Molecular Weight Analysis of Blue Shark (Prionace glauca) Collagen Hydrolysates by GPC-LS. Effect of High Molecular Weight Hydrolysates on Fibroblast Cultures: mRNA Collagen Type I Expression and Synthesis

High molecular weight (Mw) collagen hydrolysates have been demonstrated to produce a higher synthesis of collagen type I mRNA. Mw determination is a key factor maximizing the effect of collagen hydrolysates on collagen type I synthesis by fibroblasts. This work aimed to achieve a high average Mw in Blue Shark Collagen Hydrolysate, studying different hydrolysis parameters by GPC-LS analysis and testing its effect on mRNA Type I collagen expression. Analysis revealed differences in blue shark collagen hydrolysates Mw depending on hydrolysis conditions. Papain leads to obtaining a significantly higher Mw hydrolysate than Alcalase at different times of hydrolysis and at different enzyme/substrate ratios. Besides, the time of the hydrolysis factor is more determinant than the enzyme/substrate ratio factor for obtaining a higher or lower hydrolysate Mw when using Papain as the enzyme. Contrary, Alcalase hydrolysates resulted in similar Mw with no significant differences between different conditions of hydrolysis assayed. Blue shark collagen hydrolysate showing the highest Mw showed neither cytotoxic nor proliferation effect on fibroblast cell culture. Besides, it exhibited an increasing effect on both mRNA expression and pro-collagen I production.

ACE Inhibitory Peptide from Skin Collagen Hydrolysate of Takifugu bimaculatus as Potential for Protecting HUVECs Injury

Angiotensin-I-converting enzyme (ACE) is a crucial enzyme or receptor that catalyzes the generation of potent vasopressor angiotensin II (Ang II). ACE inhibitory peptides from fish showed effective ACE inhibitory activity. In this study, we reported an ACE inhibitory peptide from Takifugu bimaculatus (T. bimaculatus), which was obtained by molecular docking with acid-soluble collagen (ASC) hydrolysate of T. bimaculatus. The antihypertensive effects and potential mechanism were conducted using Ang-II-induced human umbilical vein endothelial cells (HUVECs) as a model. The results showed that FNLRMQ alleviated the viability and facilitated apoptosis of Ang-II-induced HUVECs. Further research suggested that FNLRMQ may protect Ang-II-induced endothelial injury by regulating Nrf2/HO-1 and PI3K/Akt/eNOS signaling pathways. This study, herein, reveals that collagen peptide FNLRMQ could be used as a potential candidate compound for antihypertensive treatment, and could provide scientific evidence for the high-value utilization of marine resources including T. bimaculatus.

Bioactive Collagen Hydrolysate-Chitosan/Essential Oil Electrospun Nanofibers Designed for Medical Wound Dressings

In this study, lemon balm (Melissa officinalis L.) and dill (Anethum graveolens L.) essential oils (EOs) were encapsulated into collagen hydrolysates extracted from bovine tendons and rabbit skins, both mixed with chitosan (CS) by using the coaxial electrospinning technique for potential wound dressing applications. The morphology and chemical composition of the electrospun nanofibers were investigated using scanning electron microscopy (SEM) and attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR). The antimicrobial activity of the dill EO and lemon EO, as well as the electrospun samples loaded with essential oils was determined by disk diffusion assay against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Enterococcus faecalis ATCC 29212, and Salmonella typhimurium ATCC 14028 bacterial strains; Candida albicans ATCC 10231 and Candida glabrata ATCC 90028 yeast strains; and Aspergillus brasiliensis ATCC 9642 fungal strain. In vivo biocompatibility testing of the collagen hydrolysate-chitosan/essential oil electrospun nanofibers was based on the determination of the hematological, biochemical, and immunological profile and the evaluation of the influence produced on the oxidative stress in white Swiss mice. The synergetic effect of dill and lemon balm EOs can improve the antimicrobial activity of collagen hydrolysate-chitosan nanofibers against the most important bacterial strains. The in vivo test results suggested a good biocompatibility of electrospun samples based on collagen hydrolysate extracted from bovine tendons or rabbit skin mixed with chitosan and containing dill and/or lemon balm essential oils as encapsulated bioactive compounds.

Efficacy of Chondroprotective Food Supplements Based on Collagen Hydrolysate and Compounds Isolated from Marine Organisms

Osteoarthritis belongs to the most common joint diseases in humans and animals and shows increased incidence in older patients. The bioactivities of collagen hydrolysates, sulfated glucosamine and a special fatty acid enriched dog-food were tested in a dog patient study of 52 dogs as potential therapeutic treatment options in early osteoarthritis. Biophysical, biochemical, cell biological and molecular modeling methods support that these well-defined substances may act as effective nutraceuticals. Importantly, the applied collagen hydrolysates as well as sulfated glucosamine residues from marine organisms were strongly supported by both an animal model and molecular modeling of intermolecular interactions. Molecular modeling of predicted interaction dynamics was evaluated for the receptor proteins MMP-3 and ADAMTS-5. These proteins play a prominent role in the maintenance of cartilage health as well as innate and adapted immunity. Nutraceutical data were generated in a veterinary clinical study focusing on mobility and agility. Specifically, key clinical parameter (MMP-3 and TIMP-1) were obtained from blood probes of German shepherd dogs with early osteoarthritis symptoms fed with collagen hydrolysates. Collagen hydrolysate, a chondroprotective food supplement was examined by high resolution NMR experiments. Molecular modeling simulations were used to further characterize the interaction potency of collagen fragments and glucosamines with protein receptor structures. Potential beneficial effects of collagen hydrolysates, sulfated glycans (i.e., sulfated glucosamine from crabs and mussels) and lipids, especially, eicosapentaenoic acid (extracted from fish oil) on biochemical and physiological processes are discussed here in the context of human and veterinary medicine.

