sodium metabisulfite

  • 文章类型: Journal Article
    棘阿米巴角膜炎(AK)是角膜的严重感染。由于目前可获得的化合物的无效性,预防和治疗是困难的。许多常用的化合物对棘阿米巴常规检查的影响尚未探索,但可能提供对对抗AK有用的见解。在这项研究中,我们证明了焦亚硫酸钠,眼睛护理解决方案的常见保存成分,发现浓度低于滴眼剂中常见浓度(IC500.03mg/mL)时对棘阿米巴滋养体具有活性。我们证明了焦亚硫酸钠会从生长培养基中消耗硫胺素,并且棘阿米巴是硫胺素营养缺陷型,需要硫胺素抢救才能生长。补充硫胺素可以克服焦亚硫酸钠的抑制作用。这些结果与棘阿米巴基因组中硫胺素生物合成的关键酶的缺乏相一致,使用新的或现有的化合物可能被证明是可开发的区域。的确,这项研究强调了偏亚硫酸氢钠作为一种有用的抑制剂,在体外castellanii滋养体,至少在某种程度上,通过限制可用的硫胺素。
    Acanthamoeba keratitis (AK) is a severe infection of the cornea. Prevention and treatment are difficult due to the inefficacy of currently available compounds. The impact of many commonly used compounds for routine examinations of Acanthamoeba is unexplored but might offer insight useful in combatting AK. In this study, we demonstrate that sodium metabisulfite, a common preservation constituent of eye care solutions, was found to be active against Acanthamoeba trophozoites at concentrations lower than that commonly found in eye drops (IC50 0.03 mg/mL). We demonstrate that sodium metabisulfite depletes thiamine from growth medium and that Acanthamoeba is a thiamine auxotroph, requiring thiamine salvage for growth. The inhibitory effects of sodium metabisulfite can be overcome by thiamine supplementation. These results are consistent with the lack of key enzymes for thiamine biosynthesis in the genome of Acanthamoeba, an area which might prove exploitable using new or existing compounds. Indeed, this study highlights sodium metabisulfite as a useful inhibitor of Acanthamoeba castellanii trophozoites in vitro and that it acts, at least in part, by limiting available thiamine.
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  • 文章类型: Journal Article
    鱼腥草(HC)是亚洲广泛分布的植物,广泛用于食品和药用。初步调查发现,HC在现场加工过程中经常被焦亚硫酸钠溶液漂白,导致健康风险。在这项研究中,使用挥发性和非挥发性靶向代谢组学方法综合评价了焦亚硫酸钠对HC质量的影响。结果表明,化学成分变化的程度与漂白时间呈正相关。这些显著的变化主要发生在漂白的初始阶段。随后,使用非靶向UPLC/Q-TOFMS方法来探索漂白HC中潜在的化学漂白标记。随后制备标记物1-羟基-3-氧代癸烷-1-磺酸,孤立的,并确定。市场样本验证进一步验证了该标记的准确性和有效性。
    Houttuynia Cordata (HC) is a widely distributed plant in Asia and is used extensively for both food and medicinal purposes. A preliminary investigation found that HC is often bleached with sodium metabisulfite solution during its field processing, leading to health risks. In this study, the effects of sodium metabisulfite on the quality of HC were comprehensively evaluated using volatile and non-volatile targeted metabolomic methods. The results revealed a positive correlation between the extent of chemical composition changes and the bleaching time. These notable changes mainly occurred at the initial stage of bleaching. Subsequently, an untargeted UPLC/Q-TOF MS method was used to explore the potential chemical bleaching markers in bleached HC. The marker 1-hydroxy-3-oxodecane-1-sulfonic acid was subsequently prepared, isolated, and identified. Market sample verification further validated the accuracy and effectiveness of this marker.
