CN103893816B - Method for preparing composite amniotic extracellular matrix dressing of bacterial cellulose containing plant ingredients - Google Patents
Method for preparing composite amniotic extracellular matrix dressing of bacterial cellulose containing plant ingredients Download PDFInfo
- Publication number
- CN103893816B CN103893816B CN201410061448.4A CN201410061448A CN103893816B CN 103893816 B CN103893816 B CN 103893816B CN 201410061448 A CN201410061448 A CN 201410061448A CN 103893816 B CN103893816 B CN 103893816B
- Authority
- CN
- China
- Prior art keywords
- extracellular matrix
- bacteria cellulose
- containing natural
- natural plant
- plant composition
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 title claims abstract description 101
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 title claims abstract description 101
- 210000002744 extracellular matrix Anatomy 0.000 title claims abstract description 101
- 238000000034 method Methods 0.000 title claims abstract description 15
- 239000004615 ingredient Substances 0.000 title abstract description 5
- 229920002749 Bacterial cellulose Polymers 0.000 title abstract 6
- 239000005016 bacterial cellulose Substances 0.000 title abstract 6
- 239000002131 composite material Substances 0.000 title abstract 3
- 241000196324 Embryophyta Species 0.000 claims abstract description 78
- 238000002360 preparation method Methods 0.000 claims abstract description 30
- 230000001954 sterilising effect Effects 0.000 claims abstract description 16
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 14
- 230000001580 bacterial effect Effects 0.000 claims abstract description 13
- 230000003248 secreting effect Effects 0.000 claims abstract description 10
- 206010072170 Skin wound Diseases 0.000 claims abstract description 8
- 238000009736 wetting Methods 0.000 claims abstract description 8
- 238000000746 purification Methods 0.000 claims abstract description 3
- 241000894006 Bacteria Species 0.000 claims description 120
- 239000001913 cellulose Substances 0.000 claims description 113
- 229920002678 cellulose Polymers 0.000 claims description 113
- 239000000203 mixture Substances 0.000 claims description 88
- 239000011159 matrix material Substances 0.000 claims description 55
- 150000001875 compounds Chemical class 0.000 claims description 46
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 30
- 235000015639 rosmarinus officinalis Nutrition 0.000 claims description 24
- 239000008176 lyophilized powder Substances 0.000 claims description 22
- 229940092258 rosemary extract Drugs 0.000 claims description 17
- 235000020748 rosemary extract Nutrition 0.000 claims description 17
- 239000001233 rosmarinus officinalis l. extract Substances 0.000 claims description 17
- 239000002121 nanofiber Substances 0.000 claims description 14
- 239000000427 antigen Substances 0.000 claims description 9
- 108091007433 antigens Proteins 0.000 claims description 9
- 102000036639 antigens Human genes 0.000 claims description 9
- 210000001789 adipocyte Anatomy 0.000 claims description 8
- 210000001691 amnion Anatomy 0.000 claims description 8
- 229910019142 PO4 Inorganic materials 0.000 claims description 7
- 239000001888 Peptone Substances 0.000 claims description 7
- 108010080698 Peptones Proteins 0.000 claims description 7
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 7
- 230000004913 activation Effects 0.000 claims description 7
- 229940041514 candida albicans extract Drugs 0.000 claims description 7
- 239000008367 deionised water Substances 0.000 claims description 7
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 7
- 229910000397 disodium phosphate Inorganic materials 0.000 claims description 7
- 235000019800 disodium phosphate Nutrition 0.000 claims description 7
- 230000007935 neutral effect Effects 0.000 claims description 7
- 235000019319 peptone Nutrition 0.000 claims description 7
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 7
- 239000010452 phosphate Substances 0.000 claims description 7
- 229910052700 potassium Inorganic materials 0.000 claims description 7
- 239000011591 potassium Substances 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 7
- 239000012138 yeast extract Substances 0.000 claims description 7
- 230000015572 biosynthetic process Effects 0.000 claims description 6
- 244000235858 Acetobacter xylinum Species 0.000 claims description 5
- 235000002837 Acetobacter xylinum Nutrition 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 4
- 241000590020 Achromobacter Species 0.000 claims description 3
- 241000588986 Alcaligenes Species 0.000 claims description 3
- 241000589151 Azotobacter Species 0.000 claims description 3
- 241000589180 Rhizobium Species 0.000 claims description 3
- 241000190932 Rhodopseudomonas Species 0.000 claims description 3
- 241000192023 Sarcina Species 0.000 claims description 3
- 239000000017 hydrogel Substances 0.000 claims description 2
- 241001530490 Salvia rosmarinus Species 0.000 claims 3
- 206010052428 Wound Diseases 0.000 abstract description 29
- 208000027418 Wounds and injury Diseases 0.000 abstract description 29
- 239000000463 material Substances 0.000 abstract description 12
- 241001529742 Rosmarinus Species 0.000 abstract description 7
- 230000029663 wound healing Effects 0.000 abstract description 6
- 230000001684 chronic effect Effects 0.000 abstract description 5
- 210000000416 exudates and transudate Anatomy 0.000 abstract description 5
- 230000003020 moisturizing effect Effects 0.000 abstract description 5
- 238000005516 engineering process Methods 0.000 abstract description 3
- 239000012567 medical material Substances 0.000 abstract description 3
- 238000002156 mixing Methods 0.000 abstract description 2
- 239000000843 powder Substances 0.000 abstract description 2
- 239000001963 growth medium Substances 0.000 abstract 2
- 239000012528 membrane Substances 0.000 abstract 2
- 230000003068 static effect Effects 0.000 abstract 2
- 239000000243 solution Substances 0.000 description 22
- 244000178231 Rosmarinus officinalis Species 0.000 description 21
- 208000025865 Ulcer Diseases 0.000 description 12
- 210000000265 leukocyte Anatomy 0.000 description 12
- 231100000397 ulcer Toxicity 0.000 description 12
- 208000008960 Diabetic foot Diseases 0.000 description 10
- 239000011259 mixed solution Substances 0.000 description 10
- 210000004493 neutrocyte Anatomy 0.000 description 8
- 241000700159 Rattus Species 0.000 description 6
- 235000013305 food Nutrition 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 208000005230 Leg Ulcer Diseases 0.000 description 5
- 206010053615 Thermal burn Diseases 0.000 description 5
- 210000002469 basement membrane Anatomy 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 239000003292 glue Substances 0.000 description 5
- 150000002632 lipids Chemical class 0.000 description 5
- 102000008186 Collagen Human genes 0.000 description 4
- 108010035532 Collagen Proteins 0.000 description 4
- 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 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 230000035876 healing Effects 0.000 description 4
- 210000004698 lymphocyte Anatomy 0.000 description 4
- 208000024891 symptom Diseases 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 206010007247 Carbuncle Diseases 0.000 description 3
- 206010017553 Furuncle Diseases 0.000 description 3
