Jabuticaba (Myrciaria Jaboticaba) Peel As A Sustainable Source Of Anthocyanins Thiab Ellagitannins Xa Los Ntawm Phospholipid Vesicles Rau Alleviating Oxidative Stress hauv Tib Neeg Keratinocytes Part 2

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

2.3. Hauv Vitro Studies nrog Keratinocytes

2.3.1.Biocompatibility ntawm Vesicles

Txoj kev tshawb no tau ua los soj ntsuam cov physicochemical thaj chaw ntawm kev hloov pauv, tshwj xeeb tshaj yog cov uas muaj cov khoom siv polymers, paub tseeb tias lawv cov kev ruaj ntseg ntau dua thiab zoo tagnrho rau siv rau ntawm daim tawv nqaij. Txhua qhov sib txawv tseem ceeb tau raug kuaj pom raws li kev ua haujlwm ntawm cov khoom siv polymer (1 lossis 2 mg / mL). Txhawm rau ntsuas qhov no, cov hlwv tau hloov kho nrog qhov siab tshaj plaws polymer concentration (2 mg / mL) tau siv los ua cov kev tshawb fawb tom ntej. Lawv biocompatibility raug soj ntsuam siv keratinocytes thiab cov extract hauv aqueous dispersion raug soj ntsuam thiab siv los ua kev sib piv. Keratinocytes tau raug xaiv raws li lawv yog cov neeg sawv cev tshaj plaws ntawm cov epidermis thiab txheej txheej txheej txheej tau siv txij li hauv lawv qhov tshwj xeeb sib txawv lawv tsim lub ntsiab thaiv ntawm daim tawv nqaij, uas tswj cov tawv nqaij hydration thiab tiv thaiv exogenous tshuaj los ntawm kev nkag mus rau hauv thiab dhau ntawm nws. Keratinocytes raug kho rau 48 h nrog cov extract nyob rau hauv dispersion los yog incorporated nyob rau hauv lub vesicles ntawm plaub sib txawv dilutions, tom qab uas lub cell viability yog ntsuas (Daim duab 3). Lub viability ntawm cov hlwb incubated nrog cov extract hauv dispersion yog ~ 88 feem pua ​​(p<0.05 versus="" the="" viability="" measured="" using="" extract="" loaded="" vesicles),="" indicating="" that="" the="" extract="" is="" not="" toxic.="" the="" viability="" of="" cells="" treated="" with="" extract-loaded="" transfersomes="" and="" hyaluronan-transfersomes="" at="" higher="" dilutions="" was~100%="" (p="">0.05 ntawm pawg no). Kev kho mob nrog HEcellulose-transfersomes thiab hyaluronan-transfersomes ntxiv txhim kho cell viability. Tag nrho cov formulations tau zoo heev biocompatible tsis hais cov polymer thiab dilution siv. Tseeb tiag, lub cell viability tau sib npaug los yog siab dua 100 feem pua, txawm tias nthuav tawm cov txiaj ntsig zoo cuam tshuam nrog kev thauj khoom vesicle.

Thov nias ntawm no kom paub ntxiv

2.3.2. Kev tiv thaiv cov nyhuv ntawm cov Extract, nyob rau hauv Dispersion los yog Loaded nyob rau hauv vesicles, tiv thaiv kev puas tsuaj tshwm sim nyob rau hauv Keratinocytes los ntawm Hydrogen Peroxide