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  • 文章类型: Journal Article
    这项研究评估了盐偏亚硫酸氢钠(SMB)的功效,碳酸氢铵,碳酸氢钠,和正磷酸二氢钾首先在体外对抗主要的采后果腐病真菌,交替菌,灰葡萄孢菌,意大利青霉,和指状青霉。结果表明,0.2%的SMB完全抑制了真菌菌丝的生长。碳酸氢铵和碳酸氢钠在抑制菌丝体生长的效果在0.2%时较差,从57.6%到77.6%不等。效果最差的是正磷酸二氢钾。还在接种了最致病真菌的受伤苹果上进行了体内实验,B.cinerea,并用浓度为0.2、0.5、1、2和3%的SMB处理,无论是预防性的还是治疗性的。基于衰减大小的结果表明,SMB,当用作预防性治疗时,功效降低,即使浓度最高。然而,这种盐被证明是非常有效的治疗0.5%,因为腐烂被完全阻止。我们的结果表明,作为治愈性治疗,用于收获后处理的SMB的合适浓度为0.5%。另一方面,1%剂量诱导伤口周围植物毒性的发作。为了评估植物毒性反应的程度,将较高浓度的1-4%SMB应用于受伤的果实。苹果和橙子接种或不接种灰芽孢杆菌和指状芽孢杆菌,分别。1-4%的剂量以围绕果实表皮上伤口的变色环的形式引起植物毒性;随着SMB浓度的增加,这种植物毒性增大。苹果和橙子的植物毒性特征相似。方法学程序使得可以进行SMB植物毒性的定量评估。该方法被认为是一种易于使用的技术,用于定量估计抗真菌化合物对收获后果实的植物毒性。
    This study evaluates the efficacy of the salts sodium metabisulfite (SMB), ammonium bicarbonate, sodium bicarbonate, and potassium dihydrogen orthophosphate first in vitro against the main postharvest fruit rot fungi, Alternaria alternata, Botrytis cinerea, Penicillium italicum, and Penicillium digitatum. Results showed that 0.2% SMB completely inhibited the mycelium growth of the fungal species. Ammonium bicarbonate and sodium bicarbonate were less effective at 0.2% in inhibiting mycelial growth, ranging from 57.6% to 77.6%. The least effective was potassium dihydrogen orthophosphate. Experiments were also performed in vivo on wounded apples inoculated with the most pathogenic fungus, B. cinerea, and treated with SMB at concentrations of 0.2, 0.5, 1, 2, and 3%, both preventively and curatively. Results based on the decay size showed that SMB, when used as a preventive treatment, had a reduced efficacy, even with the highest concentration. However, this salt proved to be very effective at 0.5% in curative treatment since the decay was completely blocked. Our results suggest that the appropriate concentration of SMB for post-harvest treatment is 0.5% as a curative treatment. On the other hand, the 1% dose induced the onset of phytotoxicity around the wound. To assess the extent of the phytotoxicity reaction, higher concentrations of 1-4% SMB were applied to wounded fruit. Apples and oranges were inoculated or not with B. cinerea and P. digitatum, respectively. Doses of 1-4% induced phytotoxicity in the form of a discolored ring surrounding the wound on the epidermis of the fruit; this phytotoxicity enlarged as the concentration of SMB increased. The phytotoxic features were similar on apples and oranges. The methodological procedure made it possible to carry out a quantitative assessment of SMB phytotoxicity. This method is proposed as an easy-to-use technique for quantitatively estimating the phytotoxicity of antifungal compounds on post-harvest fruit.
