CN114618008A - 一种医用丝素蛋白胶原制备方法 - Google Patents
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Abstract
本发明公开了一种医用丝素蛋白胶原制备方法。包括以下步骤:(1)制备溶液A;(2)制备溶液B;(3)制备溶液C;(4)混合。本发明的制备方法通过三层不同成分的胶原蛋白,首先是纯丝素蛋白,其次是经过甘油改性的丝素蛋白,再次是经过羟基磷灰石改性的丝素蛋白,通过分别、多次复合冻干,层与层之间相容较好;同时,采用羟基磷灰石改性,促进骨组织及血管生长;提高材料的机械强度,止血效果更佳;甘油作为改性溶剂,最后冻干之后用纯化水浸泡除去;而纯丝素蛋白胶原,后期更易吸收降解。
Description
技术领域
本发明涉及一种医用丝素蛋白胶原制备方法,属于生物医药技术领域。
背景技术
蚕丝是熟蚕结茧时分泌丝液凝固而成的连续长纤维,属蛋白质纤维。也称天然丝,是一种天然纤维,也是人类利用最早的动物纤维之一。世界上最细最柔软的天然纤维,它的细胞结构最接近人体肌肤,所以其亲肤性能是所有纤维中最好的,被誉为“纤维皇后”。
天然蚕丝由丝素蛋白与丝胶蛋白组成,其中,由于丝胶蛋白具有免疫原性,故限制了蚕丝在生物组织工程领域的广泛应用,因此,蚕丝在生物组织工程领域的应用中,通常是以蚕丝脱胶的方法制备丝素蛋白。
丝素蛋白由于具有良好的生物相容性而被广泛应用于组织工程领域,如人工血管、人工神经、人工韧带等。但是,丝素蛋白本身并不是一种十全十美的生物材料,所以,在通常情况下,丝素蛋白需要与其他材料进行复合以制备复合材料,以满足组织工程领域不同研究及应用的需要。
目前上的医用蛋白胶原大多是以牛跟腱、牛骨等牛组织为原料,通过提纯、酶解、交联等工艺制备得到。
丝素蛋白产品主要制备成膜状材料,其改性也都通过静电纺丝工艺进行制备,改性生产成本高昂,生产效率低下,并不利于进行工业化生产。
发明内容
为了克服现有技术的不足,本发明提供了一种医用丝素蛋白胶原制备方法,通过不同成分的胶原蛋白多次复合冻干得到的胶原蛋白,相比市面上的牛源胶原蛋白产品具有更低免疫性、组织亲和性强、止血性能好、组织修复功能强、生物降解性好等;是临床上使用的理想的止血修复生物材料,它可在体内自行降解吸收。
本发明是通过以下技术方案来实现的:
一种医用丝素蛋白胶原制备方法,包括以下步骤:
(1)制备溶液A:以生丝为原料,采用碳酸钠溶液在高温条件下进行处理,烘干后得到的蚕丝加入到溴化锂溶液中进行溶解,用水进行透析,离心后得到溶液A;
(2)制备溶液B:向溶液A中加入甘油,得到溶液B;
(3)制备溶液C:将溶液A与羟基磷灰石混合,得到溶液C;
(4)混合:将溶液A加入到模具中三分之一的位置,冻干;然后再加入溶液B到模具中三分之二的位置,冻干;最后加入溶液C至模具满,冻干得到成品。
所述的一种医用丝素蛋白胶原制备方法,所述碳酸钠的物质的量浓度为0.3-0.8mol/L。
所述的一种医用丝素蛋白胶原制备方法,所述高温条件为90-110℃。
所述的一种医用丝素蛋白胶原制备方法,所述溴化锂的物质的量浓度为9-10mol/L。
所述的一种医用丝素蛋白胶原制备方法,所述甘油的质量分数为40-60%。
所述的一种医用丝素蛋白胶原制备方法,所述乙醇溶液中乙醇的质量分数为40-60%。
所述的一种医用丝素蛋白胶原制备方法,羟基磷灰石为纳米级。
所述的一种医用丝素蛋白胶原制备方法,冻干得到的成品加入到过质量分数为2-4%的过氧化氢溶液中浸泡,再用水浸泡清洗,最后进行冷冻干燥。
所述的一种医用丝素蛋白胶原制备方法,冷冻干燥之后再采用钴-60辐照灭菌。
所述的一种医用丝素蛋白胶原制备方法,所述模具为圆柱形。
本发明所达到的有益效果:
