• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    Modulating intestinal mucus barrier for nanoparticles penetration by surfactants

    2019-09-10 06:52:36XinZhngWeiDongHongoChengMeixiZhngYongqingKouJinGunQioyuLiuMingyueGoXiuhuWngShiruiMo
    關(guān)鍵詞:參考文獻(xiàn)

    Xin Zhng,Wei Dong,Hongo Cheng,Meixi Zhng,Yongqing Kou,Jin Gun,Qioyu Liu,Mingyue Go,Xiuhu Wng,Shirui Mo,*

    aSchool of Pharmacy,Shenyang Pharmaceutical University,Shenyang 110016,China

    bLaboratory of Metabolic Disease Research and Drug Development,China Medical University,Shenyang 110122,China

    Keywords:Mucus barrier Mucus modulating agents Rheology PLGA nanoparticles Surfactants

    ABSTRACT Improving peroral delivery efficiency is always a persistent goal for both small-molecule and macromolecular drug development. However, intestinal mucus barrier which greatly impedes drug-loaded nanoparticles penetration is commonly overlooked.Therefore,in this study, taking fluorescent labeled PLGA (poly (lactic-co-glycolic acid)) nanoparticles as a tool, the influence of anionic and nonionic surfactants on mucus penetration ability of nanoparticles and their mucus barrier regulating ability were studied. The movement of PLGA nanoparticles in mucus was tracked by multiple particles tracking method (MPT).Alteration of mucus properties by addition of surfactants was evaluated by rheology and morphology study.Rat intestinal villus penetration study was used to further evaluate penetration enhancement of nanoparticles. The effective diffusivities of the nanoparticles in surfactants pretreated mucus were increased by 2-3 times and the mucus barrier regulating capacity was also surfactant type dependent. Sodium dodecyl sulfate (SDS) increased the complex viscosity and viscoelastic properties of mucus,but poloxamer presented a decreased trend.Tween 80 maintained the rheological property of the mucus.With the mucus barrier regulated by surfactants, the penetration of nanoparticles in intestinal villus was obviously increased.In summary,the mucus penetration ability of nanoparticles could be enhanced by altering mucus microenvironment with surfactants. Tween 80 which largely retains the original mucus rheology and morphology properties may be a promising candidate for facilitating nanoparticle penetration through the mucus barrier with good safety profile.?2018 Shenyang Pharmaceutical University.Published by Elsevier B.V.This is an open access article under the CC BY-NC-ND license.(http://creativecommons.org/licenses/by-nc-nd/4.0/)

    1. Introduction

    The safety and effective delivery of drugs to body circulation by oral administration is fraught with many challenges,which is not only hampered by epithelial barrier but also mucus barrier[1].With the presence of mucus layer,most foreign particles in the intestinal tract will be trapped by mucus to prevent them from contacting with epithelia cells.Meanwhile,the mucus barrier could significantly reduce the oral delivery efficiency of many well design nanocarriers by weakening their move ability in mucus[2].

    Permeation barrier properties of the tenacious mucus comes from its microenvironment and structure. Mucus hydrogel is mainly composed of water, 2%-5% (w/v) mucin,small amount of lipids [3]. The networks formed by entangled mucins are the main structure of mucus layer and provide various properties to mucus. Mucins are glycoproteins which are composed of peptide chain modified by hundreds of O-linked and N-linked oligosaccharides. Negative charge of glycosylation groups in mucins, hydrophobic surface of non-glycosylated protein chain and network formed by mucin fibers constitute the structure foundation for penetration barrier[2].

    Compared to the mucus penetration nanoparticles strategy,application of mucus modulating agents to overcome the mucus barrier by co-administration or co-formulation with drug carrier is easier to satisfy the industrial manufacture requirements with better applicability [4].In order to modulate the mucus barrier,mucolytic agents and mucus production inhibitors are commonly applied, but their application usually raises the safety concern[5].This is because mucus also plays a significant role in avoiding the damage of gastrointestinal environment including digestive enzymes, colonized bacterial, acid-base environment and other harmful antigens and microbes on epithelium under the mucus[1].Therefore,developing and screening potential mucus penetration modulators which are general regarded as safe(GRAS)ingredients without influencing properties of mucus are economical and of special importance.Moreover,studying the interaction between pharmaceutical excipients and mucus will facilitate the design of drug carriers with mucus penetration ability and increase the accuracy of in vivo model prediction for nanoparticles based drug delivery system.

    It has been reported that surfactants show promising potential to regulate mucus barriers,which has been widely used in pharmaceutical industry as surface tension regulator, solubility and penetration enhancing agents and stabilizers of protein and nanoparticles [6,7]. It is reported that the human cervicovaginal mucus pretreated by Pluronic F127 significantly increased the penetration of polystyrene nanoparticles without changing the pore size of mucus [8]. However,the treatment by nonionic surfactant nonoxynol-9 reduced the pore size of mucus and increased the barrier properties for 200/500 nm particles [9].This indicated that the mucus modulating property of surfactants is not only affected by mucus source but also greatly dependent on molecular structure of surfactants. For oral administration, although surfactants have shown drug permeability enhancing ability by altering the structure and microenvironment of intestinal epithelium[10],the interactions between surfactants and intestinal mucus, and the relationship between surfactant structure and its function are still unclear.It is also unknown how will the surfactants structure influence their mucus penetration enhancing capacity if this is the case. In addition, the mucus modulation function study of surfactants will provide valuable information for mucus penetration enhancing drug carrier design.

    Therefore, in this paper, using PLGA nanoparticles as a model, influence of different type of surfactants on mucus permeation efficiency of nanoparticles and the interaction between surfactants and mucus were explored. Here, surfactants with different surface charge and HLB (hydrophiliclipophilic balance) values, including sodium dodecyl sulfate(SDS,anionic,HLB 40,CMC: 8.2 mmol/l),poloxamer 188 (nonionic, HLB 29, CMC: 0.48 mmol/l), poloxamer 407 (nonionic,HLB 22,CMC:2.8 μmol/l)and Tween 80(nonionic,HLB 15,CMC:0.015 mmol/l)were selected in this study.Permeation process of the nanoparticles was tracked and calculated to further understand the interaction between surfactants and mucin.The interaction mechanism between surfactants and mucus,and the network structure change of mucus in the presence of surfactants were investigated by rheology study and morphology observation.After penetrating through the mucus barrier layer,penetration of nanoparticle in intestinal villus was also observed by confocal microscopy.

