteria> Species 0.000 description 1
- 241000588768 Providencia Species 0.000 description 1
- 101710167959 Putative UTP-glucose-1-phosphate uridylyltransferase Proteins 0.000 description 1
- 238000001069 Raman spectroscopy Methods 0.000 description 1
- 206010057190 Respiratory tract infections Diseases 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 240000000111 Saccharum officinarum Species 0.000 description 1
- 235000007201 Saccharum officinarum Nutrition 0.000 description 1
- 101000994877 Schizosaccharomyces pombe (strain 972 / ATCC 24843) Putative inorganic pyrophosphatase C3A12.02 Proteins 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- 241000607720 Serratia Species 0.000 description 1
- 208000028990 Skin injury Diseases 0.000 description 1
- 206010072170 Skin wound Diseases 0.000 description 1
- 241000295644 Staphylococcaceae Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 235000021536 Sugar beet Nutrition 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 239000005862 Whey Substances 0.000 description 1
- 108010046377 Whey Proteins Proteins 0.000 description 1
- 102000007544 Whey Proteins Human genes 0.000 description 1
- 238000002441 X-ray diffraction Methods 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 241000589651 Zoogloea Species 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 238000005299 abrasion Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 235000013334 alcoholic beverage Nutrition 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 239000013566 allergen Substances 0.000 description 1
- 230000004100 amphibolic pathway Effects 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 239000004410 anthocyanin Substances 0.000 description 1
- 229930002877 anthocyanin Natural products 0.000 description 1
- 235000010208 anthocyanin Nutrition 0.000 description 1
- 150000004636 anthocyanins Chemical class 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 210000001188 articular cartilage Anatomy 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 210000004666 bacterial spore Anatomy 0.000 description 1
- 235000015173 baked goods and baking mixes Nutrition 0.000 description 1
- RNBGYGVWRKECFJ-ARQDHWQXSA-J beta-D-fructofuranose 1,6-bisphosphate(4-) Chemical compound O[C@H]1[C@H](O)[C@@](O)(COP([O-])([O-])=O)O[C@@H]1COP([O-])([O-])=O RNBGYGVWRKECFJ-ARQDHWQXSA-J 0.000 description 1
- 229960003237 betaine Drugs 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000011942 biocatalyst Substances 0.000 description 1
- 230000000035 biogenic effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000036770 blood supply Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000004067 bulking agent Substances 0.000 description 1
- 230000009172 bursting Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 150000001722 carbon compounds Chemical class 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000035606 childbirth Effects 0.000 description 1
- 210000001612 chondrocyte Anatomy 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 230000001010 compromised effect Effects 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 208000002528 coronary thrombosis Diseases 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 239000000824 cytostatic agent Substances 0.000 description 1
- 230000001085 cytostatic effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000006114 decarboxylation reaction Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 235000021185 dessert Nutrition 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 150000001991 dicarboxylic acids Chemical class 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- UVYVLBIGDKGWPX-KUAJCENISA-N digitonin Chemical compound O([C@@H]1[C@@H]([C@]2(CC[C@@H]3[C@@]4(C)C[C@@H](O)[C@H](O[C@H]5[C@@H]([C@@H](O)[C@@H](O[C@H]6[C@@H]([C@@H](O[C@H]7[C@@H]([C@@H](O)[C@H](O)CO7)O)[C@H](O)[C@@H](CO)O6)O[C@H]6[C@@H]([C@@H](O[C@H]7[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O7)O)[C@@H](O)[C@@H](CO)O6)O)[C@@H](CO)O5)O)C[C@@H]4CC[C@H]3[C@@H]2[C@@H]1O)C)[C@@H]1C)[C@]11CC[C@@H](C)CO1 UVYVLBIGDKGWPX-KUAJCENISA-N 0.000 description 1
- UVYVLBIGDKGWPX-UHFFFAOYSA-N digitonine Natural products CC1C(C2(CCC3C4(C)CC(O)C(OC5C(C(O)C(OC6C(C(OC7C(C(O)C(O)CO7)O)C(O)C(CO)O6)OC6C(C(OC7C(C(O)C(O)C(CO)O7)O)C(O)C(CO)O6)O)C(CO)O5)O)CC4CCC3C2C2O)C)C2OC11CCC(C)CO1 UVYVLBIGDKGWPX-UHFFFAOYSA-N 0.000 description 1
- 230000003292 diminished effect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000010410 dusting Methods 0.000 description 1
- 238000002003 electron diffraction Methods 0.000 description 1
- 238000001493 electron microscopy Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000001125 extrusion Methods 0.000 description 1
- 210000000416 exudates and transudate Anatomy 0.000 description 1
- 239000003889 eye drop Substances 0.000 description 1
- 229940012356 eye drops Drugs 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 235000019985 fermented beverage Nutrition 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 238000007667 floating Methods 0.000 description 1
- PGBHMTALBVVCIT-VCIWKGPPSA-N framycetin Chemical compound N[C@@H]1[C@@H](O)[C@H](O)[C@H](CN)O[C@@H]1O[C@H]1[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](N)C[C@@H](N)[C@@H]2O)O[C@@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CN)O2)N)O[C@@H]1CO PGBHMTALBVVCIT-VCIWKGPPSA-N 0.000 description 1
- 235000015203 fruit juice Nutrition 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 235000012055 fruits and vegetables Nutrition 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 102000034356 gene-regulatory proteins Human genes 0.000 description 1
- 108091006104 gene-regulatory proteins Proteins 0.000 description 1
- 238000012252 genetic analysis Methods 0.000 description 1
- 239000000174 gluconic acid Substances 0.000 description 1
- 230000002414 glycolytic effect Effects 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000003179 granulation Effects 0.000 description 1
- 238000005469 granulation Methods 0.000 description 1
- 231100001261 hazardous Toxicity 0.000 description 1
- 238000004192 high performance gel permeation chromatography Methods 0.000 description 1
- 239000012510 hollow fiber Substances 0.000 description 1
- 235000003642 hunger Nutrition 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 150000004679 hydroxides Chemical class 0.000 description 1
- 229920003063 hydroxymethyl cellulose Polymers 0.000 description 1
- 229940031574 hydroxymethyl cellulose Drugs 0.000 description 1
- 230000007954 hypoxia Effects 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000006713 insertion reaction Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 201000002313 intestinal cancer Diseases 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 238000006317 isomerization reaction Methods 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000005461 lubrication Methods 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000012067 mathematical method Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 229940127554 medical product Drugs 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 108700005457 microfibrillar Proteins 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000001333 moisturizer Effects 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- QNILTEGFHQSKFF-UHFFFAOYSA-N n-propan-2-ylprop-2-enamide Chemical compound CC(C)NC(=O)C=C QNILTEGFHQSKFF-UHFFFAOYSA-N 0.000 description 1
- 229920005615 natural polymer Polymers 0.000 description 1
- 230000007524 negative regulation of DNA replication Effects 0.000 description 1
- 210000001640 nerve ending Anatomy 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 231100001254 ocular irritant Toxicity 0.000 description 1
- 230000009965 odorless effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 229920000620 organic polymer Polymers 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- KHPXUQMNIQBQEV-UHFFFAOYSA-L oxaloacetate(2-) Chemical compound [O-]C(=O)CC(=O)C([O-])=O KHPXUQMNIQBQEV-UHFFFAOYSA-L 0.000 description 1
- KHPXUQMNIQBQEV-UHFFFAOYSA-N oxaloacetic acid Chemical compound OC(=O)CC(=O)C(O)=O KHPXUQMNIQBQEV-UHFFFAOYSA-N 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 239000011087 paperboard Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 229920001277 pectin Polymers 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000003239 periodontal effect Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 229930029653 phosphoenolpyruvate Natural products 0.000 description 1
- DTBNBXWJWCWCIK-UHFFFAOYSA-N phosphoenolpyruvic acid Chemical compound OC(=O)C(=C)OP(O)(O)=O DTBNBXWJWCWCIK-UHFFFAOYSA-N 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 229920002338 polyhydroxyethylmethacrylate Polymers 0.000 description 1
- 239000004626 polylactic acid Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000019525 primary metabolic process Effects 0.000 description 1
- 238000004886 process control Methods 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000004537 pulping Methods 0.000 description 1