本发明的制备方法通过三层不同成分的胶原蛋白,首先是纯丝素蛋白,其次是经过甘油改性的丝素蛋白,再次是经过羟基磷灰石改性的丝素蛋白,通过分别、多次复合冻干,层与层之间相容较好;同时,采用羟基磷灰石改性,促进骨组织及血管生长;提高材料的机械强度,止血效果更佳;甘油作为改性溶剂,最后冻干之后用纯化水浸泡除去;而纯丝素蛋白胶原,后期更易吸收降解。
本发明的制备方法简单易操作,所需原料价格便宜,整体成本较低,适合进行工业化生产,同时,得到的胶原蛋白具有良好的性能。
本发明的制备方法得到的丝素胶原蛋白相比市面上的牛源胶原蛋白产品具有更低免疫性,同时,组织亲和性强、止血性能好、组织修复功能强、生物降解性好;是临床上使用的理想的止血修复生物材料,它可在体内自行降解吸收。
附图说明
图1是制备得到的丝素蛋白胶原的示意图。
具体实施方式
下面对本发明作进一步描述。以下实施例仅用于更加清楚地说明本发明的技术方案,而不能以此来限制本发明的保护范围。
实施例1
一种医用丝素蛋白胶原制备方法,包括以下具体步骤:
1. 丝蛋白粗提取(除胶)
1.1 配置0.5mol/L的碳酸钠溶液1000mL,加热煮沸,加入搅拌罐中;
1.2 搅拌加入50g的家蚕生丝,保持沸腾搅拌30min;
1.3 重复上述过程2-3次,之后在60℃电热鼓风干燥箱中烘干;
2. 丝蛋白提纯
2.1 将初步提取的蚕丝加入到9.3mol/L的LiBr溶液,加热溶解,其中,蚕丝与溴化锂溶液的重量比为1:3;
2.2 溶液装入透析袋,用纯化水透析24h;
2.3 加入冷冻离心机进行离心,重复2-3次,得到溶液A;
2.4 溶液A加入50%甘油,得到溶液B;所述溶液A和甘油的重量比为35:1;
3. 丝蛋白羟基磷灰石混合液制备
3.1 溶液A与纳米级羟基磷灰石(12μm)混合,加热搅拌均匀得到溶液C;所述溶液A和羟基磷灰石的重量比为1:100;
4. 蛋白分层冻干
4.1 溶液A加到圆柱状模具中1/3, 冻干24h;
4.2 再加入溶液B到模具2/3,冻干24h;
4.3冻干之后浸泡到纯化水中24h,隔2h换一次纯化水,之后冻干24h;
4.4 再加入溶液C,加满模具,冻干24h;
4.5 冻干之后成品浸泡到50%的乙醇溶液中24h;
4.6 纯化水浸泡清洗;
5. 病毒灭活及清洁
5.1 冻干成品加入到过3%过氧化氢溶液,浸泡24h
5.2 之后用纯化水浸泡清洗
5.3 冷冻干燥后,产品经钴-60辐照灭菌得到最终的医用丝素蛋白胶原。
对得到的医用丝素蛋白胶原进行常规检测,检测结果见表1。
表1 医用丝素蛋白胶原常规检测结果
检测项目 | 标准 | 实测值 |
外观 | 白色或浅黄色疏松的海绵 | 符合要求 |
抗拉性能 | ≥0.5N | 0.8N |
酸碱度 | 4.0~7.0 | 5.8 |
干燥失重 | ≤15.0% | 13.5% |
炽灼残渣 | ≤1.0% | 0.4% |
重金属 | ≤10μg/g | 符合要求 |
蛋白含量 | ≥90.0% | 97.6% |
羟脯氨酸含量 | ≥9.0% | 10.2% |
硫酸盐灰分 | ≤2.0% | 符合要求 |
液体吸收性 | ≥20倍 | 27倍 |
锂离子和溴离子残留 | ≤50ppm | 17ppm |
碳酸钠残留 | ≤50ppm | 23ppm |
乙醇残留 | ≤0.5% | 0.03% |
甲醇残留 | ≤0.3% | 0.02% |
DNA残留 | <100ng/mg | 符合要求 |
细菌内毒素 | <20EU/件 | 符合要求 |
可见,本实施例制备得到的丝素蛋白胶原性能优异,无毒害。应用于拔牙之后填充止血修复,促进愈合,效果好。
实施例2
一种医用丝素蛋白胶原制备方法,包括以下具体步骤:
1. 丝蛋白粗提取(除胶)
1.1 配置0.3mol/L的碳酸钠溶液1000mL,加热煮沸,加入搅拌罐中;
