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浙江大学学报（医学版）
浙江大学学报（医学版）  2021, Vol. 50 Issue (5): 633-641    DOI: 10.3724/zdxbyxb-2021-0066
原著     
一种可吸收止血纤丝的有效性和安全性评价
王玲爽1,*,童健星1,赵哲哲1,杨晓春2
1.杭州协合医疗用品有限公司，浙江 杭州 310018
2.浙江大学药学院，浙江 杭州 310058
Evaluation of self-prepared absorbable hemostatic cellulose fibrils
WANG Lingshuang1,*,TONG Jianxing1,ZHAO Zhezhe1,YANG Xiaochun2
1. Hangzhou Singclean Medical Products Co., Ltd, Hangzhou 310018, China;
2. College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China
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摘要：

目的：评估可吸收止血纤丝的有效性和安全性。方法：采用自主研发专利技术制备得到一种可吸收止血纤丝，与已上市产品速即纱进行比较，通过大体观察和傅里叶红外光谱法表征其物理形态及分子结构；电位滴定法测定纤丝的羧基含量，酸碱度计测定酸碱度值，铜乙二胺法测定相对分子质量；倒置显微镜观察纤丝与血液接触后的表现；琼脂扩散法评价纤丝的细胞毒性。建立大鼠髂外动脉出血模型和背部肌肉渗血模型，通过止血时间和血液质量考察纤丝的止血有效性，通过观察拍照、免疫器官称重、血液学和凝血功能检测及组织病理学检查分析纤丝的降解吸收情况和安全性。结果：纤丝和速即纱具有相似的分子结构。与原料再生纤维素相比，纤丝和速即纱在1727?cm–1附近均出现了典型的羧基–COOH伸缩振动吸收峰。两种材料羧基含量约为20%，酸碱度值约为3，与氧化前相比表现出明显的酸性。纤丝经氧化后的相对分子质量为4466±79，与速即纱接近(P>0.05)。纤丝与速即纱接触血液后体积膨胀，可吸收自身质量数十倍的血液。琼脂扩散实验表明纤丝无细胞毒性。动物实验结果显示，纤丝可在2?min内完成止血，止血时间和吸收血液质量与速即纱差异无统计学意义（均P>0.05）。纤丝植入肌肉渗血部位后1周即可降解，植入纤丝对动物的外观体征、体重、摄食、胸腺、脾脏、淋巴结及血液学、凝血功能等均无明显影响，组织病理学检查未见明显异常。结论：本研究自主研发的可吸收止血纤丝具有较好的生物安全性和有效性。
关键词： 纤维素;  止血材料;  纤丝;  可吸收;  安全性评价    
Abstract:

Objective:To evaluate the effectiveness and safety of self-prepared absorbable hemostatic fibrils.Methods:A kind of absorbable hemostatic fibrils were prepared by self-developed patent technique. The physical form and molecular structure of the fibrils and a marketed product Surgicel? were characterized by general observation and infrared spectroscopy; the carboxyl content, pH value and relative molecular mass of fibrils were determined by potentiometric titration method, pH meter and copper ethylenediamine method, respectively. The behavior of the fibrils and Surgicel? in contact with blood was observed by inverted microscope, the cytotoxicity was evaluated by agarose diffusion cell assay in vitro. The external iliac artery hemorrhage model and the back muscle infiltration model in rats were established. The hemostatic effectiveness of the fibrils was investigated by hemostasis time and blood weight, and the degradation and biosafety of fibrils were investigated by observation photography, immune organ weighing, hematology and coagulation index measuring, and histopathological examination. Results:The fibrils and Surgicel? had similar molecular structures. Compared with the raw material regenerated cellulose, the typical carboxyl stretching vibration absorption peak of –COOH appeared near 1727?cm–1 in both fibrils and Surgicel?. The carboxyl content of the two materials was about 20%, and the pH value was about 3. The relative molecular mass of the fibers after oxidation was 4466±79, which was close to that of Surgicel?(P>0.05). After contacting with blood, the volume of fibrils and Surgicel? expanded, and absorbed blood of dozens of times as their own weight. The results of agar diffusion test showed that the fibrils had no cytotoxicity. The results of animal experiments showed that the hemostasis completed within 2?min, and there was no significant difference in blood weight and speed of hemostasis between two products (both P>0.05). The fibrils could be degraded 1 week after being implanted to the bleeding sites of the muscle. There were no pathological effects on the appearance, body weight, food intake, immunological tissue thymus, spleen, lymph nodes, hematology and coagulation indexes of the rats, and no obvious abnormality found in the histopathological examination.Conclusion:The prepared absorbable hemostatic fibrils have excellent biological safety and effectiveness.
Key words: Cellulose    Hemostatic material    Fibril    Absorbable    Safety evaluation
收稿日期: 2021-03-12 出版日期: 2021-12-29
:  R318.08  
通讯作者: 王玲爽   
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引用本文:

王玲爽,童健星,赵哲哲,杨晓春. 一种可吸收止血纤丝的有效性和安全性评价[J]. 浙江大学学报（医学版）, 2021, 50(5): 633-641.