Cov keratinocytes tau ntxhov siab nrog hydrogen peroxide thiab tom qab ntawd kho nrog cov extract txawm nyob rau hauv dispersion los yog loaded rau hauv transfersomes. Lub cell viability tau ntsuas ntawm 4 h thiab qhov no tau siv los ntsuas qhov kev tiv thaiv ntawm cov formulations tiv thaiv oxidative kev nyuaj siab (Daim duab 4). Cov qauv no tau diluted nrog cov cell nruab nrab kom ncav cuag ob qhov sib txawv (4 thiab 0.4 ug/mL). Qhov kev ntxhov siab ntawm hydrogen peroxide ua rau muaj kev tuag ntawm tes thiab txo kev muaj peev xwm mus txog ~ 50 feem pua ​​(p<0.05 versus="" the="" viability="" of="" cells="" treated="" with="" the="" extract="" in="" dispersion="" or="" loaded="" in="" vesicles)[35].="" hydrogen="" peroxide="" is="" considered="" one="" of="" the="" most="" dangerous="" oxidative="" molecules="" among="" the="" different="" reactive="" oxygen="" species,="" capable="" of="" promoting="" apoptosis="" and="" cell="" death.="" the="" treatment="" of="" stressed="" cells="" with="" the="" aqueous="" dispersion="" of="" the="" extract="" was="" capable="" of="" reducing="" the="" damaging="" effect="" of="" hydrogen="" peroxide="" as="" the="" viability="" increased="" up=""><0.05 versus="" the="" viability="" of="" cells="" treated="" with="" the="" extract="" loaded="" in="" hecelluose-transfersomes="" at="" a="" higher="" dilution="" and="" hyaluronan-transfersomes),="" although="" the="" complete="" restoration="" of="" normal="" conditions="" was="" not="" achieved="" (figure="" 4).="" treatment="" with="" the="" extract-loaded="" transfersomes="" protected="" the="" cell="" to="" the="" same="" extent="" as="" the="" extract="" in="" dispersion="" (~82%,p="">0.05 piv rau kev muaj peev xwm ntawm cov hlwb kho nrog dispersion). Kev kho mob nrog cov txiaj ntsig zoo tshaj plaws yog cov extract loaded nrog HEcellulose-transfersomes (ntawm ntau dua dilution) thiab hyaluronan-transfersomes, uas ua tiav qhov muaj txiaj ntsig ntawm ~ 104 feem pua ​​(p<0.05 versus="" the="" values="" of="" other="" treatments).="" the="" restored="" normal="" conditions="" and="" even="" slightly="" promoted="" cell="" proliferation,="" probably="" due="" to="" the="" synergic="" effect="" of="" the="" extract="" and="" the="" polymer-immobilized="" vesicles.="" the="" vesicle="" behavior="" was="" not="" affected="" by="" the="" dilution="" levels="" of="" the="">

2.3.3. Hauv Vitro Wound Healing Effects

Kev soj ntsuam hauv vitro khawb tau ua nrog monolayer ntawm keratinocytes txhawm rau txheeb xyuas lub peev xwm ntawm cov extract hauv aqueous dispersion los yog muab tso rau hauv cov hlab ntsha los txhawb kev loj hlob thiab kev tsiv teb tsaws ntawm cov hlwb. Cov hlwv khoob tsis raug kuaj raws li hauv kev tshawb fawb yav dhau los tsis muaj kev txhim kho hauv qhov txhab kho mob thaum siv hyalurosomes khoob [37]. Qhov kaw ntawm qhov txhab ua tau raug saib xyuas rau 48 h (Daim duab 5) thiab qhov feem pua ​​​​ntawm kev kaw tau suav los ntawm kev ntsuas qhov chaw ntawm qhov txhab (Daim duab 6). Qhov txhab kaw ntawm cov hlwb tsis kho tau tshwm sim qeeb heev - 13 feem pua ​​​​ntawm 24 teev, 28 feem pua ​​​​ntawm 36 teev, thiab tsuas yog 40 feem pua ​​​​ntawm 48 teev. Kev kho nrog cov extract hauv dej dispersion me ntsis txhim kho cov txheej txheem thiab coj qhov kaw ntawm 48 h rau 50 feem pua. Kev kho mob nrog cov vesicles extract-loaded ntawm 48 h ua tiav 90 feem pua ​​​​kaw thaum siv cov hloov pauv thiab HEcellulose-transfersomes thiab nyob ib ncig ntawm 100 feem pua ​​​​kaw thaum siv hyaluronan-transfersomes. Raws li peb tuaj yeem pom, lub qhov txhab kaw yuav luag tiav, paub meej tias cov khoom siv rov qab los ntawm cov khoom siv hyaluronan-transfersomes.

Peb tag nrho cov txiaj ntsig tau lees paub qhov siab biocompatibility ntawm hyaluronan-transfersomes, uas kuj yog qhov zoo tshaj plaws ntawm kev txhawb nqa kev loj hlob thiab kev tsiv teb tsaws ntawm cov hlwb ntawm daim tawv nqaij thiab tiv thaiv kev puas tsuaj ntawm daim tawv nqaij los ntawm oxidative kev nyuaj siab.

