CN116926146B - 一种细菌纤维素及其制备方法和应用 - Google Patents
一种细菌纤维素及其制备方法和应用 Download PDFInfo
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- CN116926146B CN116926146B CN202310885884.2A CN202310885884A CN116926146B CN 116926146 B CN116926146 B CN 116926146B CN 202310885884 A CN202310885884 A CN 202310885884A CN 116926146 B CN116926146 B CN 116926146B
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- bacterial cellulose
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- glucan
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Abstract
本发明公开一种细菌纤维素及其制备方法和应用，属于生物医用材料。细菌纤维素的制备方法，包括以下步骤：将β‑葡聚糖溶液添加至木醋杆菌培养基中培养得到所述细菌纤维素。本发明还提出一种上述细菌纤维素在制备缓释敷料中的应用。本发明制得的缓释敷料，有利于创面的愈合，愈合率可高达85％以上。
Description
技术领域
本发明涉及生物医用材料技术领域，具体涉及一种细菌纤维素及其制备方法和应用。
背景技术
皮肤是人体中面积最大的一个免疫器官，具有主动的免疫防御、监视和自稳定功能。皮肤能对人体起到很强的保护作用，减少外界对人体造成的物理性伤害，帮助人体减轻酸碱类化学物质的伤害。一旦皮肤遭受意外的创伤、烧伤或受到慢性伤口的侵蚀，皮肤的保护功能随之失效，从而影响了身体机能的恢复，造成不同程度的病症。因此，需要采用敷料对创面进行保护，避免创面受到感染，同时，通过创面的功能特性辅助创面的愈合。
细菌纤维素是由微生物发酵合成的多孔性网状纳米级生物高分子聚合物，因其由细菌合成而命名为细菌纤维素。它由独特的丝状纤维组成，纤维直径在0.01～0.10μm之间，细菌纤维素具有高持水性、透气性好、良好的生物相容性及较好的力学性能，在医用敷料产业具有广阔的应用前景。由于其本身无抗菌效果，也可以通过复合技术对细菌纤维素进行修饰，赋予其良好的抗菌性能，使其在医用敷料方面有更好的应用前景。
作为皮肤损伤后的创面，尤其针对慢性创面和大面积烧伤，皮肤护理的关键三个因素为：皮肤缺损修复、伤口的抗菌及阻止外界污染物侵扰伤口、保持创面良好的湿性环境。常规敷料很难同时满足上述三个性能，尤其是针对慢性伤口的愈合。药物直接添加到敷料中，容易导致药物的直接释放；而药物可以缓慢释放时才能够持续有效的作用于创面，促进创面的自我修复和良好的愈合。如何实现创面的有效愈合是现有技术需要解决的问题。
发明内容
本发明的目的在于克服上述技术不足，提供一种细菌纤维素及其制备方法和应用，解决现有技术中如何实现创面的有效愈合的技术问题。
为达到上述技术目的，本发明的技术方案提供一种细菌纤维素的制备方法，包括以下步骤：将β-葡聚糖溶液添加至木醋杆菌培养基中培养得到所述细菌纤维素。
进一步地，所述β-葡聚糖溶液的质量浓度为0.1～1wt％；β-葡聚糖溶液加入到木醋杆菌培养基中培养12～48h，之后补加培养基后再继续培养4～8d得到所述细菌纤维素。