1.2 搅拌加入50g的家蚕生丝,保持沸腾搅拌30min;
1.3 重复上述过程2-3次,之后在60℃电热鼓风干燥箱中烘干;
2. 丝蛋白提纯
2.1 将初步提取的蚕丝加入到8mol/L的LiBr溶液,加热溶解,其中,蚕丝与溴化锂溶液的重量比为1:3;
2.2 溶液装入透析袋,用纯化水透析24h;
2.3 加入冷冻离心机进行离心,重复2-3次,得到溶液A
2.4 溶液A加入40%甘油,得到溶液B;所述溶液A和甘油的重量比为40:1;
3. 丝蛋白羟基磷灰石混合液制备
3.1 溶液A与纳米级羟基磷灰石(12μm)混合,,加热搅拌均匀得到溶液C;所述溶液A和羟基磷灰石的重量比为1:90;
3.2 加入40 %的乙醇溶液得到溶液C;
4. 蛋白分层冻干
4.1 溶液A加到圆柱状模具中1/3, 冻干24h;
4.2 再加入溶液B到模具2/3,冻干24h;
4.3冻干之后浸泡到纯化水中24h,隔2h换一次纯化水,之后冻干24h;
4.4 再加入溶液C,加满模具,冻干24h;
4.5 冻干之后成品浸泡到50%的乙醇溶液中24h;
4.6 纯化水浸泡清洗;
5. 病毒灭活及清洁
5.1 冻干成品加入到过3%过氧化氢溶液,浸泡24h
5.2 之后用纯化水浸泡清洗
5.3 冷冻干燥后,产品经钴-60 辐照灭菌得到最终的医用丝素蛋白胶原。
实施例3
一种医用丝素蛋白胶原制备方法,包括以下具体步骤:
1. 丝蛋白粗提取(除胶)
1.1 配置0.8mol/L的碳酸钠溶液1000mL,加热煮沸,加入搅拌罐中;
1.2 搅拌加入50g的家蚕生丝,保持沸腾搅拌30min;
1.3 重复上述过程2-3次,之后在60℃电热鼓风干燥箱中烘干;
2. 丝蛋白提纯
2.1 将初步提取的蚕丝加入到10mol/L的LiBr溶液,加热溶解,其中,蚕丝与溴化锂溶液的重量比为1:3;
2.2 溶液装入透析袋,用纯化水透析24h;
2.3 加入冷冻离心机进行离心,重复2-3次,得到溶液A
2.4 溶液A加入60%甘油,得到溶液B;所述溶液A和甘油的重量比为30:1;
3. 丝蛋白羟基磷灰石混合液制备
3.1 溶液A与纳米级羟基磷灰石(12μm)混合,,加热搅拌均匀得到溶液C;所述溶液A和羟基磷灰石的重量比为1:110;
3.2 加入60 %的乙醇溶液得到溶液C;
4. 蛋白分层冻干
4.1 溶液A加到圆柱状模具中1/3, 冻干24h;
4.2 再加入溶液B到模具2/3,冻干24h;
4.3冻干之后浸泡到纯化水中24h,隔2h换一次纯化水,之后冻干24h;
4.4 再加入溶液C,加满模具,冻干24h;
4.5 冻干之后成品浸泡到50%的乙醇溶液中24h;
4.6 纯化水浸泡清洗;
5. 病毒灭活及清洁
5.1 冻干成品加入到过3%过氧化氢溶液,浸泡24h
5.2 之后用纯化水浸泡清洗
5.3 冷冻干燥后,产品经钴-60 辐照灭菌得到最终的医用丝素蛋白胶原。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明技术原理的前提下,还可以做出若干改进和变形,这些改进和变形也应视为本发明的保护范围。
Claims (10)
1.一种医用丝素蛋白胶原制备方法,其特征是,包括以下步骤:
(1)制备溶液A:以生丝为原料,采用碳酸钠溶液在高温条件下进行处理,烘干后得到的蚕丝加入到溴化锂溶液中进行溶解,用水进行透析,离心后得到溶液A;
(2)制备溶液B:向溶液A中加入甘油,得到溶液B;
(3)制备溶液C:将溶液A与羟基磷灰石混合,得到溶液C;
(4)混合:将溶液A加入到模具中三分之一的位置,冻干;然后再加入溶液B到模具中三分之二的位置,冻干;最后加入溶液C至模具满,冻干得到成品。
2.根据权利要求1所述的一种医用丝素蛋白胶原制备方法,其特征是,所述碳酸钠的物质的量浓度为0.3-0.8 mol/L。