3. Cov ntaub ntawv thiab cov txheej txheem 3.1. Khoom siv

Lipoid S75 (muaj li ntawm ~ 70 feem pua ​​​​ntawm cov kua phosphatidylcholine, 9 feem pua ​​​​phosphatidylethanolamine, thiab 3 feem pua ​​​​lyso-phosphatidylcholine) tau yuav los ntawm Lipoid GmbH (Ludwigshafen, Ger-ntau).Sodium hyaluronate nrog qis molecular (k {6}) thiab polydispersity ntawm 1.4 Mw / Mn, tau yuav los ntawm DSM Nutritional Products AG Branch Pentapharm (Switzerland). Tween 80, glycerol, DPPH radical (2, 2-diphenyl-1- picrylhydrazyl), ferric chloride hexahydrate, gallic, chlorogenic, thiab ascorbic acids, catechin, tetrazolium ntsev, 3-(4, { {15}}dimethylthiazol-2-vl)-2,5-diphenyltetrazolium bromide(MTT), quercetin, Neosporin, Folin-Ciocalteu's phenol reagent, 2,4, 6-tris( 2-pyridyl)-S-triazine(TPTZ) thiab pvrocatechol violet tau yuav los ntawm Sigma-Aldrich (Milan, Ltalis). Potassium hexacyanoferrate (ⅢI) tau txais los ntawm Merck (Darmstadt, Lub Tebchaws Yelemees). Anhydrous sodium acetate, tooj liab sulfate pentahydrate thiab tooj liab chloride dihydrate tau txais los ntawm VWR Chemi-cals BDH⑧ (Frankfurt, Lub teb chaws Yelemees). Cyanidin 3-O-glucoside ( Ntau dua lossis sib npaug li 95 feem pua ​​) thiab delphinidin 3-O-glucoside ( Ntau dua lossis sib npaug li 95 feem pua ​​) tau yuav los ntawm Extrasynthese (Lyon, Fabkis). Glacial acetic acid thiab phosphoric acid (85 feem pua) tau txais los ntawm JTBaker (Mallinckrodt Baker Inc., Utrecht, Lub Netherlands). Txhua reagents thiab plastics rau kab lis kev cai ntawm tes tau yuav los ntawm Life Technologies Europe (Monza, Ltalis).

3.2. Jabuticaba Pees: Extraction, Chemical Characterization, thiab Antioxidant Activity

Jabuticaba txiv hmab txiv ntoo (Myrciaria jabuticaba (Vell.)O.Berg)cv. Sabara tau sau thaum lub sijhawm siav hauv Araucaria lub nroog, Paranå, Brazil (geographical coordinates∶25 degree 2924.7"S49 degree 2637.8"W) thaum lub Kaum Ob Hlis 2018. Txiv hmab txiv ntoo raug ntxuav thiab ntxuav (NaOCl ntawm 100 mg/L/15 min), yaug, thiab pulped manually. Peels tau qhuav ntawm 35 degree degree rau 50 h, hauv av kom ncav cuag 42 Tyler mesh, thiab muab rho tawm hauv cov kuab tshuaj nrawm nrawm (ASE{12}}, Dionex, Sunnyvale, CA, USA), ua haujlwm siab ntawm 100 bar (98.7). atm) nyob rau hauv ob lub voj voog rho tawm ntawm 50 degree . Dej acidified nrog citric acid (pH 2.10) tau siv los ua cov kuab tshuaj thiab cov extractions tau rov ua plaub zaug. Tom qab ntawd, cov extracts tau lim siv cov ntawv zoo thiab khov-qhuav hauv lub tshuab nqus tsev rau 120 teev. Tom qab ntawd, qhov kev rho tawm tawm, qhia raws li feem pua, tau suav txog cov khoom siv raw siv hauv cov txheej txheem.

Cistanche tuaj yeem txhim kho kev tiv thaiv

Tag nrho cov ntsiab lus phenolic (TPC, mg ntawm gallic acid sib npaug ib 100 g, mg GAE / 100 g) tag nrho condensed tannin (TCT, mg ntawm catechin sib npaug ib 100 g, mg CE / 100 g), tag nrho flavonoids (TF, mg CE / 100 g) thiab tag nrho cov ntsiab lus ortho-diphenols (TOD, mg ntawm chlorogenic acid sib npaug ib 100 g, mg CAE / 100 g) tau ntsuas hauv triplicate los ntawm kev siv UV-Vis spectrophotometry raws li cov txheej txheem uas tau piav qhia los ntawm Carmo li al. [20].