    2. Material and methods

    2.1. Materials

    Porcine original mucus were obtained from intestine of freshly slaughtered pigs and mucus was gently scraped from the intestinal wall(Shenyang,Liaoning).In order to minimize the influence of mucus variability on the result,mucus sample was collected and stored at -20 °C before use to make sure the mucus sample used for each compound was from the same source [11,12]. Poloxamer 407 and 188 (P407, P188) were obtained as gifts from BASF (Germany). SDS was bought from Biotopped Co.,Ltd.(China).PLGA (Resomer?RG 503) was purchased from Evonik Industries (Germany).Poly(vinyl alcohol)(PVA 205) was obtained from Kuraray China Co., Ltd.(China).Coumarin 6 were purchased from J&K chemical Ltd.(China).Tween 80 was obtained from Tianjin Bodi Chemical Co., Ltd.(Tianjin, China). DAPI (4′,6-diamidino-2-phenylindole) staining solution and antifade mounting medium were bought from Beyotime Institute of Biotechnology(China).

    2.2. Preparation of coumarin 6 labeled PLGA nanoparticles

    PLGA nanoparticles containing coumarin 6 were prepared by O/W emulsion solvent evaporation method [13,14]. In brief,20 mg PLGA were dissolved in 1 ml dichloromethane containing 150 μg coumarin 6 as oil phase in dark environment.Outer water phase,8 ml 2%PVA solution were added to the oil phase and then the mixture was sonicated by ultrasonic homogenizer for 60 s, 100 W to prepare emulsions (SCIENTZ IID, Scientz Biotechnology,Ningbo,China).The O/W emulsions were then added to 20 ml 1% PVA solution and hardened by evaporating dichloromethane under stirring for 3 h(84-1A,Shanghai Sile Instrument Co., Ltd., China). The nanoparticles were washed for 2 times by centrifugation and resuspending in the same volume of deionized water (HC-2062, USTC Zonkia Scientific Instruments Co.,Ltd.,Anhui,China) [15].The PLGA nanoparticles suspension were used for following study.

    2.3. Movement of PLGA nanoparticles in intestinal mucus

    Multiple particles tracking method(MPT)was applied to track the brownian movement of nanoparticles in mucus sample and further evaluate the mucus barrier properties for nanoparticle penetration in the presence of different surfactants [16]. Briefly, probe coumarin 6 labeled PLGA nanoparticles were gently added into mucus samples in microwells and incubate for 30 min at 37 °C.Inverted fluorescence microscope IX 71 with CCD imaging system DP 70 and 40X objective (OLYMPUS, Japan) was used to record the trajectory of coumarin 6 labeled PLGA nanoparticles at a frame rate of 15 fps for 10 s and three experiments were performed for each sample. For sample preparation, mucus samples were evenly mixed with 1% (w/w) surfactants solution including P407, P188, SDS and Tween80 at a weight ratio of 4:1. As the reference sample,native mucus was mixed with distilled water in parallel to eliminate the influence of dilution process.400 μl mucus was evenly added into 24-well cell culture plates and 10 μl nanoparticles were gently added to the mucus.Trajectory of nanoparticles were tracked by particle tracker plugin in ImageJ software[17,18].The position value of trajectory was analyzed and time-averaged mean square displacement(MSD)and effective diffusivities(Deff)were calculated according to the following equations: MSD=[x(t+τ)-x(t)]2+[y(t+τ)-y(t)]2,Deff=MSD/(4τ),where x,y is coordinates of nanoparticles in mucus and τ represents time scale[16].

    2.4. Particle size analysis

    The size and zeta potential of the nanoparticles were characterized by dynamic light scattering technology by Nano ZS90(Malvern Instruments,Worces-tershire,UK)at 25°C at a scattering angle of 90°.

    2.5. Morphology observation of mucus

    Morphology study of original mucus and mucus treated by surfactants was performed by macroscopic observation, optical microscope and atomic force microscope (AFM). In order to better distinguish the difference of mucus samples for macroscopic observation,native mucus and mucus treated by surfactants were centrifuged at 2000 r/min for 10 min.Optical microscope observation was performed by BI-2000 Image Analysis System (Chengdu Techman Software Co., Ltd) at X40 objective.Atomic force microscope(Agilent Technologies,USA)was used to study the morphology of mucus at micrometer scale.The samples were added to a clean mica plate and dried at room temperature.Then the samples were tested by tapping mode.

    2.6. Rheological measurements

    Rheology properties of mucus were determined by plate-plate model at 37 °C using controlled stress rheometer AR2000 (TA Instruments, USA) and frequency sweep of oscillation tests were performed at 2% strain that was within the linear viscoelastic regime (LVR). Volume of the sample was fixed at 0.314 ml. Mucus samples preparation process was the same with “2.3 Movement of PLGA nanoparticles in intestinal mucus”.

    Complex viscosity and viscoelastic parameters including complex modulus,elastic and viscous modulus,damping factor were used to evaluate the alteration of rheology of mucus by addition of surfactants. Complex modulus (G*) was computed based on G*=G’+i*G’’. Damping factor (tanδ) which was calculated by tanδ=G’’/G’[19].

    2.7. Intestinal villus penetration study

    Rat intestinal villus penetration study was used to further evaluate the influence of surfactants on the penetration of nanoparticles. All animal experiments followed the Principles of Laboratory Animal Care and approved by Shenyang Pharmaceutical University Ethics Committee. Male Wister rats weighing 180-230 g were fasted overnight and was anesthetized by intraperitoneal injection of 5%chloral hydrate.Abdominal cavity was opened and two ends of ileum(about 5 cm)were ligated.0.3 ml 1%surfactant were injected to the ligated ileum segment as pretreatment process for 30 min and then 0.3 ml PLGA nanoparticles were administrated to the loop.After administration of PLGA nanoparticles for 30 min,the rats were euthanized with an overdose of chloral hydrate and the loops were removed. The ileum segment was gently washed by 5 ml PBS and then treated by 4%paraformaldehyde for 2 h and 30%sucrose overnight at 4°C before frozen sections.The loops were coated by O.C.T.Compound(Tissue-Tek?,SAKURA,USA) and were rapidly frozen for section. The ileum section was stained with DAPI for nucleus dying and observed by multi-photon confocal microscope(CLSM,LSM 710,Zeiss,Germany) at 405 nm and 458 nm lasers source.The fluorescence intensity in intestinal villus was compared and quantified by Image J software.

