Handling

sites where bleeding is easy to non-treated. A two-layered gelatin sheet is possible choice for hemostasis in cases where there is a relatively large amount of bleeding.

long time residual of the material possibly make the risk of infection higher. In the present study histological examination was carried out in order to observe the tissue response to each the material, not to examine the hemostatic effects of each material. Earlier regeneration of the peritoneum and earlier absorption of the material were observed in two-layered gelatin sheet. On the other hand, TachoSil®

induced intensive inflammatory response at the implanted site and remained longer than gelatin sheet. Regarding adhesion two-layered gelatin sheet is possibly safer than TachoSil®. (24-26) In our unpublished study (under submission) the two-layered gelatin sheet has anti-adhesive effect due to early regeneration of the peritoneum and mild inflammatory response.

The two-layered gelatin sheet, which is composed of alkaline-treated gelatin, does not carr y the same risks as biomaterials (TachoSil®), such as transmission of viral infection and allergic reactions. In fact, the gelatin alkali-treatment process eliminates the risk of transmittable diseases almost completely. (23) Moreover, from the skin of animals no infectivity of Bovine Spongifor m Encephalopathy was not detected.

CONCLUSION

We showed that the two-layered gelatin sheet is a more effective, easier to handle, and safer topical hemostatic agent than TachoSil®, which is one of the most popular materials currently in use. The two-layered gelatin sheet is safer than topical agents including fibrin components and/or thrombin in terms of risk of viral transmission and inflammatory reactions. We showed the hemostatic effectiveness of a two-layered gelatin sheet in bleeding dog spleens (an organ where bleeding is difficult to non-treated). The efficacy of newly developed materials should be evaluated by application to other organs (liver, kidney, lung, and vessels) and in humans.

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Table 1. Comparison of the hemostatic effects of TachoSil®, a two-layered gelatin sheet and a gelatin sponge in bleeding dog spleens.

Hemostatic material

Positive for bleeding (A)/Total experiments

Positive for rebleeding (B)/Total experiments

Positive for bleeding

(A + B)/Total experiments Compression

time 1 min 5 min 1 min 5 min 1 min 5 min

Two-layered

gelatin sheet 0/6 0/6 0/6 0/6* 0/6* 0/6*

*

TachoSil® 3/6 0/6 3/6 5/6 6/6 5/6

*

Gelatin sponge 0/6 0/6 0/6 0/6* 0/6* 0/6*

(A) Bleeding just after 1 or 5 minutes of compression

(B) Rebleeding happened during 5-minute observation period after 1 or 5 minutes of compression

*: significantly (p<0.001) different compared to TachoSil®

Figure 1. Two-layered gelatin sheet

Figure 2. Schematic diagram of the experiment Figure 2.Schematic deiagram of the experriment

Bleeding

Detach Compression

1 or 5 minutes Re-bleeding Positive/Negative

Figure 3. Absorbency and permeability of the materials

time (sec.)

450 400 350 300 250

150 100 50

0Two-layered gelatin sheet

absorbency

Absorbency and Permerbility (canine blood)

permearbility

TachoSil® Gelatin sponge 200

Figure 4. Hemostatic agents set onto the shaved surface of the spleen.

a. Two-layered gelatin sheet; b. TachoSil®; c. Gelatin sponge

a c

b

Figure 5. Histologic findings regarding embedded materials a. Two-layered gelatin sheet; b. TachoSil®; c. Gelatin sponge

a

b

c

STUDY II-2

Anti-adhesive Effects of the Newly Developed Two-layered Gelatin Sheet in Dogs

TABLE OF CONTENTS (STUDY II-2)

INTRODUCTION

MATERIALS AND METHODS 1. Animal protocol

2. Preparation of anti-adhesive agents 2.1. Two-layered gelatin sheet

2.2. Seprafilm®

2.3. INTERCEED®

3. Surgical procedure