- 230000003078 antioxidant effect Effects 0.000 description 3
- 210000004204 blood vessel Anatomy 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000007943 implant Substances 0.000 description 3
- 230000001338 necrotic effect Effects 0.000 description 3
- 230000002980 postoperative effect Effects 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 230000008961 swelling Effects 0.000 description 3
- 206010002091 Anaesthesia Diseases 0.000 description 2
- 206010011985 Decubitus ulcer Diseases 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 206010063560 Excessive granulation tissue Diseases 0.000 description 2
- 102000001554 Hemoglobins Human genes 0.000 description 2
- 108010054147 Hemoglobins Proteins 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 2
- 208000004210 Pressure Ulcer Diseases 0.000 description 2
- 102000013275 Somatomedins Human genes 0.000 description 2
- ZSJLQEPLLKMAKR-UHFFFAOYSA-N Streptozotocin Natural products O=NN(C)C(=O)NC1C(O)OC(CO)C(O)C1O ZSJLQEPLLKMAKR-UHFFFAOYSA-N 0.000 description 2
- 241000209140 Triticum Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 206010000269 abscess Diseases 0.000 description 2
- 230000037005 anaesthesia Effects 0.000 description 2
- 230000000845 anti-microbial effect Effects 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 230000003570 biosynthesizing effect Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 230000008602 contraction Effects 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 210000000981 epithelium Anatomy 0.000 description 2
- 210000003414 extremity Anatomy 0.000 description 2
- 238000001125 extrusion Methods 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 210000002950 fibroblast Anatomy 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 210000001126 granulation tissue Anatomy 0.000 description 2
- 238000002513 implantation Methods 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 210000003141 lower extremity Anatomy 0.000 description 2
- 235000012054 meals Nutrition 0.000 description 2
- 230000005012 migration Effects 0.000 description 2
- 238000013508 migration Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000036407 pain Effects 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- ZSJLQEPLLKMAKR-GKHCUFPYSA-N streptozocin Chemical compound O=NN(C)C(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O ZSJLQEPLLKMAKR-GKHCUFPYSA-N 0.000 description 2
- 229960001052 streptozocin Drugs 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 241000228197 Aspergillus flavus Species 0.000 description 1
- 241000228245 Aspergillus niger Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 241001313855 Bletilla Species 0.000 description 1
- 206010008190 Cerebrovascular accident Diseases 0.000 description 1
- 244000035851 Chrysanthemum leucanthemum Species 0.000 description 1
- 235000008495 Chrysanthemum leucanthemum Nutrition 0.000 description 1
- 102000002734 Collagen Type VI Human genes 0.000 description 1
- 108010043741 Collagen Type VI Proteins 0.000 description 1
- 206010010904 Convulsion Diseases 0.000 description 1
- 102000016359 Fibronectins Human genes 0.000 description 1
- 108010067306 Fibronectins Proteins 0.000 description 1
- 235000019733 Fish meal Nutrition 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 244000286779 Hansenula anomala Species 0.000 description 1
- 235000014683 Hansenula anomala Nutrition 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 235000007340 Hordeum vulgare Nutrition 0.000 description 1
- 241000521257 Hydrops Species 0.000 description 1
- 208000032912 Local swelling Diseases 0.000 description 1
- 206010028851 Necrosis Diseases 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 241000228153 Penicillium citrinum Species 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 108010076039 Polyproteins Proteins 0.000 description 1
- 206010040943 Skin Ulcer Diseases 0.000 description 1
- 208000028990 Skin injury Diseases 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 208000006011 Stroke Diseases 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
- 208000033809 Suppuration Diseases 0.000 description 1
- 208000003443 Unconsciousness Diseases 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
- 230000002745 absorbent Effects 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 229920001222 biopolymer Polymers 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000009172 bursting Effects 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000036461 convulsion Effects 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 238000009795 derivation Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 210000002219 extraembryonic membrane Anatomy 0.000 description 1
- 239000004467 fishmeal Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 208000002672 hepatitis B Diseases 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 102000006495 integrins Human genes 0.000 description 1
- 108010044426 integrins Proteins 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000001365 lymphatic vessel Anatomy 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 210000002826 placenta Anatomy 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011552 rat model Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 230000000405 serological effect Effects 0.000 description 1
- 231100000019 skin ulcer Toxicity 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 208000006379 syphilis Diseases 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 235000007586 terpenes Nutrition 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 description 1
- 210000002993 trophoblast Anatomy 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
- 235000015099 wheat brans Nutrition 0.000 description 1
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Materials For Medical Uses (AREA)
Abstract
The invention discloses a method for preparing composite amniotic extracellular matrix dressing of bacterial cellulose containing plant ingredients, and relates to a technology for preparing a medical material. The method comprises the following steps: selecting a bacterial strain capable of secreting bacterial cellulose to activate and prepare seed mash; then mixing the seed mash of which the bacterial strain concentration is 30-50wt% with culture medium containing natural rosemary freeze-dried powder and then carrying out static culture in a culture container to obtain a bacterial cellulose membrane containing natural plant ingredients; taking amniotic extracellular matrix to soak into the culture medium; paving on the surface of the bacterial cellulose membrane after completely wetting; continuing to carry out static culture for 1-6 hours; carrying out purification and sterilization to obtain the composite amniotic extracellular matrix dressing of the bacterial cellulose containing the natural plant ingredients. The method is simple and feasible in preparation process, low in cost, and wide in material source, and the prepared dressing has the properties, such as sterilization, moisturizing, absorbing wound exudates, accelerating wound healing and the like, and is applicable to various chronic skin wounds.
Description
Technical field
The present invention relates to the preparing technical field of medical material, refer to a kind of preparation method of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition especially.
