Interleukin-1β Modulation of the Mechanobiology of Primary Human Pulmonary Fibroblasts: Potential Implications in Lung Repair

Pro-inflammatory cytokines like interleukin-1β (IL-1β) are upregulated during early responses to tissue damage and are expected to transiently compromise the mechanical microenvironment. Fibroblasts are key regulators of tissue mechanics in the lungs and other organs. However, the effects of IL-1β on fibroblast mechanics and functions remain unclear. Here we treated human pulmonary fibroblasts from control donors with IL-1β and used Atomic Force Microscopy to unveil that IL-1β significantly reduces the stiffness of fibroblasts concomitantly with a downregulation of filamentous actin (F-actin) and alpha-smooth muscle (α-SMA). Likewise, COL1A1 mRNA was reduced, whereas that of collagenases MMP1 and MMP2 were upregulated, favoring a reduction of type-I collagen. These mechanobiology changes were functionally associated with reduced proliferation and enhanced migration upon IL-1β stimulation, which could facilitate lung repair by drawing fibroblasts to sites of tissue damage. Our observations reveal that IL-1β may reduce local tissue rigidity by acting both intracellularly and extracellularly through the downregulation of fibroblast contractility and type I collagen deposition, respectively. These IL-1β-dependent mechanical effects may enhance lung repair further by locally increasing pulmonary tissue compliance to preserve normal lung distension and function. Moreover, our results support that IL-1β provides innate anti-fibrotic protection that may be relevant during the early stages of lung repair.     

聚乙烯醇/牛I型胶原支架材料的制备及评价

研究了一种聚乙烯醇(PVA)和胶原(COL)复合支架材料的制备方法。采用氨基硅烷对PVA海绵表面进行了氨基化修饰后，通过戊二醛溶液交联牛Ⅰ型胶原(COL)，最后通过赖氨酸溶液封闭，获得一种PVA/COL复合支架材料。采用扫描电镜(SEM)、X光电子能谱仪(XPS)、傅里叶红外光谱(FT-IR)等手段对支架材料的理化性能进行表征，并通过细胞实验对支架材料的生物学性能进行评价。结果表明，经过COL修饰的PVA孔隙率为21.33%，平均孔径为168.68 ?m且均匀分布，支架材料接触角为20.03°。对支架材料的生物学评价结果表明C3A细胞在复合材料上黏附良好，优于PVA组；CCK-8增殖检测结果表明细胞在复合材料上呈增殖生长趋势，与对照组PVA相比差异显著(P?0.01)。将PVA和COL复合制备得到的支架材料具有良好的理化及生物学特性，具有广阔的应用前景。     

The effects of washing a collagen sample prior to TEM examination

Transmission electron microscopy (TEM) is an important analysis technique to visualize (bio)macromolecules and their assemblies, including collagen fibers. Many protocols for TEM sample preparation of collagen involve one or more washing steps to remove excess salts from the dispersion that could hamper analysis when dried on a TEM grid. Such protocols are not standardized and washing times as well as washing solvents vary from procedure to procedure, with each research group typically having their own protocol. Here, we investigate the influence of washing with water, ethanol, but also methanol and 2-propanol, for both mineralized and unmineralized collagen samples via a protocol based on centrifugation. Washing with water maintains the hydrated collagen structure and the characteristic banding pattern can be clearly observed. Conversely, washing with ethanol results in dehydration of the fibrils, often leading to aggregation of the fibers and a less obvious banding pattern, already within 1 min of ethanol exposure. As we show, this process is fully reversible. Similar observations were made for methanol and propanol. Based on these results, a standardized washing protocol for collagenous samples is proposed.     

Thermal and pH dual‐responsive hydrogels based on semi‐interpenetrating network of poly(N‐isopropylacrylamide) and collagen nanofibrils

