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Bioactive Bacterial Nanocellulose Membranes Enriched Nrog Eucalyptus Globulus Labill. Leaves Aqueous Extract Rau Anti-Aging Skin Care Applications Part 2

Jun 10, 2022

Thov hu rauoscar.xiao@wecistanche.comyog xav paub ntxiv

3. Cov txiaj ntsig thiab kev sib tham

Bioactive BNC daim nyias nyias, sib xyaw cov koob tshuaj sib txawv ntawm cov tshuaj aqueous tau los ntawm hydro-distillation ntawm E.globulus nplooj, tau npaj, pom lawv daim ntawv thov raws li kev tiv thaiv kev laus ntawm daim tawv nqaij daim npog qhov ncauj. Plaub BNCmembranes muaj 7.5 mg cm-ntawm glycerol (ua raws li plasticizer thiab humectant) [39,48] thiab cov ntsiab lus sib txawv ntawm HDE, uas yog 1.0 ug cm-2 (BNC-G-HDE1) ,1.5 ug cm-2(BNC-G-HDE1.5),2.0 ug cm-2(BNC-G-HDE2), and 3.0 ug cm-7(BNC-G-HDE3), tau npaj (Table 1). Ib qho ntshiab BNC membrane (BNC) thiab BNC daim nyias nyias nrog tsuas yog glycerol (BNC-G) kuj tau npaj rau kev sib piv (Table 1). BNC-G-HDE daim nyias nyias tau tsim los ntawm txoj kev yooj yim impregnation, raws li qhov sib txawv ntawm HDE aqueous daws thiab glycerol rau hauv hydrophilic thiab porous BNC network (Daim duab 1), raws li cov kev tshawb fawb yav dhau los uas tau piav qhia txog cov txheej txheem no rau kev koom ua ke. ntawm kev sib xyaw ntawm cov nroj tsuag rho tawm [13] thiab caffeine [23].

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Nrog rau qhov sib ntxiv ntawm HDE thiab glycerol, lub thickness ntawm qhuav BNC daim nyias nyias tau nce los ntawm 50 ± 1 um rau cov ntshiab BNC mus rau 87 ± 4 um rau BNC-G, thiab rau cov nqi nyob rau hauv thaj tsam ntawm 79 ± 4 um mus rau 98. ± 8 μm rau daim nyias nyias ntim nrog HDE thiab glycerol, uas qhia txog kev ua tiav ntawm cov khoom ntawm cov extract thiab glycerol rau hauv 3D qauv ntawm BNC. Lub macroscopic nam ntawm cov tau daim nyias nyias (Daim duab 1) yog qhia txog ib homogeneous tis ntawm HDE thiab glycerol nyob rau hauv nanostructure ntawm BNC.npaum li cas cistanche cojNws kuj tseem pom tseeb tias qhov suav nrog glycerol hloov cov daim nyias nyias ntau dua, raws li tau piav qhia yav dhau los [20]. Ntxiv mus, kev koom ua ke ntawm HDE tsis ua rau pom kev hloov xim ntawm cov xim dawb BNC daim nyias nyias, uas yog nyob rau hauv txoj kab nrog cov xim tsis muaj xim ntawm HDE aqueous tov (Daim duab 1).

KSL05

Thov nias ntawm no kom paub ntxiv

Tag nrho cov tau BNC daim nyias nyias tau pom nyob rau hauv cov nqe lus ntawm morphology, mechanical kev ua tau zoo, thermal stability, thiab noo noo uptake peev xwm. Tsis tas li ntawd, kev ua haujlwm antioxidant thiab hauv vitro cytotoxicity ntawm daim nyias nyias kuj tau soj ntsuam. Rau BNC-G-HDE2 daim nyias nyias (2.0 ug cm--), qhov ruaj khov ntawm cov tshuaj tiv thaiv antioxidant tom qab 3 lub hlis ntawm kev cia ntawm 22-25 degree C thiab 52 feem pua ​​RH lossis ntawm 40 degree C thiab 75 feem pua ​​​​nrog rau kev ua haujlwm hauv vitro los tiv thaiv kev laus kuj raug soj ntsuam.