Background technology
Amnion is from cytotrophoblast derivation, is the internal layer of two-layer fetal membrane.Normal amnion is thin and bright, and not containing structures such as blood vessel, nerve and lymphatic vessels, thickness is 20 ~ 500 μm.Amnion has 5 Rotating fields and can be divided into: epithelial lining, basement membrane layer, tight zone, fibroblast layer, spongy layer.Remove in the characteristics of amniotic extracellular matrix material after cell containing Multiple components such as collagen, glycoprotein, protein-polysaccharide, integrin and laminated bodies, express the associated protein of multiple somatomedin and mRNA thereof, abundant nutritive ingredient can be provided for the propagation of cell, differentiation, be conducive to the growth and breeding of cell; Basement membrane layer is thicker, is conducive to attaching and the migration of epidermic cell, promotes epithelization, prevents its apoptosis; The collegen filament of tight zone and spongy layer form loose 3 D stereo configuration; Histocompatibility is good, does not express human leucocyte antigen, without induction rejection; Easily obtain, and plasticity-is good, is easy to processing treatment.Large quantity research confirms, amnion has good effect as biological dressing for skin injury such as treatment large-area burns, scald etc.But because amnion is thin and soft, insulation poor water retention property, the surface of a wound ooze out and be mostly that Yi Rong loses, biomechanical property is poor, and operation acquires a certain degree of difficulty, and makes it be restricted in biological dressing application simultaneously.
Bacteria cellulose is a kind of natural biopolymer, has hyperfine reticulated structure, is combined into the thick fibrous bundle of 40 ~ 60 nanometers by the fento of diameter 3 ~ 4 nanometer, and is intertwined to form flourishing hyperfine network structure.And its chemical purity is very high, due to physical strength, good fluid absorbent and performance of keeping humidity high when bacteria cellulose has good biocompatibility, a hygrometric state, make it in medical biotechnology Materials, show great advantage, can be used as desirable degradable solid support material and be applied to the numerous areas such as wound dressings, drug carrier material.The simple bacteria cellulose of external employing has been reported as moist dressing, and industrialization is used for clinical.
Rosmarinus officinalis (Rosmarinus officinatis L.) has another name called You Ancao, and another name tansy is Labiatae Rosmarinus plant, perennial evergreen undershrub.Most important in rosemary extract is rosemary antioxidant and rosemary ethereal oil.Rosemary natural anti-oxidant Stability Analysis of Structures, be not easy to decompose, resistance to 190 DEG C of high temperature, its antioxidant effect is good, and does not have toxicity.Containing the multiple compounds such as phenol, acid, ketone, terpene in rosemary plant, at food, medicine, to be industrially widely used.Rosemary extract has good antimicrobial property, all has good inhibition to streptococcus aureus, intestinal bacteria, Bacillus subtilus, Hansenula anomala bacterium, mould, aspergillus niger, flavus and Penicillium citrinum; Rosemary extract is used for the treatment of the reconstruction that chronic wounds can reduce inflammation, strengthen contraction of wounds, contributes to epithelium and granulation tissue, and can improve the formation of wound blood vessels and collagen protein.In sum, bacteria cellulose containing natural rosemary extract composition is combined with characteristics of amniotic extracellular matrix material the matrix material obtaining having three-dimensional net structure, the moist dressing having histocompatibility, antibacterial, moisturizing, absorbing wound exudate, accelerating wound healing and satisfactory mechanical property concurrently can be prepared.
This patent adopts fermentable cultural method In-situ reaction characteristics of amniotic extracellular matrix material.Bacteria cellulose material containing natural rosemary extract composition and characteristics of amniotic extracellular matrix material, to be interacted by the collegen filament of bacteria cellulose nanofiber and characteristics of amniotic extracellular matrix material tight zone and spongy layer and nanofiber runs through mutually, formed and link matrix material closely.Preparation is simple in the present invention, cost is low, material source is extensive, and the dressing of preparation has the performances such as sterilization, moisturizing, absorbing wound exudate and accelerating wound healing, is applicable to all kinds of chronic cutaneous wound.
Summary of the invention
The object of this invention is to provide a kind of preparation method of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition.Relate to a kind of technology of preparing of medical material.Preparation is simple in the present invention, cost is low, material source is extensive, the dressing of preparation has the performances such as sterilization, moisturizing, absorbing wound exudate and accelerating wound healing, is applicable to all kinds of chronic cutaneous wound such as diabetic foot ulcer, venous leg ulcers, pressure sore, skin donor site wound.
The invention discloses a kind of preparation method of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition, comprising: choose and the activation of secreting bacteria cellulosic bacterial strain can be prepared into seed mash, is then that the seed mash of 30 ~ 50wt% mixes the bacteria cellulose film being placed on quiescent culture in culture vessel and obtaining containing natural plant composition with the fermention medium containing natural Rosmarinus officinalis lyophilized powder by bacterial strain concentration; Get characteristics of amniotic extracellular matrix film to soak in the medium, after its complete wetting, be laid in bacteria cellulose film upper surface; Continue quiescent culture 1 ~ 6h, purified process, after sterilizing, obtain a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition.
As preferred technical scheme:
Wherein, a preparation method for bacteria cellulose compound characteristics of amniotic extracellular matrix dressing as above containing natural plant composition, the described cellulosic bacterial strain of energy secreting bacteria refers to one or more in acetobacter xylinum, rhizobium, Sarcina, Rhodopseudomonas, achromobacter, Alcaligenes, aerobacter or Azotobacter.
A kind of preparation method of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as above, the described fermention medium containing natural Rosmarinus officinalis lyophilized powder refers to that wherein aqueous ethanolic solution concentration is 50 ~ 70wt% containing natural Rosmarinus officinalis lyophilized powder 10 ~ 50g, Sodium phosphate dibasic 4 ~ 16g, potassium primary phosphate 4 ~ 16g, yeast extract paste 4 ~ 20g, peptone 4 ~ 20g in the aqueous ethanolic solution of every 1L.