此外，本发明还提出一种细菌纤维素，由上述制备方法制备得到。
另外，本发明还提出一种上述细菌纤维素在制备缓释敷料中的应用。
进一步地，所述应用包括以下步骤：
S1、将负载FGF的海藻酸钠溶液滴加至油相溶液中，之后添加吐温80水溶液，之后乳化，之后滴加氯化钙溶液，搅拌固化，形成FGF海藻酸钙微球；
S2、将FGF海藻酸钙微球分散于水中，之后浸润至细菌纤维素上，之后冷冻干燥得到负载FGF海藻酸钙微球的细菌纤维素；
S3、将胶原蛋白、抑菌剂和助纺剂溶于乙酸中得到混合液，采用静电纺丝法将混合液喷射在负载FGF海藻酸钙微球的细菌纤维素上，得到缓释敷料。
进一步地，在步骤S1中，所述油相溶液包括司盘80和异辛烷，所述司盘80的质量浓度为1-4wt％。
进一步地，在步骤S3中，所述胶原蛋白、所述抑菌剂和所述助纺剂的质量比为(2～4):(2～3):(3～4)，所述混合液的质量浓度为5～12wt％。
进一步地，在步骤S3中，所述静电纺丝的条件为：纺丝电压10～25Kv，接收距离为10cm～25cm，推进速率0.5mL/h～3mL/h；和/或，所述抑菌剂为壳聚糖、壳聚糖季铵盐和聚胍中的一种或者多种；和/或，所述助纺剂为聚氧化乙烯、聚乙烯醇和聚乙烯吡咯烷酮中的一种或者多种。
进一步地，在步骤S1中，所述乳化的速度为400～1200rpm，所述乳化的时间为10～150min；和/或，所述海藻酸钠溶液的浓度为0.5～2wt％，所述氯化钙溶液的质量浓度为3～12wt％。
进一步地，在步骤S1中，将负载FGF的海藻酸钠溶液滴加至油相溶液中使得油水相的体积比为(1～2):(2～1)。
与现有技术相比，本发明的有益效果包括：本发明制得的缓释敷料，负载药物的微球嵌入到敷料中，可以实现药物的缓释，同时利用敷料自身吸收渗液能力，让敷料与皮肤创面保持良好的湿性环境，有利于创面的愈合，愈合率可高达85％以上。
附图说明
图1是本发明实施例4和对比例2提出的敷料的创面愈合试验的照片。
具体实施方式
本具体实施方式提供了一种细菌纤维素的制备方法，包括以下步骤：将β-葡聚糖溶液添加至木醋杆菌培养基中培养得到所述细菌纤维素；所述β-葡聚糖溶液的质量浓度为0.1～1wt％；β-葡聚糖溶液加入到木醋杆菌培养基中培养12～48h，之后补加培养基后再继续培养4～8d得到所述细菌纤维素。
本具体实施方式还提出一种细菌纤维素，由上述制备方法制备得到。
本具体实施方式还提出一种上述细菌纤维素在制备缓释敷料中的应用，进一步地，包括以下步骤：
S1、将负载FGF的海藻酸钠溶液滴加至油相溶液中，之后添加吐温80水溶液，之后乳化，之后滴加氯化钙溶液，搅拌固化，形成FGF海藻酸钙微球；所述油相溶液包括司盘80和异辛烷，所述司盘80的质量浓度为1-4wt％；所述乳化的速度为400～1200rpm，所述乳化的时间为10～150min；和/或，所述海藻酸钠溶液的浓度为0.5～2wt％，所述氯化钙溶液的质量浓度为3～12wt％；将负载FGF的海藻酸钠溶液滴加至油相溶液中使得油水相的体积比为(1～2):(2～1)；
S2、将FGF海藻酸钙微球分散于水中，之后浸润至细菌纤维素上，之后冷冻干燥得到负载FGF海藻酸钙微球的细菌纤维素；
S3、将胶原蛋白、所述抑菌剂和所述助纺剂溶于乙酸中得到混合液，采用静电纺丝法将混合液喷射在负载FGF海藻酸钙微球的细菌纤维素上，得到缓释敷料；所述胶原蛋白、所述抑菌剂和所述助纺剂的质量比为(2～4):(2～3):(3～4)，所述混合液的质量浓度为5～12wt％；所述静电纺丝的条件为：纺丝电压10～25Kv，接收距离为10cm～25cm，推进速率0.5mL/h～3mL/h；和/或，所述抑菌剂为壳聚糖、壳聚糖季铵盐和聚胍中的一种或者多种；和/或，所述助纺剂为聚氧化乙烯、聚乙烯醇和聚乙烯吡咯烷酮中的一种或者多。
