Hesperidin Promotes Osteogenesis and Modulates Collagen Matrix Organization and Mineralization In Vitro and In Vivo

This study evaluated the direct effect of a phytochemical, hesperidin, on pre-osteoblast cell function as well as osteogenesis and collagen matrix quality, as there is little known about hesperidin’s influence in mineralized tissue formation and regeneration. Hesperidin was added to a culture of MC3T3-E1 cells at various concentrations. Cell proliferation, viability, osteogenic gene expression and deposited collagen matrix analyses were performed. Treatment with hesperidin showed significant upregulation of osteogenic markers, particularly with lower doses. Mature and compact collagen fibrils in hesperidin-treated cultures were observed by picrosirius red staining (PSR), although a thinner matrix layer was present for the higher dose of hesperidin compared to osteogenic media alone. Fourier-transform infrared spectroscopy indicated a better mineral-to-matrix ratio and matrix distribution in cultures exposed to hesperidin and confirmed less collagen deposited with the 100-µM dose of hesperidin. In vivo, hesperidin combined with a suboptimal dose of bone morphogenetic protein 2 (BMP2) (dose unable to promote healing of a rat mandible critical-sized bone defect) in a collagenous scaffold promoted a well-controlled (not ectopic) pattern of bone formation as compared to a large dose of BMP2 (previously defined as optimal in healing the critical-sized defect, although of ectopic nature). PSR staining of newly formed bone demonstrated that hesperidin can promote maturation of bone organic matrix. Our findings show, for the first time, that hesperidin has a modulatory role in mineralized tissue formation via not only osteoblast cell differentiation but also matrix organization and matrix-to-mineral ratio and could be a potential adjunct in regenerative bone therapies.     

橙皮苷酶解液中橙皮素单糖苷的树脂分离

橙皮苷酶解液组分复杂，含糖类、橙皮素和橙皮素单糖苷（hesperetin-7-O-glucoside，HMG）等。采用了6种大孔吸附树脂和半制备液相色谱分离纯化橙皮苷酶解液中的HMG，并通过红外、紫外和XRD手段表征产物结构，同时采用羟基自由基清除试验探讨其抗氧化活性。结果表明：HPD300大孔树脂最适于酶解液中HMG的分离；HPD300大孔树脂对酶解液中主要组分的静态和动态总吸附量分别为48.35 mg/g和35.82 mg/g，主要用于分离糖类杂质；以10 BV的45%乙醇为洗脱剂，洗脱流速1.2 mL/min，洗提物中HMG纯度从80.37%提高到97.83%；洗提物经半制备液相进一步分离纯化后得到单体HMG，最终纯度可达98.21%, 经紫外、红外和XRD手段验证产物为HMG；且HMG的羟基自由基清除能力略高于橙皮素，远高于橙皮苷。     

毛细管电泳法手性拆分橙皮苷对映体

通过考察电泳缓冲液pH、环糊精种类和浓度、有机溶剂浓度及操作电压和温度对分离的影响,建立了一种简便快捷的高效毛细管电泳拆分橙皮苷对映体的方法。实验结果表明,以12mmol/L的羟丙基-β-环糊精为手性选择剂,200mmol/L的硼酸/甲醇(V:V=7:3,pH9.7)为电泳缓冲溶液,23kV的分离电压,在30℃下可使橙皮苷对映体达到基线分离,分离度为1.50。分析方法快速简单。     

柑桔加工下脚料提取类胡萝卜素、橙皮苷、果胶和柠檬苦素

介绍了从柑桔加工下脚料提取类胡萝卜素、橙皮苷、果胶和柠檬苦素类产品的工艺方法 ,并简述了这些产品在医药、食品、日化等方面的应用     

秋泻灵合剂质量标准的研究

建立了秋泻灵合剂中橙皮苷的含量测定方法。采用HPLC法,Kromasil C18色谱柱,甲醇-5%冰醋酸溶液(35∶65)为流动相,检测波长为284 nm,流速1.0 m L/min。结果表明:橙皮苷对照品在0.0625~4μg范围内呈良好的线性关系(r=0.9998),平均加样回收率为97.5%,RSD=1.09%(n=6)。结论:所用方法简便、准确、重复性好,专属性强,分离效果好,可用于秋泻灵合剂的质量控制。     

陈皮最佳饮用方式探讨

陈皮是民间常备的消食导滞药品之一。通过测定陈皮中的主要活性成分橙皮甙的溢出量，对陈皮的最佳饮用方式进行探讨。实验结果表明，用矿泉水冲泡小金桔样品，并加冰糖为辅料煎煮15min左右，用温度较高的水在紫砂壶中冲泡2h后饮用较好。     

橙皮的再生利用——超临界CO2萃取橙皮精油的研究

橙皮是一种可再生利用资源,针对影响超临界CO2萃取橙皮精油的最重要因素,即萃取压力、萃取温度、萃取时间,首先设计单因素实验,然后设计正交实验。结果表明:萃取压力为16MPa,萃取温度为40℃,萃取时间为2.5h是最佳工艺组合。     

橙皮苷改性技术研究进展

橙皮苷是重要的黄酮类化合物之一,可抗氧化,防癌,抗病毒,抗细菌,还具有保护心血管的作用,但是由于其水溶性较差,使其应用受到限制。文中介绍国内外橙皮苷改性的几种方法,包括酶处理、高温下酸催化橙皮苷水解、橙皮苷甲基化、磺化和金属离子络合等。     

Solidification of hesperidin nanosuspension by spray drying optimized by design of experiment (DoE)

Objective: To accelerate the determination of optimal spray drying parameters, a “Design of Experiment” (DoE) software was applied to produce well redispersible hesperidin nanocrystals.

Significance: For final solid dosage forms, aqueous liquid nanosuspensions need to be solidified, whereas spray drying is a large-scale cost-effective industrial process.

Methods: A nanosuspension with 18% (w/w) of hesperidin stabilized by 1% (w/w) of poloxamer 188 was produced by wet bead milling. The sizes of original and redispersed spray-dried nanosuspensions were determined by laser diffractometry (LD) and photon correlation spectroscopy (PCS) and used as effect parameters. In addition, light microscopy was performed to judge the redispersion quality.

Results: After a two-step design of MODDE 9, screening model and response surface model (RSM), the inlet temperature of spray dryer and the concentration of protectant (polyvinylpyrrolidone, PVP K25) were identified as the most important factors affecting the redispersion of nanocrystals. As predicted in the RSM modeling, when 5% (w/w) of PVP K25 was added in an 18% (w/w) of hesperidin nanosuspension, subsequently spray-dried at an inlet temperature of 100?°C, well redispersed solid nanocrystals with an average particle size of 276?nm were obtained. By the use of PVP K25, the saturation solubility of the redispersed nanocrystals in water was improved to 86.81?µg/ml, about 2.5-fold of the original nanosuspension. In addition, the dissolution velocity was accelerated.

Conclusions: This was attributed to the additional effects of steric stabilization on the nanocrystals and solubilization by the PVP polymer from spray drying.