Engineering large and geometrically controlled vascularized nerve tissue in collagen hydrogels to restore large-sized volumetric muscle loss.

Developing scalable vascularized and innervated tissue is a critical challenge for the successful clinical application of tissue-engineered constructs. Collagen hydrogels are extensively utilized in cell-mediated vascular network formation because of their naturally excellent biological properties. However, the substantial increase in hydrogel contraction induced by populated cells limits their long-term use. Previous studies attempted to mitigate this issue by concentrating collagen pre-polymer solutions or synthesizing covalently crosslinked collagen hydrogels. However, these methods only partially reduce hydrogel contraction while hindering blood vessel formation within the hydrogels. To address this challenge, we introduced additional support in the form of a supportive spacer to counteract the contraction forces of populated cells and prevent hydrogel contraction. This approach was found to promote cell spreading, resist hydrogel contraction, control hydrogel/tissue geometry, and even facilitate the engineering of functional blood vessels and host nerve growth in just one week. Subsequently, implanting these engineered tissues into muscle defect sites resulted in timely anastomosis with the host vasculature, leading to enhanced myogenesis, increased muscle innervation, and the restoration of injured muscle functionality. Overall, this innovative strategy expands the applicability of collagen hydrogels in fabricating large vascularized nerve tissue constructs for repairing volumetric muscle loss (∼63 %) and restoring muscle function.

Effects of rosemary (Rosmarinus officinalis L.) extracts and dry ice on the physicochemical stability of omega-3 fatty-acid-fortified surimi-like meat products.

BACKGROUND: Lipid peroxidation entails major quality degradation in omega-3 (ω-3) fatty-acid-fortified surimi-like meat products upon storage. Currently, the use of label-friendly alternatives to synthetic antioxidants is encouraged in the industry. Hence, we aimed to examine the applicability of the hurdle-technology concept, using an 80% (v/v) ethanol solution to obtain rosemary extracts (REs) containing substantial amounts of polyphenol, and dry ice (DI) which can create a cryogenic environment, on the physicochemical stabilities of ω-3 fatty-acid (FA)-fortified meat products after manufacturing and storage periods. The polyphenolic profiles of the REs were also investigated. RESULTS: Carnosol and rosmarinic acid are major phenolic components in REs. Furthermore, DI addition during the chopping procedure increased (P < 0.05) whiteness values and hardness of products, while total ω-3 and ω-6 FAs were relatively well preserved (P < 0.05) in products with flaxseed oil premixed with RE. During 14-day storage at 4 °C, combined treatment with RE and DI decreased (P < 0.05) thiobarbituric acid reactive substance (TBARS) levels and the centrifugation loss of products. Single or combined treatment with RE and/or DI decreased (P < 0.05) TBARS levels in products after 60 days of storage at -20 °C. CONCLUSION: Due to the antioxidant-polyphenol profile of REs and a possible oxygen exclusion of DI treatment under atmospheric pressure during food manufacturing, application of the hurdle-technology concept, using treatment with both RE and DI, can reduce lipid peroxidation and maintain a greater water-holding capacity of ω-3 FA-fortified meat products upon storage. © 2019 Society of Chemical Industry.

Nutritional composition in the chia seed and its processing properties on restructured ham-like products.

Low-fat meat products always have harder texture, lower juiciness, and worse flavor. Due to their higher water-holding, water absorption, and organic molecule absorption, chia seeds (CHIA) have been applied in powders, nutrition bars, breads, and cookies. Hence, the objectives of this study were to: (1) analyze the nutritional compositions in CHIA; and (2) look for the possible application of CHIA on restructured ham-like products. CHIA has high amounts of α-linolenic acid, crude polysaccharides, and also contains essential amino acids, minerals, and polyphenols. Regarding processing properties of CHIA, a combination of CHIA and carrageenan (CA) increased (p<0.05) production yield of restructured ham-like products. A scanning electron microscope observation indicated that CHIA and CA addition can assist an emulsification in this ham-like product. Addition of 0.5% CA and 1.0% CHIA in this ham-like product showed the similar overall acceptance as products with added fat. Following storage at 4°C, higher (p<0.05) purge and centrifugation losses, as well as hardness of this ham-like product can be improved by adding CHIA and CA. CHIA addition also resulted in lower (p<0.05) lipid and protein oxidation, especially a 1.0% addition. In summary, due to both nutritional addition and improvements on physicochemical and sensorial properties of restructured ham-like products, CHIA seeds have great potential on the development of healthy and good-quality meat products.

Chicken surimi fortified by omega-3 fatty acid addition: manufacturing and quality properties.