    2.8. Statistical analysis

    The results were analyzed using ANOVA two-way and data are presented as mean value±SD (n ≥3). Probability values P <0.05 were considered to be statistically significant.

    3. Results and discussion

    3.1. Preparation and characterization of C6 labeled PLGA nanoparticles

    As a biocompatible polymer, poly (lactic-co-glycolic acid)(PLGA) was widely applied to prepare micro and nanoparticles for different administration routes due to its mature manufactory process, good encapsulation for both hydrophobic and hydrophilic drugs and controlled release behavior [20].Therefore,PLGA nanoparticles were prepared as a nanoparticles model and hydrophobic fluorescence dye coumarin 6(C6)was loaded for the ease of observation. The particle size of PLGA nanoparticles was about 241±5.2 nm, PDI: 0.294±0.04 and zeta potential value was -14.7±0.3 mV. The release of coumarin-6 from PLGA nanoparticles in PBS buffer (non-sink condition)was under detection limit in 2 h.The similar results were also reported in published literatures[21-23].Some literatures showed that PVA coating on nanoparticles could affect its mucus penetration ability and the nanoparticles could be more easily immobilized by mucus than that by PEG coating[24]. However, previous study indicated the residual amount of PVA was dependent on solvent type in oil phase and concentration of PVA. When dichloromethane was used as solvent and 5%(w/w)PVA solution were applied to prepare PLGA nanoparticles, 6.15% of the total PVA (w/w) added was adsorbed on the surface of PLGA nanopariticles after 2 times washing [25]. In this study, based on the assumption that 6.15% of the added PVA was remained on PLGA nanoparticle surface, the residual PVA on nanoparticle surface was about 4.1×10-3% (w/w),which was greatly lower than the concentration of PVA used for coating purpose(0.01%-1%),therefore,it has limited influence on the interaction between nanoparticles and mucus. In addition, although some residual PVA might exist on the surface of nanoparticles,the nanoparticles were used as nanoprobes in parallel in all the groups to show the mucus barrier properties change after treatment by different surfactants. Thus, the effect of residual PVA has been well justified.Therefore,coumarin 6 labeled PLGA nanoparticles was used as a nanoprobe to track its trajectory in intestine mucus and understand the influence of surfactants on nanoparticles penetration ability.

    3.2. Movement tracking of nanoparticles in intestinal mucus

    The diffusional barrier mainly comes from micro network structure of mucus and interactions between nanoparticles and mucus components [26]. The variation of microenvironment of mucus treated by surfactants may enhance penetration of nanoparticles.Therefore,the movement trajectories of PLGA nanoparticles in the mucus pretreated by different surfactants were tracked.The applied concentration of surfactants in pharmaceutical area as emulsifier or stabilizer was 0.3%-5%for poloxamer,0.5%-2.5%for SDS and 1%-15%for Tween 80 [27]. In preliminary experiments, the interaction strength between mucus and surfactants increased with the increase of surfactant concentration,and equilibrium was achieved when the concentration of surfactants was above 1%.Higher concentration of surfactants will also increase the risk of safety concern.Therefore,1%concentration of surfactant was selected for the following study. Mean squared displacements (MSD) (Fig.1) of PLGA nanoparticles showed that all of the surfactants investigated could improve the mobility of nanoparticles in mucus. The ensemble-average effective diffusivity of PLGA nanoparticles in mucus containing different kinds of surfactants at 5 s time scale was increased by SDS for 2.46 times (0.1308±0.03 μm2/s), P407 for 2.14 times(0.1139±0.02 μm2/s),P188 for 1.81 times(0.0965±0.023 μm2/s)and Tween 80 for 1.82 times (0.0967±0.011 μm2/s) compared with that in mucus without surfactants(0.0532±0.017 μm2/s).Among them, SDS showed the highest effective diffusivity value of nanoparticles in the mucus, indicating SDS is superior to enhance the mucus penetration of PLGA nanoparticles.For nonionic surfactant,MSD of the nanoparticles in the mucus treated by P188 was lower than that of P407,and Tween 80 exhibited a better mucus penetration enhancing ability than that of P188 at long time scale.

    Fig.1-Influence of surfactants on mucus penetrations of PLGA nanoparticles.

    3.3. Morphologic observation of mucus

    To clarify whether the improvement of mucus penetration ability of PLGA nanoparticles by surfactants is related to microstructure change of mucus,morphology of various mucus samples was observed at different scales. Since the mucus was too viscous to distinguish the change of appearance and all the samples showed similar hydrogel state,the centrifugal supernatant of different mucus samples was compared to understand the influence of different surfactants on the appearance of mucus. As shown in Fig. 2A, the supernatant of SDS treated mucus turned into a relative viscous translucent liquid,while the supernatant of P407,P188 and Tween 80 treated mucus didn’t show distinguished difference with the original mucus.

    The variation of mucus microstructure was further investigated with optical microscope.As shown in Fig.2B,based on the images observed in micrometer scale,morphology change of the mucus treated by SDS,P188 and P407 was found,with aggregation or increased gap among the components, while mucus treated by Tween 80 didn’t show obvious difference in appearance compared with the original mucus. Furthermore, the morphology was investigated in the nanometer scale (10 μm range) using AFM.As shown in Fig.2C,aggregation was observed on the mucus surface treated by SDS compared with the original mucus,while the addition of P407,P188 and Tween 80 increased the pore size of the mucus network to various degree,forming a honeycombed passages in the mucus.

    Fig.2-The morphology study of mucus at different scales.(A)Macroscopic observation(B)Optical microscope observation and(C)Atomic force microscope images.

    Since morphology study can only provide limited intuitive results and the observation process by AFM might potentially influence the mucus morphology,rheology study was further carried out to understand the interaction mechanism between surfactants and mucus.

    3.4. Influence of surfactants on the rheological properties of mucus

    The rheology property of mucus is a crucial parameter for mucus barriers,which can also reflect mucus microstructure change.In this paper, the variation of complex viscosity and viscoelastic parameters of mucus induced by surfactants were studied to better understand the way of surfactants to increase mucus penetration ability of PLGA nanoparticles and modify the mucus barrier properties.

