Brazilian essential oil of Cymbopogon martini (Poaceae): positive effects on inflammation-induced human fibroblasts and skin aging

The study evaluated the effects of essential oil from Cymbopogon martini (CMEO) on lipopolysaccharide (LPS)-stimulated human fibroblasts. Samples were collected in Monte Verde, Minas Gerais, Brazil and analyzed by gas chromatography with mass spectrometry. The fibroblasts were cultured in a monolayer using Iscove's medium and stimulated by LPS (1 μg/mL) and incubated for 24 h at 37°C. The cytotoxicity was evaluated by MTT assay and collagen concentration by Sirius red. Collagenase activity, hyaluronic acid, and the concentrations of IL-1β; IL-6; MCP-1 (CCL2), and MIP-1-α (CCL3) were evaluated. The effect of CMEO on the expression of mRNA and the secretion of enzymes in fibroblasts were evaluated by RT-qPCR and ELISA, respectively. CMEO was cytotoxic against fibroblasts, in which 10 μg/mL inhibited 50% of cell viability. When treated with CMEO, the fibroblasts produced more collagen and hyaluronic acid than control cells. When stimulated by LPS, fibroblasts exhibited higher production of IL-6, IL-1β, MCP-1, and MIP-1α than control cells. The study demonstrated the effects of CMEO on the modulation of mediators related to inflammation and decreasing the mRNA and secretion levels of metalloproteinases, revealing to be a promising candidate for anti-aging effects and wound healing treatments.

Effect Collagen Concentration Tilapia Scales on the Quality of Burn Ointment

Collagen is one of the main connective tissue animal proteins and has been widely used as a biomedical material. Collagen is divided into XIX types. Type I collagen, among others, is obtained from bone, scales and skin. Collagen derived from type I can repair tissue or accelerate tissue regeneration to heal burns. The purpose of this research was to determine the addition of fish scale collagen extract to the characteristics of the burn ointment preparation in accordance with the Indonesian National Standard (SNI) and the best quality. The method used in this research is an experimental method of Completely Randomized Design (CRD) consisting of 4 collagen addition treatments: 0%, 2%, 4% and 6% repeated 5 times. Parameters in this method include physical-chemical parameters (pH, spreadability, shelf life and homogeneity) and organoleptic parameters (appearance, aroma, texture and color). Bayes test results, the concentration of the addition of tilapia scale collagen in the ointment preparation of 4% resulted in a value close to the control treatment. The addition of 4% collagen was the best treatment compared to 2% and 6% with a pH value of 6.12, dispersion of 3.22 cm, safe ointment preparation did not change at all during 28 days of storage. Based on the results of the organoleptic test parameters, the ointment at this concentration had a homogeneous appearance, slightly yellowish white color, a distinctive smell of collagen and a semi-solid texture, this was in accordance with the quality standard of the ointment and had the best quality characteristics.

Microbial Biogas Production From Pork Gelatine.

Dark fermentation of collagen (gelatine) results are shown in this research. The concentrations of applied gelatine were of VSS (volatile suspended solids) from 10 g VSS/L to 30 g VSS/L. The initial process pH was 5.5, depending on concentration reached pH values from 7.5 to 7.8 after 55 days. Although inoculum was heat-shocked in the case of 30 g VSS/L of collagen the process was hydrogenotrophic anaerobic digestion. In collagen concentration below 30 g VSS/L, hydrogen production was dominant only in the first 5 days of experiments. Then there also changed from dark fermentation into hydrogenotrophic methane production. In the case of optimal biogas production was due to accumulative production for a concentration of collagen 20 g VSS/L: 147.2 mL of hydrogen and 57.23 L of methane. In the case of optimal biogas production was due to accumulative production for a concentration of collagen 20 g VSS/L: 147.2 mL of hydrogen and 57.23 L of methane. The optimal hydrogen and methane yields were for concentration 10 g VSS/L (7.65 mL H2 /g VSS, and 3.49 L CH4/ g VSS). In 10 g VSS/L was also the lowest accumulated emission of hydrogen sulphide (10.3 mL of H2S), while the lowest yield was for 30 g VSS/L (0.44 mL H2S /g VSS). After a lag time, the hydrogen production and hydrogen sulphide grew with a specific ratio depending on concentration. Collagen, a protein with known amounts of sulphur allowed determining the origin of hydrogen sulphide in biogas. The hydrogen sulphide emission and sulphur added analysis proved that hydrogen sulphide origins in biogas from bacteria remains more than from substrate.