- 150000003248 quinolines Chemical class 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 208000020029 respiratory tract infectious disease Diseases 0.000 description 1
- 230000000284 resting effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000001878 scanning electron micrograph Methods 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 210000004927 skin cell Anatomy 0.000 description 1
- 231100000444 skin lesion Toxicity 0.000 description 1
- 206010040882 skin lesion Diseases 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 235000010288 sodium nitrite Nutrition 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- SEEPANYCNGTZFQ-UHFFFAOYSA-N sulfadiazine Chemical compound C1=CC(N)=CC=C1S(=O)(=O)NC1=NC=CC=N1 SEEPANYCNGTZFQ-UHFFFAOYSA-N 0.000 description 1
- 229960004306 sulfadiazine Drugs 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 229920001059 synthetic polymer Polymers 0.000 description 1
- 230000009967 tasteless effect Effects 0.000 description 1
- 235000013616 tea Nutrition 0.000 description 1
- 230000028016 temperature homeostasis Effects 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 230000009772 tissue formation Effects 0.000 description 1
- 239000003860 topical agent Substances 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 230000005068 transpiration Effects 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 230000036269 ulceration Effects 0.000 description 1
- 208000019206 urinary tract infection Diseases 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000000230 xanthan gum Substances 0.000 description 1
- 235000010493 xanthan gum Nutrition 0.000 description 1
- 229940082509 xanthan gum Drugs 0.000 description 1
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Provided is an all natural microbial cellulose microparticle implantable dermal and subdermal filler derived from Nata de coco cellulose, wherein the microbial cellulose is fermented in coconut milk culture media, the active ingredient monolaurin is present in the amount of about 1 pcnt to 3 pcnt , the quality of the Nata de coco is higher than used as food in terms of cellulose content and appearance, and the size of the microbial cellulose is reduced to 300-900 nanometers after the final high pressure shearing. The final product passed the irritation, sensitization and the challenge tests, can be considered as non-irritating with low sensitization potential, and can be classified as hypoallergenic, biocompatible, biodegradable, non-toxic and non-immunogenic. Also provided are the process of producing the filler of the present invention, and the use of said filler for tissue augmentation, aesthetic correction of wrinkles or loss of facial volume and lipoatrophy correction.
Description
CHAPTER 1
INTRODUCTION
1. General introduction
Cellulose is the most abundant earth biopolymer and recognized as the major component of plant biomass. but also a representative of microbial extra- cellular polymers. T he structure of cellulose is presented in Figure 1. \
N, :
H OH \ H OH ) H \ x OH H
AY
OH H ( H
AY H
H AY OH
\
CH,0H x CH,0H \ n :
Figure 1. The structure of cellulose
Bacterial cellulose (BC) is synthesized by sgveral bacterial genera, of ; which Acetobacter strains are best known. An overview of BC wroducers is presented in Table I. BC belongs to specific products of primary metabolism. The polymer structure depends on the organism, although the pathway of biosynthesis and : mechanism of its regulation are probably common for the majority of BC-producing bacteria (Ross er al.. 1991: Jonas and Farah, 1998). >
Table 1. Bacterial cellulose producers (Jonas and Farah, 1998, modified) ;
Genus Cellulose structure
Acetobacter Extra-cellular pellicle
Achromobacter Fibrils :
Aerobacter Fibrils
Agrobacterium Short fibrils
Alcaligenes Fibrils
Pseudomonas No distinct fibrils :
Rhizobium Short fibrils E
Sarcina Amorphous cellulose
Zoogloea Not well defined
Acetobacter xylinum (A. xylinum) (synonyms A. aceti subsp. xylinum
A. xvlinus), which is the most efficient producer of cellulose, has been recently reclassified and included within the novel genus Gluconacetobacter, as ;
Gluconacetobacter xylinus (Yamada er al.. 1998, 2000) together with some other species (G. hansenii, G. europaeus, G. oboediens and G. intermedius).
A. wvlinum is a simple Gram-negative bacterium, rod to oval shape, strictly aerobe bacteria, acetic acid bacteria, A. xylimum which has been applied as model microorganisms for basis and applied studies on cellulose production (Cannon . and Anderson, 1991). It usually found on fruits and vegetables, in vinegar. fruit juices i and alcoholic beverages. Although synthesis of an extracellular gelatinous mat by A. aylinum was reported for the first time in 1896 by A.J. Brown (Brown and Saxena, g 2000). attracted more attention in the second half of the 20% century. Intensive studies .
on BC synthesis using A. xvlinum as a model bacteria were started by Hestrin er al., (1947, 1954), who proved that resting and lyophilized Acetobacter cells synthesized cellulose in the presence of glucose and oxygen. Later, Colvin (1957) detected cellulose synthesis in samples containing cell-free extract of A. xylinum, glucose and
BC has a very fine fiber structure with the average fiber diameter about 0.1 microns, with indeterminate fiber length. It is non-nutritive, non toxic on subject studied in extensive toxicological testing, neutral in sensory quality providing no taste : and little tendency to mask other flavors. The fibers show high water binding and strong hydrogen bonding (Kent er al., 1991). Cellulose fiber is an effective thickener at low concentrations and can interact synergistically with other viscosity builders such as carboxymethylcellulose (CMC), xanthan, hydrox ymethylcellulose to improve their efficiency and effectiveness (Valla er al., 1989). The cellulose synthesized by A. xylinum is identical to that made by plants in respect to molecular structure. However, the secreted polysaccharide is free from lignin, pectin and hemi-cellulose as well as other biogenic product, which are associated with plant cellulose. Additionally, extra- cellutar synthesized BC microfibrils differ from plant cellulose with respect to its ultrafine network architecture, high crystallinity. high water absorption capacity, high, mechanism strength in the wet state and mouldability during formation (Klemm et al. 2001). Because of the unique properties. resulting from the ultrafine reticulated structure, BC has found a multitude of applications in paper, textile, in food industry and as biomaterials in cosmetics and medicine as a consequence of the special i properties (Ring er al., 1986). Wider application of this polysaccharide is obviously 1 dependent on the scale of production and its cost. Therefore, basic studies run 1 tocether with intensive research on strain improvement and production process ; development. 2. Physiological functions
In natural habitats, the majority of bacteria synthesized extracellular polysaccharides, which form envelopes around the cells (Costeron, 1999). ] 2.1 To hole the cell in aerobic environment :
Cells of cellulose-producing bacteria are entrapped in the polymer : network, frequently supporting the population at the liquid-air interface (Williams and ;
Cannon, 1989). : 2.2 To colonize on [ood and substrate :
The polymer matrix takes part adhesion of the cells onto accessible surface and facilitates nutrient supply. since their concentration in the polymer lattice : is markedly enhanced due to its adsorptive properties, in comparison to the : surrounding aqueous environment (Jonas and Farah, 1998; Costeron, 1999). Some authors suppose that cellulose synthesized by A. xvlinum also plays a storage role and can be utilized by the starving microorganisms (Okamoto er al., 1994). 2.3 Prevent potential competitor
Because of the viscosity and hydrophilic properties of the cellulose layer. the resistance of producing bacterial cells against unfavorable changes (a decrease in water content, variations in pH, appearance of toxic substances, :
pathogenic organisms, etc.) in a habitat is increased and they can growth and develop inside the envelope (Ross et al., 1991). 2.4 Protect from ultraviolet radiation (UV)
BC can protect UV light because it is opacity and can protect A. xylinum from the Killing effect of UV light (Koo er al.. 1991). As much as 23% of the acetic acid bacteria cells covered with BC survived a | h treatment with UV. Removal of the protective polysaccharide brought about a drastic decrease in their viability (3% only) (Ross eral., 1991). 2.5 Help retain moisture
It may be used as moisturizer because it is very hydrophilic and aid in moisture retention to prevent dry of the nature substrates for A. xylinum while the bacterium growing on them (Ross er al., 1991). 3. Properties of bacterial cellulose :
The physicological properties of cellulose such as water holding capacity, viscosity of disintegrated cellulose suspension and the Young's modulus of dried sheet are revealed (Watanabe er al., 1998; Igushi er al., 2000).