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  • 文章类型: Journal Article
    为了提示虾的外观颜色,虾加工中经常添加偏亚硫酸氢钠,也就是说,然而,禁止在中国和许多其他国家。本研究旨在建立一种用于对虾表面焦亚硫酸钠残留的表面增强拉曼光谱(SERS)方法,以一种非破坏性的方式。使用便携式拉曼光谱仪与装载有银纳米颗粒作为基底材料的抄纸联合进行分析。焦亚硫酸钠的SERS响应分别在620(强)和927(中)cm-1处给出两个指纹峰。这使得能够明确地确认目标化学品。SERS检测方法的灵敏度确定为0.1mg/mL,等于虾表面0.31mg/kg的残留焦亚硫酸钠。建立了620cm-1峰强度与焦亚硫酸钠浓度之间的定量关系。线性拟合方程为y=2375x+8714,R2=0.985。在简单中达到理想的平衡,灵敏度,和选择性,这项研究表明,所提出的方法是理想的现场和非破坏性的筛查在海鲜中的偏亚硫酸氢钠残留。
    In order to prompt the appearance of the shrimp color, sodium metabisulfite is frequently added in shrimp processing, which is, however, prohibited in China and many other countries. This study aimed to establish a surface-enhanced Raman spectroscopy (SERS) method for screening sodium metabisulfite residues on shrimp surfaces, in a non-destructive manner. The analysis was carried out using a portable Raman spectrometer jointly with copy paper loaded with silver nanoparticles as the substrate material. The SERS response of sodium metabisulfite gives two fingerprint peaks at 620 (strong) and 927 (medium) cm-1, respectively. This enabled unambiguous confirmation of the targeted chemical. The sensitivity of the SERS detection method was determined to be 0.1 mg/mL, which was equal to residual sodium metabisulfite on the shrimp surface at 0.31 mg/kg. The quantitative relationship between the 620 cm-1 peak intensities and the concentrations of sodium metabisulfite was established. The linear fitting equation was y = 2375x + 8714 with R2 = 0.985. Reaching an ideal balance in simplicity, sensitivity, and selectivity, this study demonstrates that the proposed method is ideally suitable for in-site and non-destructive screening of sodium metabisulfite residues in seafood.
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  • 文章类型: Journal Article
    雌性猪对DON和ZEN反应敏感,伴有厌食和内分泌干扰,分别,当超过临界饮食浓度时。因此,DON和ZEN对饲料的频繁共污染需要它们的平行灭活。添加剂ZenA水解ZEN,而SBS通过磺化使DON失活。同时添加两种补充剂(+,2.5克SBS和100UZenA/kg)与对照饮食(CON-,0.04mgDON和<0.004mgZEN/kg;CON+,0.03mgDON和<0.004mgZEN/kg)和镰刀菌毒素污染的饮食(FUS-,2.57mgDON和0.24mgZEN/kg;FUS+,2.04mgDON和0.24mgZEN/kg)。将4种日粮饲喂20只雌性断奶仔猪(初始体重6kg)35天;此后处死仔猪以收集样品。补充剂改善了性能,改善了代谢和血液学,而与饮食DON污染无关。这些变化背后的机制无法澄清,需要进一步考虑。SBS将饲料中的DON浓度降低了约20%,并在血浆和尿液中降低了相同程度,这表明在上消化道吸收DON之前,消化道中没有进一步形成DON磺酸盐,或者另外形成的DON磺酸盐逃脱了吸收。在粪便中检测到DON磺酸盐,表明未吸收的DON磺酸盐到达粪便和/或未吸收的DON在后肠中被磺化。观察到的20%的减少率被评估为不足以用于喂养实践。添加到干饲料中的SBS的Galenic形式需要改进以支持近端消化道中的DON磺化。ZenA在消化道中具有活性,其水解的非雌激素反应产物水解的ZEN(HZEN)和脱羧和水解的ZEN(DHZEN)在粪便中均存在,全身循环,FUS+组尿液与FUS-组相比。这些水解产物的存在伴随着高雌激素ZEN浓度的显着降低,反过来,与FUS组相比,与子宫和卵巢的相对重量降低有关。因此,ZenA被证明是有效的;在生物标志物和生物效应方面。