Hydrogels with environment‐sensitive properties have great potential applications in the controlled drug release field. In this paper, hybrid hydrogels with semi‐interpenetrating polymer networks (semi‐IPNs), composed of poly(N‐isopropylacrylamide) (PNIPAM) as the thermo‐sensitive component by in situ polymerization and self‐assembled collagen nanofibrils as the pH‐sensitive framework, were prepared for controlled release of methyl violet as a model drug. From Fourier transform infrared spectroscopy and scanning electron microscopy, it was indicated that the crosslinking of PNIPAM in the presence of collagen nanofibrils led to the formation of semi‐IPNs with homogeneous porous structure, and the semi‐IPNs showed improved thermal stability and elastic properties compared with the native collagen as determined using differential scanning calorimetry and rheologic measurements. Furthermore, the semi‐IPNs possessed swelling behaviors quite different from those of neat collagen or PNIPAM hydrogel under various pH values and temperatures. Correspondingly, as expected, the drug release behavior in vitro for semi‐IPNs performed variously compared with that for single‐component semi‐IPNs, which revealed the tunable performance of semi‐IPNs for release ability. Finally the thermo‐ and pH‐responsive mechanism of the semi‐IPNs was illuminated to provide guidance for the application of the thermo‐ and pH‐sensitive collagen‐based hybrid hydrogels in controlled drug delivery systems. © 2019 Society of Chemical Industry     

Recent trends for the employment of jumbo squid (Dosidicus gigas) by-products as a source of bioactive compounds with nutritional,functional and preservative applications: a review

Only 50%–60% of total seafood catch is used for human consumption, seafood processing being considered as one of the main sources of by-products. Among marine species, jumbo squid (JS; Dosidicus gigas) represents the most important squid fishery, showing an increasing economic interest in many countries. As for any other marine species, the regular cleaning, dressing and processing produce high quantities of by-products (skin, head, fins, viscera, tentacles, unclaimed mantle, etc.) that are rich in many nutrients (proteins, lipids, minerals, vitamins, enzymes, biopolymers, etc.). This review compiles information about extraction and employment of JS by-products with the aim of enhancing their economic value and reduce environmental drawbacks. A special emphasis is given to the relevance in developing methods susceptible to transform by-products into useful and profitable products susceptible to be applied in several industries such as food, medicine, agrochemical or pharmaceutical. Future possible trends for widening this profitable use are mentioned.     

胶原蛋白-壳聚糖膜的制备及其对猪肉的保鲜作用

为了延缓猪肉的腐坏变质,延长猪肉的保质期,保持猪肉良好的品质,本文研究了没食子酸(GA)改性超声辅助制备的胶原蛋白-壳聚糖复合膜对冷藏猪肉的保鲜效果。在4℃冷藏条件下,采用未经过膜包裹(对照组)和GA改性超声膜包裹(膜包裹组)2种方式处理猪肉样品,并贮藏10d,检测其p H值、挥发性盐基氮(TVB-N)值、细菌总数、质构、色差ΔE值,以及感官质量。结果表明,对照组和膜包裹组猪肉的p H值、TVB-N、细菌总数、色差ΔE均随着贮存时间的增加而上升,但是膜包裹组的数值比对照组分别低了8.39%、26.18%、17.54%和8.23%,得出膜包裹处理可以显著降低碱性含氮物质和细菌产生的数量以及减缓颜色的变差(p<0.05)。对照组和膜包裹组猪肉的硬度、弹性和感官指标均随着贮存时间的增加而下降,但是膜包裹组的数值比对照组分别高了80.01%、18.66%和160.38%,得出膜包裹处理可以显著改善猪肉的质构和感官指标(p<0.05)。由此说明,膜包裹处理可以通过抑制p H值、TVB-N和细菌总数的增高,有效延缓猪肉的腐坏变质,延长猪肉的保质期。而且,经膜包裹处理10d后的猪肉仍然具有良好的质构、颜色和感官品质。     

Hydrolysed collagen from Lates calcarifer skin: its acute toxicity and impact on cell proliferation and collagen production of fibroblasts

Acute toxicity in Wistar rat and the impact of hydrolysed collagen (HC) from seabass skin on in vitro cell proliferation and collagen production were studied using L929 fibroblasts. HC was rich in glycine (326 residue/1000 residue) and imino acids (196 residue/1000 residue). MALDI mass spectrum of HC showed several low MW peptides with MW range of 1050–1330 Da as the major components. Based on acute oral cytotoxicity test in Wistar rat, HC was considered as safe with LD₅₀ value higher than 5000 mg kg^?1 body. HC could promote L929 cell growth, especially when used in combination with vitamin C (VitC) at a ratio of 2:1. HC/VitC (2:1) mixture also exhibited the higher enhancement effect on collagen production of L929 cells, compared with HC or VitC alone. Thus, HC could be a promising candidate for biological applications, especially in combination with VitC, as nutraceuticals for skin care.