One of the most important features of BC is its chemical purity, which distinguishes this cellulose from plants, usually associated with hemicelluloses. lignocellulose (Fiedler, 1989) and waxy aromatic substance removal of which is mherently difficult (Ross er al.. 1991). BC can be purified using less energy or chemical-intensive processes without hazardous by-products (Hong ez al.. 2005), high water-binding capacity due to a large surface area can be hold large amount of water (up to 200 times of its dry mass). Majority of water is not bound with the polymer and can be squeezed out by gentle pressing (Czaja er al., 2005). High tensile strength, extremely insoluble and elasticity, durability and shape retention, high crystallinity index due to BC has a reticulated structure, in which numerous ribbon-shaped fibrils, are composed of highly crystalline and highly uniaxially oriented cellulose subfibrils.
This three dimensional structure, not found in the plant originating cellulose, brings about higher crystallinity index (60-70%) of BC (Wananabe et al., 1998). The BC can be molded into any shape and size during it synthesized, depending on the fermentation technique and conditions used (Bielecki er al., 2002). Highly nano- porous material that allows for the potential transfer of antibiotics or other medicines : into wound. while at the same time serving as an efficient physical barrier against any external infection (Czaja et al., 2005). Ougiya er al., (1997) showed that bacterial cellulose from A. xylinum subsp. sucrofermentans has the highest emulsion-stabilizing effect among the examined cellulose materials such as microcrystalline cellulose, microfibrinated cellulose, xanthan gum and sorbitan monolurate. Extensive toxicologic evaluations on BC from A. aceri subsp. xvlinum have shown no adverse effects on the subjects studied. Schmitt er al., (1991) were tested for ocular irritant and not a dermal irritant in rabbit. 4. Structure of bacterial cellulose
Cellulose is an unbranched polymer of B-1.4-linked glucopyranose residues. Extensive research on BC revealed that it is chemically identical to plant i cellulose (PC), but its macromolecular structure and properties differ from the latter.
Nascent chains of BC aggregate to form subfibrils, which have a width of approximately 1.5 nm and belong to the thinnest naturally occurring fibers, comparable only to subelemental fibers of cellulose detected in the cambium of some plants and in quinee mucous (Kudlicka, 1989). BC subfibrils are crystallized into microfibrils (Jonas and Farah, 1998), these into bundles, and the latter into ribbons (Yamanaka er al., 2000). Dimensions of the ribbons are 3-4 (thickness) x 70-80 nm (width), according to Zaar, (1977) or 3.2 x 133 nm, according to Brown er al., (1976) or 4.1 x 117 nm, according to Yamanaka et al., (2000) whereas the width of cellulose fibers produced by pulping of birch or pine wood is two orders of magnitude larger (1.4-4.0 x 102 and 3.0-7.5 x 102 mm, respectively). The ultrafine ribbons of microbial cellulose, the length of its ranges from 1-9 pm, form a dense reticulated structure, stabilized by extensive hydrogen bonding. The crystallinity index and crystalline size are calculated based on X-ray diffraction measurements (Watanabe et al., 1998). BC is also distinguished from its plant counterpart by a high crystallinity index (above 60%). The degree of polymerization (DP) of cellulose and the DP distribution are determined by high-performance gel permeation chromatography (Watanabe er al., ] 1998) and DP usually between 2,000 and 6,000 (Jonas and Farah, 1998). but in the some cases reaching even 16,000 or 20.000 (Watanabe et al., 1998), whereas the average DP of plant polymer varies from 13,000 to 14,000 (Teeri, 1997).
Macroscopic morphology of BC strictly depends on culture conditions.
In static conditions. bacteria accumulate cellulose mats on the surface of the nutrient broth, at the oxygen-rich (air-liquid interface). The subfibrils of cellulose are continuously extruded from linearly ordered pores at the surface of the bacterial cell, i crystallized into microfibrils and forced deeper into the growth medium. Therefore,
the leather-like pellicle, supporting the population of A. xylinum cells, consists of overlapping and intertwisted cellulose ribbons, forming parallel but disorganized planes (Jonas and Farah, 1998). The adjacent static-BC strands branch and interconnect less frequently than these in BC produced in agitated culture, in a form of irregular granules and fibrous strands, well-dispersed in culture both (Vandamme er al.. 1998). The strands of reticulated agitated-BC interconnect to form a grid like pattern. and have both roughly perpendicular and roughly parallel orientations (Watanabe er al., 1998).
Differences in three-dimensional structure of agitated-BC and static-
BC fibrils are noticeable in their scanning electron micrographs. The static-BC fibrils are more extended and piled above one another in a criss-crossing manner. Strands of agitated-BC are entangled and curved (Johnson and Neogi, 1989). Besides, they have a larger cross-sectional width (0.1-0.2 pm) than static-BC fibrils (usually 0.05-0.10 um). Morphological differences between static-BC and agitated-BC contribute to varying degrees of crystallinity, different crystallite size and la cellulose content.
Two common crystalline forms of cellulose, designated as 1 and II, are distinguishable by X-ray, nuclear magnetic resonance (NMR), Raman spectroscopy, and infrared analysis (Johnson and Neogi, 1989). It is known that in the metastable cellulose I. which is synthesized by the majority of plants and also by A. xvlinum in static culture, parallel B8-1,4-glucan chains are arranged uniaxially, whereas B-1.4- glucan chains of cellulose 11 are arranged in a random manner. They are mostly anti- parallel and linked with a lager number of hydrogen bonds that result in higher } thermodynamic stability of the cellulose 11. Agitated-BC has a lower crystallinity : dex and a smaller crystallite size than static-BC (Watanabe er al.. 1998). It was also ;
observed that a significant portion of cellulose II occurred in BC synthesized in agitated culture. In nature, cellulose 11 is synthesized by few organisms only (some algae, moulds and bacteria, such as Sarcina ventriculi) (Jonas and Farah, 1998). 5. Biochemistry of bacterial cellulose synthesis 5.1 Carbon metabolism in Acetobacter xylinum
Two main amphibolic pathways are operative in A. xylinum bacteria (Figure 2) the pentose phosphate cycle for oxidation of carbohydrates and the Krebs cycle for the oxidation of organic acids and related compounds (Ross et al., 1991).
Phosphofructokinase is absent or weakly absent (De Ley er al., 1984), resulting in no or only a weak glycolytic activity, indicating that glucose cannot be metabolized anaerobically. However, Tonouchi et al (1996) detected phosphofructokinase activity in A. xvlinum subsp. sucrofermentans BPR 2001. These researchers also investigated several specific enzyme activities involved in cellulose synthesis, and demonstrated the existence of a phosphotransferase system for fructose uptake in cellulose- producing Acetobacter cells. Gluconeogenesis can occur from oxaloacetate via pyruvate by means of the enzymes oxaloacetate decarboxylase and pyruvate phosphate dikinase. : 5.2 Cellulose synthesis in Acetobacter xylinum
J
A. xvlintm converts various carbon compounds, such as hexoses, glycerol. dihydroxyacetone, pyruvate and dicarboxylic acids into cellulose, usually with about 50% efficacy (Cannon and Anderson, 1991). The latter compounds enter the Krebs cycle and due to oxalacetate decarboxylation to pyrovate undergo conversion to hexoses via gluconeogenesis, similarly to glycerol, dihydroxyacetone and intermediates of the pentose phosphate cycle as shown in Figure 3.