司盘80为表面活性剂和乳化剂，异辛烷为油相，海藻酸钠溶液和氯化钙溶液为水相，以吐温80水溶液调整溶液HLB值，海藻酸钠是先分散到乳化剂中，通过异辛烷混合形成油相；通过本发明提出的乳化法制备的微球粒径可以达到纳米级。
壳聚糖具有抑菌功能，这在创面的初期愈合阶段比较重要；聚氧化乙烯为助纺剂，胶原蛋白和壳聚糖自身比较难以纺丝，需要通过具有较好纺丝性能的助纺剂来助纺。
本具体实施方式提出的缓释敷料，敷料外层为细菌纤维素层，敷料内层为含有胶原蛋白、壳聚糖和聚氧化乙烯的纳米纤维层，所述辅料内层为接触皮肤的面。
需要说明的是，FGF为成纤维细胞生长因子，FGF包括碱性成纤维细胞生长因子(bFGF)或酸性成纤维细胞生长因子(aFGF)。
β-葡聚糖能激活免疫系统中的基础细胞—巨噬细胞，巨噬细胞产生表皮生长因子，从而促进伤口愈合所必需的胶原蛋白产生，同时血管生成因子可促进伤口愈合必需的新血管形成。β-葡聚糖可增加受损皮肤细胞的再生能力，提高角质层的再生速率，具有帮助伤口复原的特性。
本发明中所使用的原料都具有良好的生物相容性，且制备过程中没有引入化学交联剂，不会对损伤的皮肤细胞产生毒副作用。所用的细菌纤维素/β-葡聚糖外层为木醋杆菌原位合成细菌纤维素/β-葡聚糖敷料层，能够有效提高细菌纤维素在保湿、抗炎及抗菌方面的不足；所用的药物为FGF，是一种能促进中胚层和神经外胚层细胞分裂的多肽，具有强烈的血管生成作用。在体外，能刺激细胞增殖、迁移，诱导纤溶酶原激活物及胶原酶活性，促进细胞再生。所用的负载FGF海藻酸钙微球是通过乳化法实现了对药物的包裹，进而达到FGF的缓释。而通过高压静电纺丝技术制备的胶原蛋白/壳聚糖/聚氧化乙烯纳米纤维层具有良好的纤维网状结构，可以吸附渗液，同时，由于外源性胶原蛋白的加入，使得创面皮肤内的基质金属酶不再主动降解真皮层内的胶原蛋白和弹性蛋白，因而能够有效维持基质金属酶的浓度，从而加速创面的止血和愈合。
下述实施例中，所述抑菌剂采用壳聚糖，所述助纺剂采用聚氧化乙烯。在其他实施例中也可以采用其他的抑菌剂和助纺剂。下述实施例中，bFGF混匀到溶解的海藻酸钠溶液中使得其浓度为1μg/ml，在其他实施例中，bFGF混匀到溶解的海藻酸钠溶液中使得其浓度还可以是为1-5μg/ml。
为了使本发明的目的、技术方案及优点更加清楚明白，以下结合附图及实施例，对本发明进行进一步详细说明。应当理解，此处所描述的具体实施例仅仅用以解释本发明，并不用于限定本发明。
实施例1
本实施例提出一种细菌纤维素，由以下步骤制得：
将β-葡聚糖预先溶于纯化水中，配制成质量百分比为0.5％的β-葡聚糖溶液，加入到木醋杆菌培养基中培养24h，β-葡聚糖溶液和培养基的体积比为1:1，期间定期补加培养基使得培养基混合液的总液面高度不变，补加培养基后再继续培养4d，将得到的上层细菌纤维素薄膜取出，置于0.2mol/L的NaOH溶液中处理40min，除去残留的培养基、菌体后，将细菌纤维素薄膜从NaOH溶液中取出，用纯化水洗涤，冷冻干燥，得到细菌纤维素/β-葡聚糖敷料外层。
实施例2
本实施例提出一种细菌纤维素，由以下步骤制得：
将β-葡聚糖预先溶于纯化水中，配制成质量百分比为1％的β-葡聚糖溶液，加入到木醋杆菌培养基中培养48h，β-葡聚糖溶液和培养基的体积比为1:1，期间定期补加培养基使得培养基混合液的总液面高度不变，补加培养基后再继续培养6d，将得到的上层细菌纤维素薄膜取出，置于0.2mol/L的NaOH溶液中处理50min，除去残留的培养基、菌体后，将细菌纤维素薄膜从NaOH溶液中取出，用纯化水洗涤，冷冻干燥，得到细菌纤维素/β-葡聚糖敷料外层。