3.根据权利要求1或2所述的一种医用丝素蛋白胶原制备方法,其特征是,所述高温条件为90-110℃。
4.根据权利要求1所述的一种医用丝素蛋白胶原制备方法,其特征是,所述溴化锂的物质的量浓度为9-10mol/L。
5.根据权利要求4所述的一种医用丝素蛋白胶原制备方法,其特征是,所述甘油的质量分数为40-60%。
6.根据权利要求1所述的一种医用丝素蛋白胶原制备方法,其特征是,所述乙醇溶液中乙醇的质量分数为40-60%。
7.根据权利要求1所述的一种医用丝素蛋白胶原制备方法,其特征是,羟基磷灰石为纳米级。
8.根据权利要求1所述的一种医用丝素蛋白胶原制备方法,其特征是,冻干得到的成品加入到过质量分数为2-4%的过氧化氢溶液中浸泡,再用水浸泡清洗,最后进行冷冻干燥。
9.根据权利要求8所述的一种医用丝素蛋白胶原制备方法,其特征是,冷冻干燥之后再采用钴-60辐照灭菌。
10.根据权利要求1所述的一种医用丝素蛋白胶原制备方法,其特征是,所述模具为圆柱形。
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Citations (4)
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---|---|---|---|---|
CN101891962A (zh) * | 2010-07-22 | 2010-11-24 | 苏州大学 | 丝素蛋白多孔三维材料的制备方法 |
US20120015003A1 (en) * | 2009-01-23 | 2012-01-19 | Royal College Of Surgeons In Ireland | Layered Scaffold Suitable for Osteochondral Repair |
CN105327401A (zh) * | 2015-11-17 | 2016-02-17 | 上海纳米技术及应用国家工程研究中心有限公司 | 丝素蛋白双层仿骨膜材料的制备方法 |
CN107789668A (zh) * | 2017-11-03 | 2018-03-13 | 华中科技大学同济医学院附属协和医院 | 具有多层结构的仿生胶原蛋白骨修复材料及其制备方法 |
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Publication number | Priority date | Publication date | Assignee | Title |
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US20120015003A1 (en) * | 2009-01-23 | 2012-01-19 | Royal College Of Surgeons In Ireland | Layered Scaffold Suitable for Osteochondral Repair |
CN101891962A (zh) * | 2010-07-22 | 2010-11-24 | 苏州大学 | 丝素蛋白多孔三维材料的制备方法 |
CN105327401A (zh) * | 2015-11-17 | 2016-02-17 | 上海纳米技术及应用国家工程研究中心有限公司 | 丝素蛋白双层仿骨膜材料的制备方法 |
CN107789668A (zh) * | 2017-11-03 | 2018-03-13 | 华中科技大学同济医学院附属协和医院 | 具有多层结构的仿生胶原蛋白骨修复材料及其制备方法 |
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