Effect of the Addition Tilapia Skin Collagen Concentration to Lotion Characteristics

The purpose of this research was to determine the best concentration of the tilapia skin collagen addition to obtain the characteristics of the lotion in accordance with the established standards. The research was carried out at the Fishery Product Processing Technology Laboratory, Faculty of Fisheries and Marine Sciences, Padjadjaran University, Central Laboratory of Padjadjaran University and Faculty of Mathematics and Natural Sciences Laboratory of Padjadjaran University between March 2021 - June 2021. The method used in this research was experimental with a completely randomized design consisted of 5 treatments with 4 replications added the concentration of the tilapia skin collagen (0%, 4%, 5%, 6% and 7%) based on lotion formulations. Parameters observed included pH, viscosity, spreadability, weight loss, homogeneity and the organoleptic (appearance, color, scent, texture). The conclusion from this research was that the addition of 7% the tilapia skin collagen concentration was the best treatment with a homogeneous appearance, a slightly yellowish white color, a slightly lavender scent and a slightly thick texture. The lotion had pH value of 6.35, viscosity of 7,197 cP, 5.74 cm spreadability and a weight loss of 2.8%.

pH Modification of High-Concentrated Collagen Bioinks as a Factor Affecting Cell Viability, Mechanical Properties, and Printability

The 3D bioprinting of cell-incorporated gels is a promising direction in tissue engineering applications. Collagen-based hydrogels, due to their similarity to extracellular matrix tissue, can be a good candidate for bioink and 3D bioprinting applications. However, low hydrogel concentrations of hydrogel (<10 mg/mL) provide insufficient structural support and, in highly concentrated gels, cell proliferation is reduced. In this study, we showed that it is possible to print highly concentrated collagen hydrogels with incorporated cells, where the viability of the cells in the gel remains very good. This can be achieved simply by optimizing the properties of the bioink, particularly the gel composition and pH modification, as well as by optimizing the printing parameters. The bioink composed of porcine collagen hydrogel with a collagen concentration of 20 mg/mL was tested, while the final bioink collagen concentration was 10 mg/mL. This bioink was modified with 0, 5, 9, 13, 17 and 20 μL/mL of 1M NaOH solution, which affected the resulting pH and gelling time. Cylindrical samples based on the given bioink, with the incorporation of porcine adipose-derived stromal cells, were printed with a custom 3D bioprinter. These constructs were cultivated in static conditions for 6 h, and 3 and 5 days. Cell viability and morphology were evaluated. Mechanical properties were evaluated by means of a compression test. Our results showed that optimal composition and the addition of 13 μL NaOH per mL of bioink adjusted the pH of the bioink enough to allow cells to grow and divide. This modification also contributed to a higher elastic modulus, making it possible to print structures up to several millimeters with sufficient mechanical resistance. We optimized the bioprinter parameters for printing low-viscosity bioinks. With this experiment, we showed that a high concentration of collagen gels may not be a limiting factor for cell proliferation.

The interplay between matrix deformation and the coordination of turning events governs directed neutrophil migration in 3D matrices

Neutrophils migrating through extravascular spaces must negotiate narrow matrix pores without losing directional movement. We investigated how chemotaxing neutrophils probe matrices and adjust their migration to collagen concentration ([col]) changes by tracking 20,000 cell trajectories and quantifying cell-generated 3D matrix deformations. In low-[col] matrices, neutrophils exerted large deformations and followed straight trajectories. As [col] increased, matrix deformations decreased, and neutrophils turned often to circumvent rather than remodel matrix pores. Inhibiting protrusive or contractile forces shifted this transition to lower [col], implying that mechanics play a crucial role in defining migratory strategies. To balance frequent turning and directional bias, neutrophils used matrix obstacles as pivoting points to steer toward the chemoattractant. The Actin Related Protein 2/3 complex coordinated successive turns, thus controlling deviations from chemotactic paths. These results offer an improved understanding of the mechanisms and molecular regulators used by neutrophils during chemotaxis in restrictive 3D environments.

Effect of Aging on the Quality of Breast Meat from Broilers Affected by Wooden Breast Myopathy

This study aimed to evaluate the effects of aging on the quality of breast meat from broilers affected of wooden breast. Samples that were classified as normal (control), moderate (hardness verified only in one region of breast fillet), and severe (hardness verified in all the extension of breast fillet) were evaluated fresh and after three and seven days of aging. Normal samples and samples with a moderate degree of myopathy showed greater water-holding capacity, which may benefit the processing industry of poultry meat. During the aging process, increase was observed in total collagen concentration (from 0.41% in normal samples to 0.56% in samples with severe degree). Samples of chicken breast affected by moderate degree showed higher myofibril fragmentation index (MFI = 115) than normal chicken samples (95.65). Although chicken samples affected with severe degree of wooden breast myopathy are more tender than normal chicken breasts, they produce more exudate, which can be detrimental to the processing of poultry meat. The aging process may improve the reduction of cooking weight loss and protein loss in exudation of broilers’ breasts affected by wooden breast myopathy.

