Isolation, Characterization and Evaluation of Collagen from Jellyfish Rhopilema esculentum Kishinouye for Use in Hemostatic Applications

PLOS ONE, Dec 2019

Hemostat has been a crucial focus since human body is unable to control massive blood loss, and collagen proves to be an effective hemostat in previous studies. In this study, collagen was isolated from the mesoglea of jellyfish Rhopilema esculentum Kishinouye and its hemostatic property was studied. The yields of acid-soluble collagen (ASC) and pepsin-soluble (PSC) were 0.12% and 0.28% respectively. The SDS-PAGE patterns indicated that the collagen extracted from jellyfish mesoglea was type I collagen. The lyophilized jellyfish collagen sponges were cross-linked with EDC and interconnected networks in the sponges were revealed by scanning electron microscope (SEM). Collagen sponges exhibited higher water absorption rates than medical gauze and EDC/NHS cross-linking method could improve the stability of the collagen sponges. Compared with medical gauze groups, the blood clotting indexes (BCIs) of collagen sponges were significantly decreased (P < 0.05) and the concentration of collagen also had an influence on the hemostatic property (P < 0.05). Collagen sponges had an improved hemostatic ability compared to the gauze control in tail amputation rat models. Hemostatic mechanism studies showed that hemocytes and platelets could adhere and aggregate on the surface of collagen sponge. All properties make jellyfish collagen sponge to be a suitable candidate used as hemostatic material and for wound healing applications.

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Isolation, Characterization and Evaluation of Collagen from Jellyfish Rhopilema esculentum Kishinouye for Use in Hemostatic Applications

January Isolation, Characterization and Evaluation of Collagen from Jellyfish Rhopilema esculentum Kishinouye for Use in Hemostatic Applications Xiaochen Cheng 0 1 Ziyu Shao☯ 0 1 Chengbo Li☯ 0 1 Lejun Yu☯ 0 1 Mazhar Ali Raja☯ 0 1 Chenguang Liu 0 1 0 College of Marine Life Sciences, Ocean University of China , Qingdao , P. R. China 1 Editor: Ming Dao, Massachusetts Institute of Technology , UNITED STATES Hemostat has been a crucial focus since human body is unable to control massive blood loss, and collagen proves to be an effective hemostat in previous studies. In this study, collagen was isolated from the mesoglea of jellyfish Rhopilema esculentum Kishinouye and its hemostatic property was studied. The yields of acid-soluble collagen (ASC) and pepsin-soluble (PSC) were 0.12% and 0.28% respectively. The SDS-PAGE patterns indicated that the collagen extracted from jellyfish mesoglea was type I collagen. The lyophilized jellyfish collagen sponges were cross-linked with EDC and interconnected networks in the sponges were revealed by scanning electron microscope (SEM). Collagen sponges exhibited higher water absorption rates than medical gauze and EDC/NHS cross-linking method could improve the stability of the collagen sponges. Compared with medical gauze groups, the blood clotting indexes (BCIs) of collagen sponges were significantly decreased (P < 0.05) and the concentration of collagen also had an influence on the hemostatic property (P < 0.05). Collagen sponges had an improved hemostatic ability compared to the gauze control in tail amputation rat models. Hemostatic mechanism studies showed that hemocytes and platelets could adhere and aggregate on the surface of collagen sponge. All properties make jellyfish collagen sponge to be a suitable candidate used as hemostatic material and for wound healing applications. - Data Availability Statement: All relevant data are within the paper and its Supporting Information files. Funding: The authors received no specific funding for this work. Competing Interests: The authors have declared that no competing interests exist. 1. Introduction Uncontrolled hemorrhage after trauma and in surgical procedures has associated with the increased mortality rate, and emergency hemostatic management has been a crucial focus [1± 3]. The body's natural responses to an injury are comprised of hemostatic process and healing of the wound site [ 4 ]. However, the body's natural mechanism is unable to control massive hemorrhaging caused by major trauma or surgery. There is a medical need to develop an effective hemostat for emergency circumstances [ 5 ]. Traditional hemostasis techniques (cautery and suture ligation) used in the operating room can always cause problems such as oozing bleeding [ 3 ] and damaging of capillaries resulting in tissue necrosis [ 6 ]. For these reasons, a number of hemostatic agents that can arrest bleeding and promote hemostasis have been developed. Either natural or synthetic polymers have been employed for the construction of hemostatic agents. The configurations of these agents are mostly sheet [ 7 ], sponge [8±9] and glue [ 10 ]. Natural polymers have been widely used as topical hemostatic agents for their excellent properties such as biodegradability and biocompatibility. Natural hemostatic agents such as oxidized cellulose [11±12], chitosan [9,13±14], gelatin [ 15 ], thrombin [ 16 ], collagen [17±18] and fibrin [ 19 ] can be divided in active and passive agents [ 20 ]. For example, fibrin glue has been widely used as active hemostatic agents which can effectively prevent the postoperative complications such as bleeding, hematoma formation, seroma, edema, and prolonged drainage [21±23]. However, plasma derived fibrin sealant/ hemostatic products can cause viral contamination and anaphylaxis [24±25]. Horowitz and Busch have reported that the fibrin sealants have some risks of transmission of HIV, hepatitis virus and other parvovirus [ 26 ]. Passive hemostatics, including collagen, gelatin and oxidized cellulose, are not biologically active; their mechanism of action is to provide platelet activation and aggregation [ 3 ]. Among them, collagen has been reported as useful hemostatic agent [27± 30]. Collagen is the most abundant protein (approximately 30% by weight of total protein) in human's body which consists in extracellular matrix [ 31 ]. The triple helical structures exist in all collagen molecules and form three parallel, left-handed helical polypeptide α-chains [ 32 ]. Each α-chain has a characteristic [Gly-X-Y] domain repeat, in which X and Y mostly represent the proline (Pro) and hydroxyproline (HyP) [ 33 ]. Moreover, it has been proved that collagen has better biocompatibility, higher biodegradability, lower antigenicity and cell-binding properties as a natural protein, which can be degraded into physiologically tolerable compounds in vivo. [34±36]. The role of collagen in regulation of hemostasis is that (...truncated)


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Xiaochen Cheng, Ziyu Shao, Chengbo Li, Lejun Yu, Mazhar Ali Raja, Chenguang Liu. Isolation, Characterization and Evaluation of Collagen from Jellyfish Rhopilema esculentum Kishinouye for Use in Hemostatic Applications, PLOS ONE, 2017, Volume 12, Issue 1, DOI: 10.1371/journal.pone.0169731