Kaigen Pharma Co., Ltd.

Various types of hemostatic agents are used to manage bleeding in surgery. Many such agents are animal products, which carry the risk of secondary infection. The aim of this study is to develop a novel hemostatic agent from a non-animal source that quickly stops bleeding, is easy to use, and has no risk of infection. In this study, we synthesized calcium ion-crosslinked sodium alginate (Alg-Na/Ca) by partial substitution of Ca ions for Na ions in sodium alginate. We prepared 12 kinds of Alg-Na/Ca powders with different Ca mass ratios, molecular weights, M/G ratios and particle size distributions and measured their swelling ratio and the burst pressure generated. We found that Alg-Na/Ca began to swell immediately after contact with saline, especially Alg-Na/Ca at Ca mass ratios of 74.1-77.0 % showed a high swelling ratio after 2 min and a high burst pressure, over 200 % and 500 mmHg respectively. Also, there is a correlation between the swelling ratio after 2 min and the burst pressure. Our results suggest that, by optimizing the composition conditions, Alg-Na/Ca may be an effective hemostatic agent that could act as a tamponade by absorbing and swelling at a bleeding site to quickly achieve primary hemostasis.

The demand for nonbiological hemostatic materials has increased in surgical practice; however, few have sufficient hemostatic efficacy. This study investigated whether optimizing the composition of sodium calcium alginate, which forms a stable gel, improves hemostatic efficacy.

The sodium calcium alginate was changed for each composition, including powder size, calcium substitution rate, molecular weight, and mannuronic/guluronic acid ratio. The hemostasis in liver punch-out injury and bleeding was evaluated using swine. For the study, sodium calcium alginate with different compositions was randomly applied to bleeding sites. Previous hemostatic materials, including cellulose and sodium alginate, were used as a control. We investigated hemostasis at 2 and 5 minutes after application.

Each powder size (53-150 μm, 150-250 μm, 53-250 μm) had a similar hemostatic effect. However, the hemostatic rate was significantly higher in sodium calcium alginate than in previous hemostatic materials (P value < .01). A 74.1% calcium substitution rate had the highest hemostasis at 2 and 5 minutes after application (2 minutes: 100%, 5 minutes: 95.2%). Regarding molecular weight, the hemostatic rate was significantly higher in 333,000 than in 50,000 (2 minutes: 95% vs 70%, 5 minutes: 95% vs 70%, P = .038). Moreover, a 1.9 mannuronic/guluronic acid ratio had the highest hemostasis compared with the 0.4 mannuronic/guluronic acid ratio (2 minutes: 58.8% vs 11.8%, 5 minutes: 58.8% vs 0%, P = .033) CONCLUSION: The composition of sodium calcium alginate with a high hemostatic effect was optimized. This new hemostatic material demonstrated potential usefulness for bleeding during surgery.

N ε-(Carboxymethyl)lysine (CML), which is formed by the glycation of collagen, is a skin-accumulating advanced glycation end product and has been shown to be deeply involved in diabetic osteopenia and skin aging. In this study, we prepared the phlorotannins of marine algal polyphenols from Japanese Lessoniaceae ( Ecklonia cava, Ecklonia kurome, cultured E. kurome, Ecklonia stolonifera, Eisenia nipponica, and Eisenia bicyclis) and evaluated their inhibitory activities against CML formation in a collagen-glyoxal environment. The level of CML formed from the glycation of collagen by glyoxal was detected using an enzyme-linked immunosorbent assay. Except for E. stolonifera, the level of CML formation in the treatment with crude phlorotannins at 0.16 µg/mL was found to be comparable to that in the treatment with 0.40 mM aminoguanidine hydrochloride (AG) which is a typical antiglycation agent. In the test using phloroglucinol and isolated eckols (eckol, fucofuroeckol A, phlorofucofuroeckol A, dieckol, and 8,8’-bieckol) at a concentration of 0.80 µg/mL, the level of CML formed was lower for each compound, except for phlorofucofuroeckol A, than the data obtained with the addition of 2.0 mM AG. The mass concentration of 0.80 µg/mL was converted to 6.3 µM for phloroglucinol, 2.2 µM for eckol, 1.7 µM for fucofuroeckol A, 1.3 µM for phlorofucofuroeckol A, and 1.1 µM for dieckol and 8,8’-bieckol. From a comparison of the molar concentrations, it was found that phloroglucinol and the eckols inhibited the formation of CML resulting from glycation of collagen by glyoxal at concentrations of approximately 317 to 1818 times lower than AG.