The direct cellulose precursor is uridine-diphosphoglucose (UDPGlc), which is a product of conventional pathway, common of many organisms, including plants and involving glucose phosphorylation to glucose-6-phosphate (Glc-6-P), catalyzed by glucokinase (GK), followed by isomerization of this intermediate to glucose-1-phosphate (Gle-1-P), catalyzed by phosphoglucomutase (PGM) and conversion of the latter metabolite to UDPGlc by uridine-diphosphoglucose pyrophosphorylase (UDPGlc pyrophosphorylase) which catalyzing the synthesis of
UDPGlc is the key enzyme in the cellulose biosynthesis (Tonouchi, 1996). They display cellulose synthase (CS) activity that was confirmed in vitro by mean of observation of cellulose synthesis, catalyzed by cell-free extracts of Cel strains (Saxena et al., 1989). Some Acetobacter strains produce extracellular cellulose fibrils as a part of its normal metabolic activity. UDPGIlc which produced from UTP and glucose-1-phosphate is the direct precursor in the synthetic pathway of cellulose (Ishikawa, 1997). Furthermore, the pyrophosphorylase activity varies between different A. xvlinum strains and the highest activity was detected in the most effective cellulose producers, such as A. xylinum subsp. sucrofermentants BPR 2001. The latter : strain prefers fructose as a carbon source, displays high activity of phosphoglucoisomerase and processes a system of phosphotransferases, dependent on phosphoenolpyruvate. The system catalyses conversion of fructose (Fru) to fructose-
I-phosphate (Fru-1-P) and further to fructose-1,6-biphosphate (Fru-1,6-bisP) as shown in Figure 3. ] i ll 5.3 Cellulose synthase (CS)
Cellulose synthase of A. xylinum is a typical anchored membrane protein, having molecular mass of 400-500 kDa and CS is localized on the cytoplasmic side of the cell membrane. Because of this localization, purification of
CS was extremely difficult, and isolation of the membrane fraction, before CS solubilization and purification was necessary (Lin and Brown, 1989). Furthermore, A. xylinum CS appeared to be a very unstable protein (Lin and Brown, 1989). CS isolation from membranes was carried out using digitonin (Mayer er al., 1989), or detergents (Triton X-100) and treatment with trypsin (Saxena et al., 1989), followed by CS entrapment on cellulose. Mayer er al., (1989), the purified CS preparation contained 3 different types of subunits, having molecular mass of 90, 67 and 54 kDa.
Saxena er al., (1989) found only 2 types of polypeptides (83 and 93 kDa). 1 5.4 Mechanism of bacterial cellulose biosynthesis
Formation of BC is catalyzed by the cellulose synthase complexes aligned linearly in the A. xylinum cytoplasmic membrane. Synthesis of the metastable cellulose in A. xvlinum and other cellulose-producing organisms, including at least ] (wo steps such as i - polymerization of glucose molecules to the linear [,4-B-glucan, - assembly and crystallization of individual nascent polymer chains into 3 supramolecular structures, characteristic for each cellulose-producing ; organism. 1
: 5.4.1 Mechanism of B-1,4-glucan polymerization
Han and Robyt (1998) proposed that BC biosynthesis involved 3 enzymes embedded in the cytoplasmic membrane: cellulose synthase (CS), lipid pyrophosphate (LP: UDPGlc-PT) and lipid pyrophosphate phosphohydrolase (LPP).
The reaction mechanism, called by the authors the insertion reaction, is presented in :
Figure 4.
Reaction 1: The first enzyme transfers Glc-1-P from UDPGlc onto the : lipidmonophosphate (Lip-P). thus giving the lipid pyrophosphate-o0-D-Glc (LipPP-a-
Gle). The a configuration on the anomeric carbon, involved in the Glc phosphoester 3 bond, remains the same as in the substrate molecule. The product second product of this reaction is UMP (according to the Brown's model, UDP is released).
Reaction 2: To catalyze by CS, the glucose residue is transferred from one LipPP-0-Glc molecule onto another one, and the B-1,4-glycosidic linkage between the two glucose residues is formed, due to the attack of the C-4 hydroxyl group of one of them onto C-1 hydroxyl group of the second Gle¢ (from the second
LipPP-¢-Gle).
Reaction 3: The hydrolysis of the lipid pyrophosphate formed in the previous step occurs. and another Gle-a-1P from UDPGlc can be attached to the LipP, J released in this reaction. The cycle of these three reactions (as shown in Figure 3) is : continued to give the B-1,4-glucans chain of an appropriate length. 5.4.2 Assembly and crystallization of cellulose chains
A. xylinum is a simple Gram-negative bacterium which has an ability to synthesize a large quantity of high-quality cellulose organized as twisting ribbons of i microfibrillar bundles (Brown er al., 1976). During the process of actual bacteria, then polymerized into single, linear B-1.4-glucan chains and finally secreted outside the ] cells through a linear row of pores located on their outer membrane (Haigler, 1980).
The subsequence assembly of the B-1.4-glucan chains outside of the cell is a precise, hierarchical process. Initially, they form subfibrils (consisting of 10-15 nascent 8-1,4- glucan chains). then latter microfibrils, and finally bundles of microfibrils consisting of a loosely wound ribbon, which is comprised of about 1,000 individual glucan chains (Figure 5) (Ross er al., 1991).
Synthesis of BC is a precisely and specifically regulated multi-step process, involving a large number of both individual enzymes and complexes of catalytic and regulatory proteins, whose supramolecular structure has not yet been well defined. The process includes the synthesis of uridinediphosphoglucose (UDPGlc), which is the cellulose precursor, followed by glucose polymerization into
B-1.4-glucan chains, and nascent chain association into characteristic ribbon-like ! structure, formed by hundreds or even thousands of individual cellulose chains.
Pathways and mechanisms of UDPGlc synthesis are relatively well known, whereas molecular mechanisms of glucose polymerization into long and unbranched chains, their extrusion outside the cell. and self-assembly into fibrils (Brown er al., 1976).
6. Bacterial cellulose productions
The choice of a cultivation technique is dependent on further biopolymer commercial destination, considering that cellulose ultrastructure and its physical and mechanical properties are strictly influenced by the culture method (Galas er al.. 1999). Cellulose can be produced in the several techniques for BC production have been reported, some of which demonstrate a potential tool for economic and commercial BC production. 6.1 Stationary culture conditions
In stationary culture method, gelatinous membrane of BC is accumulated on the surface of a culture medium and BC is produced and forms a ] pellicle. The productivity of BC is dependent on the surface area of the medium.
Therefore a system in which a pellicle is formed on an oxygen-permeable membrane was developed to increase the surface area of the pellicle (Yoshino et al., 1996). BC yield in static cultures is mostly dependent on the surface: volume ratio (s/v).
Optimum s/v ratio protects from either too high (unnecessary) or too low aeration (cell growth and BC synthesis termination). Krystynowicz et al., (1997) reported values of s/v ratio vary from 2.2 em” to 0.7 em. However, the stationary culture method has disadvantages such as high labor cost and lower productivity (Zuo ef al., ] 2006). synthesis of the polymer only in the form of a sheet and relatively low productivity, contributed to the development of new fermentation process (Masaoka et al.. 1993). The conventional method of BC production by static cultivation is not ] applicable 10 large-scale industrial production; thus, the establishment of an 3 economical production process is urgently required (Toyosaki et al., 1995). The i control of BC synthesis in static culture is very difficult since the pellicle limits an access to the liquid medium. The particularly important parameter, which requires continuous control is pH. Accumulation of keto-gluconic acids in culture broth brings about a decrease in pH much below its value that is optimum for growth of bacteria and the polysaccharide synthesis because conventional methods of pH adjustment can not be used in static cultures. Vandamme et al., (1998) applied an in situ pH control J via an optimized fermentation medium design, based on introducing acetic acid as an additional substrate for Acetobacter sp. LMG 1518. Products of acetic acid catabolism counteracted the pH decrease caused by keto-gluconic formation and provided constant pH of the growth medium, equal to 5.5, throughout the whole process. 6.2 Agitated culture conditions
Cellulose can be produced in the form of a fibrous suspension, irregular masses, pellets or spheres. Screening of strains suitable and the influence of the culture medium components on the BC productivity have been reported (Dudman, 1960). Since the productivity of BC in agitated culture is dependent on the growth of : the BC-producing organisms increasing the cell density is essential for increasing BC productivity. Since the growth of Acetobacter (a typical aerobic BC producer) is i dependent on oxygen supply, oxygen supply is directly associated with BC productivity in an aerated and agitated culture of Acetobacter. However, excessive oxygen supply is reported to result in a decrease in BC productivity because of a loss of substrate by direct oxidation (Yamanaka er al.. 1989). Agitated culture is ] considered more suitable for the commercial cellulose production of BC mainly due to the higher production rates that potential can be achieved (Yamanaka er al., 1989).
