蠶蛹?xì)ぞ厶菑?fù)合止血材料的制備及性能研究
發(fā)布時(shí)間：2018-06-04 14:50
本文選題：蠶蛹?xì)ぞ厶?/strong> + 止血��； 參考：《西南大學(xué)》2012年碩士論文
【摘要】：在戰(zhàn)傷急救及日常手術(shù)治療中,過度失血是致死的主要原因。目前國內(nèi)廣泛使用的止血材料都有各自的不足之處,而國外進(jìn)口的高效止血材料因其昂貴的價(jià)格限制了其在日常止血中的大范圍應(yīng)用。殼聚糖具有良好的抗菌、止血性能和生物相容性,是制備醫(yī)用敷料的理想材料。而中國蠶業(yè)資源豐富,繅絲副產(chǎn)物蠶蛹中蘊(yùn)含著大量的殼聚糖資源,且蠶蛹?xì)ぞ厶沁�具有某些優(yōu)于蝦蟹殼聚糖的性能,因此,本研究以蠶蛹中提取出的殼聚糖為主要原料,輔加以明膠作為復(fù)合劑,并以氯化鈣作為促凝劑、戊二醛作為交聯(lián)劑、甘油作為抗凍塑性劑,通過冷凍干燥法研制出一種新型的海綿狀傷口止血材料。本文采用單因素試驗(yàn)及正交試驗(yàn)對蠶蛹?xì)ぞ厶菑?fù)合止血材料的制備工藝進(jìn)行了優(yōu)化,并詳細(xì)探討了其體外凝血性能和動(dòng)物活體止血性能,以及材料的促傷口愈合性能、抑菌性能和毒理學(xué)生物安全性能,研究結(jié)果如下： 1、從蠶蛹中提取甲殼素進(jìn)而制備殼聚糖,且通過控制反應(yīng)時(shí)間能夠得到不同脫乙酰度的蠶蛹?xì)ぞ厶?最高脫乙酰度可達(dá)到93.13%,適合作為醫(yī)藥級產(chǎn)品原料。將從蠶蛹中提取出的殼聚糖粉末,與明膠共混溶解并添加適當(dāng)助劑后,通過冷凍真空干燥法制備蠶蛹?xì)ぞ厶菑?fù)合止血材料,制備出的樣品呈海綿狀,表面及內(nèi)部孔隙豐富、分布均勻,且具有一定彈性材料觸感。 2、通過單因素試驗(yàn)研究了蠶蛹?xì)ぞ厶菑?fù)合止血材料的制備工藝對凝血指數(shù)的影響,實(shí)驗(yàn)表明,最佳制備工藝參數(shù)為：預(yù)凍溫度-40℃、預(yù)凍時(shí)間1h。正交試驗(yàn)分析表明,以質(zhì)量分?jǐn)?shù)為2.5%的蠶蛹?xì)ぞ厶恰?.1%的氯化鈣、0.005%的戊二醛、3.0%的明膠、0.05%的甘油制備的蠶蛹?xì)ぞ厶菑?fù)合止血材料的凝血效果最佳,該復(fù)合止血材料中各組分濃度對體外凝血指數(shù)的影響大小順序是：蠶蛹?xì)ぞ厶锹然}戊二醛明膠甘油。 3、殼聚糖是主導(dǎo)蠶蛹?xì)ぞ厶菑?fù)合止血材料凝血性能的關(guān)鍵性因素,樣品凝血性能隨著蠶蛹?xì)ぞ厶菨舛鹊纳叨行岣�；與蝦蟹殼商品殼聚糖相比,蠶蛹?xì)ぞ厶侵苽涞臉悠纺Ч�。�?yōu)化配方下制備的蠶蛹?xì)ぞ厶菑?fù)合止血.材料在體外及動(dòng)物活體上均具有優(yōu)良的止血效果：與醫(yī)用紗布相比較,其體外凝血指數(shù)減小了62.71%,使家兔耳動(dòng)脈出血模型中的出血量減少了92.08%,止血時(shí)間縮短了80.55%：與醫(yī)用明膠海綿組比較,其體外凝血指數(shù)減小了35.61%,使家兔耳動(dòng)脈出血模型中的出血量減少了88.35%,止血時(shí)間縮短了57.01%。 4、以家兔燙傷模型研究了蠶蛹?xì)ぞ厶菑?fù)合止血材料對傷口面積和愈合率的影響。結(jié)果表明,敷加了蠶蛹?xì)ぞ厶菑?fù)合止血材料的創(chuàng)面在3d、7d、14d及21d時(shí),傷口面積均小于空白對照組,愈合率均高于空白對照組,促傷口愈合效果明顯。對燙傷創(chuàng)面組織進(jìn)行病理切片檢查的結(jié)果顯示,敷加了蠶蛹?xì)ぞ厶菑?fù)合止血材料的創(chuàng)面組織生長活躍,炎性細(xì)胞浸潤少,表皮無潰瘍,與空白對照相比表現(xiàn)出了優(yōu)良的促傷口愈合性能。 5、以大腸桿菌為革蘭氏陰性菌的代表、以金黃色葡萄球菌為革蘭氏陽性菌的代表,采用抑菌率和抑菌直徑兩種指標(biāo)對蠶蛹?xì)ぞ厶菑?fù)合止血材料的抑菌性能進(jìn)行了測試。