Anthocyanins (cyanidin {{0}}}O-glucoside thiab delphinidin 3-O-glucoside) raug ntsuas los ntawm kev ua haujlwm siab ua kua chromatography (HPLC) siv Agilent 110{{37} }(Agilent Technologies Inc., Espoo, Finland) ntaus ntawv nruab nrog diode array detection (DAD). Kev sib cais tau ua tiav siv Gemini C18 kem (150 mm × 4.6 mm, 5 μm) nrog gradient elution ntawm acetonitrile acidified nrog formic acid ntawm 5 feem pua. Tag nrho cov ntsiab lus ellagitannin tau txiav txim siab tom qab cov kua qaub hydrolysis, raws li Mattila thiab Kumpulainen [38], thiab gallic acid tau txiav txim siab siv Inertsil ODS{13}} kem (150 mm × 4.0 mm , 3 μm) nrog gradient elution ntawm acetonitrile rau hauv 50 mmol / L H3PO4 (pH 2.5). Kev tshuaj xyuas tau ua nyob rau hauv triplicate thiab cov txiaj ntsig tau qhia tias mg / 100 g. Ultra-high performance kua chromatography (UHPLC) ua ke nrog high-resolution mass spectrometry (MS) tau thov rau cov yam ntxwv ntawm loj ellagitannins thiab anthocyanins. Ib qho Acquity UPLC-Xevo G2 QTOF mass spectrometer (Waters, Milford, MA, USA) tau nruab nrog Waters Acquity BEH C18 (1.7 μm, 2.1 mm × 150 mm) kem thiab kev sib cais tau ua los ntawm kev siv gradient ntawm acetonitrile rau hauv dej acidified. nrog 0.1 feem pua ​​​​ formic acid, raws li Santos li al. [39] ndlwg npaum li cas yog 0.55 mL / min, qhov kub ntawm lub qhov cub yog 45 degree thiab qhov ntim txhaj tshuaj yog 1.0 μL. Ib qho electrospray interface (ESI) hauv qhov tsis zoo. thiab hom zoo tau siv nrog capillary voltages ntawm -1 kV thiab ntxiv rau 0.5 kV, ntsig txog. Argon tau siv los ua cov roj sib tsoo. MS kev tshuaj xyuas tau ua los ntawm cov ntaub ntawv ywj pheej acquisition (MSE) centroid cov ntaub ntawv qauv nyob rau hauv tag nrho scan m /z 50-1500 nrog 0.2 s scan lub sij hawm. Nyob rau hauv lub MSE muaj nuj nqi, lub precursor ions ntawm MS tau fragmented siv high collision zog ramped li ntawm 25 mus rau 45 V.

Kev ua haujlwm antioxidant ntawm cov extract tau txheeb xyuas hauv triplicate los ntawm kev siv cov kev ntsuas sib txawv: kev ua haujlwm dawb-radical scavenging hauv kev sib raug zoo rau DPPH radical, ferric txo antioxidant zog (FRAP), cupric-ion txo antioxidant peev (CUPRAC), txo hwj chim, thiab Cu2. ntxiv rau chelate muaj peev xwm. Cov txiaj ntsig tau qhia tias mg GAE / 100 g (txo lub zog), mg ntawm ascorbic acid sib npaug rau 100 g, mg AAE / 100 g (DPPH, FRAP, thiab CUPRAC), thiab mg EDTA sib npaug / 100 g (hlau chelation), feem. . Txhua txoj kev siv tau piav qhia tob dhau los ntawm kev ua Carmo li al. thiab Fidelis et al. [9, 20].

3.3. Kev npaj Vesicle

Transfersomes, hydroxyethylcellulose enriched transfersomes (HEcellulose-thansferomes), thiab sodium hyaluronate enriched transfersomes (hyaluronan-transfersomes) tau npaj los ntawm dispersing phospholipid (S75, 180 mg / mL), Tween 80 (20 mg / mL), (40 mg / mL) thiab polymer (hydroxyethylcellulose los yog sodium hyaluronate 1 thiab 2 mg / mL), thaum tsim nyog, hauv dej thiab tawm hauv qhov sib tov kom hydrate rau ob peb teev. Tom qab no, cov dispersions tau sonicated (4 cycles 2 ntawm 5 tawm 15.0μ amplitude), tos 5 min ntawm txhua lub voj voog los txhawb kom txias thiab tsis txhob overheating ntawm tus qauv. Ib qho kev ua tau zoo Soniprep 150 sonicator (MSE Crowley, London, UK) tau siv los sonicate tag nrho cov dispersions thiaj li yuav tau ib tug homogeneous system nrog ib tug me me [15]. Cov txheej txheem no zam qhov poob ntawm cov khoom siv rau kev npaj, ua rau kev tsim cov vesicles ua tau zoo uas muaj peev xwm suav nrog cov nyiaj ntau ntawm cov extract.