3.1. Morphology

Lub morphology ntawm BNC, BNC-G, thiab BNC-G-HDE membranes tau soj ntsuam los ntawm SEM tsom xam, raws li depicted nyob rau hauv daim duab 2. Nyob rau hauv lub nto micrograph ntawm cov ntshiab BNC, nws cov yam ntxwv 3D nanofibrillar network yog kom meej meej pom; Txawm li cas los xij, qhov microstructure no tsis tshua pom muaj nyob ntawm BNC-Gand BNC-G-HDE daim nyias nyias, tshwj xeeb tshaj yog nrog kev nce ntxiv ntawm cov extract, uas, ua ke nrog glycerol, npog cov cellulose nanofibrils. Cov nyhuv no twb tau pom nyob rau hauv BNC daim nyias nyias koom nrog tsuas yog glycerol [49] lossis glycerol thiab cov khoom xyaw tshuaj nquag (xws li caffeine, diclofenac, thiab lidocaine) [22]. Nws tseem tuaj yeem pom tau tias cov membranes tso tawm cov qauv qis dua li cov BNC pristine vim yog qhov nkag ntawm glycerol (nrog rau cov khoom siv extract) hauv BNC network. Kev koom ua ke ntawm glycerol molecules ntawm BNC nanofibrils txo cov intermolecular hydrogen bonds ntawm cellulose nanofibrils [49], thiab yog li ntawd, diminishes lub cev qhuav dej ntawm BNC qauv thaum ziab, ua li no nyob rau hauv daim nyias nyias nrog nce yooj [23]. Hauv cov duab hla ntu, qhov ntim ntawm lamellar qhov chaw ntawm BNC nrog glycerol thiab cov extract kuj pom meej meej. Cov txiaj ntsig zoo sib xws kuj tau tshaj tawm rau BNC-raws li cov ntaub ntawv ntim nrog ethanolic tov ntawm propolis extract [50]Scrophularia striata Boiss extract [51], los yog hyaluronic acid [52]. Yog li, cov txiaj ntsig no tau lees paub qhov ua tiav kev koom ua ke ntawm glycerol thiab HDE rau hauv BNC daim nyias nyias, tsis muaj pov thawj extract agglomerates tsim.dab tsi yog cistancheQhov kev soj ntsuam no yog ib qho kev qhia txog kev sib raug zoo ntawm cov khoom siv extract (phenolic compounds) nrog cov cellulose fibrils thiab glycerol.

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3.2. Mechanical Performance

Kev ua tau zoo ntawm txhua BNC daim nyias nyias tau soj ntsuam los ntawm kev ntsuas tensile thiab piv nrog cov khoom lag luam BNC-raws li lub ntsej muag lub ntsej muag. Xav tias daim ntawv npog ntsej muag feem ntau yog siv rau hauv lub xeev ntub dej, qhov kev ua haujlwm ntawm cov ntaub ntawv npaj tau raug soj ntsuam hauv ob qho tib si qhuav thiab ntub dej (80 feem pua ​​​​cov dej noo) hais. Kev ntsuam xyuas ntawm qhov kev tiv thaiv thiab elasticity ntawm daim ntawv npog ntsej muag yog qhov tseem ceeb vim tias nws yuav tsum muaj peev xwm tiv taus thiab, tib lub sijhawm, elastic txaus los pab kom haum zoo meej thiab nruj ua raws li daim tawv nqaij, yog li tso cai rau kev nkag mus zoo dua ntawm lub ntsej muag. active compound nyob rau hauv daim tawv nqaij [13] Cov txiaj ntsig ntawm kev sim tensile ntawm cov tawv nqaij qhuav (Daim duab 3A) qhia tau tias kev sib xyaw ntawm glycerol cuam tshuam rau cov khoom ntawm BNC daim nyias nyias los ntawm kev txhim kho, mus rau qhov siab, lawv elasticity. Qhov no tshwm sim los ntawm qhov tseem ceeb ntawm tus nqi ntawm qhov elongation ntawm qhov tawg ntawm 3.6 ± 0.6 feem pua ​​​​rau BNC rau 15.5 ± 1.4 feem pua ​​​​rau BNC-G, nrog rau qhov txo qis hauv Young's modulus (los ntawm 1. 0.1 ± 1.2 GPa rau BNC rau 0.5 ± 0.1 GPa rau BNC-G) thiab hauv lub zog tensile (los ntawm 237 ± 34 MPa rau BNC rau 40 ± 9 MPa rau BNC-G). Cov txiaj ntsig no pom zoo nrog cov kev tshawb fawb yav dhau los hais txog BNC-raws li cov ntaub ntawv sib xyaw glycerol ua cov yas yas [39,49,53]. Raws li tau piav qhia saum toj no, kev koom ua ke ntawm glycerol rau hauv BNC network ua rau txo qis ntawm cov rog hauv nruab nrab ntawm cov cellulose nanofibrils, yog li ua rau qhov chaw inter-fiber ntau ntxiv thiab lawv cov kev txav mus los, ua rau cov tawv nqaij elastic dua [49]. Hais txog kev coj tus cwj pwm ntawm cov ntaub ntub dej (Daim duab 3B), cov nyhuv ntawm qhov sib ntxiv ntawm glycerol ntawm qhov elongation ntawm daim nyias nyias yog tsis pom tseeb.cistanche แอมเวQhov tseeb, BNC qhia qhov zoo tib yam elongation tus nqi rau ntawm BNC-G (BNC: 15.2 ± 1.3 feem pua, BNC-G: 20.8 ± 3.5%), uas tej zaum yuav txuam nrog cov dej molecules. nyob rau hauv BNC network uas tseem yuav ua rau tsis muaj zog interfibrillar bonds [24], ua rau BNC ntub dej stretchable. Txawm li cas los xij, ib yam li hauv daim nyias nyias, qhov txo qis hauv Young's modulus (BNC:0.3±{14}}.0 GPa vs.BNC-G:0.02 ± 0.01 GPa) thiab tensile lub zog (BNC: 26 ± 1 MPa vs. BNC-G: 2 ± 1 MPa) yog pom tom qab glycerol loading.