    To evaluate the influence of surfactants on the overall viscosity of mucus, change of complex viscosity of mucus was determined.Complex viscosity of original mucus and mucus containing different surfactants were tested under oscillation model at a fixed strain of 2%.The results,as shown in Fig.3,showed that the addition of surfactants modulated the complex viscosity of mucus with different extent. SDS showed a viscosity increase effect on mucus.In contrast,Tween 80 and poloxamer series surfactants decreased the viscosity of mucus.Among these,P407 exhibited a stronger ability to decrease mucus complex viscosity. No statistical difference between P188 and Tween80 was found.Viscoelastic property of mucus were further studied to understand the interaction between mucus and surfactants.

    Fig.3-Complex viscosity of the original and surfactants treated samples.

    The change of mucus viscoelastic properties in terms of elastic,viscous modulus,complex modulus and damping factor were studied. Fig. 4A showed the influence of different surfactants on elastic modulus (G’) of mucus. The addition of anionic surfactants,SDS,enhanced the elastic property of mucus.For nonionic surfactant, Tween 80, P188 and P407 reduced the elastic modulus (G’) of mucus, and the value can be decreased to a lower degree by P407. Similar trend was also observed in the results of viscous modulus(G’’)and complex modulus (G*) (Fig. 4B and C). The comparison of damping factor (tan δ) of mucus containing different surfactants showed that all the mucus sample exhibited a lower damping factor (tan δ <1), indicating the mucus tended to exhibit higher elastic properties rather than the viscous property(Fig.4D).Among all the surfactants investigated,the damping factor is similar at frequency less than 1 rad/s. With the increase of frequency, P407 exhibited a stronger ability to increase the damping factor of mucus in comparison with other surfactants.

    Fig.4-Viscoelastic modulus of the original and surfactants treated mucus samples:(A)elastic modulus(G'),(B)viscous modulus(G''),(C)complex modulus(G*)and(D)damping factor(tan δ).

    The observed aggregation of mucus induced by SDS(Fig.2)is also in good agreement with the increase of its viscoelastic properties, implying that SDS may induce denaturation and aggregation process [28]. The denaturation process may be due to the insertion of SDS in mucin which may change the molecular conformation. SDS, as a linear anionic surfactant,has shown a strong potential to bind to the protein molecular.It has been reported that longer hydrophobic alkyl group and ionic group will lead to stronger interactions with proteins[29-31].Thus,the anionic head group of SDS,which provides high hydrophilicity (HLB 40) and hydrocarbon chains,might be the essential component to provide the high strength of interaction. Although both the hydrophilic head of SDS and mucin are negatively charged,the electrostatic repulsion among them were not strong enough to prevent their associations which are mainly driven by hydrophobic interaction[29]. Thus, the binding of SDS to mucin will weaken the interaction among mucin and decrease the hydrophobic property of mucus and then increase the mucin-water interaction[29-31]. The extended conformational structure and the improved water solubility of mucin fibers eventually facilitate the aggregation of mucin, leading to increased complex viscosity and viscoelastic properties of mucus.

    As liner nonionic surfactants composed of polyoxyethylene and polyoxypropylene segment, poloxamer, showed a good ability to decrease the viscoelastic properties of mucus,indicating the mucus network structure strength was reduced(Fig.4).However,a report indicated that in human cervicovaginal mucus (CVM) pretreated by P407,the penetration ability of nanoparticles increased, but the pore structure of mucus was not changed [32]. This may be due to the mucus source difference, and the contents of native intestinal mucus was more complex, which contains not only mucin but also DNA, proteins and lipids mixture [18,33]. The lipids exited in the intestinal mucus, which greatly influence the mucus properties, might also be sensitive to surfactants. Although P188 and P407 own similar structure, P407 showed a greater ability to reduce the mucus rheology properties.This can probably be attributed to the variation of P407 and P188 in HLB value, ratio of polyoxyethylene to polyoxypropylene segment and molecular weight, which may influence the interaction process between surfactant and mucus components(P188:HLB:29;MW:7.7-9.5 kDa;ratio:2.96.P407:HLB:22;MW:9.8-14.6 kDa; ratio: 1.80.). For poloxamer, higher ratio of hydrophilic parts and HLB value have a higher tendency to adsorb on the surface of mucus membrane rather than insertion[34,35]. As the most hydrophilic surfactant among poloxamers,P188 may tend to adsorb on the mucus surface,with slight change of mucus structure.In contrast,the higher molecular weight of P407, which can provide better steric stabilization,may contribute to the decrease of the viscoelastic properties of mucus.For branched nonionic surfactant Tween 80,alteration of mucus rheology was comparable to that of P188, which was consistent with the limited change of mucus morphology(Fig.2).

    Fig.5-Intestinal villus penetration study.(A)CLSM images of intestinal villi of rat ileum segment after administration of coumarin 6 labeled PLGA nanoparticles suspension,(coumarin 6 and DAPI exhibited green and red color,respectively).(B)Average fluorescence intensity of coumarin 6 labeled PLGA nanoparticles in intestinal villus(n=3).(For interpretation of the 參考文獻(xiàn) to color in this figure legend,the reader is referred to the web version of this article.)

    3.5. Intestinal villus penetration study

    The nanoparticles which penetrated through mucus barrier would reach intestinal epithelium[36,37].

    For the same PLGA nanoparticles,the amount variation of nanoparticles in intestinal villus would depend on the penetration ability of nanoparticles in the mucus layer treated by surfactants.The penetration of PLGA nanoparticles labeled by coumarin 6 in villus was observed by confocal microscope and the improvement of mucus penetration induced by surfactants was evaluated intuitively. Fig. 5 shows that, except for P188, the coumarin 6 fluorescence intensity in the intestinal villus was obviously increased after pretreatment with surfactants, but no significant difference was found between SDS,P407 or Tween 80 groups.