3.4. Characterization ntawm Vesicles

Qhov nruab nrab txoj kab uas hla thiab polydispersity Performance index (ib tug dimensionless ntsuas ntawm qhov dav ntawm qhov loj me me) ntawm cov hlab ntsha tau ntsuas los ntawm photon correlation spectroscopy siv Zetasizer Ultra (Malvern Instrument, UK). Tib cov cuab yeej siv los ntsuas qhov zeta muaj peev xwm ntawm vesicles los ntawm kev ntsuas cov electrophoretic txav ntawm cov khoom [40]. Cov qauv tsim nyog tau diluted ua ntej ntsuas kom pom qhov pom tseeb thiab zam kom tsis txhob poob ntawm lub teeb tawg uas tuaj yeem kuaj pom. Txhawm rau ntsuas tus nqi ntawm cov extract tau muab tso rau hauv vesicles, cov dispersions tau purified los ntawm cov extract tsis koom nrog los ntawm kev lim ntshav. Vesicle dispersions (1 mL) tau muab tso rau hauv polycarbonate dialysis raj (Spectra/Pormembranes:12-14 kDa MW txiav tawm, nrog pores 3 nm; Spectrum Laboratories Inc., Rancho Dominguez, CA, USA) thiab muab tso rau hauv dej distilled. (2 L) ntawm 25 degree rau 2 teev nyob rau hauv stirring. Cov dej tau rov ua dua tom qab ib teev, yog li siv 4 L ntawm cov dej los solubilize cov bioactive molecules tsis suav nrog hauv 1 mL ntawm vesicle dispersion (40 mg). Tus nqi ntawm cov extract nyob rau hauv lub vesicle suspensions ua ntej thiab tom qab kev lim ntshav tau ntsuas ntsuas lawv cov tshuaj tiv thaiv antioxidant siv DPPH assay. Qhov kev ua haujlwm zoo ntawm cov extract hauv cov hlab ntsha tau suav nrog qhov feem pua ​​​​ntawm cov kev ua haujlwm antioxidant ntawm cov qauv ua ntej thiab tom qab cov txheej txheem purification [41-43].

3.5. Stability ntawm Vesicles ntawm Cia

Kev ruaj ntseg ntawm cov hlab ntsha hauv qhov tawg tau raug soj ntsuam los ntawm kev saib xyuas lawv qhov nruab nrab qhov loj me, qhov ntsuas polydispersity, thiab tus nqi saum npoo rau 90 hnub thaum khaws cov dispersions ntawm chav tsev kub (25 ± 1 degree).

3.6.Measurement of Antioxidant Activity of Samples Siv DPPH Colorimetric Test Cov tshuaj tua kab mob antioxidant ntawm jabuticaba tev extract loaded rau hauv vesicles tau ntsuas raws li kev ua haujlwm ntawm nws lub peev xwm los khawb DPPH. Cov dispersions (10 μL) tau diluted (1∶100) nrog cov tshuaj methanolic ntawm DPPH (0.4 ug / mL). Cov qauv diluted tau muab khaws cia rau hauv chav sov thiab hauv qhov tsaus ntuj rau 30 min, tom qab ntawd qhov nqus ntawm cov tshuaj tau ntsuas ntawm 517 nm siv lub UV spectrophotometer. Cov kev ua antioxidant ntawm cov formulations raug xam raws li qhov sib npaug (1): (1)Antioxidant kev ua feem pua ​​=[(ABSDPPH-ABSample)/ABSpPPH] × 100

3.7.Biocompatibility thiab tiv thaiv cov nyhuv ntawm cov qauv tiv thaiv Oxidative Stress hauv Keratinocytes