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Kev koom ua ke ntawm HDE rau hauv BNC daim nyias nyias tsis hloov, hauv txoj hauv kev tseem ceeb, feem ntau ntawm cov khoom siv ntawm cov tawv nqaij qhuav thaum piv nrog BNC-G counterpart. Qhov tshwj xeeb yog qhov nce ntawm tensile lub zog thiab Young's modulus rau BNC-G-HDE2 thiab qhov nce ntawm qhov elongation ntawm qhov tawg ntawm BNC-G-HDE3. Tag nrho BNC-G-HDE membranes qhia siab elongations (11-18 feem pua ​​) ua ke nrog cov nqi me me ntawm Young's modulus ({{10}}}7-1.9 GPa) thiab tensile zog qhov tseem ceeb ntawm 51 thiab 116 MPa. Ib yam li ntawd, hauv lub xeev ntub, nws kuj tau lees paub tias qhov sib ntxiv ntawm HDE tsis cuam tshuam rau feem ntau ntawm cov khoom siv kho tshuab ntawm BNC-G (Daim duab 3B). Cov ntaub ntub dej BNC-G-HDE tseem qhia tau tias muaj qhov siab elongations (19-27 feem pua), qis tus nqi ntawm Young's modulus (0.03-0.06 GPa), thiab tensile zog qhov tseem ceeb ntawm 4 thiab 5 MPa.

KSL06

Muab tag nrho ua ke, cov khoom siv kho tshuab tau tham saum toj no paub meej tias BNC-G-HDE daim nyias nyias yog pliable txaus los tswj thiab moldable rau ntawm daim tawv nqaij, raws li qhov xav tau rau daim ntawv thov npaj. Qhov tseeb, cov txiaj ntsig tau pom tias cov membranes no tau txais txiaj ntsig tiag tiag, tshwj xeeb tshaj yog xav txog kev sib piv nrog cov lag luam BNC daim npog ntsej muag. Qhov qhuav BNC-G-HDE daim nyias nyias muaj qhov zoo ib yam lossis tseem ceeb heev (p<0.05)superior mechanical="" properties="" (e.g.,="" elongation="" at="" break="" of="" dry="" bnc-g-hde3)="" to="" those="" of="" the="" commercial="" bnc="" counterpart="" when="" evaluated="" in="" a="" wet="" state,="" the="" commercial="" bnc="" shows="" higher="" values="" of="" elongation="" at="" break="" and="" tensile="" strength="" than="" those="" observed="" in="" the="" bnc-g-hde="" membranes,="" whereas="" values="" of="" young's="" modulus="" are="" identical.="" differences="" in="" the="" content="" of="" plasticizers="" and="" humectants="" (e.g.,="" glycerol,="" hyaluronic="" acid,="" peg-450)="" can="" explain="" the="" increased="" elasticity="" of="" wet="" commercial="" bnc="" compared="" to="" the="" bnc-g-hde="" membranes="" developed="" in="" the="" present="">

3.3. Thermal stability

Rau kev pom rau hauv thermal stability ntawm BNC-G-HDE qhuav daim nyias nyias, thermogravimetric tsom xam tau ua nyob rau hauv ib qho inert (N,) atmosphere. BNC-G membrane tau soj ntsuam raws li kev tswj hwm. Cov degradation profiles thiab cov derivatives sib raug tau piav qhia hauv daim duab 4. Txhua daim nyias nyias qhia qhov kev poob qis hauv qab 100 degree, suav nrog cov evaporation ntawm cov dej adsorbed los ntawm polymeric matrix, uas lees paub qhov hydrophilicity ntawm BNC thiab hygroscopic xwm ntawm glycerol. Sib nrug los ntawm cov kauj ruam no lub cev qhuav dej, tag nrho cov mem-branes tso saib ob-kauj ruam poob qhov profile. Lub thermal profile ntawm BNC-G membrane qhia ib kauj ruam ntawm kev poob phaus nrog thawj zaug thiab siab tshaj plaws decomposition kub ntawm txog 125 degree thiab 233 degree, feem, feem ntau yog txuam nrog degradation ntawm glycerol, nyob rau hauv kev pom zoo nrog yav dhau los cov txiaj ntsig tau txais lwm qhov nrog plasticized BNC. [49]. Qhov thib ob qhov hnyav-poob theem tshwm sim ntawm thawj zaug thiab siab tshaj plaws decomposition kub ntawm txog 295 degree thiab 355 degree, feem, thiab yog ntaus nqi rau decomposition ntawm cellulose glycosyl units [30].