实施例3
本实施例提出一种细菌纤维素，由以下步骤制得：
将β-葡聚糖预先溶于纯化水中，配制成质量百分比为1.5％的β-葡聚糖溶液，加入到木醋杆菌培养基中培养48h，β-葡聚糖溶液和培养基的体积比为1:1，期间定期补加培养基使得培养基混合液的总液面高度不变，补加培养基后再继续培养4d，将得到的上层细菌纤维素薄膜取出，置于0.2mol/L的NaOH溶液中处理60min，除去残留的培养基、菌体后，将细菌纤维素薄膜从NaOH溶液中取出，冷冻干燥，得到细菌纤维素/β-葡聚糖敷料外层。
实施例4
本实施例提出一种缓释辅料，由以下步骤制得：
S1、将海藻酸钠预先溶于纯化水中，配制成质量百分比为1wt％的海藻酸钠溶液，再将bFGF混匀到溶解的海藻酸钠溶液中使得其浓度为1μg/ml；将司盘80和异辛烷搅拌混匀后，形成质量浓度为4wt％的乳化剂，置于机械搅拌器中搅拌，搅拌速度为800rpm。搅拌均匀后，用恒流泵将负载bFGF的海藻酸钠溶液缓慢滴加到油相中，使得油水相体积比为1:1，恒流泵速度为1mL/min，高度为30cm，搅拌20min。加入微量的吐温80水溶液调节HLB值至3，搅拌乳化1h，用恒流泵将10wt％的氯化钙溶液缓慢滴入海藻酸钠乳液中，恒流泵速度为1mL/min，高度为30cm，搅拌固化2h，形成bFGF海藻酸钙微球；
S2、将bFGF海藻酸钙微球过滤，加纯化水分散微球后，浸润到实施例1冻干的细菌纤维素/β-葡聚糖敷料上，冷冻干燥，得到负载bFGF海藻酸钙微球的细菌纤维素/β-葡聚糖层；
S3、将胶原蛋白、壳聚糖和聚氧化乙烯按照质量比为1:1:2溶解于0.5％的乙酸中得到混合液，使得混合液的质量浓度为10wt％，采用静电纺丝法将胶原蛋白/壳聚糖/聚氧化乙烯混合液喷射在负载bFGF海藻酸钙微球的细菌纤维素/β-葡聚糖层上，纺丝电压15Kv，接收距离为20cm，推进速率1mL/h得到细菌纤维素复合纳米纤维缓释功能敷料。
实施例5
本实施例提出一种缓释辅料，由以下步骤制得：
S1、将海藻酸钠预先溶于纯化水中，配制成质量百分比为2wt％的海藻酸钠溶液，再将aFGF混匀到溶解的海藻酸钠溶液中使得其浓度为1μg/ml；将司盘80和异辛烷搅拌混匀后，形成质量浓度为3wt％的乳化剂，置于机械搅拌器中搅拌，搅拌速度为600rpm。搅拌均匀后，用恒流泵将负载aFGF的海藻酸钠溶液缓慢滴加到油相中，使得油水相体积比为1:1，恒流泵速度为1mL/min，高度为30cm，搅拌20min。加入微量的吐温80水溶液调节HLB值至2，搅拌乳化1h，用恒流泵将8wt％的氯化钙溶液缓慢滴入海藻酸钠乳液中，恒流泵速度为1mL/min，高度为30cm，搅拌固化1h，形成aFGF海藻酸钙微球。
S2、将aFGF海藻酸钙微球过滤，加纯化水分散微球后，浸润到实施例2冻干的细菌纤维素/β-葡聚糖敷料上，冷冻干燥，得到负载aFGF海藻酸钙微球的细菌纤维素/β-葡聚糖层；
S3、将胶原蛋白、壳聚糖和聚氧化乙烯按照质量比为2:1:2溶解于0.5％的乙酸中得到混合液，使得混合液的质量浓度为12wt％，采用静电纺丝法将胶原蛋白/壳聚糖/聚氧化乙烯混合液喷射在负载aFGF海藻酸钙微球的细菌纤维素/β-葡聚糖层上，纺丝电压15Kv，接收距离为20cm，推进速率1mL/h得到细菌纤维素复合纳米纤维缓释敷料。
实施例6
本实施例提出一种缓释辅料，由以下步骤制得：