BC production in agitation and aeration encounters many problems such as spontaneous appearance of Cel” mutans (cellulose non-producers), which contributes to a decline in polymer synthesis (Ross ef al., 1991). Optimized agitation and aeration ! prevent turbulence, which negatively effects cellulose polymerization and crystallization, thus reducing the polysaccharide yield (Laboureur, 1988). Laboureur (1988). who applied the rate of agitation equal to 60 rpm and aeration of 0.6 vvm were optimum for A. aceti subsp. xylinum ATCC 2178 strain cultured in 300 litre fermentor for 45 h at 30°C and 10 g of BC per litre a day was obtained. Recently investigations showed that in agitated culture, high oxygen supply and high volumetric agitated power are required for increased of BC productivity (Kouda et al., 1998). Other factors such as agitator configuration, effects of oxygen and carbon dioxide pressure on BC productivity have been investigated (Ring er al., 1989).
Agitated culture method often leads to morphological and structural abnormalities of bacterial cellulose due to high shear stress induced by agitation (Zuo et al., 2006). 6.3 Horizontal fermentors
Cellulose production in horizontal fermentor is the combination of ] stationary and submerged cultures. The polymer is deposited on the surface of rollers or discs. rotating around the long axis. A part of their surface temporarily dips in the liquid medium or is above its surface (in the air). The advantages of this method ] include a large polymer surface, synthesis of cellulose in a form of hollow fibers, different in diameter, as well as good process control, easy scale enlargement, i appropriate accessible surface for adhesion of bacteria and product deposition, higher rate of cellulose production (Sattler and Fiedler, 1990). i
7. Factor effecting growth and cellulose production
The rate of cellulose production depended proportionally on the surface-area of the culture medium and was unaffected by the depth and volume of the medium. The optimum pH for cellulose production was 4.0 to 6.0. Glucose, fructose and glycerol were preferred carbon sources for cellulose production.
Embuscado et al., (1994) selected that the strain of A. xvlinum for optimum cellulose production. The A. xylinum will be grown on shaker and statically. Optimal fermentation of non-agitated production of cellulose by selected strains will be determined. Four factors such as carbon-sources concentration, pH, temperature of incubation and their relationship on cellulose yield will be determined. 7.1 Isolation from natural sources and improvement of bacterial cellulose producing strains
The genetic analysis of cellulose biosynthesis in A. xvlinum has included the isolation of mutants that effect cellulose production, characterization of indigenous plasmid species, and the cloning of gene in the process. A number of researchers, beginning with Schramm and Hestrin (Schramm and Hestrin, 1954), have ] described the isolation of cellulose-negative (cel’) mutants. Both spontaneous and mutagen-induced variants have been isolated. Many reports have indicated that apparent spontaneous celluloseless mutants arise at a high rate when wild-type cells are grown in aerated liquid culture (Smith, 1990). The frequency with which these mutants occur also increases with the age of the culture. The spontaneous mutants ] have a mucoid appearance on solid agar like true cel’ mutants, but the majority reverts back to wild-type when grown statically in broth culture (Cannon and Anderson, :
1991). A variety of mutagens have been used in an effort to induce mutation in A. ] xvlinum i.e. nitrosoguanidine, nitrous acid are very effective whereas hydroxylamine and ultraviolet light are relatively ineffective (Valla er al, 1989; Williams and ]
Cannon. 1989). The characteristics of A. xylinum strains are shown in Table 2. J
One of the methods enabling selection of proper A. xylinum strain, is ] the screening for strains, which cannot oxidize glucose via gluconic acid (Winkelman . and Clark, 1984; Johnson and Neogi: 1989; De Wulf et al., 1996; Vandamme, 1998) ] to 2.-5-or 2.5-ketogluconate. 1
Table 2. Characteristics of Acetobacter xylinum strains (Cannon and Anderson, 1991) ]
Characteristic Wild type Cel-negative ~~ Overproducer ; - Colony morphology Small-rough Large, mucoid ~~ Small-rough - Fluorescence with Bright Dull Bright ;
Tinopal - Pellicle production Pellicle No pellicle Thick pellicle 1 - Relative cellulose 1.0 0.0-0.1° 5.0-6.0 . production (wet weight) ; a The trace amounts of cellulose produced in these pellicle-deficient cultures are : thought to be cellulose 11. : 7.2 Carbon and nitrogen sources :
The factors affecting the BC production yield, many attentions was paid to carbon sources. Numerous mono-, di- and polysaccharides, alcohols, organic E acids and other compounds were compared by Jonas and Farah (1998), who found out that the preferred carbon sources were D-arabitol and D-mannitol. which presence resulted in 6.2 or 3.8 higher cellulose production, respectively, in comparison to glucose. Both sugars alcohols provided stabilization of pH throughout the culture, since were not converted to gluconic acids. Tonouchi er al., (1996) used a strain of A. xvlinum to obtain cellulose from glucose and fructose, found out that fructose stimulated the activity phosphoglucose isomerase and UDPGlc pyrophosphorylase, thus enhancing cellulose yield. Matsuoka et al., (1996) investigated BC synthesis by
A. xylinum subsp. sucrofermentous BPR 2001 in agitated culture and found out that the presence of lactate in the growth medium stimulated bacterial growth and enhanced 4-5 times of cellulose yield. The preferred nitrogen sources are yeast extract and peptone, which are basic components of the model medium developed by Hestrin and Schramm (1954). It was also found out that significant part of the expensive medium components, i.e. yeast extract and bactopeptone, can be placed with corn steep liquor. Waste plant materials such as sugar beet molasses, spent liquors after glucose separation from starch hydrolysates, as well as whey and some pharmaceutical industry wastes (e.g. spent liquors after dextran precipitation with 1 ethanol) were appropriate medium components (Krystynowicz er al., 2000).
The satisfying microbial cellulose production can be achieved using i optimum growth medium composition, designed by mathematical methods and computer analysis (Joris et al., 1990, Embuscado er al., 1994, Galas er al.. 1999). ;
Some other compounds, strongly stimulating cellulose production by A. xvlinum ] strains, like derivatives of choline. betaine and fatty acids (salts and esters). were also 1 selected (Hikawa et al.. 1996). ;
wo 7.3 Effect of pH and temperature
Analysis of the influence of pH on A. xylinum cellulose yield and properties, indicates that optimum pH depends on a strain, and usually varies between 4.0-7.0 (Johnson and Neogi, 1989; Galas ef al., 1999). For instance, Ishikawa et al., (1995) and Tahara er al., (1997) who applied for studies two different A. xylinum strains, observed the highest polymer yield at pH 5.0. Except pH of a nutrient broth, also temperature influenced BC yield and properties. Krystynowicz et al., (1997) reported that the temperature range from 28 to 30°C and its variations caused changes of cellulose degree of polymerization and water-binding capacity. Tahara et al., (1997) revealed that pH 5.0, optimum pH for the strain growth and BC synthesis, the activity of both cellulases is several times higher than at pH 4.0, at which the BC production is only slightly declined. 7.4 Fermentor types
Synthesis of BC is run either in static culture or in submerged conditions, providing proper agitation and aeration, necessary for medium ] homogeneity and effective mass transfer. The choice of culture conditions strictly depends on polymer destinstion. : 8. Bacterial cellulose purifications
BC obtained through stationary or agitated culture is not pure and contains some impurities such as culture broth components and A. xylinum cells. Prior ] to use in medicine, food production or paper industry, all these impurities must be i removed. One of the most widely used purification methods is based on treatment of i
BC with solutions of hydroxides (mainly sodium and potassium), sodium chlorate and hypochlorate, H,0,, diluted acids, organic solvent or hot water. The reagents can be used alone or in combinations (Yamanaka er al., 1990).