A kind of preparation method of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as above, described quiescent culture refers to that seed mash mixes with the fermention medium containing natural Rosmarinus officinalis lyophilized powder and is placed on quiescent culture 5 ~ 7d in culture vessel, wherein seed mash mix with the fermention medium containing collagen solution after liquid level in culture vessel be 0.5 ~ 3cm.Under normal circumstances, bacteria cellulose film floats on the top of nutrient solution, and bacteria cellulose can consume a part of substratum in fermentation culture process, and a part of substratum is lost by evaporation simultaneously.Method described in this patent is by controlling nutrient solution consumption at zone of reasonableness, and substratum consumption crosses the biosynthesizing that major general affects bacteria cellulose, and affecting characteristics of amniotic extracellular matrix material at most is effectively laid in bacteria cellulose film upper surface excessively.
A kind of preparation method of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as above, the described bacteria cellulose film containing natural plant composition refers to the hydrogel bacteria cellulose film that quiescent culture obtains, thickness is 1 ~ 3cm, the three-dimensional network nanofiber of wherein bacteria cellulose is evenly attached with the chemical substance containing natural rosemary extract composition.Bacterium a large amount of amplification also biosynthesizing Mierocrystalline cellulose fento first in the medium in fermenting process, chemical substance containing natural rosemary extract composition is attached on Mierocrystalline cellulose fento, along with longer fermentation times is formed cellulose nano-fibrous, and evenly adhere to rosemary extract composition on nanofiber; Final formation contains the gluey bacteria cellulose film of water-setting of natural rosemary extract composition.
A kind of preparation method of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as above, described characteristics of amniotic extracellular matrix material refers to safe source, treated removal cell, fat and soluble antigen and the de-cell biological amnion of dry sterilization.Fresh amnion takes from the placenta of healthy Cesarean esction puerpera, and hepatitis B, the third liver, syphilis and adaptive immune deficit syndrome are got rid of in antenatal Serological testing, and process of drawing materials all completes under aseptic technique; Treated removal cell, fat, soluble antigen, general adopt freeze-drying to carry out drying and through irradiation sterilization.
A preparation method for bacteria cellulose compound characteristics of amniotic extracellular matrix dressing as above containing natural plant composition, the described bacteria cellulose film upper surface be laid on containing natural plant composition refers to that the spongy layer of characteristics of amniotic extracellular matrix material contacts with the bacteria cellulose film upper surface containing natural plant composition.Characteristics of amniotic extracellular matrix material has 5 Rotating fields and can be divided into: epithelial lining, basement membrane layer, tight zone, fibroblast layer, spongy layer.Wherein the collegen filament of tight zone and spongy layer form loose 3 D stereo configuration, are conducive to the cellulose nano-fibrous network structure mutually run through with its formation in bacteria cellulose film.Basement membrane layer contains IV type and V Collagen Type VI, ln, the various chelating albumen of fibronectin bletilla, can promote the differentiation of seed cell, propagation and migration.Therefore, when preparing dressing, the spongy layer of characteristics of amniotic extracellular matrix material is contacted with bacteria cellulose film upper surface, be conducive to the cellulose nano-fibrous loose collegen filament inside entering spongy layer of bacteria cellulose film, maintain the biological activity of basement membrane layer simultaneously.
A kind of preparation method of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as above, described purification process refers to that matrix material soaks 12 ~ 24h at the aqueous sodium hydroxide solution 30 ~ 60 DEG C of 1 ~ 3wt%, and by washed with de-ionized water to neutral.The sodium hydroxide of low concentration is selected to process matrix material at low temperatures in this patent, sodium hydroxide solution soaks the residual media thoroughly can removed tropina and stick on bacteria cellulose film, guarantee that the intracellular toxin of matrix material meets the requirement of medical implanting material, simultaneously low-concentration sodium hydroxide can prevent from affecting in treating processes all polyproteins and somatomedin that contain in characteristics of amniotic extracellular matrix material.
A kind of preparation method of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as above, the described bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition refers to the matrix material of bacteria cellulose film containing natural plant composition and characteristics of amniotic extracellular matrix, wherein characteristics of amniotic extracellular matrix is positioned at the upper strata of dressing, bacteria cellulose film containing natural plant composition is positioned at the lower floor of dressing, and the collegen filament in the tight zone of characteristics of amniotic extracellular matrix material and spongy layer run through mutually with containing the nanofiber in the bacteria cellulose film of natural plant composition, formation links matrix material closely.
A kind of preparation method of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as above, in use, the characteristics of amniotic extracellular matrix being positioned at dressing upper strata contacts with skin wound in described a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition.
Another object of the present invention is to provide a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition, it is characterized in that: comprise bacteria cellulose, Rosmarinus officinalis and characteristics of amniotic extracellular matrix.
Compared with prior art, the invention has the beneficial effects as follows:
This patent adopts fermentable cultural method In-situ reaction characteristics of amniotic extracellular matrix material.Bacteria cellulose material containing natural rosemary extract composition and characteristics of amniotic extracellular matrix material, to be interacted by the collegen filament of bacteria cellulose nanofiber and characteristics of amniotic extracellular matrix material tight zone and spongy layer and nanofiber runs through mutually, formed and link matrix material closely.Ensureing, under the histocompatibility of matrix material, bioactive prerequisite, effectively to enhance the mechanical property of material, anti-microbial property; The reconstruction that contained natural rosemary extract composition can reduce wound inflammation, strengthen contraction of wounds, contributes to epithelium and granulation tissue, and the formation of wound blood vessels and collagen protein can be improved.Preparation is simple in the present invention, cost is low, material source is extensive, the dressing of preparation has the performances such as sterilization, moisturizing, absorbing wound exudate and accelerating wound healing, is applicable to all kinds of chronic cutaneous wound such as diabetic foot ulcer, venous leg ulcers, pressure sore, skin donor site wound.
Embodiment
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.In addition should be understood that those skilled in the art can make various changes or modifications the present invention, and these equivalent form of values fall within the application's appended claims limited range equally after the content of having read the present invention's instruction.
Embodiment 1:
Choosing and the cellulosic acetobacter xylinum activation of secreting bacteria can be prepared into seed mash, is then that the seed mash of 50wt% mixes with the fermention medium containing natural Rosmarinus officinalis lyophilized powder by bacterial strain concentration.The described fermention medium containing natural Rosmarinus officinalis lyophilized powder refers to that wherein aqueous ethanolic solution concentration is 70wt% containing natural Rosmarinus officinalis lyophilized powder 30g, Sodium phosphate dibasic 16g, potassium primary phosphate 16g, yeast extract paste 20g, peptone 20g in the aqueous ethanolic solution of every 1L.Mixed solution is placed in culture vessel, and the liquid level of mixed solution in culture vessel is 3cm, and quiescent culture 5d obtains the bacteria cellulose film of water-setting glue containing natural rosemary extract composition that film thickness is 3cm.