Hais txog cov membranes sib txuas nrog HDE, cov kauj ruam poob phaus tau muab rau kev puas tsuaj ntawm glycerol pom qhov kub siab tshaj plaws decomposition ntawm ca. 232 degree rau BNC-G-HDE1,224 degree rau BNC-G-HDE1.5,226 degree rau BNC-G-HDE2, thiab 213 degree rau BNC-G-HDE3. Zoo ib yam li BNC-G, qhov kev poob phaus thib ob yog pom ntawm qhov siab tshaj plaws decomposition kub txog 355 degree C rau BNC-G-HDE1, 353 degree C rau BNC-G-HDE1.5, 343 degree rau BNC-G-HDE2, thiab 350 degree rau BNC-G-HDE3, tshuav rau decomposition ntawm BNCpolymeric chains.cistanche tubulosa noj redditLos ntawm cov thermograms, nws tsis tuaj yeem txheeb xyuas qhov kev poob phaus vim qhov kev puas tsuaj ntawm HDE phenolic tebchaw, uas raws li cov ntaub ntawv yuav tshwm sim nyob rau hauv qhov ntau ntawm 100-300 degree [54,55]. Txawm li cas los xij, muab qhov qis qis (<0.1%, w/w)of="" the="" hde="" in="" the="" bnc="" membranes,="" the="" weight="" loss="" associated="" with="" the="" extract="" components="" is="" probably="" not="">

Cov txiaj ntsig no qhia tau tias tag nrho cov membranes ruaj khov mus txog ca. 125 degree, uas yog piv nrog cov txiaj ntsig yav dhau los tau tshaj tawm rau glycerol-plasticized BNC daim nyias nyias [49] thiab tsim nyog rau kev siv lub ntsej muag lub ntsej muag, txiav txim siab nws siv ntawm lub cev kub.

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3.4.Moisture Uptake Capacity

Kev tuav dej noo yog ib qho khoom tsim nyog ntawm daim ntawv npog ntsej muag lub hom phiaj kom tsis txhob muaj lub cev qhuav dej ntawm cov qauv polymeric thiab ua kom lub ntsej muag ua raws li daim tawv nqaij thaum lub sijhawm kho [24]. Nyob rau tib lub sijhawm, qhov o ntawm cov qauv nanoporous ntawm BNC, vim yog dej khi, tseem yuav pab txhawb kev xa khoom ntawm cov khoom sib xyaw ua ke. Yog li, lub peev xwm ntawm cov av noo ntawm cov ntaub ntawv qhuav tau txiav txim siab los ntsuas lawv lub peev xwm los nqus cov av ib puag ncig. Yog li ntawd, cov ntaub so ntswg tau muab tso rau hauv lub desiccator nrog tswj cov av noo (RH ca.52 feem pua) ntawm chav tsev kub, thiab cov av noo tau saib xyuas lub sijhawm, los ntawm 30 min mus txog 48 h (Daim duab 5). RHof txog 52 feem pua ​​​​ntawm kev siv hauv qhov kev ntsuam xyuas yog ze rau tus nqi ntawm 45 ± 5 feem pua, qhia raws li cov lus qhia dav dav rau kev tshawb fawb txog dej nrog cov tshuaj pleev ib ce moisturizing [56].


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All the prepared BNC membranes can absorb environmental humidity, showing an incremental uptake over time and reaching a maximum at 48 h, which is in agreement with a previous work with BNC loaded with silk-sericin [24]. As expected, the incorporation of glycerol, a highly hygroscopic compound, resulted in BNC membranes with a higher aptitude to uptake environmental humidity, even for short periods of time, such as 1 h, with BNC-G membranes absorbing 5.3±0.3% of moisture after this time, and pristine BNC absorbing only 1.3±0.2%. Similar behavior was observed for the BNC-Gmembranes loaded with the HDE, presenting a moisture uptake of 4.4± 0.6% for BNC-G-HD,4.4± 0.3% for BNC-G-HDE1.5,4.2±0.3% for BNC-G-HDE2, and 3.9±0.6% for BNC-G-HDE3 after 1 h. The amount of the HDE seems to have no influence on the moisture uptake capacity of the membranes since the differences are not significant(p>0.05). Considering that the time of treatment of a beauty mask is generally between 20 min and 1 h, the capacity of the BNC-G-HDE membranes to rapidly absorb environmental humidity is fundamental to guarantee the adherence of the BNC sheet to the skin during the entire treatment and favor the release of the active compounds. It is also noteworthy that for the short period of 1 h, the moisture uptake capacity of BNC-G-HDE membranes is identical (p>{{0}}.05) rau qhov uas tau txais rau kev lag luam BNC daim npog qhov ncauj (3.2 ± 1.0 feem pua).