Immortalized human keratinocytes (HaCaT) tau loj hlob raws li monolayers ntawm 37 degree, nrog 100 feem pua ​​​​cov av noo thiab 5 feem pua ​​​​CO2 siv Dulbecco's Modified Eagle Medium (DMEM) cov qabzib siab ntxiv nrog fetal bovine serum, penicillin thiab streptomycin ua ib qho kev loj hlob nruab nrab. Txhawm rau ntsuas qhov biocompatibility ntawm formulations, cov hlwb tau muab noob rau hauv 96- cov phiaj zoo ntawm qhov ntom ntawm 7.5 × 10 degree cell/well. Tom qab 24 teev, hlwb raug kho rau 48 teev nrog jabuticaba extract nyob rau hauv aqueous dispersion los yog loaded nyob rau hauv vesicles kom diluted nrog DMEM kom ncav cuag txawv concentrations (40, 4, 0.4, thiab 0.04 ug / mL). Thaum kawg ntawm qhov incubation, MTT [3(4,{18}}dimethyl thiazolyl-2)-2, 5-diphenyltetrazolium bromide] (100 μL, 0.5 mg / mL zaum kawg concentration ) tau ntxiv rau txhua qhov dej, thiab, tom qab peb teev, cov ntaub ntawv tsim formazan tau yaj nrog dimethyl sulfoxide. Qhov nqus ntawm txhua qhov dej tau ntsuas ntawm 570 nm siv tus nyeem ntawv microplate (Synergy 4 Reader, BioTek Instruments, AHSI SpA, Bernareggio, Ltalis). Txhua qhov kev sim tau rov ua dua tsawg kawg peb zaug, txhua zaus hauv triplicate. Cov txiaj ntsig tau pom raws li feem pua ​​​​ntawm cov cell viability nyob rau hauv kev sib piv nrog cov kev tswj tsis tau kho (100 feem pua ​​​​ntawm kev muaj sia nyob).

Cov kev tiv thaiv hauv vitro ntawm cov tshuaj tiv thaiv kev puas tsuaj los ntawm oxidative kev nyuaj siab tau raug soj ntsuam thiab. Cells tau muab noob rau hauv {{0}}cov phiaj zoo ntawm qhov ceev ntawm 7.5 × 10cells/well. Tom qab 24 teev ntawm incubation, cov hlwb raug kev nyuaj siab nrog hydrogen peroxide (30 feem pua ​​​​ diluted 1:40, 000 v/o nrog PBS) thiab kho nrog cov extract nyob rau hauv aqueous dispersion los yog loaded nyob rau hauv vesicles thiab diluted mus txog ob qhov sib txawv concentrations (4 thiab 0.4 ug / ml,). Cov cell uas muaj kev ntxhov siab nrog hydrogen peroxide tsuas yog siv los ua kev tswj tsis zoo, thaum cov hlwb tsis kho tau siv los ua kev tswj xyuas zoo. Tom qab 4 teev ntawm incubation, hlwb raug ntxuav nrog ib tug tshiab nruab nrab thiab lawv muaj peev xwm raug txiav txim los ntawm MTT assay. Cov txiaj ntsig tau tshaj tawm raws li feem pua ​​​​ntawm cov hlwb tsis kho (100 feem pua ​​​​ntawm kev muaj sia nyob).

3.8. Hauv Vitro Wound Healing Properties

Lub peev xwm ntawm jabuticaba tev extract loaded rau hauv vesicles los kho cov kab mob ntawm daim tawv nqaij thiab txhawb lawv txoj kev kho tau raug soj ntsuam los ntawm kev ntsuas lub cell expansion ntawm qhov txhab ntawm lub cell monolayer. Cov hlwb raug coj mus rau hauv 6 qhov zoo daim hlau kom txog thaum ua tiav thiab homogeneous monolayer tau mus txog. Lub qhov txhab linear yog tsim los ntawm kev siv lub tshuab tsis muaj menyuam yas pipette taub. Cov tawg tawg tawg ntawm cov hlwb raug tshem tawm los ntawm kev ntxuav maj mam nrog ib qho nruab nrab tshiab. Extract aqueous dispersion los yog extract-loaded vesicles tau diluted nrog cell nruab nrab mus txog 4 ug / mL ntawm extract thiab siv los kho cov kab mob cell monolayers. Cov kab mob ntawm tes tau pom, thiab cov duab raug ntes ntawm 24, 36, thiab 48 h ntawm incubation siv lub tshuab ntsuas qhov muag nrog lub hom phiaj 10 ×. Cov hlwb tsis kho tau siv los ua kev tswj tsis zoo. Cov kab mob hauv cheeb tsam hauv cov duab ntes tau ntsuas los ntawm Java's duab J software (http://rsb.info.nih.gov, nkag mus rau lub Plaub Hlis-Lub Rau Hli 2021). Qhov kaw ntawm qhov txhab raug suav nrog Kev Sib Txuas (2):

qhov twg ao yog qhov chaw raug mob tam sim ntawd tom qab khawb, thiab a yog qhov chaw raug mob ntsuas ntawm 24, 36, thiab 48 h [44].