By safe source, treated removal cell, fat and soluble antigen and the characteristics of amniotic extracellular matrix material of dry sterilization soak in the medium, are laid in bacteria cellulose film upper surface after its complete wetting.Wherein, the spongy layer of characteristics of amniotic extracellular matrix material contacts with the bacteria cellulose film upper surface containing natural plant composition.Continue quiescent culture 1h, the aqueous sodium hydroxide solution 60 DEG C that the matrix material obtained is placed in 1wt% soaks 12h, and by washed with de-ionized water to neutral, obtains a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition.Wherein characteristics of amniotic extracellular matrix is positioned at the upper strata of dressing, bacteria cellulose film containing natural plant composition is positioned at the lower floor of dressing, and the collegen filament in the tight zone of characteristics of amniotic extracellular matrix material and spongy layer run through mutually with containing the nanofiber in the bacteria cellulose film of natural plant composition, formed and link matrix material closely.In use, the characteristics of amniotic extracellular matrix being positioned at dressing upper strata contacts with skin wound in this dressing.
Embodiment 2:
Choose can the cellulosic rhizobium of secreting bacteria and Rhodopseudomonas activation be prepared into seed mash, be then that the seed mash of 40wt% mixes with the fermention medium containing natural Rosmarinus officinalis lyophilized powder by bacterial strain concentration.The described fermention medium containing natural Rosmarinus officinalis lyophilized powder refers to that wherein aqueous ethanolic solution concentration is 70wt% containing natural Rosmarinus officinalis lyophilized powder 20g, Sodium phosphate dibasic 14g, potassium primary phosphate 14g, yeast extract paste 18g, peptone 18g in the aqueous ethanolic solution of every 1L.Mixed solution is placed in culture vessel, and the liquid level of mixed solution in culture vessel is 2cm, and quiescent culture 5d obtains the bacteria cellulose film of water-setting glue containing natural rosemary extract composition that film thickness is 2.5cm.
By safe source, treated removal cell, fat and soluble antigen and the characteristics of amniotic extracellular matrix material of dry sterilization soak in the medium, are laid in bacteria cellulose film upper surface after its complete wetting.Wherein, the spongy layer of characteristics of amniotic extracellular matrix material contacts with the bacteria cellulose film upper surface containing natural plant composition.Continue quiescent culture 2h, the aqueous sodium hydroxide solution 50 DEG C that the matrix material obtained is placed in 2wt% soaks 12h, and by washed with de-ionized water to neutral, obtains a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition.Wherein characteristics of amniotic extracellular matrix is positioned at the upper strata of dressing, bacteria cellulose film containing natural plant composition is positioned at the lower floor of dressing, and the collegen filament in the tight zone of characteristics of amniotic extracellular matrix material and spongy layer run through mutually with containing the nanofiber in the bacteria cellulose film of natural plant composition, formed and link matrix material closely.In use, the characteristics of amniotic extracellular matrix being positioned at dressing upper strata contacts with skin wound in this dressing.
Embodiment 3:
Choose can the cellulosic Sarcina of secreting bacteria and Alcaligenes activation be prepared into seed mash, be then that the seed mash of 30wt% mixes with the fermention medium containing natural Rosmarinus officinalis lyophilized powder by bacterial strain concentration.The described fermention medium containing natural Rosmarinus officinalis lyophilized powder refers to that wherein aqueous ethanolic solution concentration is 60wt% containing natural Rosmarinus officinalis lyophilized powder 10g, Sodium phosphate dibasic 12g, potassium primary phosphate 12g, yeast extract paste 16g, peptone 16g in the aqueous ethanolic solution of every 1L.Mixed solution is placed in culture vessel, and the liquid level of mixed solution in culture vessel is 1cm, and quiescent culture 6d obtains the bacteria cellulose film of water-setting glue containing natural rosemary extract composition that film thickness is 1cm.
By safe source, treated removal cell, fat and soluble antigen and the characteristics of amniotic extracellular matrix material of dry sterilization soak in the medium, are laid in bacteria cellulose film upper surface after its complete wetting.Wherein, the spongy layer of characteristics of amniotic extracellular matrix material contacts with the bacteria cellulose film upper surface containing natural plant composition.Continue quiescent culture 3h, the aqueous sodium hydroxide solution 30 DEG C that the matrix material obtained is placed in 3wt% soaks 16h, and by washed with de-ionized water to neutral, obtains a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition.Wherein characteristics of amniotic extracellular matrix is positioned at the upper strata of dressing, bacteria cellulose film containing natural plant composition is positioned at the lower floor of dressing, and the collegen filament in the tight zone of characteristics of amniotic extracellular matrix material and spongy layer run through mutually with containing the nanofiber in the bacteria cellulose film of natural plant composition, formed and link matrix material closely.In use, the characteristics of amniotic extracellular matrix being positioned at dressing upper strata contacts with skin wound in this dressing.
Embodiment 4:
Choosing and the activation of the cellulosic acetobacter xylinum of secreting bacteria, aerobacter and Azotobacter can be prepared into seed mash, is then that the seed mash of 50wt% mixes with the fermention medium containing natural Rosmarinus officinalis lyophilized powder by bacterial strain concentration.The described fermention medium containing natural Rosmarinus officinalis lyophilized powder refers to that wherein aqueous ethanolic solution concentration is 60wt% containing natural Rosmarinus officinalis lyophilized powder 50g, Sodium phosphate dibasic 10g, potassium primary phosphate 10g, yeast extract paste 10g, peptone 10g in the aqueous ethanolic solution of every 1L.Mixed solution is placed in culture vessel, and the liquid level of mixed solution in culture vessel is 0.5cm, and quiescent culture 6d obtains the bacteria cellulose film of water-setting glue containing natural rosemary extract composition that film thickness is 2cm.