WANG Lingshuang,TONG Jianxing,ZHAO Zhezhe,YANG Xiaochun. Evaluation of self-prepared absorbable hemostatic cellulose fibrils. J Zhejiang Univ (Med Sci), 2021, 50(5): 633-641.

链接本文:

https://www.zjujournals.com/med/CN/10.3724/zdxbyxb-2021-0066    或    https://www.zjujournals.com/med/CN/Y2021/V50/I5/633

图 1  纤丝的外观
图 2  再生纤维素、速即纱和纤丝的红外光谱图

材料

n

酸碱度

相对分子质量

再生纤维素

3

5.35±0.11

34?699±188

速即纱

3

2.94±0.01*

4203±142*

纤丝

3

2.99±0.03*

4466±79*
表 1  再生纤维素、速即纱和纤丝的酸碱度及相对分子质量
图 3  速即纱及纤丝在显微镜下吸血前后纤维直径对比纤丝单根纤维素短纤维吸收血液前直径约70~90 μm（A），吸收血液后膨胀至近200 μm（B）； 速即纱单根纤维素短纤维吸收血液前直径约100 μm（C），吸收血液后膨胀至200 μm以上（D）.标尺=200 μm.

材料

吸血前质量（g）

吸血后质量（g）

吸血量（g）

吸血倍数

纤丝

0.11

2.75

2.64

24.0

0.14

4.41

4.27

30.5

速即纱

0.10

2.55

2.45

24.5

0.10

2.93

2.83

28.3
表 2  速即纱和纤丝的吸血量及吸血倍数
图 4  大鼠术后肌肉渗血部位材料降解情况术后第1周，速即纱和纤丝在肌肉渗血部位基本被降解吸收，组织表面未见明显残留，各脏器外观未见明显异常；术后第2、3、4周，植入部位各脏器的外观未见明显异常. 黄色框区域为材料植入部位.
图 5  各组摄食量和体重增长情况
图 6  术后第4周各组胸腺、脾脏和肠系膜淋巴结组织病理学表现各组各脏器组织结构均正常，未见明显差异.标尺=50 μm.

组别

n

胸腺（g）

脾脏（g）

胸腺系数

脾脏系数

无菌纱布组第1周

39

0.576 ± 0.159

0.912 ± 0.157

0.220 ± 0.040

0.354 ± 0.052

第4周

10

0.504 ± 0.060

0.728 ± 0.190

0.154 ± 0.029

0.216 ± 0.032

速即纱组第1周

39

0.524 ± 0.149

0.807 ± 0.272

0.206 ± 0.054

0.314 ± 0.091

第4周

8

0.480 ± 0.091

0.728 ± 0.145

0.147 ± 0.034

0.226 ± 0.066

纤丝组第1周

30

0.416 ± 0.147

0.757 ± 0.185

0.168 ± 0.052

0.300 ± 0.048

第4周

7

0.411 ± 0.066*

0.835 ± 0.178

0.120 ± 0.013*

0.254 ± 0.127
表 3  各组术后第1周和第4周胸腺、脾脏称重结果比较

组别

白细胞数（×109/L）

红细胞数（×1012/L）

血红蛋白量（g/L）

红细胞容积（%）

平均红细胞容积（fL）

第1周

第4周

第1周

第4周

第1周

第4周

第1周

第4周

第1周

第4周

无菌纱布组

9.0 ± 3.3

10.0 ± 3.3

5.6 ± 0.5

7.3 ± 0.6

115 ± 8

136 ± 10

37.2 ± 2.6

42.8 ± 2.5

66.4 ± 5.3

59.1 ± 3.5

速即纱组

10.0 ± 3.3

9.5 ± 3.6

5.6 ± 0.3

6.6 ± 1.1

112 ± 6

124 ± 21

36.5 ± 2.2

38.9 ± 6.1

65.6 ± 5.2

58.6 ± 2.5

纤丝组

10.2 ± 5.7

10.0 ± 2.5

5.6 ± 0.4

6.4 ± 1.7

111 ± 7

118 ± 31

36.0 ± 2.0

37.3 ± 8.7

64.1 ± 3.1

59.5 ± 4.5

组别

平均血红蛋白量（pg）

平均血红蛋白浓度（g/L）

血小板数（×109/L）

活化部分凝血活酶时间（s）

凝血酶原时间（s）

第1周

第4周

第1周

第4周

第1周

第4周

第1周

第4周

第1周

第4周

无菌纱布组

20.5 ± 1.5

18.8 ± 0.7

308 ± 5

318 ± 10

1487 ± 165

1221 ± 184

11.7 ± 1.4

11.3 ± 2.5

8.7 ± 0.4

8.9 ± 0.9

速即纱组

20.2 ± 1.3

18.7 ± 0.8

309 ± 11

319 ± 5

1497 ± 260

1249 ± 283

11.8 ± 1.4

10.8 ± 2.0

8.7 ± 0.4

8.5 ± 0.7

纤丝组

19.7 ± 0.6

18.6 ± 1.1

308 ± 7

313 ± 12

1398 ± 410

1233 ± 153

13.0 ± 2.1

12.8 ± 1.6

8.8 ± 0.5

8.8 ± 0.5
表 4  各组术后第1周和第4周血液学和凝血功能检测结果比较
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