    Taking into consideration of rheology and mucus penetration results, the mechanism of improving mucus penetration of nanoparticles by surfactant was predicted and discussed. The steric barrier of mucus network and interaction between the mucus substances and nanoparticles greatly limit the penetration movement of nanoparticles in mucus.As common components used in drug delivery system, surfactants could modify the penetration ability of mucus by decreasing the interaction strength, changing the mucus structure and increasing hydrophilicity of the microenvironment. The adsorption of nanoparticles to mucin will also be reduced because of the adsorbed surfactant layers.The interaction between SDS and mucin could change the mucin conformation, and adsorption of SDS on mucin and lipids will increase hydrophilicity of the microenvironment, this would also facilitate the penetration ability of nanoparticles. However, the increased viscosity and viscoelastic properties induced by aggregate of mucin would also show adverse impact on penetration of nanoparticles.For nonionic surfactant P407/188 and Tween 80,they could reduce the interaction between mucin fibers and increase the network pores, which can eventually facilitate nanoparticles penetration by increasing the pore size of mucus. Compared with P188, the better mucus penetration enhancing ability of P407 may attribute to its stronger ability to decrease the viscoelastic properties and therefore morphology change of the mucus. Although modification of mucus structure induced by Tween 80 was similar with P188, it showed better promotion effect on the penetration of nanoparticles. It is predicted that branched hydrophobic segment may promote the adsorption of surfactant on mucin and increase the nanoparticle penetration,although the change of morphology and rheology of mucus is not as obvious as P407 and SDS treated mucus.

    The intestinal villus penetration enhancing effect was in good agreement with the mucus penetration ability of different surfactants (Fig. 5). The binding of surfactants to mucin will increase the mesh space of mucus and modify the mucus microenvironment to increase the penetration possibilities of nanoparticles. The arrangement of surfactant on the surface of the lipid and mucin will form a hydrophilic layer to greatly reduce the interactions.Thus,more nanoparticles will contact with intestinal villus and increase its penetration possibility.Although M cells in intestinal loop models will potentially affect the uptake of hydrophobic nanoparticles,the low number ratio of M cells in total intestinal epithelial cells(1 in 107)and higher affinity to hydrophobic nanoparticles limit its influence on the uptake of hydrophilic nanoparticles [38,39].In our study, the hydrophilic surface of PLGA nanoparticles will reduce the influence of M cells. The slices also showed nanoparticles were evenly distributed in the villus,nanoparticles uptake by M cells were not found.In addition,the membrane permeability which was enhanced by surfactants may also facility the nanoparticle penetration[10].

    4. Conclusions

    Mucus, as viscoelastic hydrogel, showed inevitable penetration barrier for peroral delivery of nanoparticles. Mucus penetration enhancing ability of surfactants for PLGA nanoparticles were investigated and compared in this paper.The results showed that both ionic and nonionic surfactant can increase the mucus permeation ability of PLGA nanoparticles.Although all surfactants can increase the hydrophilicity of mucus microenvironment,mechanism of different surfactants to facilitate the movement of nanoparticles is structure dependent. As foremost consideration of its application in drug administration, safety of surfactants which would not change the mucus properties is important. Thus, Tween 80 which partly retains the original mucus rheology and morphology properties,may be a promising candidate for facilitating nanoparticle penetration through the mucus barrier.

    Conflicts of interest

    The authors declare that there is no conflicts of interest.

    Acknowledgment

    This project is financially supported by the National Natural Science Foundation of China(Grant No.31870987).