BC was immersed in their solutions for 14-18 h, in some cases up to 24 h at elevated temperature (55-65°C), markedly reduces the number of cells and coloration degree. BC was boiled in 2% NaOH solution after preliminary running tap water treatment (Yamanaka er al., 1989). Above, it was immersed in 0.1% NaOH at 80°C for 20 minutes, and next washed it with distilled water (Watanabe er al., 1998).
The processes of purification of BC was developed, crude BC was washed in running tap water for overnight, followed by boiling in 1% NaOH solution for 2 h, washed its in tap water to accomplish NaOH removal for 24 h, neutralized with 5% acetic acid (Krystynowicz et al., 1997). In medical application of BC requires special processes to remove bacterial cells and toxins, which can cause pyrogenic reaction. One of the most effective protocols begins with gentle pressing of BC pellicle and immersed in J 3% NaOH for 12 h (repeat 3 times) and after that incubated in 3% HCI solution, pressed and thoroughly washed in distilled water. The purified pellicle is sterilized in autoclave or by cobalt-60 radiation (Ring et al., 1986). Dilute alkaline solutions are capable of hydrolyzing and removing impurities present in the cellulose pellicle. After alkaline treatment and washing in distilled water, the cellulose mass can be dried and processed into pure cellulose membranes (Supaphol and Spruviell, 2000). Cellulose consists of amorphous and crystalline regions existing together. The degree of ; crystallinity is known to vary depending on the origin and mode of chemical treatment of the material. Attalla and Vanderhart (1984) showed that native cellulose i is 4 composite of two or more distinct crystalline forms mainly comprising lo and 18 i phases. Sugiyama er al., (1991) confirmed the existence of two different crystalline forms co-existing within single cellulose micro fibrils through electron microscopy and diffraction studies. The hydrothermal annealing treatment using alkaline solutions can cause transformation of metastable la phase, which occurs predominantly in bacterial cellulose, to more thermally stable 18 phase (Yamamoto et al., 1989). ; 9. Applications of bacterial cellulose
Commercial application of this polymer results from its unique properties and developments in effective technology of production, based on growth of improved microbes on cheap waste materials. The advantage of BC is its chemical purity and the absence of substance usually assisting the plant polysaccharide, which requires laborious purification. Besides, the shape of BC sheet, its area and thickness can be tailored by means of culture conditions. Relatively easy BC modification during its biosynthesis, enables regulation of such properties as molecular mass, elasticity. resilience, water holding capacity, crystallinity index, etc. 9.1 Filter membranes
Filter membranes are categorized as symmetric, asymmetric, porous or non-porous. depending on their filtration behavior, materials used and manufacturing processes. Normally, they are manufactured from polymers composite materials between two polymers, or between polymer and ceramic. Related theories, membrane type and application can be studies in the handbook provided by Ho and Sirkar 3 (1992). It has been well known that bacteria, such as Acetobacter. Rhizobium, :
Argrobacterium and Sarcina. synthesize bio-polymers. Among these. gram-negative
A. xylinum is claimed to be an effective cellulose-producing bacterium and is widely used (Jonas and Farah, 1998; Yang et al., 1998). It can be simply grown in a shallow tray with a culture medium such as coconut juice, sugarcane juice, vinegar, and fermented beverage, which are plentiful locally. Cellulose network formed as a sheet floating on the medium surface has been proved to have high tensile strength, elasticity, resilience, durability, shape-retention, high water binding capacity, non- toxic and non-allergen (Schmitt et al., 1991). 9.2 Paper industries
BC is an excellent component of papers, providing better mechanical properties. Microfibrils of the bacterial polymer from a great number of hydrogen bonds when the paper is subjected to drying, thus giving improved chemical adhesion and tensile strength (higher value of the Young's modulus). BC containing paper show better retention of solid additive such as filler and pigments, but are also more elastic, air-permeable, resistant to tearing and bursting forces, and bind more water (Iguchi er al., 2000). A good effect of BC containing paper such as improved ageing : resistance, was achieved by adding of BC to cotton fibers to obtain handmade paper, use as information and document paper, pressboard, paperboard and bookbinding, use as old documents repairing, appropriate ink receptivity and specific snap (Krystynowicz et al., 1997). BC was added to improve the surface coating for specific paper such as to improve gross, brightness, smoothness, ink receptivity and tensile strength. Substitution of BC for paper and organic polymers could greatly reduce pollution of the environment (Jenelten, 1998).
9.3 Food industries
Chemically pure and metabolically inert, BC has been applied as non caloric bulking and stabilizing agent in processes food. The first successful commercial application of BC in food production is nata de coco (Sutheland, 1998). It is a traditional dessert from Philippines, prepared from coconut milk and coconut water with sucrose. which serve as a growth medium for BC producing bacteria.
Consumption of the pellicle is believed to protect against bowel cancer, artheriosclerosis and coronary thrombosis and prevent sudden rise of glucose in the urine. BC-containing food product is Chinese Kombucha (Teakvass or tea-fungus), obtamed by growing yeast and acetic acid bacteria on tea and sugar extract. The pellicle formed on the surface contains both cellulose and enzymes healthy for humans. Their abiotic activity is especially stimulating for large bowel and the whole alimentary tract. Kombucha is believed to protect from some cancers (Iguchi et al., 2000). The preparations of bioactive anthocyanin enriched in dietary fiber are excellent for functional food production. BC also appeared to be an attractive component of bakery products, since it plays a role of dietary fiber, is taste and odorless and prolongs the shelf-life. 9.4 Miscellaneous uses
BC can be applied as a carrier for immobilization of biocatalysts because the large surface area, high durability and superior adsorptive properties as well as possibility of modification by mean of physical or chemical methods.
Cellulose gel containing immobilized animal cells was used for their breeding, in production of interferon, interleukin-1, cytostatics and monoclonal antibodies (Iguchi et al., 2000). 9.5 Medical applications 9.5.1 Bacterial cellulose as a potential scaffold for tissue engineering of cartilage
The main purposes of articular cartilage, which contains a small number of cells (chondrocytes) in an extra cellular matrix (ECM) mainly composed of water, collagen type II and proteoglycans, are to cover the ends in bones in joins to provide frictionless movement and to distribute loads (Wilkins et al, 2000).
Ostheoarthritis is a disease of synovial joints resulting in pain and loss of function for the patient (Freeman, 1973). Damaged cartilage has limited regenerative capacity and therefore over | million patients in the United States require treatment for cartilage defects each year. However, presently these treatments result in limited pain relief and restorative tissue function. ]
Thus, tissue engineering has potential to provide a supply of functional cartilage for the repair and regeneration of compromised native soft tissues (Schreiber et al., 1999). The use of scaffolds in the tissue engineering of cartilage is essential in order to support cell proliferation and maintain their differentiate function in addition to definition of the shape of new growing tissue. A variety of scaffold materials have been evaluated including natural polymers like collagens, alginate, hyaluronic acid, fibringlue and chitosan and synthetic polymers including polyglycolic acid (PGA), polylactic acid (PLA), polyvinyl alcohol (PVA), polyhydroxyethylmethacrylate
(pHEMA) and poly N-isopropylacrylamide (pNIPAA). However, tissue constructs with native mechanical properties have not yet been described in the literature. It is also desirable to utilize a scaffold material that has the porosity necessary to support cell in growth and effective mass transport while also supplementing the mechanical properties of engineered tissue. This scaffold must also be biocompatible and support native ECM and biopolymer production, with degradation rates commensurate with the rate of the new tissue formation.
BC is secreted by A. xvlinum which has unique properties including high water holding capacity, high crystallinity, a fine fiber network, and high tensile strength (Wilkins and Browning, 2000). BC has potential to be used as a substrate for tissue engineering of cartilage due to its high strength in the wet state as well as its moldability in situ, biocompatibility and relatively simple, cost-efficient production (Schreiber er al., 1999), 9.5.2 Bacterial cellulose as a potential wound dressing
Biofilm from modified bacterial cellulose displays several advantages such as biological dressing when applied on exudation or bloody tissue. It is valuable as a temporary skin substitute in the treatment of skin wounds such as burns, ulcers, grafts and as an adjuvant in all abrasions (Fontana er al., 1990).
The cellulose biomembrane originating from bacterial fulfils a function similar that of the epidermis in relation to the dermis due of its physicochemical and electrostatic properties. The cellulose biomembrane establishes a suitable microenvironment, creating optimum physiological conditions necessary for correct : wound healing, which indeed reduces the cost of treatment especially in cases of burns and venous leg ulceration (Slezak et al., 2004). Selective permeability with regard to liquids and gases allows transpiration and permits gas exchange and thermoregulation. Moreover, it improves the wound’s granulation, which allows rapid and complete wound healing. Due to its electrostatic properties, the membrane adheres firmly to the wound site and protects the wound and nerve endings from mechanical stimulation and reduces pain. One type of bacterial alkali-cellulose biomembrane was obtained through biosynthesis from the cells of A. xvlinum. In contrast to cellulose obtained from wood, bacterial cellulose has a microfibrous structure and its hypo-allergic, non-toxic, non-irritant, biodegradable, non-pyrogenic, highly hydrophilic and biocompatible (Ulmer er al., 2002).
The small size of BC fibrils seems to be a key factor that determines its remarkable performance as a wound healing system. Furthermore, the never dried cellulose membrane is a highly nano-porous material that allows for the potential transfer of antibiotics or other medicine into the wound, while at the same time serving as an efficient physical barrier against any external infection. BC produced in the form of gelatinous membrane can be molded into any shape and size during its synthesis, depending on the fermentation technique and conditions used (Bielecki er : al, 2002). Unlike celluloses of plant origin, BC is entirely free from lignin and hemicelluloses. A vigorous treatment with strong bases at high temperatures allows the removal of cells embedded in the cellulose net and it is possible to achieve a non- pyrogenic, non-toxic and fully biocompatible biomaterial dressing material (Alvarez et al., 2004). Cellulose pad from static culture is a ready-to-use, usually wound dressing material that meets standards for modern wound dressing. It is sterilizable, biocompatible, porous, elastic. easy to handle and store, adsorbs exudation, provide optimum humidity, which is essential for fast wound healing, protects from secondary infection and mechanical injury, does not stick to the newly regenerated tissue and alleviates a pain by heat adsorption from bums. BC sheets are also excellent carrier for immobilization of medicine preparations, which speed up the healing process.
Wound dressing produced from bacterial cellulose
There have been several publications and reports on the successful use of BC as a medical product such as Biofill® is product of BC with wide applications.
It has been used for several skin injury treatments such as basal cell carcinoma/skin grall, severe body burns, facial peeling, sutures, dermabrasion, skin lesions, chronic ulcer, and both donor and receptor sites in skin grafts (Fontana er al., 1990). Cases of second and third degree burns, ulcers and others could be treated successfully with
BioFill® as tempolary substitute for human skin. The authors documented the following advantages for Biofill® in more than 300 treatments such as immediate pain relief, close adhesion to the wound bed, diminished postsurgery discomfort, reduced infection rate, easiness of wound inspection (transparency), faster healing, improved exudates retention, spontaneous detachment following reepithelization and reduced treatment time and costs. Only one disadvantage was mentioned limited elasticity in areas of great mobility (Klemm er al. 2001). Gengiflex® was developed to recover periodontal tissues. Novaes (1992) who described a complete restoration of an osseus defect around and IMZ implant in association with a Gengiflex® therapy. Gengiflex® ] is biocompatible, inert and hypoallergenic (Gottlow er al.. 1994),
10. Topical antimicrobial agents used in burn wounds
A cutaneous bum denudes the skin of keratin layer and a number of layers of essential skin cells, as well as inducing localized reduction of blood supply, thus resulting in tissue hypoxia (Hettich, 1986). Loss of this protective integuum results in bacterial penetration into the underlying tissue, which can result in fatal burn sepsis. Prevention of burn wound sepsis is dependent on optimum wound management, including the use of topical antimicrobial and prompt closure of the wound defect. The advent of topical antimicrobials in the 1960s reduced the mortality from burns wound sepsis from 60% to 28% (Pruitt er al., 1968). However, burn wound sepsis remains and important and potentially remediable cause of significant mortality and morbidity (Pruitt ez al., 1998). The advent of multiresistant bacteria has hastened the almost exclusive use of silver-base topical antimicrobial agents (TAAs) for the prevention of burn wound sepsis (Klasen, 2000).
During the late 1960s, silver sulphadiazine was the gold standard topical anti-microbial used in burns patients around the world. An outbreak of resistant Staphylococcus aureus in the burns unit in Royal Melbome Hospital in 1971 led Dr. M. Clarke to introduce 0.2% chlorhexidine digluconate to 1% silver sulphadiazine for attempt to control this outbreak. The trial was successful and the resultant dressing was introduced as Silvazine™ (1% silver sulphadiazine and 0.2% chlorhexidine digluconate). The transition from standard 1% silver sulphadiazine to 19% silver sulphadiazine with 0.2% chlorhexidine digluconate resulted in a 50% reduction of burn wound sepsis (Georage et al.. 1997).
Silver sulphadiazine is an excellent of activity, low toxicity and ease of application with minimal pain, it is still the most frequently used topical agent. Silver
:sulphadiazine is thought to act via inhibition of DNA replication and modifications of the cell membrane and cell wall. This drug is bactericidal against species of both
Gram-negative and Gram-positive organisms, but resistance has occasionally been reported (Fox et al., 1968). 11. Chlorhexidine
Chlorhexidine is a biguanide compound, widely used in clinical practice as a skin and mucous membrane antiseptic and disinfectant. Chlorhexidine is a disinfectant which is effective against a wide range of vegetative Gram-positive and
Gram-negative bacteria; it is ineffective against acid fast bacteria, bacterial spores, fungi and viruses (Bondar, 2000). It disrupts the plasma membrane of the bacterial cell and cellular contents are lost and it is more effective against Gram-positive than
Gram-negative bacteria, some species of Pseudomonas and Proteus being relatively less susceptible (Kuyyakanond et al., 1992). Chlorhexidine is most active at a neutral or slightly alkaline pH, but its activity is reduced by blood and other organic matter ; (Sawada, 1985). As local chlorhexidine treatment has proved to be effective in the prevention of nosocomial respiratory tract infections, urinary tract infections in transurethral surgery, reducing material and neonatal infections morbidity during childbirth, and decreasing mortality in experimental intra-abdominal sepsis (Bondar er al., 2000). As a handwash or surgical scrub, 4% chlorhexidine causes a greater initial decrease in the number of cutaneous bacteria than does either 7.5% povidone iodine or 3% hexachlorophene and it has a persistent effect equal to or greater than that of hexachlorophene (Smylie er al., 1973). 0.5% solution in 95% ethanol experts a greater effect than 4% chlorhexidine emulsion, 3% hexachlorophene solution (Lowbury et al., 1971). A 1% aqueous solution is more erratic and has a less persistent action than do the other preparations. In experiments in which gloved hands were inoculated withlarge number of Providencia, Serratia, Pseudomonas and Escherichia, 4% chlorhexidine appeared to be somewhat less effective than 7.5% povidone-iodine (Dineen, 1978). : 11.1 Chlorhexidine gluconate
Chlorhexidine gluconate is used in disinfectant solutions, creams and gels. A 0.5% solution in alcohol (70%) is used for the pre-operative disinfection of the skin and 0.05% aqueous solution is used as a wound disinfectant. A 0.05% solution in glycerol is used for urethral disinfection and catheter lubrication, and a 0.02% solution for bladder irrigation. A 1% cream is used in obstetrics. A 0.02% solution containing sodium nitrite 0.1% is used for the storage of sterile instrument (Martindale, 1993). 11.2 Chlorhexidine acetate
Chlorhexidine acetate is used for skin disinfection and in concentration of 0.01% as a preservative for eye-drops (Martindale, 1993). 11.3 Chlorhexidine hydrochloride
Chlorhexidine hydrochloride has been used in creams and dusting powders: a cream containing 0.1% of chlorhexidine hydrochloride and 0.5% of neomycin sulphate is used in the prophylaxis and treatment of nasal carriers of
Staphylococci (Martindale, 1993), ;
11.4 Chlorhexidine digluconate
Chlorhexidine digluconate is commercially available as a 20% w/v aqueous solution since the substance cannot be isolated as a solid. Its molecular structure is shown in Figure 6. It is soluble in water to at least 50% w/v but high viscosity makes such concentrated solutions inconvenient to use. Chlorhexidine digluconate solution has been used as a topical antiseptic and disinfectant effective against a wide range of bacteria, some fungi and some viruses (Martindale, 1993).