Skeletal Muscle progenitors are sensitive to collagen architectural features of fibril size and crosslinking

Muscle stem cells (MuSCs) are essential for the robust regenerative capacity of skeletal muscle. However, in fibrotic environments marked by abundant collagen and altered collagen organization, the regenerative capability of MuSCs is diminished. MuSCs are sensitive to their extracellular matrix environment, but their response to collagen architecture is largely unknown. The present study aimed to systematically test the effect of underlying collagen structures on MuSC functions. Collagen hydrogels were engineered with varied architectures: collagen concentration, crosslinking, fibril size, and fibril alignment, and the changes were validated with second harmonic generation imaging and rheology. Proliferation and differentiation responses of primary mouse MuSCs and immortal myoblasts (C2C12s) were assessed using EdU assays and immunolabeling skeletal muscle myosin expression, respectively. Changing collagen concentration and the corresponding hydrogel stiffness did not have a significant influence on MuSC proliferation or differentiation. However, MuSC differentiation on atelocollagen gels, which do not form mature pyridinoline crosslinks, was increased compared to the crosslinked control. In addition, MuSCs and C2C12 myoblasts showed greater differentiation on gels with smaller collagen fibrils. Proliferation rates of C2C12 myoblasts were also higher on gels with smaller collagen fibrils, while MuSCs did not show a significant difference. Surprisingly, collagen alignment did not have significant effects on muscle progenitor function. This study demonstrates that MuSCs are capable of sensing their underlying ECM structures and enhancing differentiation on substrates with less collagen crosslinking or smaller collagen fibrils. Thus, in fibrotic muscle, targeting crosslinking and fibril size rather than collagen expression may more effectively support MuSC-based regeneration.

Gastrocnemius Muscle Structural and Functional Changes Associated with Domestication in the Turkey

Selection for increased muscle mass in domestic turkeys has resulted in muscles twice the size of those found in wild turkeys. This study characterizes muscle structural changes as well as functional differences in muscle performance associated with selection for increased muscle mass. We compared peak isometric force production, whole muscle and individual fiber cross-sectional area (CSA), connective tissue collagen concentration and structure of the lateral gastrocnemius (LG) muscle in wild and adult domestic turkeys. We also explored changes with age between juvenile and adult domestic turkeys. We found that the domestic turkey’s LG muscle can produce the same force per cross-sectional area as a wild turkey; however, due to scaling, domestic adults produce less force per unit body mass. Domestic turkey muscle fibers were slightly smaller in CSA (3802 ± 2223 μm2) than those of the wild turkey (4014 ± 1831 μm2, p = 0.013), indicating that the absolutely larger domestic turkey muscles are a result of an increased number of smaller fibers. Collagen concentration in domestic turkey muscle (4.19 ± 1.58 μg hydroxyproline/mg muscle) was significantly lower than in the wild turkeys (6.23 ± 0.63 μg/mg, p = 0.0275), with visible differences in endomysium texture, observed via scanning electron microscopy. Selection for increased muscle mass has altered the structure of the LG muscle; however, scaling likely contributes more to hind limb functional differences observed in the domestic turkey.

Curcumin Loading on Alginate Nano-Micelle for Anti-Infection and Colonic Wound Healing

Despite the antibacterial, and anti-inflammatory properties of curcumin (C), its effect on wound healing, especially in the colorectal, is ambiguous. Moreover, due to the hydrophobic properties of C, its use is limited. Therefore, to reduce the bioavailability challenge and improve the transfer to colon area, we designed a C-alginate-based nano-micelle (C-A-NM). After fabrication of C-A-NM (55.5 nm) and physicochemical studies with the TEM, DLS and XRD, the C release rate based on gastrointestinal state was evaluated. Furthermore, the effects of C-A-NM on the survival of HCT-8 cells at 24 and 48 hours by MTT method and its antibacterial effects were also evaluated. To explain the effects of wound healing in rats, in addition to colonoscopy on the 14th-day, the repaired tissue on the 7th and 14th days were examined by Hematoxylin and Eosin method. Also, for evaluating wound healing in the colon, the protein/collagen concentration, and TGFβ1/NFκB gene expression were determined. The results of C cumulative release showed that the NM allows the drug to be loaded in the colon in a favourable manner. Also, the toxicity outputs revealed that C-A-NM at a concentration of 7.5 mg had no negative effects on cell viability. While the activity of Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli, bacteria decreased based on the minimum inhibitory concentration value with 153, 245 and 319 (μg/mL). The use of C-A-NM not only increases protein and collagen in damaged sites, but also increases TGFβ1 expression in contrast to NFκB. Based on these results, and the results of histopathology and colonoscopy, it was found that C-A-NM accelerates the healing of damaged areas. Overall, the results show that the use of C-A-NM can significantly accelerate the healing of wounds in the gastrointestinal tract based on collagen induction and reduced bacterial activity.