試驗(yàn)結(jié)果表明,目前廣泛使用的明膠海綿止血材料對上述兩種菌種均無抑菌作用,而蠶蛹?xì)ぞ厶菑?fù)合止血材料對上述兩種菌種在72h內(nèi)的抑菌率均達(dá)到了100%,且對大腸桿菌出現(xiàn)了直徑為1.8-2.1cm的抑菌圈,對金黃色葡萄球菌出現(xiàn)了直徑為1.6-1.8cm的抑菌圈。 6、根據(jù)國家標(biāo)準(zhǔn)對醫(yī)用生物材料的要求,對蠶蛹?xì)ぞ厶菑?fù)合止血材料進(jìn)行了包括溶血試驗(yàn)、原發(fā)性皮膚刺激試驗(yàn)、皮內(nèi)反應(yīng)試驗(yàn)、熱源試驗(yàn)、急性全身毒性試驗(yàn)等一系列毒理學(xué)研究。研究結(jié)果表明,蠶蛹?xì)ぞ厶菑?fù)合止血材料具有不溶血、對完整和破損皮膚及皮內(nèi)組織均無刺激性、不含致熱源、無急性全身毒性等特點(diǎn),作為傷口敷料使用具有安全可靠性,是一種組織相容性良好的止血材料。該研究結(jié)果,為蠶蛹?xì)ぞ厶菑?fù)合止血材料的臨床應(yīng)用提供了理論依據(jù)。
[Abstract]:Excessive bleeding is the main cause of death in the first aid and routine surgical treatment. At present, the widely used hemostat has its own shortcomings, and the high efficient hemostat imported from abroad restricts its wide application in the daily hemostasis because of its expensive price. Chitosan has good antibacterial, hemostatic performance and birth. The material compatibility is an ideal material for preparation of medical dressings. However, the Chinese sericulture resources are rich, and the silk reeling by-products contain a lot of chitosan resources, and the silkworm pupa chitosan has some properties superior to the shrimp and crab chitosan. Therefore, the chitosan as the main raw material and the gelatin as the compound, and the gelatin as the compound, are used as the compound, and the gelatin is used as the compound. A new type of spongy wound hemostat was developed with calcium chloride as a coagulant and glutaraldehyde as a crosslinker and glycerol as a freeze-resistant plastic agent. A single factor test and orthogonal test were used to optimize the preparation process of the compound hemostat of silkworm chrysalis chitosan, and the blood coagulation in vitro was discussed in detail. The results are as follows: the ability of hemostasis in vivo, and the wound healing, antibacterial and toxicological biological safety of materials.