BACKGROUND: The meat of spent hens is hard to use owing to its small amount and poor quality. A washing process to remove sarcoplasmic proteins and other impurities can prolong the shelf life of surimi-like products. Owing to the benefits of omega-3 polyunsaturated fatty acids (ω-3 PUFAs), functional foods fortified with ω-3 PUFAs are increasingly being marketed. Hence, in this study, ω-3 FA-fortified chicken surimi was manufactured, and how to ameliorate its lipid peroxidation during frozen storage was investigated. RESULTS: A 0.10% (w/v) solution of sodium chloride (NaCl) instead of distilled water in the third washing step decreased (P < 0.05) myofibrillar protein loss and moisture content of spent hen breast protein recoveries. Oil droplets in fish, flaxseed or soybean oil-added chicken surimi were well distributed. Moreover, flaxseed oil addition increased (P < 0.05) total ω-3 FAs and ω-3/ω-6 FA ratio, while only fish oil provided long-chain PUFAs. Oil addition decreased (P < 0.05) hardness and gumminess of chicken surimi, while flaxseed oil resulted in more (P < 0.05) yellow surimi than fish and soybean oil. Fish oil-added samples showed higher (P < 0.05) lipid oxidation than flaxseed or soybean oil-added samples under -15 to -10 °C storage, but α-tocopherol addition ameliorated it. CONCLUSION: A novel semi-manufactured chicken surimi product with nutritional benefits could be developed by fortification with fish or flaxseed oil.

Micellar electrokinetic chromatography profiles of human high-density lipoprotein phospholipids.

A simple and fast micellar electrokinetic chromatography (MEKC) method was developed to investigate phospholipids isolated from human high-density lipoproteins (HDL). To optimize the MEKC conditions, several factors including bile salt concentration and organic modifier concentration in the separation buffer as well as temperature have been examined. The optimal separation buffer chosen was a mixture of 50 mM bile salts, 30% v/v 1-propanol and 10 mM sodium phosphate (pH 8.5). The applied voltage and temperature selected were 25 kV and 40°C, respectively. Meanwhile, high-salt stacking has been performed for sample pre-concentration to enhance peak sensitivity. Several factors including organic modifier concentration and salt concentration in the sample matrix as well as sample injection time have been optimized. The optimal sample buffer selected was a mixture of 100 mM NaCl and 20% 1-propanol, and the optimal sample injection time selected was 32 s under a pressure of 0.5 psi. Several phospholipid standards including lysophosphatidyl choline, phosphatidyl choline (PC), sphingomyelin, phosphatidyl ethanolamine, phosphatidyl inositol, phosphatidyl serine and phosphatidic acid have been studied using the optimal MEKC method. The MEKC profile of the mixed phospholipid standards showed good separation and reproducibility. The linear ranges for PC and sphingomyelin were 0.025-1.2 and 0.025-2.0 mg/mL, respectively. The concentration limits of detection of PC and sphingomyelin were 0.0156 and 0.0199 mg/mL, respectively. Using phosphatidic acid as an internal standard, precision and accuracy have been measured for PC and sphingomyelin. The intraday and interday quantitative analysis showed good results. The new MEKC method has been used to characterize native, in vitro oxidized and glycated human HDL phospholipids within 16 min. At absorbance 200 nm, two similar peaks were observed for native and oxidized HDL phospholipids, but three peaks were observed for glycated HDL phospholipids. Interestingly, at absorbance 234 nm, distinctively different MEKC profiles were observed for the three HDL phospholipids.

Antioxidant and anti-inflammatory effects of Orthosiphon aristatus and its bioactive compounds.

Orthosiphon aristatus (Blume) Miq., which can be used as a food ingredient, is grown throughout Southeast Asia and Australia. O. aristatus is frequently used for the treatment of renal inflammation, kidney stones and dysuria. The focus of the current work was to study the antioxidant and anti-inflammatory effects of methanol, ethanol and water extracts from O. aristatus (abbreviated as MEOA, EEOA and WEOA, respectively). The evaluation of antioxidant activity was determined by total phenolics, Trolox equivalent antioxidant capacity (TEAC), oxygen-radical absorbance capacity (ORAC) and cellular antioxidant activity (CAA) assays. These assays demonstrated a relatively high antioxidant activity for MEOA and EEOA. These results revealed that EEOA had the most prominent inhibitory effect on lipopolysaccharide (LPS)-stimulated nitric oxide (NO), prostaglandin E(2) (PGE(2)) and intracellular reactive oxygen species (ROS) production in RAW 264.7 cells. A high performance liquid chromatography profile indicated that MEOA and EEOA contained both ursolic acid and oleanolic acid. Moreover, ursolic acid significantly reduced NO production in LPS-stimulated RAW 264.7 cells. Both EEOA and ursolic acid inhibited LPS-stimulated protein and mRNA expression of both inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in these cells. These results demonstrate that EEOA and its bioactive compound, ursolic acid, suppress LPS-induced NO and PGE(2) production by inhibiting ROS generation, along with reducing expression of iNOS and COX-2 in RAW 264.7 cells.