    Female pigs respond sensitive both to DON and ZEN with anorexia and endocrine disruption, respectively, when critical diet concentrations are exceeded. Therefore, the frequent co-contamination of feed by DON and ZEN requires their parallel inactivation. The additive ZenA hydrolyzes ZEN while SBS inactivates DON through sulfonation. Both supplements were simultaneously added (+, 2.5 g SBS and 100 U ZenA/kg) to a control diet (CON-, 0.04 mg DON and < 0.004 mg ZEN/kg; CON+, 0.03 mg DON and < 0.004 mg ZEN/kg) and a Fusarium toxin contaminated diet (FUS-, 2.57 mg DON and 0.24 mg ZEN/kg; FUS+, 2.04 mg DON and 0.24 mg ZEN/kg). The 4 diets were fed to 20 female weaned piglets each (6 kg initial body weight) for 35 days; the piglets were sacrificed thereafter for collecting samples. Supplements improved performance and modified metabolism and hematology independent of dietary DON contamination. The mechanisms behind these changes could not be clarified and require further consideration. SBS reduced DON concentration in feed by approximately 20% and to the same extent in blood plasma and urine suggesting that no further DON sulfonate formation occurred in the digestive tract before absorbing DON in the upper digestive tract or that additionally formed DON sulfonates escaped absorption. DON sulfonates were detected in feces suggesting that unabsorbed DON sulfonates reached feces and/or that unabsorbed DON was sulfonated in the hindgut. The observed reduction rate of 20% was evaluated to be insufficient for feeding practice. Galenic form of SBS added to dry feed needs to be improved to support the DON sulfonation in the proximal digestive tract.ZenA was active in the digestive tract as demonstrated by the presence of its hydrolyzed none-estrogenic reaction products hydrolyzed ZEN (HZEN) and decarboxylated and hydrolyzed ZEN (DHZEN) both in feces, systemic circulation, and urine of group FUS+ compared to group FUS-. The presence of these hydrolysis products was paralleled by a significant decrease in high-estrogenic ZEN concentrations which, in turn, was related to a decrease in relative weights of uteri and ovaries when compared to group FUS-. Thus, ZenA was proven to be effective; both in terms of biomarkers and biological effects.
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  • 文章类型: Journal Article
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  • 文章类型: Journal Article
    二氧化硫-亚硫酸盐(E220-228)在2016年进行了重新评估,导致临时ADI设定为0.7mgSO2当量/kgbw/天。在欧盟委员会要求提供数据之后,本后续意见评估了感兴趣的企业经营者(IBO)提供的数据以及公开文献中确定的其他证据.IBO没有提交新的生物学或毒理学数据,以解决重新评估中描述的数据空白。考虑到文献检索中确定的数据,小组的结论是,先前在重新评估中确定的不确定性没有大大减少。因此,小组认为现有的毒性数据库不足以得出ADI,因此撤回了目前临时组的每日可接受摄入量(ADI).暴露边际(MOE)方法被认为适合评估这些食品添加剂的风险。基准剂量的置信下限为每天38mgSO2当量/kgbw,低于以前的参考点70mgSO2当量/kgbw/天,是根据延长的视觉诱发电位潜伏期估计的。评估因子为80用于教育部的评估。在估计的饮食暴露中,当使用精细曝光场景(数据集D)时,除青少年外,所有人群的MOE在第95百分位数范围的最大值均低于80。使用最大允许水平估计的饮食暴露将导致所有人口群体的MOE低于80,在平均值的最大范围内,对于大多数人口群体,在第95百分位数的范围的最小值和最大值。专家小组的结论是,这引起了对两种饮食暴露情况的安全关注。小组还对二氧化硫-亚硫酸盐中存在的有毒元素进行了风险评估(E220-228),根据IBO提交的数据,得出的结论是,欧盟规范中砷的最大限值,铅和汞应降低,并应引入镉的最大限值。