    猜你喜歡
    參考文獻(xiàn)
    Eurydice’s Face:the Paradox of Mallarmé’s Musical Poetics*
    Kidney health for everyone everywhere—from prevention to detection and equitable access to care
    Effect of low high-density lipoprotein levels on mortality of septic patients: A systematic review and meta-analysis of cohort studies
    SINO-EUROPE SYMPOSIUM ON TRADITIONAL CHINESE MEDICINE & HERBAL MEDICINE-MARKET OVERVIEW ®ULATION POLICY
    A prediction method for the performance of a low-recoil gun with front nozzle
    The Muted Lover and the Singing Poet:Ekphrasis and Gender in the Canzoniere*
    Where Does Poetry Take Place? On Tensions in the Concept of a National Art* #
    Chinese Cultural Influence on Hannah Jelkes in The Night of the Iguana*
    The serum and breath Raman fingerprinting methodfor early lung cancer and breast cancer screening
    Study on the physiological function and application of γ—aminobutyric acid and its receptors
    東方教育(2016年4期)2016-12-14 13:52:48
    亚洲第一电影网av| 又爽又黄a免费视频| 亚洲国产精品久久男人天堂| 国产免费男女视频| 亚洲精品乱码久久久久久按摩| 美女国产视频在线观看| 国产精品女同一区二区软件| 99在线视频只有这里精品首页| 亚洲美女搞黄在线观看| 国产精品1区2区在线观看.| 91av网一区二区| 亚洲内射少妇av| 美女cb高潮喷水在线观看| 高清毛片免费看| 国产成人freesex在线| 成人特级黄色片久久久久久久| 亚洲欧美日韩无卡精品| 日韩欧美一区二区三区在线观看| 日韩av在线大香蕉| 男女视频在线观看网站免费| 国产精品蜜桃在线观看 | 夜夜看夜夜爽夜夜摸| 亚洲国产精品sss在线观看| 变态另类丝袜制服| 国产精品美女特级片免费视频播放器| 又黄又爽又刺激的免费视频.| 三级经典国产精品| 少妇丰满av| 一级毛片久久久久久久久女| 中出人妻视频一区二区| 日韩,欧美,国产一区二区三区 | 国产视频首页在线观看| 欧美丝袜亚洲另类| 真实男女啪啪啪动态图| 在线天堂最新版资源| 国产不卡一卡二| eeuss影院久久| 国产精品av视频在线免费观看| 国产精品,欧美在线| 久久精品国产清高在天天线| 久久精品综合一区二区三区| 超碰av人人做人人爽久久| 国产人妻一区二区三区在| 亚洲人成网站高清观看| 日韩一区二区视频免费看| 91狼人影院| 国产三级中文精品| 欧美性感艳星| 国产单亲对白刺激| 麻豆成人av视频| 最好的美女福利视频网| 欧美zozozo另类| 欧美成人精品欧美一级黄| 搞女人的毛片| 日韩制服骚丝袜av| 99久久精品国产国产毛片| 国产综合懂色| 国产亚洲精品久久久久久毛片| 久久精品久久久久久噜噜老黄 | 啦啦啦啦在线视频资源| 黄色一级大片看看| 国产高清激情床上av| 亚洲国产精品成人综合色| 国产v大片淫在线免费观看| 内射极品少妇av片p| 美女xxoo啪啪120秒动态图| 国产淫片久久久久久久久| 国产综合懂色| 又粗又爽又猛毛片免费看| 男女边吃奶边做爰视频| 非洲黑人性xxxx精品又粗又长| 日韩一区二区视频免费看| 天天躁夜夜躁狠狠久久av| 精品久久国产蜜桃| 欧美丝袜亚洲另类| h日本视频在线播放| 热99在线观看视频| 老司机影院成人| 国产日韩欧美在线精品| 亚洲激情五月婷婷啪啪| 欧美一区二区亚洲| 免费黄网站久久成人精品| 免费无遮挡裸体视频| 深夜a级毛片| 国产视频首页在线观看| 中文字幕制服av| 久久久久国产网址| 久久精品国产清高在天天线| 国产人妻一区二区三区在| 小蜜桃在线观看免费完整版高清| av天堂在线播放| 18禁在线播放成人免费| 亚洲欧美精品综合久久99| 欧美激情久久久久久爽电影| 婷婷六月久久综合丁香| 麻豆国产av国片精品| 麻豆一二三区av精品| 久久精品人妻少妇| 嫩草影院新地址| 99热这里只有精品一区| 在线观看免费视频日本深夜| 色哟哟·www| 国产精品美女特级片免费视频播放器| 日本熟妇午夜| av免费在线看不卡| 欧美xxxx性猛交bbbb| 久久亚洲国产成人精品v| 亚洲成人精品中文字幕电影| 色播亚洲综合网| 亚洲不卡免费看| 在线观看免费视频日本深夜| 亚洲无线在线观看| 亚洲美女搞黄在线观看| 伊人久久精品亚洲午夜| 1000部很黄的大片| 婷婷色综合大香蕉| 亚洲熟妇中文字幕五十中出| a级毛片免费高清观看在线播放| av在线播放精品| 99久国产av精品国产电影| 99热这里只有精品一区| 日日摸夜夜添夜夜添av毛片| av在线蜜桃| 男女下面进入的视频免费午夜| 亚洲国产欧美人成| 亚洲国产精品成人综合色| 久久精品夜色国产| 久久人妻av系列| 看十八女毛片水多多多| 亚洲激情五月婷婷啪啪| 亚州av有码| videossex国产| 99久国产av精品| 在线播放无遮挡| 国内精品美女久久久久久| 国产精品日韩av在线免费观看| 国产精品一二三区在线看| 国产伦在线观看视频一区| 18禁黄网站禁片免费观看直播| av视频在线观看入口| 乱码一卡2卡4卡精品| 12—13女人毛片做爰片一| 最近视频中文字幕2019在线8| 精品日产1卡2卡| 日本成人三级电影网站| 日韩强制内射视频| 高清在线视频一区二区三区 | 波多野结衣高清无吗| 最近中文字幕高清免费大全6| 成人亚洲精品av一区二区| av天堂中文字幕网| 