3.9. Kev txheeb cais

Cov txiaj ntsig tau qhia raws li qhov nruab nrab ± tus qauv sib txawv thiab qhov tseem ceeb tau sim ntawm 0.05 qib ntawm qhov yuav tshwm sim (p). Rau qhov loj me, zeta muaj peev xwm, viscosity, tsub zuj zuj ntawm cov tshuaj, thiab cytotoxicity, ib txoj kev tsom xam ntawm qhov sib txawv (ANOVA) tau siv los txheeb xyuas qhov sib txawv ntawm cov pab pawg ua raws li Tukey qhov kev xeem, thaum Tub Kawm Ntawv t-test tau siv rau kev sib piv ntawm ob tus qauv siv XLStatistic. rau Excel.

4. Cov lus xaus

Lub lyophilized jabuticaba tev extract tau muab tso rau hauv kev hloov pauv thiab polymer (hydroxyethylcellulose thiab sodium hyaluronate) enriched transfersomes nrog lub hom phiaj ntawm stabilizing cov extract thiab txhim kho nws cov kev kho mob. Cov vesicles tau txais yog me me nyob rau hauv loj thiab homogeneously dispersed. Qhov sib ntxiv ntawm ob qho tib si polymers tsuas yog ua rau tsim cov vesicles loj me ntsis yam tsis muaj qhov sib txawv ntawm cov khoom siv polymeric. Polymer-enriched vesicles zoo li zoo tagnrho rau cov tshuaj pleev ib ce thiab muaj peev xwm koom nrog cov bioactive-nplua nuj extract nyob rau hauv siab. Hauv particular, kev sib xyaw ua ke ntawm phospholipid, Tween 80, thiab sodium hyaluronate kom tau Hyaluronan-transfersomes, tau raug xaiv los ua cov qauv zoo tshaj plaws ntawm kev ruaj ntseg thiab muaj peev xwm cuam tshuam nrog keratinocytes, vim tsuas yog cov hlwv no tuaj yeem tiv thaiv kev puas tsuaj. induced nyob rau hauv cov hlwb thaum siv hydrogen peroxide thiab txhawb kev kho mob hauv tib neeg keratinocytes. Zuag qhia tag nrho, peb cov txiaj ntsig tau qhia tias hyaluronan-transfersomes tuaj yeem sawv cev rau kev cog lus rau kev kho cov kab mob ntawm daim tawv nqaij lossis daim tawv nqaij qhov txhab txuas nrog oxidative kev nyuaj siab. Tsis tas li ntawd, peb qhia thawj zaug ntawm kev siv jabuticaba tev extract nyob rau hauv ib tug dermatological xa system.

Tus sau kev koom tes:DG, MLM, MM: conceptualization, formal analysis, data curation, sau ntawv, thiab tshuaj xyuas; IC, MA, JH: kev txheeb xyuas thiab sau ntawv. Txhua tus kws sau ntawv tau nyeem thiab pom zoo rau cov ntawv luam tawm ntawm cov ntawv sau.

Nyiaj txiag:Qhov kev tshawb fawb no tsis tau txais nyiaj txiag sab nraud. Institutional Review Board Statement: Tsis siv tau. Cov Lus Qhia Txog Kev Pom Zoo: Tsis siv tau.

Cov ntaub ntawv muaj nyob:Cov ntaub ntawv nyoos muaj thiab tuaj yeem nug ncaj qha rau cov kws sau ntawv.

Kev tsis sib haum xeeb ntawm kev txaus siab:Cov kws sau ntawv tshaj tawm tsis muaj kev sib cav txog kev txaus siab.

Qauv Muaj:Cov qauv ntawm jaboticaba tev extract yog muaj los ntawm cov kws sau ntawv.

Kab lus no yog muab rho tawm los ntawm Molecules 2021, 26, 6697. https://doi.org/10.3390/molecules26216697 https://www.mdpi.com/journal/molecules