1, chitin is extracted from silkworm pupae and chitosan is prepared and chitosan is obtained by controlling reaction time. The highest deacetylation degree can be obtained by controlling reaction time. The highest deacetylation degree can reach 93.13%. It is suitable to be used as a pharmaceutical product. The chitosan powder extracted from silkworm pupae is dissolved and added with appropriate additives. The compound hemostat of silkworm chrysalis chitosan was prepared by air drying method. The prepared samples were sponge like, the surface and internal pores were rich, distributed evenly, and had a certain elastic material touch.
2, the effect of the preparation process of the silkworm chrysalis composite Hemostat on the blood coagulation index was studied by a single factor test. The experiment showed that the optimum preparation parameters were prefreezing temperature -40 C, and the prefreezing time 1h. orthogonal test showed that the silkworm pupa shell polysugar, 0.1% calcium chloride, 0.005% glutaraldehyde, 3% gelatin, 0 gelatin, 0 were analyzed by the prefreezing time. The blood coagulation effect of the silkworm chrysalis composite hemostat prepared by.05% glycerin is the best. The effect of the concentration of each component in the compound Hemostat on the external coagulation index is: silkworm chrysalis chitosan gelatin glycerol.
3, chitosan is the key factor to lead the clotting performance of the silkworm chrysalis composite hemostat. The coagulation performance of the sample is improved effectively with the increase of the concentration of the silkworm chrysalis chitosan. Compared with the chitosan chitosan, the sample prepared by the silkworm pupa chitosan has better coagulation effect. Both in vitro and in animal living body have excellent hemostatic effect: compared with medical gauze, the blood coagulation index in vitro decreased by 62.71%, which reduced the amount of bleeding in the model of rabbit ear artery bleeding by 92.08% and the time of hemostasis by 80.55%. Compared with the medical gelatin sponge group, the coagulation index in vitro decreased by 35.61%, and the rabbit ear artery was made. The bleeding volume in the hemorrhage model was reduced by 88.35%, and the hemostasis time was shortened by 57.01%.
4, the effect of the compound hemostat of silkworm chrysalis on the wound area and healing rate was studied with the scald rabbit model. The results showed that the wound area was less than that of the blank control group at 3D, 7d, 14d and 21d, and the healing rate was higher than that of the blank control group, and the wound healing effect was obvious. The pathological sections of the facial tissue showed that the wound tissue was active, the infiltration of inflammatory cells was less and the epidermis was not ulcerated, and the wound healing performance was better than that of the blank control.
5, on behalf of the Gram-negative bacteria of Escherichia coli and Staphylococcus aureus as the representative of Gram-positive bacteria, the bacteriostasis performance of the compound hemostat of silkworm chrysalis chitosan was tested by the two indexes of bacteriostasis rate and bacteriostasis diameter. The results showed that the two kinds of bacteria were widely used in the Gelfoam hemostat. There was no bacteriostasis, and the antibacterial rate of the silkworm chrysalis compound hemostat was 100% in 72h, and the diameter of the Escherichia coli was 1.8-2.1cm, and the Bacteriostasis of Staphylococcus aureus was 1.6-1.8cm in diameter.
6, according to the requirements of national standard for medical biomaterials, a series of toxicological studies, including hemolytic test, primary skin irritation test, intradermal reaction test, heat source test, acute systemic toxicity test, were carried out on the silkworm chrysalis composite hemostatic material. The results showed that the sericulture chitosan composite hemostat had no hemolysis, and the results showed that the silkworm chrysalis composite hemostat had no hemolysis and was not hemolytic. The complete and damaged skin and intradermal tissues have no irritation, no heat source and no acute systemic toxicity. As a wound dressing, it is safe and reliable. It is a kind of hemostatic material with good histocompatibility. The result of this study provides a theoretical basis for the clinical application of the compound hemostat of silkworm chrysalis chitosan.
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R318.08
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