3.5. Hauv Chemico Antioxidant Activity

TPC ntawm E.globulus HDE tau txiav txim siab siv txoj kev Folin-Ciocalteu, ac- suav rau 329 ± 9 mg ntawm GAEg-I ntawm lyophilized extract. Qhov txiaj ntsig no tau lees paub tias muaj cov ntsiab lus siab ntawm phenolic tebchaw hauv cov extract no, piv nrog TPC cov txiaj ntsig yav dhau los qhia rau phenolic extracts ntawm E.globulus nplooj (311 ± 20 mg GAEg-Iof qhuav extract) [57].

KSL07

Phenolic tebchaw yog paub zoo rau lawv cov antioxidant zog uas tshwm sim los ntawm cov scavenging ntawm dawb radicals, uas yog ib tug ntawm cov mechanisms tiv thaiv cellular oxidative kev nyuaj siab [29]. Yog li, cov kev ua haujlwm antioxidant ntawm HDE thiab ntawm BNC daim nyias nyias (BNC, BNC-G, BNC-G-HDE1, BNC-G-HDE1.5, BNC-G-HDE2, thiab BNC-G-HDE3) tau soj ntsuam. los ntawm DPPH dawb radical scavenging method (Daim duab 6). Cov tshuaj tiv thaiv antioxidant tau saib xyuas txog li 2.5 teev, raws li lub sijhawm luv luv ntawm kev siv lub ntsej muag lub ntsej muag thiab cov txheej txheem piav qhia lwm qhov [44,58].

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Cov ntshiab BNC daim nyias nyias tsis muaj qhov cuam tshuam loj heev los tiv thaiv kev ua si, uas pom zoo nrog cov ntaub ntawv qhia hauv kev tshawb fawb yav dhau los [51,59]. Cov txiaj ntsig zoo sib xws tau txais rau BNC-G daim nyias nyias vim glycerol tsis yog cov tshuaj antioxidant. Hais txog HDE aqueous cov kev daws teeb meem, lawv qhia txog cov tshuaj tiv thaiv kab mob antioxidant, uas tuaj yeem raug ntaus nqi rau lawv cov ntsiab lus hauv phenolics, zoo ib yam li cov txiaj ntsig tau tshaj tawm yav dhau los rau aqueous E.globulus nplooj extract [57]. Qhov siab tshaj plaws DPPH scavenging kev ua si tau ua tiav thaum kawg ntawm 2.5h los ntawm cov tshuaj HDE3 (7.5 ug mL-) (75.1 ± 5.8 feem pua). Cov txiaj ntsig no qhia tau hais tias HDE muaj cov tshuaj tiv thaiv antioxidant ntau dua thaum piv rau ethanolic extract ntawm cov khoom lag luam qhuav E.globulus biomass, uas tsuas yog qhia txog DPPHscavenging kev ua ub no ntawm 65 feem pua ​​(ntawm 0.5 h incubation) rau cov extract concentration. Ntawm 250 ug mL-I 【37】 Hais txog BNC-G-HDE daim nyias nyias, cov txiaj ntsig tau pom tias txhua daim nyias nyias muaj cov tshuaj tiv thaiv antioxidant nrog cov lus teb ntawm koob tshuaj uas nce ntxiv raws sijhawm.cistanche salsa cov txiaj ntsigQhov siab tshaj plaws DPPH scavenging kev ua si tau mus txog qhov kawg ntawm 2.5 h, nrog 14.2 ± 1.6 feem pua, 28.1 ± 6.3 feem pua, 48.9 ± 6.1 feem pua, thiab 53.3 ± 5.7 feem pua ​​​​rau BNC-G-HDE1, BNC-G-HDE1.5, BNC-G-HDE2, thiab BNC-G-HDE3 membranes, feem. Yog li ntawd, cov txiaj ntsig tau pom meej meej tias kev koom ua ke ntawm HDE rau hauv BNC network tau ua tiav cov BNC ntshiab nrog kev ua haujlwm antioxidant, zoo ib yam li qhov tau ua tiav hauv kev ua haujlwm dhau los nrog cov extracts ntawm S.striata Boiss [51] lossis Epilobium Angustifolia L.[60] ].