By safe source, treated removal cell, fat and soluble antigen and the characteristics of amniotic extracellular matrix material of dry sterilization soak in the medium, are laid in bacteria cellulose film upper surface after its complete wetting.Wherein, the spongy layer of characteristics of amniotic extracellular matrix material contacts with the bacteria cellulose film upper surface containing natural plant composition.Continue quiescent culture 3h, the aqueous sodium hydroxide solution 30 DEG C that the matrix material obtained is placed in 1wt% soaks 24h, and by washed with de-ionized water to neutral, obtains a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition.Wherein characteristics of amniotic extracellular matrix is positioned at the upper strata of dressing, bacteria cellulose film containing natural plant composition is positioned at the lower floor of dressing, and the collegen filament in the tight zone of characteristics of amniotic extracellular matrix material and spongy layer run through mutually with containing the nanofiber in the bacteria cellulose film of natural plant composition, formed and link matrix material closely.In use, the characteristics of amniotic extracellular matrix being positioned at dressing upper strata contacts with skin wound in this dressing.
Embodiment 5:
Choose can the cellulosic acetobacter xylinum of secreting bacteria and achromobacter activation be prepared into seed mash, be then that the seed mash of 40wt% mixes with the fermention medium containing natural Rosmarinus officinalis lyophilized powder by bacterial strain concentration.The described fermention medium containing natural Rosmarinus officinalis lyophilized powder refers to that wherein aqueous ethanolic solution concentration is 50wt% containing natural Rosmarinus officinalis lyophilized powder 40g, Sodium phosphate dibasic 4g, potassium primary phosphate 4g, yeast extract paste 4g, peptone 4g in the aqueous ethanolic solution of every 1L.Mixed solution is placed in culture vessel, and the liquid level of mixed solution in culture vessel is 2cm, and quiescent culture 7d obtains the bacteria cellulose film of water-setting glue containing natural rosemary extract composition that film thickness is 2cm.
By safe source, treated removal cell, fat and soluble antigen and the characteristics of amniotic extracellular matrix material of dry sterilization soak in the medium, are laid in bacteria cellulose film upper surface after its complete wetting.Wherein, the spongy layer of characteristics of amniotic extracellular matrix material contacts with the bacteria cellulose film upper surface containing natural plant composition.Continue quiescent culture 6h, the aqueous sodium hydroxide solution 40 DEG C that the matrix material obtained is placed in 3wt% soaks 18h, and by washed with de-ionized water to neutral, obtains a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition.Wherein characteristics of amniotic extracellular matrix is positioned at the upper strata of dressing, bacteria cellulose film containing natural plant composition is positioned at the lower floor of dressing, and the collegen filament in the tight zone of characteristics of amniotic extracellular matrix material and spongy layer run through mutually with containing the nanofiber in the bacteria cellulose film of natural plant composition, formed and link matrix material closely.In use, the characteristics of amniotic extracellular matrix being positioned at dressing upper strata contacts with skin wound in this dressing.
Embodiment 6:
The subcutaneous implantation experiment of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition
Experimental technique: get healthy SD rat 30, male or female, be divided into experimental group and control group, sterilization, adopts ether inhalation anesthesia, and normal sterile operates, and cut skin of back, 51 × 1cm are implanted at experimental group back
2dressing (the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition respectively prepared by Example 1-5), namely control group is sewed up after cutting.Observe the postoperative generalized case of animal and wound healing situation.Within postoperative 1,2,4 week, put to death each treated animal, take out implantation dressing glutaraldehyde and fix, conventional embedded section, HE dyes, om observation implant peripheral lymphocyte, macrophages infiltration situation.
Experiment shows: can normally take food after operation animal on the same day is clear-headed, wound is without Inflammatory responses such as redness, sepages, and all surface of a wound all can well heal, and operative incision infects without 1 example, animal is without the toxic reaction such as death, convulsions, apoplexy, implant surrounding soft tissue has no the performances such as blackout, necrosis, suppuration, hydrops, and postoperative 1 week of histological observation, has large amount lymphocyte around implant, when 2 weeks, only a small amount of lymphocyte, when 4 weeks, exists without lymphocyte substantially.Confirm dressing abiology toxicity of the present invention, have good histocompatibility.
Embodiment 7
Bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition is to the Experiment on therapy of diabetic foot ulcer rat
Use high lipid food to coordinate the rats with type 2 diabetes of streptozotocin induction, use scald apparatus to make dark II degree of scald induced skin ulcer and make diabetic foot ulcer rat model.Rat is SD male rat, SPF level, body weight (180 ± 10) g, totally 30, often organize 5, totally 6 groups, wherein one group is contrast, another 5 groups of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition used embodiment 1-5 respectively and prepare.The preparation method of high lipid food is: first, by barley 15%, and wheat 31%, corn 15%, wheat bran 15%, after fish meal 6% is uniformly mixed in proportion, after adding dregs of beans, oil, salt mixing, basal feed is made in extrusion forming, then, by basal feed 73%, lard 20%, milk powder 2%, cholesterol 1%, sucrose 4% is uniformly mixed rear extrusion forming in proportion and makes high lipid food, and finished product is that cylinder is block.After high lipid food feeds 8 weeks, the citric acid-sodium citrate damping fluid that 1.5% streptozotocin is dissolved in pH value 4.5 is injected by the dosage left lower quadrant of 35mg/kg.Use SQL-5Q scald apparatus, temperature 90 DEG C, duration of contact 10S, pressure 9.8N, area 4cm
2, scald in back part skin after anesthesia.After modeling success, the surface of a wound is cleaned with Iodophor, with the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing external application surface of a wound containing natural plant composition of the present invention, then wrap up with sterile gauze, within 1 day, change dressings 1 time, add up to indicator-specific statistics the average healing with ulcer area and blood leukocytes, the results are shown in Table 1.
The average healing of table 1 pair diabetic foot ulcer rat compares (n=5)
| Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 | Embodiment 5 | |
| The average healing/sky | 18.5±1.4 | 17.8±0.8 | 18.4±1.2 | 19.2±1.0 | 17.0±1.2 |
Embodiment 8:
The hospital clinical trial of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition
Bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition prepared by the embodiment of the present invention 1 is through hospital clinical trial treatment totally 102 examples.Wherein relate to diabetic foot ulcer 37 example, venous leg ulcers 21 example, sore or carbuncle furuncle 44 example, its result for the treatment of refers to table 2.
In table 2, the curative effect determinate standard of various diabetic foot ulcer, venous leg ulcers, sore or carbuncle furuncle is:
1. fully recover: constitutional symptom disappears, local swelling is dissipated, or fester absorbs dissipation, or sore face healing after bursting, and blood leukocytes sum normally.