亚洲最大成人av| 欧美丝袜亚洲另类| 精品熟女少妇av免费看| 有码 亚洲区| 1024手机看黄色片| 乱码一卡2卡4卡精品| 丝袜喷水一区| 国产成人freesex在线| 国产又黄又爽又无遮挡在线| 亚洲美女搞黄在线观看| 精品少妇黑人巨大在线播放 | 久久99蜜桃精品久久| 亚州av有码| 毛片女人毛片| 午夜精品国产一区二区电影 | 大香蕉久久网| 国产乱人视频| 99热网站在线观看| 日韩成人av中文字幕在线观看| av女优亚洲男人天堂| 亚洲在线自拍视频| 伦理电影大哥的女人| 亚洲乱码一区二区免费版| 久久精品综合一区二区三区| 欧美色视频一区免费| 国产高清三级在线| 黄色日韩在线| 少妇丰满av| av在线播放精品| 中出人妻视频一区二区| 欧美日韩国产亚洲二区| 只有这里有精品99| 国产黄a三级三级三级人| 一级黄片播放器| 色综合色国产| 激情 狠狠 欧美| 内地一区二区视频在线| 亚洲av中文av极速乱| 久久综合国产亚洲精品| 午夜精品一区二区三区免费看| 国产精品福利在线免费观看| 久久久国产成人免费| 久久久久久久久久成人| 欧美高清性xxxxhd video| 免费观看精品视频网站| 国产精品无大码| 午夜福利在线观看吧| 国产精品野战在线观看| 久久中文看片网| 桃色一区二区三区在线观看| av专区在线播放| 午夜福利视频1000在线观看| 两个人视频免费观看高清| 久久久国产成人免费| 久久精品国产鲁丝片午夜精品| 亚洲,欧美,日韩| 亚洲激情五月婷婷啪啪| АⅤ资源中文在线天堂| 美女cb高潮喷水在线观看| 99久久人妻综合| 国产黄片美女视频| 看非洲黑人一级黄片| av在线天堂中文字幕| 中文字幕精品亚洲无线码一区| 18+在线观看网站| 中文字幕av成人在线电影| 欧美性猛交黑人性爽| 99热6这里只有精品| 少妇的逼水好多| 精品人妻偷拍中文字幕| 久久人人爽人人爽人人片va| 在线免费观看不下载黄p国产| 美女被艹到高潮喷水动态| 亚洲成人久久性| 97超视频在线观看视频| 日韩强制内射视频| 欧美又色又爽又黄视频| 国产精品免费一区二区三区在线| 好男人视频免费观看在线| 国产亚洲欧美98| 波多野结衣巨乳人妻| 精品久久久久久久久久免费视频| 午夜福利成人在线免费观看| 国产精品乱码一区二三区的特点| 欧美日本亚洲视频在线播放| 少妇裸体淫交视频免费看高清| 国产精品一区www在线观看| 日本在线视频免费播放| 久久久久久久久久久免费av| 国产精品无大码| 国产黄色小视频在线观看| 亚洲国产精品成人久久小说 | 免费无遮挡裸体视频| 黄片无遮挡物在线观看| 一级二级三级毛片免费看| 亚洲国产精品成人久久小说 | 欧美精品国产亚洲| 给我免费播放毛片高清在线观看| 国产在线精品亚洲第一网站| 夜夜看夜夜爽夜夜摸| 国产成年人精品一区二区| 老司机影院成人| 国产激情偷乱视频一区二区| 2022亚洲国产成人精品| a级毛片a级免费在线| 免费在线观看成人毛片| 成人永久免费在线观看视频| 日日啪夜夜撸| 中文精品一卡2卡3卡4更新| 国产亚洲av嫩草精品影院| 精品人妻一区二区三区麻豆| 你懂的网址亚洲精品在线观看 | 99热6这里只有精品| 99视频精品全部免费 在线| 搡老妇女老女人老熟妇| 97超碰精品成人国产| 97超视频在线观看视频| 国内久久婷婷六月综合欲色啪| 村上凉子中文字幕在线| 青春草国产在线视频 | 嫩草影院入口| 不卡视频在线观看欧美| 亚洲av一区综合| 国产欧美日韩精品一区二区| 国产成人精品婷婷| 乱码一卡2卡4卡精品| 哪个播放器可以免费观看大片| 国产黄色视频一区二区在线观看 | 日韩欧美三级三区| 国产精品永久免费网站| 国产一区二区在线观看日韩| 极品教师在线视频| 2021天堂中文幕一二区在线观| 国产片特级美女逼逼视频| 在线a可以看的网站| 校园人妻丝袜中文字幕| 国产成人一区二区在线| 成年女人永久免费观看视频| 国产精品电影一区二区三区| 亚洲精品色激情综合| 别揉我奶头 嗯啊视频| 久久人人精品亚洲av| 国产成人aa在线观看| 亚洲中文字幕日韩| 亚洲精品国产av成人精品| 大香蕉久久网| 亚洲七黄色美女视频| 亚洲人成网站在线观看播放| 又粗又硬又长又爽又黄的视频 | 免费观看a级毛片全部| 99热全是精品| 日韩av不卡免费在线播放| 美女cb高潮喷水在线观看| 亚洲经典国产精华液单| 免费搜索国产男女视频| 久久久久久久久久久免费av| 久久韩国三级中文字幕| 岛国毛片在线播放| 欧美在线一区亚洲| 大香蕉久久网| 一个人观看的视频www高清免费观看| 亚洲国产日韩欧美精品在线观看| 久久久久久大精品| 赤兔流量卡办理| 国产精品av视频在线免费观看| 国产亚洲精品久久久com| 国产人妻一区二区三区在| 黄色视频,在线免费观看| 丝袜喷水一区| 99国产极品粉嫩在线观看| 国产精品久久视频播放| 成人一区二区视频在线观看| 国产 一区精品| 国产亚洲av嫩草精品影院| 麻豆成人av视频| 中文欧美无线码| 亚洲欧美日韩高清专用| kizo精华| 亚洲综合色惰| 插阴视频在线观看视频| 日本色播在线视频| 成人漫画全彩无遮挡| 少妇人妻精品综合一区二区 | 成年版毛片免费区| 免费电影在线观看免费观看| 乱系列少妇在线播放| 黄色配什么色好看| 我的女老师完整版在线观看| 久久人人爽人人片av| 99久久九九国产精品国产免费| 97超视频在线观看视频| 日韩一区二区三区影片| 国产亚洲精品久久久久久毛片| 日韩人妻高清精品专区| 男插女下体视频免费在线播放| 亚洲最大成人中文| 国产成人精品婷婷| 国产精品一区www在线观看| 亚洲,欧美,日韩| 一本—道久久a久久精品蜜桃钙片 精品乱码久久久久久99久播 | 久久午夜福利片| 国产高清三级在线| 国产极品天堂在线| 亚洲高清免费不卡视频| 亚洲精华国产精华液的使用体验 | 观看免费一级毛片| 国产真实伦视频高清在线观看| 男女视频在线观看网站免费| 久久久久久久久中文| 国产一区二区三区av在线 | 国产精品久久久久久久电影| 亚洲最大成人av| 联通29元200g的流量卡| 日本与韩国留学比较| 亚洲国产精品成人综合色| 欧美成人一区二区免费高清观看| 人人妻人人看人人澡| 热99在线观看视频| 人妻少妇偷人精品九色| 亚洲国产精品合色在线| 哪里可以看免费的av片| 三级毛片av免费| 国产精华一区二区三区| 国产激情偷乱视频一区二区| 亚洲精品日韩在线中文字幕 | 精品99又大又爽又粗少妇毛片| 免费看日本二区| 国产欧美日韩精品一区二区| 免费不卡的大黄色大毛片视频在线观看 | 嫩草影院精品99| 亚洲人成网站在线播| 