Anticancer effects of Alpinia pricei Hayata roots.

The leaves and roots of Alpinia pricei Hayata are used as a traditional wrapping for food and as a cooking substitute for fresh ginger. Our previous study showed that ethanol extracts from the roots of A. pricei Hayata (EEAP) and its phenolic compounds have anti-inflammatory effects. The aims of this work were to further study the in vitro anticancer activity of EEAP and its active compounds with respect to various cancer cells. The results from an MTT assay demonstrated that EEAP decreased the cell population growth of CH27, HL-60, and A549 cells. Flow cytometric analysis of HL-60 cells exposed to EEAP showed that the number of apoptotic cells increased in a time- and dose-dependent manner. Western blot data revealed that EEAP stimulated an increase in the level of protein expression of Fas, FasL, caspase-8, and tBid. Moreover, the ratio of the expression levels of pro- and anti-apoptotic Bcl-2 family members was changed after treatment with EEAP. EEAP-induced apoptosis involved the release of mitochondrial cytochrome c and subsequently induced the activation of caspase-9 and caspase-3, which were followed by the cleavage of poly(ADP-ribose) polymerase (PARP). The results also demonstrated that phenolic compounds (caffeic acid, apigenin, curcumin, and pinocembrin) from EEAP decreased the rate of population growth of HL-60 cells. Treatment of HL-60 cells with these phenolic compounds caused the loss of mitochondrial membrane potential. Our finding could provide critical information regarding the chemopreventive potential of ethanol extracts from A. pricei Hayata. These results also demonstrate that the EEAP-induced apoptotic ability in HL-60 cells might be related to the phenolic compounds.

Anti-inflammatory effects of the roots of Alpinia pricei Hayata and its phenolic compounds.

Alpinia pricei Hayata is cultivated throughout Asia and is an endemic plant in Taiwan. The leaf and root of this plant are used for traditional wrapping of food and as a cooking substitute for fresh ginger. The aim of this work was to study the in vitro anti-inflammatory effects of ethanol extracts from A. pricei Hayata (EEAP) and its phenolic compounds. High-performance liquid chromatography (HPLC) profiling indicated that EEAP contained caffeic acid, chlorogenic acid, ferulic acid, p-hydroxybenzoic acid, rutin, apigenin, curcumin and pinocembrin. EEAP and its phenolic compounds, apigenin, curcumin, and pinocembrin, inhibited lipopolysaccharide (LPS)-stimulated nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production in RAW 264.7 cells. Furthermore, EEAP, apigenin, curcumin, and pinocembrin decreased LPS-mediated induction of protein and mRNA expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in RAW 264.7 cells. In addition, EEAP and its major active compound pinocembrin inhibited LPS-induced nuclear translocation of nuclear factor-kappaB (NF-kappaB) and NF-kappaB-mediated reporter gene expression. EEAP and pinocembrin also significantly inhibited LPS-induced intracellular reactive oxygen species (ROS) production in RAW 264.7 cells. When these results are taken together, they indicate that EEAP and pinocembrin suppressed LPS-induced NO and PGE(2) production by inhibition of NF-kappaB nuclear translocation and ROS generation.

Lucidenic acid B induces apoptosis in human leukemia cells via a mitochondria-mediated pathway.

Ganoderma lucidum is known as a medicinal mushroom used in traditional Chinese medicine. In the present study, the effect of lucidenic acids (A, B, C, and N) isolated from a new G. lucidum (YK-02) on induction of cell apoptosis and the apoptotic pathway in HL-60 cells were investigated. The results demonstrated that lucidenic acids decreased cell population growth of HL-60 cells, assessed with the MTT assay. The cell cycle assay indicated that treatment of HL-60 cells with lucidenic acid A, C, and N caused cell cycle arrest in the G 1 phase. Lucidenic acid B (LAB) did not affect the cell cycle profile; however, it increased the number of early and late apoptotic cells but not necrotic cells. Treatment of HL-60 cells with LAB caused loss of mitochondria membrane potential. Moreover, the ratio of expression levels of pro- and antiapoptotic Bcl-2 family members was changed by LAB treatment. LAB-induced apoptosis involved release of mitochondria cytochrome c and subsequently induced the activation of caspase-9 and caspase-3, which were followed by cleavage of poly(ADP-ribose) polymerase (PARP). Pretreatment with a general caspase-9 inhibitor (Z-LEHD-FMK) and caspase-3 inhibitor (Z-DEVD-FMK) prevented LAB from inhibiting cell viability in HL-60 cells. Our finding may be critical to the chemopreventive potential of lucidenic acid B.