KSL08

These results also demonstrate that the HDE is gradually released from the membranes, and after 2.5 h, the DPPH scavenging activities obtained for all membranes, except for BNC-G-HD3, are identical (p>0.05) rau cov ua tiav rau cov kev daws teeb meem HDE. Qhov txawv (p<0.05)between bnc-g-hde3="" and="" hde3="" samples="" may="" be="" related="" to="" the="" kinetics="" of="" the="" hde="" release="" from="" the="" membrane.="" these="" are="" promising="" results="" regarding="" the="" use="" of="" these="" membranes="" for="" the="" delivery="" of="" the="" active="" compounds="" of="" the="" hde.="" in="" particular,="" the="" bnc-g-hde2="" membrane="" seems="" preferable="" for="" further="" studies,="" because="" it="" shows="" similar="" results,="" in="" terms="" of="" antioxidant="" activity,="" to="" the="" membrane="" with="" the="" highest="" dose="" of="" hde,="" even="" though="" a="" lower="" dose="" of="" hde="" is="" used,="" which="" is="" especially="" relevant="" when="" future="" scale-up="" is="">

3.6.Eoaluation ntawm Stability ntawm BNC-G-HDE2 Membrane nyob rau hauv Cia

Cov yam ntxwv ib puag ncig (xws li, kub, lub teeb, thiab oxygen) paub tias muaj kev cuam tshuam rau cov qauv kev ruaj ntseg ntawm phenolic tebchaw thiab ntawm lawv cov kev ua haujlwm antioxidant 61. Qhov chaw cia kub thiab lub teeb yog ib qho ntawm cov ntsiab lus tshaj tawm uas cuam tshuam nrog kev txo qis. antioxidant kev ua ntawm extracts muaj phenolic tebchaw [62,63]. Li no, raws li cov txheej txheem thoob ntiaj teb ntawm kev soj ntsuam kev ruaj ntseg ntawm cov khoom siv tshuaj pleev ib ce [46], kev tshawb nrhiav kev ruaj ntseg tau ua los ntsuas cov kev hloov pauv hauv cov tshuaj tiv thaiv antioxidant ntawm BNC-G-HDE2 daim nyias nyias nyob rau hauv ib txwm cia tej yam kev mob, xws li ntawm 22-25 degree thiab 52 feem pua ​​​​RH (ib yam mob kuv) thiab nyob rau hauv ceev ceev tej yam kev mob (viz. exaggerated cia tej yam kev mob), uas yog nyob rau ntawm 40 degree thiab 75 feem pua ​​RH ( mob II) rau 3 lub hlis.

Kev soj ntsuam ntawm macroscopic nam ntawm daim nyias nyias (Daim duab 7) qhia tau hais tias tsis muaj kev hloov pauv tseem ceeb hauv lawv cov homogeneity nyob rau 3 lub hlis ntawm kev cia nyob rau hauv ob qho tib si cia.

Results from the antioxidant activity of the BNC-G (control) and BNC-G-HDE2 mem-branes over the 3 months of storage in each condition are depicted in Figure 8. As in the assay of the antioxidant activity of BNC-G-HDE membranes(Section 3.5), the values obtained for the antioxidant activity of BNC-G are not significant. Regarding the BNC-G-HDE2 membrane, when it is stored at 22-25°C and 52% RH (Figure8A), the antioxidant activity is kept unchanged over the entire assay(p>0.05), showing values of 37.0±6.9%in the beginning of the assay and 39.6± 2.4% after 3 months. Similar results were obtained after storage at accelerated conditions(Figure 8B) of 40°C and 75%RH, in which the variation in the antioxidant activity after 1 month (31.7±7.5%)and after 3 months (31.0±5.2%)is not significantly different(p>0.05) los ntawm qhov pib ntawm qhov kev xeem. Yog li, cov txiaj ntsig no qhia tau hais tias cov tshuaj tiv thaiv antioxidant ntawm BNC-G-HDE2 yog qhov ruaj khov rau tsawg kawg 3 lub hlis nyob rau hauv cov xwm txheej khaws cia, txhais tau hais tias tsis muaj qhov pom tseeb degradation ntawm phenolic tebchaw los ntawm HDE tshwm sim hauv daim nyias nyias. Ntxiv mus, nrog rau cov txiaj ntsig tau zoo nyob rau hauv cov xwm txheej ceev, nws tau kwv yees tias kev ruaj ntseg ntawm daim nyias nyias yuav ntev dua 3 lub hlis nyob rau hauv cov xwm txheej cia.


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3.7. Hauv Vitro Biological Assay

3.7.1 ib. Cytotoxicity Assay

Kev ntsuam xyuas ntawm kev nyab xeeb ntawm BNC-raws li daim nyias nyias yog qhov tseem ceeb rau lawv daim ntawv thov ua lub ntsej muag lub ntsej muag vim qhov no qhia txog lawv qhov kev sib cuag ncaj qha rau ntawm daim tawv nqaij. Yog li ntawd, cytotoxicity ntawm BNC-G (rau kev tswj hwm) thiab plaub BNC-G-HDE daim nyias nyias tau soj ntsuam siv MTT kev soj ntsuam, nrog rau kev rho tawm ntawm daim nyias nyias, rau tib neeg keratinocytes (HaCaT) thiab nas fibroblasts (NIH/3T3) hlwb. . Cov kab ntawm tes no tau raug xaiv los ua tus sawv cev ntawm ob qho tseem ceeb ntawm cov qauv ntawm tes ntawm daim tawv nqaij. Keratinocytes yog cov hlwb tseem ceeb ntawm cov epidermis (cov tawv nqaij saum toj kawg nkaus), thaum fibroblasts yog ib feem ntawm dermis (thib ob ntawm daim tawv nqaij) [64].