婷婷色综合大香蕉| 国产美女午夜福利| h日本视频在线播放| 亚洲国产欧洲综合997久久,| 中文字幕人妻熟人妻熟丝袜美| 国产亚洲精品av在线| 最好的美女福利视频网| 99热全是精品| 夜夜看夜夜爽夜夜摸| 国产精品女同一区二区软件| 高清毛片免费看| 岛国毛片在线播放| 欧美日韩乱码在线| 午夜视频国产福利| 成人三级黄色视频| 亚洲人与动物交配视频| 久久精品91蜜桃| 日日干狠狠操夜夜爽| 麻豆一二三区av精品| 亚州av有码| 内地一区二区视频在线| 国产av在哪里看| 午夜免费激情av| 国产精品人妻久久久久久| 国产私拍福利视频在线观看| 免费看日本二区| 大型黄色视频在线免费观看| av卡一久久| 在线播放无遮挡| 又粗又硬又长又爽又黄的视频 | 国产精品99久久久久久久久| 长腿黑丝高跟| 九九爱精品视频在线观看| 久久99精品国语久久久| 1024手机看黄色片| 在线观看66精品国产| 少妇裸体淫交视频免费看高清| 欧美潮喷喷水| 在线免费观看的www视频| 91精品国产九色| 最好的美女福利视频网| 成年av动漫网址| 国产私拍福利视频在线观看| 国产欧美日韩精品一区二区| 美女xxoo啪啪120秒动态图| 亚洲欧美日韩东京热| 欧美在线一区亚洲| 日本-黄色视频高清免费观看| 一本久久中文字幕| 久久热精品热| 久久精品久久久久久久性| 亚洲成人中文字幕在线播放| 午夜久久久久精精品| 长腿黑丝高跟| 国产午夜精品论理片| 丝袜美腿在线中文| 国产精品一区二区三区四区免费观看| av在线天堂中文字幕| 欧美高清成人免费视频www| 免费不卡的大黄色大毛片视频在线观看 | h日本视频在线播放| 国产黄色小视频在线观看| 久久久欧美国产精品| 一边亲一边摸免费视频| 久久九九热精品免费| 亚洲最大成人中文| 亚洲最大成人av| 老女人水多毛片| 成人三级黄色视频| 少妇熟女欧美另类| 成人毛片60女人毛片免费| 18禁裸乳无遮挡免费网站照片| 亚洲色图av天堂| 日韩成人av中文字幕在线观看| 国产精品野战在线观看| 欧美精品国产亚洲| 99热这里只有是精品50| 国产亚洲91精品色在线| 久久久a久久爽久久v久久| 久久久久久久久大av| 国产在视频线在精品| 免费观看a级毛片全部| 亚洲在线观看片| 少妇高潮的动态图| 免费大片18禁| 亚洲人成网站在线观看播放| 国产成人精品婷婷| 高清在线视频一区二区三区 | av.在线天堂| 日韩欧美精品免费久久| 成年版毛片免费区| 国产黄片视频在线免费观看| 尾随美女入室| 日本爱情动作片www.在线观看| 看免费成人av毛片| 综合色丁香网| 在线播放无遮挡| 乱人视频在线观看| 中文字幕av在线有码专区| 亚洲自偷自拍三级| 丝袜喷水一区| 一本—道久久a久久精品蜜桃钙片 精品乱码久久久久久99久播 | 国产单亲对白刺激| 亚洲乱码一区二区免费版| 久久精品国产亚洲网站| 国产精品久久久久久亚洲av鲁大| 国产精华一区二区三区| 欧美最新免费一区二区三区| 日日摸夜夜添夜夜爱| 国产成人freesex在线| 国产一区二区三区在线臀色熟女| 国产精品野战在线观看| 亚洲内射少妇av| 亚洲三级黄色毛片| 亚洲熟妇中文字幕五十中出| 十八禁国产超污无遮挡网站| 男人舔女人下体高潮全视频| 特大巨黑吊av在线直播| 黄片无遮挡物在线观看| 别揉我奶头 嗯啊视频| 床上黄色一级片| 免费看光身美女| 99久久精品国产国产毛片| 日本五十路高清| 高清毛片免费看| 麻豆国产97在线/欧美| 伊人久久精品亚洲午夜| 亚洲av免费高清在线观看| 欧美日本亚洲视频在线播放| 国产亚洲av片在线观看秒播厂 | 此物有八面人人有两片| 久久精品国产亚洲av天美| 亚洲内射少妇av| 人妻制服诱惑在线中文字幕| 好男人在线观看高清免费视频| 最好的美女福利视频网| 国产亚洲av片在线观看秒播厂 | 久久99精品国语久久久| 天堂影院成人在线观看| 激情 狠狠 欧美| 变态另类成人亚洲欧美熟女| 一级黄片播放器| 国产蜜桃级精品一区二区三区| 波多野结衣巨乳人妻| 国产成人a∨麻豆精品| 男的添女的下面高潮视频| 一卡2卡三卡四卡精品乱码亚洲| 亚洲18禁久久av| 国产探花极品一区二区| 好男人在线观看高清免费视频| 亚洲精品日韩av片在线观看| 黄色欧美视频在线观看| 少妇裸体淫交视频免费看高清| 国产精品久久久久久久电影| 欧美丝袜亚洲另类| 国产精品,欧美在线| 尾随美女入室| 99久久精品国产国产毛片| 亚洲成人精品中文字幕电影| 精品久久久久久久久久免费视频| 搡老妇女老女人老熟妇| 我的老师免费观看完整版| 蜜桃亚洲精品一区二区三区| 国产成人精品婷婷| 欧美激情久久久久久爽电影| 乱系列少妇在线播放| 亚洲欧洲国产日韩| 看黄色毛片网站| 六月丁香七月| 99国产极品粉嫩在线观看| 中文字幕人妻熟人妻熟丝袜美| 中国国产av一级| 成人特级黄色片久久久久久久| 干丝袜人妻中文字幕| 国产高清视频在线观看网站| 国内精品一区二区在线观看| 国产成人aa在线观看| 99久久精品热视频| 久久亚洲国产成人精品v| 男插女下体视频免费在线播放| 亚洲欧美日韩无卡精品| 国产探花在线观看一区二区| 男女视频在线观看网站免费| 日日啪夜夜撸| 男女那种视频在线观看| a级毛片a级免费在线| 亚洲成人av在线免费| a级毛色黄片| 国产一区亚洲一区在线观看| 国产成人freesex在线| 99在线视频只有这里精品首页| 午夜a级毛片| 国产精品人妻久久久久久| 一本久久中文字幕| 美女大奶头视频| 亚洲丝袜综合中文字幕| 一个人观看的视频www高清免费观看| 22中文网久久字幕| 精品人妻一区二区三区麻豆| 日本免费a在线| 精品一区二区三区人妻视频| 亚洲久久久久久中文字幕| 亚洲最大成人av| 免费观看在线日韩| 午夜福利成人在线免费观看| 久久久久久久午夜电影| 插逼视频在线观看| 久久久国产成人精品二区| 中文精品一卡2卡3卡4更新| 老熟妇乱子伦视频在线观看| 日韩一区二区三区影片| 国产高清视频在线观看网站| 亚洲色图av天堂| 99热这里只有是精品在线观看| 国产高清视频在线观看网站| 三级经典国产精品| 桃色一区二区三区在线观看| 亚洲av成人av| 看十八女毛片水多多多| 桃色一区二区三区在线观看| 久久精品影院6| 在线观看66精品国产| 成人二区视频| 97在线视频观看| 中文亚洲av片在线观看爽| 国产精品蜜桃在线观看 | 久久久精品大字幕| 国产成人a∨麻豆精品| 自拍偷自拍亚洲精品老妇| 大又大粗又爽又黄少妇毛片口| 精品久久久久久久末码| 久久国产乱子免费精品| 国产午夜精品一二区理论片|