    Sulfur dioxide-sulfites (E 220-228) were re-evaluated in 2016, resulting in the setting of a temporary ADI of 0.7 mg SO2 equivalents/kg bw per day. Following a European Commission call for data, the present follow-up opinion assesses data provided by interested business operators (IBOs) and additional evidence identified in the publicly available literature. No new biological or toxicological data addressing the data gaps described in the re-evaluation were submitted by IBOs. Taking into account data identified from the literature search, the Panel concluded that there was no substantial reduction in the uncertainties previously identified in the re-evaluation. Therefore, the Panel considered that the available toxicity database was inadequate to derive an ADI and withdrew the current temporary group acceptable daily intake (ADI). A margin of exposure (MOE) approach was considered appropriate to assess the risk for these food additives. A lower confidence limit of the benchmark dose of 38 mg SO2 equivalents/kg bw per day, which is lower than the previous reference point of 70 mg SO2 equivalents/kg bw per day, was estimated based on prolonged visual evoked potential latency. An assessment factor of 80 was applied for the assessment of the MoE. At the estimated dietary exposures, when using a refined exposure scenario (Data set D), MOEs at the maximum of 95th percentile ranges were below 80 for all population groups except for adolescents. The dietary exposures estimated using the maximum permitted levels would result in MOEs below 80 in all population groups at the maximum of the ranges of the mean, and for most of the population groups at both minimum and maximum of the ranges at the 95th percentile. The Panel concluded that this raises a safety concern for both dietary exposure scenarios. The Panel also performed a risk assessment for toxic elements present in sulfur dioxide-sulfites (E 220-228), based on data submitted by IBOs, and concluded that the maximum limits in the EU specifications for arsenic, lead and mercury should be lowered and a maximum limit for cadmium should be introduced.
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  • 文章类型: Journal Article
    在一项为期35天的研究中,总共使用了4,318头猪(337×1,050,PIC;最初为6.5±0.08kg),以评估饮食中霉菌毒素控制策略对保育猪性能和血液指标的影响。猪在大约21日龄断奶,并随机分配给5种饮食治疗中的1种,采用随机完整的区组设计,具有包括母猪农场来源的阻塞结构,进入设施的日期,和平均笔BW。共有160支钢笔与80支双面5孔不锈钢围栏线馈线一起使用,用喂食器作为实验单元。对于每个馈线,1笔包含27只母猪,1笔包含27只手推车。每个饮食处理有16次重复。在处理饮食之前,将常见的1期饮食以颗粒形式饲喂给所有猪7天。实验治疗在断奶后第7至42天(研究的第0至35天)喂养,包括低脱氧雪腐镰刀菌烯醇(DON)饮食(1.12±0.323mg/kg),高DON饮食(2.34±1.809mg/kg),高DON+0.50%焦亚硫酸钠(SMB),高DON+两种缓解产品之一;0.30%技术1,或0.30%技术1+。技术1和1+由粘土组成,酵母细胞壁成分,和植物提取物的混合物。Technology1+还包含SMB。总体上(第0至35天),饲喂高DON的猪最终BW降低(P<0.05),ADG,和ADFI与低DON相比。此外,与所有其他处理相比,饲喂高DON+SMB的猪ADG增加(P<0.05)。在饲喂高DON+SMB或高DON+技术1+的猪中观察到G:F的改善(P<0.05),与低DON或高DON+技术1日粮相比。与饲喂具有高DON和高DON技术1中间体的低DON日粮的猪相比,饲喂高DONSMB或高DON技术1日粮的猪的总去除量和死亡率降低(P<0.05)。在第35天收集的循环血液的液相色谱/质谱分析显示,与具有高DON+SMB和高DON+技术1+中间体的低DON相比,饲喂高DON或高DON+技术1的猪的DON浓度增加(P<0.05)。总之,与饲喂低DON的猪相比,饲喂高DON日粮的猪的表现降低。高DON日粮中的焦亚硫酸钠在ADG和G:F的生长性能方面具有优势,超过了低DON日粮中的生长性能,而G:F比值的改善以及与Technology1+相关的其他免疫代谢变化(γ-谷氨酰转移酶和肌酸激酶)值得进一步研究.