Characterization, Antioxidant and α-Glucosidase Inhibitory Activity of Collagen Hydrolysate from Lamuru (Caranx ignobilis) Fishbone

The Lamuru fish (Caranx ignobilis) is mostly found in tropical waters of the indo-pacific region, namely Indonesia. It is believed to contain collagen and this study aims to isolate collagen from its bone and determine the collagen’s antioxidant and α-glucosidase inhibitory activity. In our study, the collagen was extracted using acetic acid which was hydrolyzed by collagenase enzyme from Clostridium histolyticum at a temperature of 37 °C, and pH 7.0. During hydrolysis, the degree of hydrolysis (DH) was calculated and collagen hydrolysates were characterized by SDS -PAGE, UV-Visible spectroscopy and FT-IR spectroscopy. After characterization, the collagen hydrolisate of lamuru ( CHL) fish was analyzed for its antioxidant properties and α-glucosidase inhibitory activity. The result shows that a higher percentage degree of hydrolysis was obtained, 31.17%, at 120 min of hydrolysis. The CHL characterization by SDS -PAGE showed its molecular weight ranging from 35,000-180,000 Daltons and identified the collagen as type I. The UV-Vis analysis of CHL provided a maximum absorbance at a wavelength of 233 nm. At the same time, the FT-IR analysis showed the presence of amides I, II, and III, which confirms the formation of the collagen triple helix. For its bioactivity assay, the CHL shows that CHL provided DPPH radical reduction activity reaching 51.45±1.24% (IC50 at 485.9 µg/mL). The ferric reduction antioxidant power of CHL (FRAP value) showed a significant reduction of Fe3+ to Fe2+ with a value of 711.27 µM/g. The CHL inhibition activity of α-glucosidase enzyme IC50 was determined to be 574 µg/mL. Based on the antioxidant bioactivity and α-glucosidase inhibition, the collagen peptide enables its use as a therapeutic development for a variety of disorders caused by oxidative stress, such as diabetes mellitus.

EVALUATION OF ANTIOXIDANT FUNCTIONALITY OF FISH COLLAGEN ENZYMIC HYDROLYSATE

This study aimed to investigate the anti- oxidative function of catfish collagen hydrolysate in beef minced meat samples through 10 days storage at 4 o C. Collagen hydrolysate were added to minced meat sample at two different concentrations (50 & 100 mg/100gm) . The experiment consist from control sample (C without additives) , BHT sample ( B with 200ppm butylated hydroxytoluene ) , collagenase hydrolysate samples C.C.H1, C.C.H2 (100&50 mg/100gm minced meat) respectively, and collagenase-trypsin hydrolysate samples C.H.T1 & C.H.T2 respectively (50,100mg/100gm minced meat). Peroxide values (PV), thiobarbutyric acid TBA, free fatty acid (FFA) , pH , total volatile nitrogen TVN values and total plate count assay at 0., 3, 7., 10 days were determined . PV of the BHT, C.C.H2 , C.H.T2, C.C.H1, C.H.T1 groups were significantly (p<0.05) lower than control during storage period .Meanwhile C.C.H2 and C.H.T2 groups gave the lowest TBA values through the storage period . There were no significant difference found between the C.C.H2 , C.H.T2 groups and BHT added group in all studied factors. In conclusion, the fish collagen enzymatic hydrolysates are a promising natural antioxidant and good alternative for synthetic antioxidant.

A Review of the Literature Relating to Collagen Hydrolysate and Its Potential Clinical Applications

The increasing commercial necessity to valorise commodities such as gelatine has led to significant developments in its processing and the outcome of these refinements has resulted in new applications in fields such as pharmaceuticals, medical devices, cosmetics, food and nutraceuticals. This in turn has led to the investigation of alternative sources of compounds with collagen-like properties, other than the conventionally used raw materials from mammalian species. Moreover, the current desire to seek natural, rather than synthetic compounds-especially regarding oral consumption and/or topical application-combined with the ability of gelatine derived products to form gels with varying degrees of flexibility and hydroplasticity has also accelerated research into previously unexplored applications. In the food sector, these include:- use of gelatine derivatives as an encapsulating agent (including the development of micro-beads as carriers of active compounds) foaming agents, emulsifiers, biodegradable films, colloid stabilizers and as nutraceuticals. The latter sector has especially benefitted from developments in enzymatic hydrolysis processes, where specific and highly characterised bioactive peptides often containing the amino acid hydroxyproline are end-products which have been identified to be orally bioavailable and metabolised and hence likely to deliver potential clinical benefits. This review examines manufacturing processes employed to typically produce hydrolysed collagen, evaluates studies examining bioavailability, metabolism and likely health benefits as well as potential clinical applications as a nutraceutical.