Cov txiaj ntsig tau txais rau BNC-G daim nyias nyias (Daim duab 9) yog nyob rau hauv txoj kab nrog cov paub zoo uas tsis yog-cytotoxicity ntawm ntshiab BNC rau HaCaT [52,65] thiab NIH/3T3 [65,66] hlwb thiab ntawm BNC plasticized nrog glycerol ntawm HaCaT hlwb [39]. Hais txog cov txiaj ntsig ntawm cytotoxicity ntawm BNC daim nyias nyias ntim nrog HDE, nws yog qhov pom tau tias, ntawm nws tus kheej ntawm HDE koob, cov membranes tsis muaj qhov cuam tshuam loj rau kev muaj peev xwm ntawm HaCaT lossis NIH / 3T3 hlwb. Txhua qhov txiaj ntsig ntawm cell viability tau rau BNC-G-HDE daim nyias nyias yog siab dua 70 feem pua ​​​​thiab yog li suav tias tsis yog-cytotoxic rau cov kab ntawm tes, raws li tus txheej txheem thoob ntiaj teb ISO10993-5 rau kev soj ntsuam lom ntawm cov khoom siv kho mob (Part 5 : Kev kuaj rau hauv vitro cytotoxicity). Yog li, cov txiaj ntsig no yog qhov kev lees paub tias kev koom ua ke ntawm cov koob tshuaj no ntawm E.globulus HDE rau hauv BNC yog kev nyab xeeb kiag li rau daim ntawv thov dermal. Tseeb, cov ntaub ntawv no tau pom zoo nrog cov txiaj ntsig tau tshaj tawm nyob rau lwm qhov uas qhia qhov tsis yog-cytotoxicity ntawm ethanolic extract los ntawm E.globulus biomass (1, 10, thiab 100 ug mL-1) kuaj ntawm tib neeg dermal fibroblasts [37].

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3.7.2 ib. Anti-Senescence Activity

Cov txheej txheem kev laus ntawm daim tawv nqaij tau cuam tshuam tsis tau tsuas yog nrog kev hloov pauv hauv cov khoom sib xyaw thiab kev ncaj ncees ntawm cov kab mob dermal extracellular matrix (ECM) [67] tab sis kuj nrog kev nce ntawm senescent hlwb hauv dermal txheej [68]. Kev sib sau ntawm senescent dermal fibroblasts tam sim no tau lees paub tias ua lub luag haujlwm tseem ceeb hauv cov txheej txheem ntawm daim tawv nqaij laus, ua rau kev poob ntawm cov ntaub so ntswg ua haujlwm [69,70].

Hauv qhov kev nkag siab no, kev ua haujlwm los tiv thaiv kev ua haujlwm ntawm BNC-G-HDE2 daim nyias nyias tau soj ntsuam ntawm nas fibroblasts (NIH / 3T3 cell kab). BNC-G-HDE2 daim nyias nyias raug xaiv rau qhov kev ntsuam xyuas no vim, ua ke nrog BNC-G-HDE3, nws qhia tau hais tias ib qho ntawm cov kev ua haujlwm antioxidant siab tshaj plaws, thiab vim tias qhov tsawg dua ntawm HDE yog siv hauv BNC-G-HDE2, nws yog preferable hais txog kev siv nyiaj txiag yav tom ntej.

