Effects of chitin and chitosan on blood coagulation
Introduction
The first step in the early wound healing process is hemostasis due to blood coagulation. Platelets are the most important component in blood coagulation. Furthermore, platelets release some cytokines that enhance the healing process during blood coagulation (Beatriz, Porras-Reyes, & Mustoe, 1992). We have investigated the mechanism of wound healing acceleration by chitin and chitosan (Kojima et al., 1998, Kojima et al., 2001, Minami et al., 1996, Minami et al., 1997, Mori et al., 1997, Okamoto et al., 1993a, Okamoto et al., 1993b, Okamoto et al., 1995a, Okamoto et al., 1995b, Okamoto et al., 2002, Suzuki et al., 1997, Usami et al., 1994a, Usami et al., 1994b, Usami et al., 1997). However, we could not elucidate all evidence of wound healing acceleration by chitin and chitosan including the effects of blood coagulation. Some reports indicate that chitosan accelerate blood coagulation in vivo (Brandenberg et al., 1984, Klokkevold et al., 1991, Klokkevold et al., 1992, Klokkevold et al., 1999, Malette et al., 1983). These reports suggest that chitosan influence platelets. To our knowledge, there are no reports regarding the direct effects of chitin and chitosan on platelets.
In this paper, the effects of chitin and chitosan on blood coagulation were evaluated, particularly in the function of platelets.
Section snippets
Reagents
Chitin and chitosan: Chitin (MW, 300 kD) and chitosan (MW, 80 kD) were supplied by Sunfive Co., Ltd (Japan). The mean particles of chitin and chitosan were 2.8 and 6.9 μm, 2.8 and 6.2 μm, respectively. They were each sterilized using ethylene oxide gas, and suspended in phosphate-buffered solution (PBS, pH 7.2) at a concentration of 30 mg/ml. The chitin and chitosan had degrees of deacetylation of <10% and >80%, respectively. The molecular weight and degree of deacetylation were determined using
Effect of chitin and chitosan on blood coagulation time (BCT)
The effect of the samples on BCT is shown in Fig. 1. Chitin and chitosan reduced BCT significantly in a dose-dependent manner. In the final concentration of 0.1 mg/ml, BCTs in the chitin and chitosan groups shortened by 3.7 and 4.7 min, respectively (control BCT was 12 min). Latex, which was used as a physical control, also reduced BCT significantly but not in a dose-dependent manner. Cellulose, which was used as a chemical control, also reduced BCT. However, the effect of cellulose was weak
Conclusion
Chitin and chitosan was found to enhance blood coagulation. This phenomenon was be more effective in chitosan than chitin. On the other hand, chitin was found to aggregate platelets more than chitosan. This means that the platelets aggregation does not directly reflect blood coagulation. Therefore, the shortening of BCT by chitosan may be related to not only platelets aggregation, but also erythrocyte aggregation. The present results also indicate that chitin and chitosan enhance the release of
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