Hemostasis and in vivo degradation of thermosensitive chitosan hemostatic film

doi:10.3969/j.issn.2095-4344.2014.34.009

Abstract

Abstract:

BACKGROUND: The thermosensitive chitosan is a kind of chitosan, its hemostatic effect, tissue compatibility and in vivo absorption need further investigations.

OBJECTIVE: To investigate the hemostasis, in vivo degradation and tissue compatibility of thermosensitive chitosan hemostatic film.

METHODS: A total of 48 Sprague-Dawley rats were randomly divided into four groups, and carried out two experiments at the same time. (1) The incisions of the liver in three groups were covered with the thermosensitive chitosan hemostatic film, cellulose hemostatic cotton and gelatin sponge, respectively. Blank control group received no treatment. The bleeding time and bleeding amount were recorded. (2) The incisions of the quadriceps femoris muscle of rats in the above three groups were embedded with the same hemostatic materials respectively. Blank control group was not embedded. At 1, 2, 3, 4, 6 weeks, the incision tissues of the liver and the quadriceps femoris muscle were harvested for observation. After 4 weeks, the incisions were observed with hematoxylin- eosin staining and transmission electron microscopy.

RESULTS AND CONCLUSION: The bleeding time and bleeding amount of thermosensitive chitosan hemostatic film and cellulose hemostatic cotton groups were significantly lower than those of gelatin sponge and blank control groups (P < 0.05). After 6 weeks, the thermosensitive chitosan hemostatic film was absorbed completely. After 3 weeks, the cellulose hemostatic cotton was absorbed completely. After 2 weeks, the gelatin sponge was absorbed completely. The liver lobules of thermosensitive chitosan hemostatic film were complete, the liver cell were normal structure, showing light swelling and little inflammatory cell infiltration. Under transmission electron microscopy, the liver cells had integral structure, cell nucleus and organelles remained intact. The muscle fibers showed complete structure and little inflammatory cell infiltration. Under transmission electron microscopy, the muscle fibers ranked tidily, with integral cell nucleus and organelles. The thermosensitive chitosan hemostatic film has good hemostasis effect and tissue compatibility.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: chitosan, cellulose, gelatin sponge, degradation

CLC Number:

R318

Yin Gang, Chen Hui-hao, Guo Xing-feng, Wei Chang-zheng, Hou Chun-lin. Hemostasis and in vivo degradation of thermosensitive chitosan hemostatic film[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(34): 5461-5465.

Figures/Tables 2

2.1 实验动物数量分析 48只大鼠均进入结果分析。 2.2 各组大体观察肝创面止血实验：①温敏性壳聚糖止血膜能快速聚集血液，形成一层凝胶保护膜，快速止血。术后1-6周，腹腔内的肝脏、小肠及大肠周围粘连轻，缺损的肝创面生长良好。②纤维素止血较快，但纤维素止血绵结构松散，吸附血液能力较温敏性壳聚糖止血膜弱，出血较多。术后1-6周，腹腔有一定的粘连，肝脏与周围肠道粘连一起，但较容易分开，肝创面愈合良好，无明显炎性反应。③明胶海绵止血效果较差，出血量较多。术后1-6周，腹腔内粘连较严重，肝脏与肠道组织粘连一起，并且较难分开，肝创面愈合良好，无明显炎症反应。④空白对照组4 min后创面血液继续流出，创面不能止血。术后1-6周，腹腔粘连较重，肝创面愈合良好。股四头肌实验：①术后1-6周，温敏性壳聚糖组股四头肌粘连轻，原切口周围无明显粘连，肌肉间隙少瘢痕形成，较容易分离，肌肉组织愈合良好。②术后1-6周，纤维素组股四头肌有一定的粘连，肌肉间隙粘连，不易分开，股四头肌创面愈合良好，无明显炎性反应。③术后1-6周，明胶海绵组股四头肌内粘连较严重，较难分开，股四头肌创面愈合良好，无明显炎症反应。④术后1-6周，空白对照组股四头肌内粘连较严重，较难分开。 2.3 各组出血时间及出血量测定通过统计软件分析，温敏性壳聚糖组、纤维素组、明胶海绵组止血时间和出血量均显著低于空白对照组(P < 0.05)。温敏性壳聚糖组、纤维素组出血量和出血时间显著少于明胶海绵组(P < 0.05)，温敏性壳聚糖组、纤维素组组间出血量和出血时间比较差异均无显著性意义(P > 0.05)，见表1。 2.4 各组止血材料代谢及组织形态学观察止血材料代谢观察：①肝创面：温敏性壳聚糖止血膜2周时少量吸收，4周时大部分吸收，6周后基本吸收，未见明显的残留；纤维素止血棉2周时少量残留，3周后基本降解吸收；2周时明胶海绵基本降解吸收。②股四头肌：温敏性壳聚糖止血膜2周时少量吸收，4周时大部分吸收，6周后基本吸收；2周时纤维素棉少量残留，3周时基本降解吸收；2周时明胶海绵基本降解吸收，无明显残留。苏木精-伊红染色光镜观察：①肝组织：温敏性壳聚糖组肝小叶结构完整，肝细胞结构基本正常，肿胀轻，变性坏死少，炎症细胞浸润程度轻(图1A)；纤维素组肝小叶基本完整，肝细胞肿胀情况轻，变性坏死的肝细胞稍多，炎症细胞浸润程度重(图1B)；明胶海绵组肝小叶基本完整，肝细胞肿胀情况轻，变性坏死的肝细胞多，炎症细胞浸润程度重(图1C)；空白对照组基本正常肝组织结构(图1D)。②股四头肌：温敏性壳聚糖组肌肉纤维结构完整，变性坏死的肝细胞少，炎症细胞浸润程度轻(图2A)；纤维素组肌细胞结构基本完整，炎性细胞少量浸润(图2B)；明胶海绵组肌细胞结构基本完整，炎性细胞较多(图2C)；空白对照基本正常肌肉组织结构(图2D)，见表1。透射电镜观察：①肝组织：温敏性壳聚糖组整个肝细胞外形结构清楚，细胞核无破损，细胞核内物质分布均匀，细胞质内线粒体、高尔基体丰富，糖原含量较多(图3A)；纤维素组肝细胞肿胀不明显，大体外形结构清楚，细胞核无破损，细胞核内物质分布均匀，细胞质内线粒体、高尔基体丰富，糖原含量较多，滑面内质网增生，扩张不明显(图3B)；明胶海绵组肝细胞肿胀，大体结构紊乱，细胞核不规则或破损，细胞核内的物质分布不均匀，细胞内的线粒体分布少及糖原较少，滑面内质网增生严重，扩张明显(图3C)；空白对照组基本正常肝组织结构(图3D)。②股四头肌：温敏性壳聚糖组肌细胞内纤维走向整齐，肌细胞核形态正常，肌细胞核内物质分布均匀，高尔基体丰富，糖原含量较多(图4A)；纤维素组肌细胞内纤维走向整齐，细胞核形态基本正常，肌细胞核内物质分布均匀，高尔基体较多，糖原含量较多(图4B)；明胶海绵组肌细胞内纤维走向稍紊乱，细胞核破损，不规则，肌细胞核内物质分布不均匀，内质网少，糖原含量较少(图4C)；空白对照组基本正常肌肉组织结构(图4D)。"

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