S1、将海藻酸钠预先溶于纯化水中，配制成质量百分比为1.5wt％的海藻酸钠溶液，再将bFGF混匀到溶解的海藻酸钠溶液中使得其浓度为1μg/ml；将司盘80和异辛烷搅拌混匀后，形成质量浓度为2wt％的乳化剂，置于机械搅拌器中搅拌，搅拌速度为1000rpm。搅拌均匀后，用恒流泵将负载bFGF的海藻酸钠溶液缓慢滴加到油相中，使得油水相体积比为1:1，恒流泵速度为1mL/min，高度为30cm，搅拌20min。加入微量的吐温80水溶液调节HLB值至4，搅拌乳化1h，用恒流泵将10wt％的氯化钙溶液缓慢滴入海藻酸钠乳液中，恒流泵速度为1mL/min，高度为30cm，搅拌固化2h，形成bFGF海藻酸钙微球。
S2、将bFGF海藻酸钙微球过滤，加纯化水分散微球后，浸润到实施例3冻干的细菌纤维素/β-葡聚糖敷料上，冷冻干燥，得到负载bFGF海藻酸钙微球的细菌纤维素/β-葡聚糖层；
S3、将胶原蛋白、壳聚糖和聚氧化乙烯按照质量比为1:2:2溶解于0.5％的乙酸中得到混合液，使得混合液的质量浓度为10wt％，采用静电纺丝法将胶原蛋白/壳聚糖/聚氧化乙烯混合液喷射在负载bFGF海藻酸钙微球的细菌纤维素/β-葡聚糖层上，纺丝电压20Kv，接收距离为20cm，推进速率1.5mL/h得到细菌纤维素复合纳米纤维缓释敷料。
对比例1
本对比例与实施例1的区别在于，没有添加β-葡聚糖，具体地，
本对比例提出一种细菌纤维素，由以下步骤制得：
在木醋杆菌培养基中培养24h，期间定期补加培养基，补加培养基后再继续培养4d，将得到的上层细菌纤维素薄膜取出，置于0.2mol/L的NaOH溶液中处理，除去残留的培养基、菌体后，将细菌纤维素薄膜从NaOH溶液中取出，于常温下处理40min，除去残留的培养基、菌体后，用纯化水洗涤，冷冻干燥，得到细菌纤维素。
对比例2
本对比例提出的缓释辅料中，与实施例4的区别仅在于bFGF海藻酸钙微球不同，本对比例中的bFGF海藻酸钙微球通过高压静电液体法制得，具体地，保持针尖与氯化钙液面的距离为15cm，推进速率1mL/h，电场电压为15KV。
细菌纤维素外层敷料吸水率测试
将实施例1中原位法制备的细菌纤维素/β-葡聚糖敷料外层和对比例1的细菌纤维素敷料进行吸水率的测试。分别取5片样品，裁剪成正方形(5cm×5cm)，将样品在-20℃冰箱预冻后，放置于冷冻干燥箱中冷冻干燥48h。取出样品，称量重量，再分别在纯化水浸泡中24h后，用滤纸吸干敷料表面水分后继续称量其重量。计算样品的吸水率，见表1。
表1不同方法制备的细菌纤维素吸水率测试结果
通过吸水率的测试结果可知，在原位法制备细菌纤维素过程中添加β-葡聚糖可以将细菌纤维素的吸水率提高10.18％，这是因为β-葡聚糖有较高的高分子量，同时分子链中拥有大量的亲水基团。吸水率的提高有利于吸收创面的渗液，同时β-葡聚糖还可以起到抗炎、辅助修复的作用。
在微球粒径、包封率和药物释放方面进行了试验对比。采用动态光散射仪和体式显微镜分别对实施例4和对比例2制得的bFGF海藻酸钙微球进行粒径的测量；分别准确称取两种不同方法制备、经冷冻干燥后的微球各10mg，加入20mLPBS缓冲溶液(pH＝7.4)中，采用酶联免疫吸附试验(ELISA)对测定bFGF的含量；分别准确称取两种不同方法制备、经冷冻干燥后的微球各10mg，模拟体内环境，加入20mLPBS缓冲溶液(pH＝7.4)中，放置于37℃水浴摇床100rpm振荡。分别在6h、12h、24h、36h、48h取出，10000rpm离心3min。吸取上清液20mL，重新加入20mLPBS缓冲液，继续放置于37℃水浴摇床振荡。用标准曲线法测定上清液中bFGF含量。通过计算结果发现，见表2和表3。
表2不同制备法制得载bFGF海藻酸钙微球的粒径和包封率
| 对比项目/制备方法 | 实施例4 | 对比例2 |
| 微球粒径 | 395.2±0.8nm | 196.4±5.2um |
| 包封率 | 26.86％ | 14.65％ |
表3不同制备法制得载bFGF海藻酸钙微球的药物释放速率
| 制备方法/释放时间 | 6h | 12h | 24h | 36h | 48h |
| 实施例4 | 30.52％ | 46.58％ | 60.82％ | 72.25％ | 82.07％ |
| 对比例2 | 58.21％ | 65.42％ | 78.05％ | 92.36％ | 96.59％ |
实施例4乳化法制备的负载bFGF海藻酸钙微球为纳米级且包封率更高，对比例2高压静电液体法制备的微球为微米级、包封率较低；实施例4的bFGF海藻酸钙微球粒径更小，生长因子的缓释效果越好。
创面愈合试验
实验动物为240～260g的SD大鼠，实验前先将大鼠在实验环境下饲养一周。采用10mg/mL的戊巴比妥钠注射麻醉大鼠，待其生理反应有明显降低后，将大鼠四肢固定，然后用电动剃须刀除去其背部的大部分毛，再用8％的硫化钠进行脱毛处理。带脱毛处理完后，使用90℃的水处理过的铜块烫伤大鼠背部，烫伤面积的直径为1.5cm。采用手术剪在大鼠背部剪开四个伤口，生理盐水清洗创面，脱脂棉吸干残留液。将实施例4的细菌纤维素复合纳米纤维缓释敷料和对比例2的缓释辅料分别敷在创面上，采用硅胶膜将材料固定，缝线，用纱布包扎好。实验大鼠在25℃环境中分笼饲养，让其自由进水、进食，观察大鼠的生长情况，分别在7、14天后取出实验样品和对照样品，观察伤口的愈合情况。见附图1。结果显示，实施例4的细菌纤维素复合纳米纤维缓释功能敷料覆盖的创面已经基本愈合完全，缺损皮肤的愈合率为85.12％，皮下毛发也接近生长完整；而对照组细菌纤维素覆盖的创面仍然有较多真皮裸露，缺损皮肤的愈合率为57.33％，愈合效果欠佳。大鼠肤缺损修复实验表明实施例4制备的细菌纤维素复合纳米纤维缓释功能敷料生物相容性好，可以有效降低伤口的感染，并可以局部止血，同时能够加速创面的愈合。
其他有益效果：
1)本发明中所使用的原料都具有良好的生物相容性，且制备过程中没有引入化学交联剂，不会对损伤的皮肤细胞产生毒副作用，可以有效的保护好创面，为创面提供良好的愈合环境。
2)本发明中针对细菌纤维素敷料单一组分在抗菌、保湿和促修复方面的不足，引入β-葡聚糖，原位形成细菌纤维素/β-葡聚糖敷料层，能够增加细菌纤维素敷料的保湿性能，同时能一定程度上增加敷料的抗炎、抗菌和促进创面愈合的功能。
3)本发明中通过乳化法制备的负载FGF海藻酸钙微球，嵌入到细菌纤维素/β-葡聚糖层后能够实现药物的缓释作用，持续有效的促进成损伤皮肤中纤维细胞的再生。
4)本发明中与皮肤贴合的胶原蛋白/壳聚糖/聚氧化乙烯纳米纤维层，纳米纤维层中的胶原蛋白主要起到止血、抑制创面内胶原蛋白降解和促进伤口愈合的作用，壳聚糖主要起到止血、抗菌作用。复合的纳米纤维层为创面的愈合提供良好的基础，有利于创面的修复。
以上所述本发明的具体实施方式，并不构成对本发明保护范围的限定。任何根据本发明的技术构思所做出的各种其他相应的改变与变形，均应包含在本发明权利要求的保护范围内。
Claims (5)
1.一种细菌纤维素在制备缓释敷料中的应用，其特征在于，包括以下步骤：
S1、将负载FGF的海藻酸钠溶液滴加至油相溶液中，之后添加吐温80水溶液，之后乳化，之后滴加氯化钙溶液，搅拌固化，形成FGF海藻酸钙微球；所述海藻酸钠溶液的浓度为0.5～2wt％，所述氯化钙溶液的质量浓度为3～12wt％；
S2、将FGF海藻酸钙微球分散于水中，之后浸润至细菌纤维素上，之后冷冻干燥得到负载FGF海藻酸钙微球的细菌纤维素；
S3、将胶原蛋白、抑菌剂和助纺剂溶于乙酸中得到混合液，采用静电纺丝法将混合液喷射在负载FGF海藻酸钙微球的细菌纤维素上，得到缓释敷料；所述胶原蛋白、所述抑菌剂和所述助纺剂的质量比为(2～4):(2～3):(3～4)，所述混合液的质量浓度为5～12wt％；
所述细菌纤维素由以下步骤制得：将β-葡聚糖溶液添加至木醋杆菌培养基中培养得到所述细菌纤维素；所述β-葡聚糖溶液的质量浓度为0.1～1wt％；β-葡聚糖溶液加入到木醋杆菌培养基中培养12～48h，之后补加培养基后再继续培养4～8d得到所述细菌纤维素。
2.根据权利要求1所述的应用，其特征在于，在步骤S1中，所述油相溶液包括司盘80和异辛烷，所述司盘80的质量浓度为1-4wt％。
3.根据权利要求1所述的应用，其特征在于，在步骤S3中，所述静电纺丝的条件为：纺丝电压10～25Kv，接收距离为10cm～25cm，推进速率0.5mL/h～3mL/h；和/或，所述抑菌剂为壳聚糖、壳聚糖季铵盐和聚胍中的一种或者多种；和/或，所述助纺剂为聚氧化乙烯、聚乙烯醇和聚乙烯吡咯烷酮中的一种或者多种。
4.根据权利要求1所述的应用，其特征在于，在步骤S1中，所述乳化的速度为400～1200rpm，所述乳化的时间为10～150min。
5.根据权利要求1所述的应用，其特征在于，在步骤S1中，将负载FGF的海藻酸钠溶液滴加至油相溶液中使得油水相的体积比为(1～2):(2～1)。
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| CN105524956A (zh) * | 2016-01-13 | 2016-04-27 | 山东贝诺医药生物科技有限公司 | 一种医用细菌纤维素凝胶 |
| CN109350762A (zh) * | 2018-11-23 | 2019-02-19 | 北京诺康达医药科技股份有限公司 | 一种应用于慢性创面的医用敷料及其制备方法 |
| CN110193090A (zh) * | 2019-05-31 | 2019-09-03 | 盐城工学院 | 一种具有抗菌抑菌功能的细菌纤维素敷料的制备方法 |
| CN112522345A (zh) * | 2020-12-29 | 2021-03-19 | 山东纳美德生物科技有限公司 | 一种快速发酵、工业化生产细菌纤维素的方法 |
| CN116217969A (zh) * | 2022-09-07 | 2023-06-06 | 四川大学 | 细菌纤维素复合水凝胶及其制备方法 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105524956A (zh) * | 2016-01-13 | 2016-04-27 | 山东贝诺医药生物科技有限公司 | 一种医用细菌纤维素凝胶 |
| CN109350762A (zh) * | 2018-11-23 | 2019-02-19 | 北京诺康达医药科技股份有限公司 | 一种应用于慢性创面的医用敷料及其制备方法 |
| CN110193090A (zh) * | 2019-05-31 | 2019-09-03 | 盐城工学院 | 一种具有抗菌抑菌功能的细菌纤维素敷料的制备方法 |
| CN112522345A (zh) * | 2020-12-29 | 2021-03-19 | 山东纳美德生物科技有限公司 | 一种快速发酵、工业化生产细菌纤维素的方法 |
| CN116217969A (zh) * | 2022-09-07 | 2023-06-06 | 四川大学 | 细菌纤维素复合水凝胶及其制备方法 |
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