Preparation and properties of biodegradable plant polysaccharide hemostatic microspheres

doi:10.12307/2022.245

Abstract

Abstract: BACKGROUND: Hemostasis is an important part of clinical treatment. Fast and effective hemostasis is necessary to ensure the safety of surgical patients.
OBJECTIVE: To prepare a kind of plant polysaccharide hemostatic microsphere with porous structure and its performance evaluation.
METHODS: Porous hemostatic microspheres were prepared from potato starch by gelatinization, emulsification crosslinking, cleaning, and drying technique. The hemostatic performance of the hemostatic microspheres was improved by improving the gelatinization and crosslinking processes of the starches, setting the grouping: group A (gelatinization for 1 hour, crosslinking for 6 hours, group B (gelatinization for 1 hour, crosslinking for 12 hours), group C (gelatinization for 1 hour, crosslinking for 18 hours), group D (gelatinization for 2 hours, crosslinking for 6 hours), group E (gelatinization for 2 hours, crosslinking for 12 hours), and group F (gelatinization for 2 hours and crosslinking for 18 hours). The listed absorbable hemostatic microspheres were used as controls to examine the microtopography, pH, porosity and in vitro degradation of microspheres in each group. According to the above experimental results, suitable microspheres were selected for hemostasis of liver and spleen in New Zealand rabbits, comparing the sample dosage, hemostasis amount with hemostasis time.
RESULTS AND CONCLUSION: (1) Scanning electron microscope showed that group B and group E formed porous microspheres with a stable structure of 50-200 μm in diameter, and the spherical morphology and porosity were better than those of the control group. Except for groups C and E, the pH values of the other samples were in the range of 6.5-7.5, which met the pH requirements. The same quality of samples, the cross-linking time growth and degradation time increased significantly (P < 0.01). Based on the examination of morphological structure, porosity, pH and in vitro degradation in the previous stage, groups A, B, E and the control group were selected for hemostasis experiments. (2) Under the same hemostatic effect, the hemostatic time of microspheres in group E was shorter and less used compared with that in the control group, and the hemostatic effect in group B was not significantly different from that in the control group. (3) The results showed that the porous starch microspheres prepared after gelatinization at 60 °C for 2 hours, emulsification for 45 minutes, and cross-linking for 12 hours had the best hemostatic effect and its degradation time was also relatively short, and its performance was superior to that of listed products.

Key words: porous microspheres, structural analysis, preparation method, gelatinization, crosslinking, in vitro degradation, hemostasis, rabbit

CLC Number:

Yang Xue, Wang Baoqun, Jiang Xiaowen, Zou Shengcan, Ming Jinfa, Lin Shasha. Preparation and properties of biodegradable plant polysaccharide hemostatic microspheres[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(16): 2487-2491.

Figures/Tables 4

2.2 多孔止血微球的孔隙率、pH值及体外降解由表3可以看出，除A、E组孔隙率接近外，其余各组孔隙率和pH值两两对比均差异均有显著性意义(P < 0.05)，其中糊化时间增长可以适当增大孔隙率，随着交联时间延长成球时间增长，其聚合结构过于紧实，致使孔隙率降低；虽然 A、D组的孔隙率较高，但是其未交联出规则形态，致使样品密度过小、飘浮性强，而B、E组成球性较好的同时其孔隙率也保持在较高水平。成球性较好的B、E组样品孔隙率均高于上市产品 (P < 0.05)，与图1中得到的结论相同。表3同时反映出糊化和交联时间均与pH值呈正比，即糊化时间相同，交联时间越长pH值越大，当交联时间达到18 h时pH值达到最高，为7.8。交联时间相同，糊化时间适当增长，pH也略有增大，糊化2 h样品D、E、F组的pH值均略高于对应相同交联时间的A、B、C组。除 C、E组外，其他样品的pH值均在6.5-7.5之间，符合pH值要求。淀粉在糊化后形成有一定弹性和强度的半透明凝胶，凝胶的黏性和强度都会影响淀粉产品的性能，且在碱性条件下马铃薯淀粉的热稳定性、凝沉性和凝胶性能增加，糊化过程中加入氢氧化钠可以增加糊化后淀粉乳的黏度[14]；马铃薯淀粉在过量的水中加热到糊化温度，淀粉颗粒膨胀，晶体结构逐渐被破坏[15]，不同糊化时间使得晶体结构破坏程度不同，糊化淀粉乳黏度不同[16-17]。在搅拌条件下，将淀粉乳分散到含有乳化剂的有机溶剂中形成反相悬浮乳液，加入环氧氯丙烷，淀粉分子之间彼此聚合交联成球[18-19]。随着交联时间增长，聚合结构更加紧实，微球内化学残留不易随无水乙醇的清洗析出，不但会影响pH值，更加会增加临床应用的风险，故而不宜交联时间过长。另外，表3中的多孔微球体外降解时间结果显示，在活性为60 U/mL的α-淀粉酶溶液中进行酶解，相同质量的样品，交联时间增长降解时间明显增长(P < 0.01)，交联时间为18 h时降解时间长达6 h，明显高于交联时间为12 h、酶解时间为4.5 h。α-淀粉酶主要水解微球内部结构中的α-1，4糖苷键[20-21]，且水解酶分子可进入颗粒内部，具有天然多孔表面的淀粉更容易被消化[22]，最终产物以麦芽糖为主，还有少量葡萄糖和糊精，这些分解产物都是机体既有物质，会参与到正常的生理效应，或通过肝脏、肾脏排出体外[20,23]。随着交联时间增长，微球结构稳固，但并没有得到更加圆润多孔的微球结构，故而交联时间只能适度增长，既要得到稳定的微球结构，也要易于被淀粉酶水解[24-25]。"

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