2. effective: constitutional symptom disappears, and swelling, abscess or the open sore part reduce more than 70%, blood leukocytes sum is normal.
3. effective: constitutional symptom alleviates, swelling, abscess or the open sore part reduce more than 30%, and less than 70%, blood leukocytes sum is close to normal.
4. invalid: not reach effective standard.
The clinical experiment of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing of table 2 containing natural plant composition
| Case type | Case load | Recovery from illness | Effective | Effectively | Invalid | Curative ratio |
| Diabetic foot ulcer | 37 | 36 | 1 | 0 | 0 | 97.3% |
| Venous leg ulcers | 21 | 19 | 1 | 1 | 0 | 90.5% |
| Sore or carbuncle furuncle | 44 | 42 | 1 | 1 | 0 | 95.5% |
Wherein concrete case has:
Week certain, man, 68 years old, diabetic history 20 years, fasting plasma glucose 14 mnol/l, early 2 hours blood glucose 17.9 mmol/L, glycolated hemoglobin 11.2 mg/dl after the meal, diabetic foot ulcer three grades, red swelling of the skin, fester, deep ulcer, forms ulcer chamber, ulcer area 145.5 mm
2, blood leukocytes sum 1.8 × 10
9/ l, neutrophil leucocyte number 77%.The surface of a wound is cleaned with Iodophor, remove necrotic tissue and purulent secretion, clog in chamber and the external application surface of a wound with the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition of the present invention, then wrap up with sterile gauze, lifting affected limb, does not make foot pressurized, within 1 day, changes dressings 1 time, 12 days courses for the treatment of of continuous treatment one, ulcer area reduces to 64.5 mm
2, blood leukocytes sum is down to 0.7 × 10
9/ l, neutrophil leucocyte number 55%.Continued treatment to 24 day, muscle groups tissue layer, lipid layer, skin corium, epidermal area recover original weave construction, cure completely.
Wang, female, 47 years old, fasting plasma glucose 8.1 mmol/L, early 2 hours blood glucose 10.4 mmol/L, glycolated hemoglobin 8.7 mg/dl after the meal, left sufficient diabetic foot ulcer one-level, ulcer area 123.3 mm
2, wound festers, blood leukocytes sum 1.6 × 10
9/ l, neutrophil leucocyte number 70%.The surface of a wound is cleaned with Iodophor, remove necrotic tissue and purulent secretion, with the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing external application surface of a wound containing natural plant composition of the present invention, then wrap up with sterile gauze, lifting affected limb, does not make foot pressurized, within 1 day, changes dressings 1 time, continuous treatment 7 days local symptoms disappear, and blood leukocytes sum is down to 0.5 × 10
9/ l, neutrophil leucocyte number 48%, ulcer surface heals completely.
Jiao, female, 67 years old, sore, intractable ulcer patient, left lower extremity ulcer 5 wheat harvesting period, local presents swollen, hard tubercle, and risen root bundle, sore look red, there is purulent core on sore top, and pain is obvious, and blood leukocytes sum and neutrophil leucocyte number increase, blood leukocytes sum 1.4 × 10
9/ l, neutrophil leucocyte number 73%.The surface of a wound is cleaned with Iodophor, remove necrotic tissue and purulent secretion, adopt of the present invention containing on the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing external application ulcer surface of natural plant composition, then wrap up with sterile gauze, within 1 day, change dressings 1 time, medication is after 7 days, and redness obviously alleviates, ulcer surface reduces, and blood leukocytes sum is down to 0.7 × 10
9/ l, neutrophil leucocyte number 60%.Continue medication to the 14th day, left lower extremity is without red, swollen, and without pain, ulcer wound surface heals, and blood leukocytes sum is down to 0.6 × 10
9/ l, neutrophil leucocyte number 53%, does not stay obvious scar, treatment recovery from illness.
Claims (9)
1. the preparation method of the bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition, it is characterized in that: choose and the activation of secreting bacteria cellulosic bacterial strain can be prepared into seed mash, is then that the seed mash of 30 ~ 50wt% mixes the bacteria cellulose film being placed on quiescent culture in culture vessel and obtaining containing natural plant composition with the fermention medium containing natural Rosmarinus officinalis lyophilized powder by bacterial strain concentration; Get characteristics of amniotic extracellular matrix film to soak in the medium, after its complete wetting, be laid in bacteria cellulose film upper surface; Continue quiescent culture 1 ~ 6h, purified process, after sterilizing, obtain a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition.
2. the preparation method of a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as claimed in claim 1, is characterized in that: the described cellulosic bacterial strain of energy secreting bacteria refers to one or more in acetobacter xylinum, rhizobium, Sarcina, Rhodopseudomonas, achromobacter, Alcaligenes, aerobacter or Azotobacter.
3. the preparation method of a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as claimed in claim 1, it is characterized in that: the described fermention medium containing natural Rosmarinus officinalis lyophilized powder refers to that wherein aqueous ethanolic solution concentration is 50 ~ 70wt% containing natural Rosmarinus officinalis lyophilized powder 10 ~ 50g, Sodium phosphate dibasic 4 ~ 16g, potassium primary phosphate 4 ~ 16g, yeast extract paste 4 ~ 20g, peptone 4 ~ 20g in the aqueous ethanolic solution of every 1L.
4. the preparation method of a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as claimed in claim 1, it is characterized in that: the bacteria cellulose film containing natural plant composition refers to the hydrogel bacteria cellulose film that quiescent culture obtains, thickness is 1 ~ 3cm, the three-dimensional network nanofiber of wherein bacteria cellulose is evenly attached with the chemical substance containing natural rosemary extract composition.
5. the preparation method of a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as claimed in claim 1, it is characterized in that: described characteristics of amniotic extracellular matrix material refers to safe source, treated removal cell, fat and soluble antigen and the de-cell biological amnion of dry sterilization.
6. the preparation method of a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as claimed in claim 1, is characterized in that: the described bacteria cellulose film upper surface be laid in containing natural plant composition refers to that the spongy layer of characteristics of amniotic extracellular matrix material contacts with the bacteria cellulose film upper surface containing natural plant composition.