    A total of 4,318 pigs (337 × 1,050, PIC; initially 6.5 ± 0.08 kg) were used in a 35-day study to evaluate dietary mycotoxin control strategies on nursery pig performance and blood measures. Pigs were weaned at approximately 21 d of age and randomly allotted to 1 of 5 dietary treatments in a randomized complete block design with blocking structure including sow farm origin, date of entry into facility, and average pen BW. A total of 160 pens were used with 80 double-sided 5-hole stainless steel fence line feeders, with feeder serving as the experimental unit. For each feeder, 1 pen contained 27 gilts and 1 pen contained 27 barrows. There were 16 replications per dietary treatment. A common phase 1 diet was fed to all pigs in pelleted form for 7 day prior to treatment diets. Experimental treatments were fed from days 7 to 42 after weaning (days 0 to 35 of the study) and included a low deoxynivalenol (DON) diet (1.12 ± 0.623 mg/kg), high DON diet (2.34 ± 1.809 mg/kg), high DON+ 0.50% sodium metabisulfite (SMB), high DON+ one of two mitigating products; 0.30% Technology1, or 0.30% Technology1+. Technology1 and 1+ are comprised of clays, yeast cell wall components, and a blend of plant extracts. Technology1+ also contains SMB. Overall (days 0 to 35), pigs fed high DON had decreased (P < 0.05) final BW, ADG, and ADFI compared with low DON. Additionally, pigs fed high DON+SMB had increased (P < 0.05) ADG compared with all other treatments. An improvement (P < 0.05) in G:F was observed in pigs fed high DON + SMB or high DON + Technology1+ compared with the low DON or high DON + Technology1 diets with high DON diets intermediate. Pigs fed high DON + SMB or high DON + Technology1 diets had reduced (P < 0.05) total removals and mortality compared with pigs fed low DON diets with high DON and high DON + Technology1+ intermediate. Liquid chromatography/mass spectrometry analysis of circulating blood collected on day 35 revealed that pigs fed high DON or high DON + Technology1 had increased (P < 0.05) DON concentrations compared to low DON with high DON + SMB and high DON + Technology1+ intermediate. In summary, pigs fed high DON diets had reduced performance compared with pigs fed low DON. Sodium metabisulfite in high DON diets provided a benefit in growth performance with ADG and G:F exceeding growth performance in the low DON diet while, the improved G:F ratio combined with other immunometabolic changes (gamma glutamyltransferase and creatine kinase) associated with Technology1+ warrant further investigation.
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  • 文章类型: Journal Article
    背景:防腐剂广泛用于化妆品中,家庭,和工业产品,以防止微生物生长和产品变质。最近发生了对甲基异噻唑啉酮(MI)的接触过敏。
    目的:建立随着MI被替换的防腐剂接触过敏的新趋势。
    方法:我们于2011年1月至2019年12月在我们中心对连续斑贴试验的患者进行了回顾性研究。
    结果:在2011年1月至2019年12月期间,共包括7846名连续斑贴测试的患者。对MI过敏的患病率从2013年的9.39%的峰值下降到2019年的1.98%。MI/甲基氯异噻唑啉酮(MCI)遵循类似的趋势,2014年的峰值患病率为7.85%,2018年降至1.39%。对苯并异噻唑啉酮(BIT)的接触过敏的患病率从2014年的0.26%(置信区间[CI]0.03-0.93)稳步上升至2019年的3.42%(CI2.43-4.66)。
    结论:我们的数据显示,在过去6年中,对BIT的接触过敏的患病率显着增加,可能是家用产品使用量增加的结果。必须继续寻找接触过敏的新趋势,以便及时采取预防措施。
    BACKGROUND: Preservatives are used widely in cosmetic, household, and industrial products to prevent microbial growth and spoiling of the products. There has been a recent epidemic of contact allergy to methylisothiazolinone (MI).
    OBJECTIVE: To establish emerging trends in preservative contact allergy as MI is replaced.
    METHODS: We performed a retrospective study on consecutively patch-tested patients at our center from January 2011 to December 2019.