Cellular senescence raug ntxias nrog genotoxic tus neeg sawv cev etoposide, thiab cov kev ua ntawm -gal, ib tug dav siv biomarker rau senescent hlwb [71], raug ntsuas. Raws li cov ntaub ntawv nthuav tawm hauv daim duab 10, tsis yog BNC-G lossis BNC-G-HDE2 daim nyias nyias ua kom muaj kev sib raug zoo-gal ntawm NIH/3T3 hlwb. Ntawm qhov tsis sib xws, raws li kev kho mob etoposide, tus naj npawb ntawm senescence-associated-gal-positive (xiav staining) NIH/3T3 hlwb signifi-cantly nce piv rau kev tswj (44.1 ± 2.7 feem pua ​​​​vim 8.9±1.0 feem pua ​​) . Qhov no kuj pom nyob rau hauv qhov tau txais morphology ntawm NIH / 3T3 hlwb, uas qhia cov yam ntxwv senescent phenotype nrog loj thiab flattened hlwb (Daim duab 10A). Pom zoo, tom qab 24 teev ntawm kev kho mob nrog BNC-G-HDE2 extract, muaj qhov txo qis ntawm cov senescent NIH / 3T3 hlwb (32.2 ± 4.0 feem pua) (Daim duab 10B). Qhov txiaj ntsig zoo no muaj tseeb rau cov khoom ntawm HDE, uas yog cov tshuaj phenolic uas tau tso tawm los ntawm daim nyias nyias. Qhov tseeb, phenolic tebchaw twb tau pom tias muaj txiaj ntsig zoo ntawm kev tiv thaiv cellular senescence los ntawm suppressing senescence-sociated -gal, feem ntau yog vim lawv lub luag haujlwm paub tias yog inhibitors ntawm oxidative puas tsuaj [72,73]. Lwm cov piv txwv nyob rau hauv txoj kab nrog cov txiaj ntsig tau txais ntawm no yog cov kev tshawb pom ntawm cov teebmeem ntawm Sonchus oleraceus L. nplooj extract (muaj caftaric acid, chlorogenic acid, thiab chicoric acid) ntawm H2O2-induced cell senescence hauv WI{{45 }} tib neeg lub ntsws diploid fibroblast hlwb [74], los yog ntawm cov txiv ntseej phenols (hydroxytyrosol thiab oleuropein) ntawm pre-senescent tib neeg dermal thiab ntsws fibroblasts [72]Txawm li cas los xij, raws li peb paub, qhov no yog thawj zaug uas qhov haujlwm no tau ua. qhia rau BNC daim nyias nyias enriched nrog cov nroj tsuag rho tawm. Yog li, cov no yog cov txiaj ntsig tau pom zoo qhia txog qhov muaj peev xwm loj ntawm BNC-G-HDE daim nyias nyias rau kev siv tshuaj tiv thaiv kev laus ntawm daim tawv nqaij.

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4. Cov lus xaus

Lub hom phiaj ntawm kev txhim kho cov khoom siv bio-raws li tag nrho nrog cov khoom siv bioactive thiab cov khoom tsim nyog rau kev siv tshuaj tiv thaiv kev laus ntawm daim tawv nqaij, BNC mem-branes loaded nrog glycerol thiab ntau koob tshuaj (1-3 ug cm-2) ntawm ib qho aqueous E.globulus extract tau los ntawm hydro-distillation (HDE) ntawm nplooj uas muaj phenolic tebchaw (329 ± 9 mg GAE gI ntawm lyophilized extract) tau npaj los ntawm txoj kev impregnation. Tag nrho BNC-G-HDE daim nyias nyias tau pom tias muaj kev tiv thaiv tab sis tseem muaj kev ywj pheej ntawm cov neeg kho tshuab, zoo ib yam (ntawm ob lub xeev qhuav thiab ntub dej) rau ntawm kev lag luam BNC lub ntsej muag lub ntsej muag. Lub peev xwm ntawm BNC-G-HDE daim nyias nyias los ua kom cov dej noo ib puag ncig yog siab dua li ntawm BNC ntshiab thiab kuj piv rau cov lag luam BNC daim npog ntsej muag. BNC-G-HDE daim nyias nyias tau pom tias muaj thermally ruaj khov txog li 125 degree. Tsis tas li ntawd, kev koom ua ke ntawm HDE muab cov tshuaj tiv thaiv kab mob antioxidant (DPPH radical scavenging assay) rau BNC. Kev ruaj ntseg ntawm cov tshuaj tiv thaiv antioxidant ntawm daim nyias nyias nrog 2 ug cm -2 ntawm HDE tau ua pov thawj tom qab cia ntau tshaj 3 lub hlis ntawm qhov xav tau ib txwm (22-25 degree / 52 feem pua ​​RH) thiab nrawm (40C / 75 feem pua ​​RH ) khaws cia tej yam kev mob. Tag nrho BNC-G-HDE daim nyias nyias tau pom tias tsis yog-cytotoxic rau HaCaT thiab NIH/3T3 cell kab. Ntxiv mus, kev kho mob nrog BNC-G-HDE2 daim nyias nyias tau pom tias ua rau txo qis ntawm feem pua ​​​​ntawm cov hlwb senescent (ca. 32 feem pua) piv rau qhov pom hauv cov hlwb kho nrog etoposide (ca. 44 feem pua). Cov kev tshawb pom tau tshaj tawm ntawm no kom meej meej qhia txog lub peev xwm ntawm kev npaj HDE-loaded BNC daim nyias nyias raws li daim ntawv npog lub ntsej muag zoo rau kev saib xyuas tawv nqaij.

Kab lus no yog muab rho tawm los ntawm Cov Ntaub Ntawv 2022, 15, 1982. https://doi.org/10.3390/ma15051982 https://www.mdpi.com/journal/materials






















































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