7. the preparation method of a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as claimed in claim 1, it is characterized in that: described purification process refers to that matrix material soaks 12 ~ 24h at the aqueous sodium hydroxide solution 30 ~ 60 DEG C of 1 ~ 3wt%, and by washed with de-ionized water to neutral.
8. the preparation method of a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as claimed in claim 1, it is characterized in that: the described bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition refers to the matrix material of bacteria cellulose film containing natural plant composition and characteristics of amniotic extracellular matrix, wherein characteristics of amniotic extracellular matrix is positioned at the upper strata of dressing, bacteria cellulose film containing natural plant composition is positioned at the lower floor of dressing, and the collegen filament in the tight zone of characteristics of amniotic extracellular matrix material and spongy layer run through mutually with containing the nanofiber in the bacteria cellulose film of natural plant composition, formation links matrix material closely.
9. the preparation method of a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition as claimed in claim 1, it is characterized in that: in use, the characteristics of amniotic extracellular matrix being positioned at dressing upper strata contacts with skin wound in described a kind of bacteria cellulose compound characteristics of amniotic extracellular matrix dressing containing natural plant composition.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410061448.4A CN103893816B (en) | 2014-02-24 | 2014-02-24 | Method for preparing composite amniotic extracellular matrix dressing of bacterial cellulose containing plant ingredients |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410061448.4A CN103893816B (en) | 2014-02-24 | 2014-02-24 | Method for preparing composite amniotic extracellular matrix dressing of bacterial cellulose containing plant ingredients |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103893816A CN103893816A (en) | 2014-07-02 |
| CN103893816B true CN103893816B (en) | 2015-06-10 |
Family
ID=50985594
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410061448.4A Active CN103893816B (en) | 2014-02-24 | 2014-02-24 | Method for preparing composite amniotic extracellular matrix dressing of bacterial cellulose containing plant ingredients |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103893816B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108888799A (en) * | 2018-06-25 | 2018-11-27 | 东莞市联洲知识产权运营管理有限公司 | A kind of enhanced compound cellulose medical dressing and preparation method thereof with disinsectization performance |
| CN111481735A (en) * | 2019-01-25 | 2020-08-04 | 华中科技大学同济医学院附属协和医院 | Medical antibacterial wound-protecting hydrogel dressing and preparation method thereof |
| CN111790000A (en) * | 2020-06-23 | 2020-10-20 | 洛阳巴库生物科技有限公司 | Stem cell amniotic membrane dressing structure for wound repair and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1618954A (en) * | 2003-05-01 | 2005-05-25 | 四川大学华西医院 | Bioderived amniotic membrane, composite bioderived amniotic membrane and preparation method thereof |
| CN102250378A (en) * | 2011-03-30 | 2011-11-23 | 东华大学 | Bacterial cellulose/polymer composite film and preparation method thereof |
| CN103385896A (en) * | 2013-07-23 | 2013-11-13 | 钟春燕 | Preparation method of bacterial cellulose composite flavonoid compound |
| CN103481600A (en) * | 2012-06-12 | 2014-01-01 | 钟春燕 | Preparation method of bacterial cellulose composite membrane material |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8871016B2 (en) * | 2011-08-03 | 2014-10-28 | The Johns Hopkins University | Cellulose-based hydrogels and methods of making thereof |
-
2014
- 2014-02-24 CN CN201410061448.4A patent/CN103893816B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1618954A (en) * | 2003-05-01 | 2005-05-25 | 四川大学华西医院 | Bioderived amniotic membrane, composite bioderived amniotic membrane and preparation method thereof |
| CN102250378A (en) * | 2011-03-30 | 2011-11-23 | 东华大学 | Bacterial cellulose/polymer composite film and preparation method thereof |
| CN103481600A (en) * | 2012-06-12 | 2014-01-01 | 钟春燕 | Preparation method of bacterial cellulose composite membrane material |
| CN103385896A (en) * | 2013-07-23 | 2013-11-13 | 钟春燕 | Preparation method of bacterial cellulose composite flavonoid compound |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103893816A (en) | 2014-07-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Rosenbaum et al. | Advances in wound management | |
| US4414202A (en) | Composition for treatment of wounds | |
| CN103893825B (en) | Method for preparing bacterial cellulose compounded amnion extracellular matrix material containing collagen | |
| US4778679A (en) | Method and composition for treatment of wounds | |
| CN103893816B (en) | Method for preparing composite amniotic extracellular matrix dressing of bacterial cellulose containing plant ingredients | |
| Lo et al. | Wound healing after cultured epithelial autografting in patients with massive burn injury: a cohort study | |
| CN110623792B (en) | Medical dressing and preparation method thereof | |
| CN104474573A (en) | Biological mask and preparation method thereof | |
| CN105169464B (en) | A kind of natural honey wound dressing and preparation method and application | |
| Sikka et al. | Modern developments in burn wound dressing | |
| JP2009269904A (en) | Herbal medicinal composition for curing external wound and bedsore and method of manufacturing medicine from the composition | |
| TWI825362B (en) | Use of lactobacillus fermentation product in preparation of external composition for enhancing skin wound healing | |
| CN105664226A (en) | Medical composite dressing and preparation method thereof | |
| US20080102106A1 (en) | Wound Healing Composition Comprising Substances From Diptera Larvae | |
| CN101797376A (en) | Preparation method of modified collagen film | |
| CN109432483B (en) | Medical dressing for accelerating wound healing and preparation method and application thereof | |
| EP0034504B1 (en) | Combinations for the treatment of wounds | |
| SEID et al. | A review on equine wound management and healing process | |
| Furtado et al. | Wound healing concepts: contemporary practices and future perspectives | |
| Arbab et al. | Latest advance anti-inflammatory hydrogel wound dressings and traditional Lignosus rhinoceros used for wound healing agents | |
| CN107468715A (en) | Bacillus cercus CGMCC0601 zymotic fluids are preparing the application in treating diabetes trauma Wound medicine | |
| CN107583101A (en) | A kind of biological membrane preparations for promoting wound healing and preparation method thereof | |
| Turriziani et al. | Dressing: Indications on Applications | |
| CN112245653B (en) | Protein film dressing assisting wound healing based on cooperation of three bioactive scaffold materials and cell trophic factors | |
| CN110801464A (en) | Sanhua Ju-Neng trauma rescue medicated oil and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |