A role for specific collagen motifs during wound healing and inflammatory response of fibroblasts in the teleost fish gilthead seabream (original) (raw)
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Collagen regulates the activation of professional phagocytes of the teleost fish gilthead seabream
Molecular Immunology, 2009
The innate immune system mediates the initial inflammatory response that follows infection or injury. Although the innate immune response of fish to infection has been relatively well characterized during recent years at both cellular and molecular levels, no studies have examined the role of extracellular matrix (ECM) in the regulation of innate immunity and inflammation. We report here that collagen and gelatin in vitro were able to prime the respiratory burst of phagocytes from the bony fish gilthead seabream. In addition, collagen and gelatin induced a specific set of immune-related and ECM remodelling enzymes that substantially differed from that induced by pathogen-associated molecular patterns. Notably, both collagen and gelatin induced the expression of interleukin-1 , chemokine (C-C motif) ligand 4 and matrix metalopeptidases (MMP) 9 and 13 in acidophilic granulocytes and macrophages but were unable to significantly increase the expression of other pro-inflammatory genes. Furthermore, it was found that the MMP2/MMP9 inhibitor V had a dose-dependent inhibitory effect on seabream phagocyte activation by either collagen or gelatin. In contrast, pre-treatment of collagen and gelatin by collagenase resulted in a higher stimulatory capacity compared to non-digested proteins.
Developmental and Comparative Immunology, 2010
Extracellular matrix (ECM) components, in addition to their structural functions, interact with cell surface receptors and intracellular components to modulate the transduction of signals for cell growth, differentiation, migration, proliferation, polarization, apoptosis and inflammation. Our previous findings in the gilthead seabream (Sparus aurata L.), a marine seasonal hermaphrodite teleost fish, have shown that both endocrine and immune stimuli modulate the expression of matrix metalloproteases (MMPs) and tissue inhibitors of MMP (TIMPs). In addition, collagen type I (COL1) induces the expression of some pro-inflammatory cytokines and MMPs in professional phagocytes. Consequently, in this study we use real-time RT-PCR to analyze the gene expression profile of several ECM-related molecules (MMP-2, -9 and -13, TIMP-2a, and -2b, COL1A1, and integrin 1a) in different organs of adult specimens as well as in response to innate immune challenges. Our results showed that liver had the lowest basal levels of them, although they were clearly modulated during injury and infection. In the same way, ECM-related molecules seem to participate in pro-inflammatory processes, being of particular interest COL1 which is synthesized by immune cells and is able to act as autocrine/paracrine stimulus for them. Lastly, we propose that the observed correlations between ECM-related molecules during the inflammatory response should be considered to obtain a more accurate picture of their roles in this process.
Advances in Clinical and Experimental Medicine, 2016
(Poland) using collagen extracted from the skin of silver carp [7]. These ulcers are wounds that are very difficult to heal. A few effects were seen: wound closure, accelerated granulation, anti-inflammatory effect, decreased evaporation of water from the wound. The collagen gel also protected the wound edges from the effects of toxic substances produced by the infected wound (proteolytic enzymes, etc.) [7]. Figure 1 shows examples of the treatment effects of collagen from the skin of silver carp-the subject of present article-on difficult-to-heal wounds in leg ulceration. The ulceration remained For a few years now, low-temperature techniques that prevent protein denaturation has been used to extract collagen from fish skin for cosmetic purposes. These extracts contain collagen with its triple helix structure preserved, as well as a number of other proteins. An interesting question concerning composition of those extracts is the presence of small-molecule peptides. Preparations of collagen are known to have a stimulatory influence on some cellular and tissuespecific processes-effects that are caused only by active compounds of low molecular weight, which are capable of penetrating into the skin and deeper tissues [1]. Reports [2-6] on the beneficial effects of collagen-containing products on wound healing
An Estimation of the Biological Properties of Fish Collagen in an Experimental In Vitro Study
Advances in Clinical and Experimental Medicine, 2015
Background. The principal sources of medical collagen are pork, calf skin and bone. There are now more studies on a much safer, alternative source of active collagen, mainly from aquatic life. Active collagen and its peptides FCP (fish collagen peptides) have already been extracted from the skin of salmon, cobia, hoki, tilapia, zebrafish, ling, shark, silver carp and also jellyfish. Objectives. The aim of the study is to evaluate the effect of fish collagen on human fibroblasts from gingiva. The cytotoxicity of the new formulation and induction of endogenous collagen was estimated by means of the collagen derived from fish skin. Material and Methods. Fish collagen was extracted from the skin of silver carp at 16 degrees Celsius. To compare the biocompatibility and endogenous collagen production Geistlich Bio-Gide ® membrane was ordered in Geistlich Biomaterials (Geistich AG, Wolhusen, Switzerland). The culture of human fibroblasts was performed acc. to Saczko et al. The fibroblasts were treated 96 hours with 1.0%, 0.5% and 0.1% experimental collagen formulation to induce endogenous collagen production. The Sircol collagen assay was done to measure amount of collagen. Cell viability was assessed by measuring mitochondrial activity in MTT assay after 24 h followed by 24 h of incubation with experimental collagen formulation. Qualitative analysis was performed by immunocytochemically staining of collagen type I and III. Results. Preparations of fish collagen are not cytotoxic at concentrations below 1%. Cells cultured in the presence of this product are characterized by a large number of endogenous collagen, which is comparable to the control. In case of porcine collagen membrane was noticed decreased to 83% production of endogenous collagen and reduction of cell viability to 69%. Conclusions. Our study showed that experimental fish collagen is an innovative product which may induce expression of endogenous collagen in fibroblasts (Adv Clin Exp Med 2015, 24, 3, 385-392).
Fish & Shellfish Immunology, 2013
Modulation of the tissue regenerative process in fish by ß-glucans Immune modulators are compounds capable of interacting with the immune system and thereby modifying the host response. This interaction enhances non-specific defense mechanisms, improving health and promoting survival. are glucose polysaccharides present in sea weed, bacteria, fungi and cereal but not in animals. are commonly used as immune modulators, but the mechanisms through which the modulation is achieved remains to be understood. Wound healing and tissue regeneration are essential mechanisms to ensure the survival and health of any organism. Studies from the mammalian systems have shown the importance of fibroblasts, macrophages, reactive oxygen species (especially hydrogen peroxide) and certain cytokines during wound healing processes. In fish however, only a few studies have been devoted tissue regeneration and modulation of cell proliferation during wound healing, even though mechanical injury as well as numerous diseases can severely damage fish tissues. The work presented examines for the first time the immunomodulatory effects of during wound healing processes in fish. Experiments have been conducted both in vivo and in vitro and results clearly show the immunomodulatory effects of during the wound healing process. The wound healing process was monitored using image analysis, real time PCR and proteomics. The study showed that treatment enhanced wound closure in fish, probably due to the enhancement of a localized inflammatory response. The modulatory effect of on wound healing seems to be orchestrated by the immune system, since as direct effect on fibroblast proliferation were observed. Furthermore, production of ROS may influence the fate of tissue regeneration, and differences in ROS patterns could be one of the possible ways in which fish alert the immune system to drive the immune response towards pathogen eradication or tissue repair.
PLoS ONE, 2010
Fibroblasts have shown to be an immune competent cell type in mammals. However, little is known about the immunological functions of this cell-type in lower vertebrates. A rainbow trout hypodermal fibroblast cell-line (RTHDF) was shown to be responsive to PAMPs and DAMPs after stimulation with LPS from E. coli, supernatant and debris from sonicated RTHDF cells. LPS was overall the strongest inducer of IL-1b, IL-8, IL-10, TLR-3 and TLR-9. IL-1b and IL-8 were already highly up regulated after 1 hour of LPS stimulation. Supernatant stimuli significantly increased the expression of IL-1b, TLR-3 and TLR-9, whereas the debris stimuli only increased expression of IL-1b. Consequently, an in vivo experiment was further set up. By mechanically damaging the muscle tissue of rainbow trout, it was shown that fibroblasts in the muscle tissue of rainbow trout contribute to electing a highly local inflammatory response following tissue injury. The damaged muscle tissue showed a strong increase in the expression of the immune genes IL-1b, IL-8 and TGF-b already 4 hours post injury at the site of injury while the expression in non-damaged muscle tissue was not influenced. A weaker, but significant response was also seen for TLR-9 and TLR-22. Rainbow trout fibroblasts were found to be highly immune competent with a significant ability to express cytokines and immune receptors. Thus fish fibroblasts are believed to contribute significantly to local inflammatory reactions in concert with the traditional immune cells.
A simple method has been developed for the isolation of Acid Soluble Collagen (ASC) and Pepsin Digestible Collagen (PDC) from the skin of Albacore tuna (Thunnus alalunga), Dog shark (Scoliodon sorrakowah), and one among Indian Major Carps ie, Rohu (Labeo rohita). Biochemical characterization of the collagen extracted was carried out. On wet weight basis the yields of ASC and PDC from shark skin were 8.96% & 7.68% respectively and that from Rohu was ASC 4.13% & PDC 3.68% respectively. The yield of collagen from Tuna skin was ASC 13.97%. No PDC was obtained for tuna skin. Proximate analysis showed all collagens had protein as a major constituent with trace amount of ash and fat. Amino acid analysis revealed that they contained glycine as a major amino acid with high contents of alanine, proline and hydroxyproline. Based on sodium dodecyl sulfate -polyacrylamide gel electrophoretic patterns and subunit compositions, all were identified to be type 1 collagens. A comparison of these collagens with calf skin type 1 collagen indicated the same and α 1 ,α 2 and β chains were the major components of these collagens. γ components were also found in lesser amounts in these collagens. The results of the present study indicated that comparing the three species Dog shark skin had good yield of collagen and it could be served as an alternative source of collagen for different biomedical applications.
European Food Research and Technology, 2013
Australasia has a large fish industry, and fish skin by-products from the processing industry could be used for the commercial production of fish collagen. The aim of this study was to characterize collagen extracted from the Asian sea bass (Australian barramundi) (Lates calcarifer) and snapper (Pagrus auratus) skin as an alternative to mammalian-derived collagen in gelatin products. The acidsoluble fractions of collagen from Asian sea bass and snapper skin were extracted and yielded about 8 and 7.5 % collagen (on a dry weight basis), respectively. The electrophoretic and chromatography patterns indicated that both collagens comprise of a1, a2, a3, and b chains, corresponding to the properties of calf skin collagen type I. Amino acid analysis and peptide mapping of digested collagen suggested differences in their amino acid sequences and collagen primary structure. Fourier transform infrared spectroscopy demonstrated that the helical structure of collagen was completely maintained in Asian sea bass and partially in snapper. Transition temperatures for the completion of the melting process in the two collagen networks were confirmed with differential scanning calorimetry and dynamic oscillatory rheology to be about 29°C. Zeta potential analysis identified the isoelectric points (pI values) of collagen from Asian sea bass and snapper skin at pH 6.90 and 7.75, respectively. Thus, Asian sea bass and snapper skin could be an important alternative source of collagen to replace mammalian collagen for industrial applications.
Animals
Fraser’s dolphins (Lagenodelphis hosei) possess great healing abilities. Their skin composition can be restored after wounding, including collagen spacing, orientation, and bundle thickness. However, it remains unclear how collagens are involved in the wound-healing process and eventually regain normality in Fraser’s dolphins. Learned from the other two scarless healing animals, changes in type III/I collagen composition are believed to modulate the wound healing process and influence the scarring or scarless fate determination in human fetal skin and spiny mouse skin. In the current study, Herovici’s, trichrome, and immunofluorescence staining were used on normal and wounded skin samples in Fraser’s dolphins. The results suggested that type I collagens were the main type of collagens in the normal skin of Fraser’s dolphins, while type III collagens were barely seen. During the wound healing process, type III collagens showed at early wound healing stages, and type I collagen increa...
Food Research International, 2013
Acid-solubilized collagen (ASC) and pepsin-solubilized collagen (PSC) were extracted from the skin of sail fish (Istiophorus platypterus) with yields of 5.76% and 2.11% respectively, on the basis of wet weight. According to the electrophoretic pattern, ASC and PSC consisted of two different α chains (α1 and α2) and were characterized as Type I collagen. Peptide maps of ASC and PSC hydrolysed by Achromopeptidase from Achromobacter lyticus exhibited similar banding patterns with human-placenta collagen suggesting, sailfish collagen is a Type I collagen. Fourier transform infrared (FTIR) spectra of ASC and PSC were quite similar and the regions of amides A, I, II and III were 3422 and 3337, 1654 and 1646, 1560 and 1559 and 1240 cm −1 respectively. FTIR results suggest that the pepsin hydrolysis did not affect the secondary structure of collagen, especially triple-helical structure. 1 H NMR analysis revealed the presence of water molecules along with their corresponding singlet medium chemical shift of about 4.6 to 4.8 ppm in ASC and PSC. Scanning electron microscopy (SEM) studies confirmed the presence of collagen in the isolated, as fine globular filaments. Thus, possibility of sail fish collagen as an alternative source for mammalian collagen and also it could be used in nutraceutical and pharmaceutical industries.