The addition of chlorhexidine hydrochloride or digluconate 0.2% w/v of silver sulphadiazine cream used in the treatment of burns reduced the isolation rate of
Staphylococcus aureus in 51 patients to 9.2% compared with 34% in 91% patients treated with silver sulphadiazine alone (Clarke, 1975).
Q 2
NH
H H H
N N N PY :
H H H
NH NH : a HO HO O O OH OH
OH HO
HO HO OH OH
Figure 6. Molecular structure of chlorhexidine digluconate
12. Aims and scope of this thesis
The aims of this study were to evaluate the properties of BC produced from different carbon sources and to prepare the film of BC from different carbon sources containing chlorhexidine digluconate for studying the release of chlorhexidine digluconate.
Scope of this thesis;
I. To evaluate the properties of BC produced from different carbon sources 2. To prepare the film of BC containing chlorhexidine digluconate for studying the release of chlorhexidine digluconate from BC
Claims (1)
- 5-ketogluconate !glucose gluconate 2-ketogluconate 2.5-ketogluconate j GDH GADH 2-KGDH GDH KGR glucose ra gluconate NADP ” NADP NADPH’ : Glucose-6-P a. 6-PG ——————————— > Enter-Doudoroff route NADP NADPH TT glycolysis Pentosé phosphate cycleGlucose-1-P . biomass Krebs :cycle respiration ; UDP-Glucose ; Cellulose’ Figure 2. Pathways of carbon metabolism in Acetobacter xvlinum (De Wulf et al., 1996) Cellulose : CS UDPGilc Glucose ATP GK uGP ADP Gle-1.p +——————— Glc-6-P —————————» PGA PGI FK Fructose TS Fru-6-P ¢——— ATP ADP Sn I-PFK yd Fru-1-P ——— Fru-bisP : Figure 3. Pathways of carbon metabolism in Acetobacter xylinum (Ross er al., 1991: i Tonoucht er al.. 1996) 3. 9 Vibe 2 UMP a a 8 ; Lip—P’ Lib—b—p~@ cs Lip—P—P—@—@ ] RL Lip—p; "HUPPEHT LL Lib—p—p—@ Lib—P—p" 0 v a LpP i a ao 8 UMP a 8 Lip—P—P’ ¢ —Lip—p_P-0@F — Lip—P—P-0-0. “a Lie oN gto Lip—P—P—@ —@—@ Lib—P—P—@LFUDPG PTL) jb p" a B 8 a # 1 U—Rib—P—P—@ d H,0 V a : LPP Pp, xX) a —Rib—P—'P_@ { U Ty x , CS LPP LPUDPGPT A. 0 1 LP-UDPG PT i LPF 8 —p , , f Lip—P—P—-@ 0] o Lip—P—P—@—0—@ rye Lip—b" of R 8 7] B 8 3 Figure 4. Mechanism of bacterial cellulose biosynthesis involving lipid ; intermediated (Han and Robyt. 1998) ;Subel ibril microfibrils ” Subelementary fibrils ribbon : : f 1 : vy, pore —» \/ \/ ra ] ro ;! . 1 | 3 Lo Cytoplasma membrane i LNG \/ Cellulose synthase j Figure 5. Scheme for the formation of bacterial cellulose (Vandamme er al., 1997)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PH12014000281A PH12014000281A1 (en) | 2014-10-08 | 2014-10-08 | Injectable coco cellulose microparticle for tissue augmentation dermal and subdermal filler |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PH12014000281A PH12014000281A1 (en) | 2014-10-08 | 2014-10-08 | Injectable coco cellulose microparticle for tissue augmentation dermal and subdermal filler |
Publications (1)
Publication Number | Publication Date |
---|---|
PH12014000281A1 true PH12014000281A1 (en) | 2016-05-02 |
Family
ID=57904423
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PH12014000281A PH12014000281A1 (en) | 2014-10-08 | 2014-10-08 | Injectable coco cellulose microparticle for tissue augmentation dermal and subdermal filler |
Country Status (1)
Country | Link |
---|---|
PH (1) | PH12014000281A1 (en) |
-
2014
- 2014-10-08 PH PH12014000281A patent/PH12014000281A1/en unknown
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Zhong | Industrial-scale production and applications of bacterial cellulose | |
Ullah et al. | Synthesis, structure, and properties of bacterial cellulose | |
Liu et al. | Enhanced mechanical properties of bacterial cellulose nanocomposites produced by co-culturing Gluconacetobacter hansenii and Escherichia coli under static conditions | |
US8951551B2 (en) | Multiribbon nanocellulose as a matrix for wound healing | |
EP1660670A1 (en) | A method for the production of bacterial cellulose | |
Aramwit et al. | The characteristics of bacterial nanocellulose gel releasing silk sericin for facial treatment | |
Bagewadi et al. | Statistical optimization and characterization of bacterial cellulose produced by isolated thermophilic Bacillus licheniformis strain ZBT2 | |
Mohammad et al. | An overview of biocellulose production using Acetobacter xylinum culture | |
Avcioglu | Bacterial cellulose: Recent progress in production and industrial applications | |
Cherian et al. | Bacterial nanocellulose for medical implants | |
Fontana et al. | New insights on bacterial cellulose | |
US20180216148A1 (en) | Composite cellulose hydrogels and methods of making and use thereof | |
Sukara et al. | Potential values of bacterial cellulose for industrial applications | |
Foresti et al. | Bacterial nanocellulose: Synthesis, properties and applications | |
Basmaji et al. | Novel antimicrobial peptides bacterial cellulose obtained by symbioses culture between polyhexanide biguanide (PHMB) and green tea | |
Amr et al. | Bacterial Cellulose: Biosynthesis and Applications | |
Keshk et al. | Natural bacterial biodegradable medical polymers: Bacterial cellulose | |
Trovatti | Bacterial cellulose | |
PH12014000281A1 (en) | Injectable coco cellulose microparticle for tissue augmentation dermal and subdermal filler | |
Nicomrat | Silver nanoparticles impregnated biocellulose produced by sweet glutinous rice fermentation with the genus Acetobacter | |
JPS6283877A (en) | Inhibitor of growth and proliferation of bacteria | |
Nasresfahani et al. | Recent research on economic production and water absorption improvement of bacterial cellulose: a review | |
KR20180119282A (en) | Biocellulose comprising solid original ingredient, composition of culture medium for manufacturing the same, and method of manufacturing the same | |
CN102991037B (en) | Bacteria cellulose controlled-release carrier material of sandwich structure and preparation method thereof | |
Ul-Islam et al. | Production of bio-cellulose from renewable resources: Properties and applications |