    RESULTS: A total of 7846 consecutively patch-tested patients were included who were patch tested between January 2011 and December 2019. The prevalence of allergy to MI fell from a peak of 9.39% in 2013 to 1.98% in 2019. MI/methylchloroisothiazolinone (MCI) followed a similar trend, with a peak prevalence of 7.85% in 2014 and falling to 1.39% in 2018. The prevalence of contact allergy to benzisothiazolinone (BIT) has increased steadily increased in 2014 from 0.26% (confidence interval [CI] 0.03-0.93) to 3.42% (CI 2.43-4.66) in 2019.
    CONCLUSIONS: Our data show a significant increase in the prevalence of contact allergy to BIT over the last 6 years, probably as a consequence of increased use in household products. It is essential to continue to look for emerging trends in contact allergy to enable prompt preventative measures to be taken.
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  • 文章类型: Journal Article
    胶原蛋白支架,角膜组织工程中最常用的生物材料之一,经常交联以改善机械性能,酶耐受性,和热稳定性。交联剂如1-乙基-3-(3-二甲基氨基丙基)碳二亚胺盐酸盐(EDC)与组织相容,但提供低交联密度和降低的机械性能。相反,交联剂如戊二醛(GTA)可以产生机械上更坚固的支架;然而,它们也可以引起更大的毒性。在这里,我们评估了EDC和GTA双重交联的有效性,以及偏亚硫酸氢钠(SM)和硼氢化钠(SB)在交联后中和毒性并恢复生物相容性的能力。用不同浓度的GTA处理EDC交联的胶原支架。为了中和游离的未反应的醛基,支架用SM或SB处理。评估了这些反应中涉及的化学以及胶原支架的机械和功能特性。使用不同的角膜细胞类型研究了在支架上生长的细胞的活力。评价每种类型的支架处理对人单核细胞分化的影响。单因素方差分析用于统计分析。GTA作为双交联剂的加入显著提高了EDC交联胶原支架的力学性能和酶稳定性。GTA降低了细胞生物相容性,但用SB或SM处理可以逆转这种作用。这些试剂不会影响机械性能,酶稳定性,或双交联支架的透明度。单核细胞与不同支架的接触不会触发它们分化成活化的巨噬细胞。我们的结果表明,GTA以剂量依赖的方式提高了EDC交联支架的机械性能,随后用SB或SM处理部分恢复了生物相容性。这种新颖的制造方法将有助于将基于胶原蛋白的人造角膜转化为临床环境。
    Collagen scaffolds, one of the most used biomaterials in corneal tissue engineering, are frequently crosslinked to improve mechanical properties, enzyme tolerance, and thermal stability. Crosslinkers such as 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) are compatible with tissues but provide low crosslinking density and reduced mechanical properties. Conversely, crosslinkers such as glutaraldehyde (GTA) can generate mechanically more robust scaffolds; however, they can also induce greater toxicity. Herein, we evaluated the effectivity of double-crosslinking with both EDC and GTA together with the capability of sodium metabisulfite (SM) and sodium borohydride (SB) to neutralize the toxicity and restore biocompatibility after crosslinking. The EDC-crosslinked collagen scaffolds were treated with different concentrations of GTA. To neutralize the free unreacted aldehyde groups, scaffolds were treated with SM or SB. The chemistry involved in these reactions together with the mechanical and functional properties of the collagen scaffolds was evaluated. The viability of the cells grown on the scaffolds was studied using different corneal cell types. The effect of each type of scaffold treatment on human monocyte differentiation was evaluated. One-way ANOVA was used for statistical analysis. The addition of GTA as a double-crosslinking agent significantly improved the mechanical properties and enzymatic stability of the EDC crosslinked collagen scaffold. GTA decreased cell biocompatibility but this effect was reversed by treatment with SB or SM. These agents did not affect the mechanical properties, enzymatic stability, or transparency of the double-crosslinked scaffold. Contact of monocytes with the different scaffolds did not trigger their differentiation into activated macrophages. Our results demonstrate that GTA improves the mechanical properties of EDC crosslinked scaffolds in a dose-dependent manner, and that subsequent treatment with SB or SM partially restores biocompatibility. This novel manufacturing approach would facilitate the translation of collagen-based artificial corneas to the clinical setting.
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