CN116196329A - 一种透明质酸-脱细胞基质制剂及其制备方法与应用 - Google Patents
一种透明质酸-脱细胞基质制剂及其制备方法与应用 Download PDFInfo
- Publication number
- CN116196329A CN116196329A CN202310213104.XA CN202310213104A CN116196329A CN 116196329 A CN116196329 A CN 116196329A CN 202310213104 A CN202310213104 A CN 202310213104A CN 116196329 A CN116196329 A CN 116196329A
- Authority
- CN
- China
- Prior art keywords
- hyaluronic acid
- preparation
- acellular matrix
- targeting
- acellular
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
- A61K31/728—Hyaluronic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
- A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/04—Drugs for skeletal disorders for non-specific disorders of the connective tissue
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Rheumatology (AREA)
- Physical Education & Sports Medicine (AREA)
- Dermatology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Neurosurgery (AREA)
- Pain & Pain Management (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
Abstract
本发明公开了一种透明质酸‑脱细胞基质制剂及其制备方法与应用,属于生物医用材料领域。所述制剂包括透明质酸、软骨脱细胞基质和靶向高分子,所述透明质酸可以保持关节部位的润滑状态,脱细胞基质以消化液形式与透明质酸结合提供了二型胶原、糖胺聚糖等天然软骨组织成分,可以为关节软骨修复提供较好的微环境;所述靶向高分子能够在透明质酸之间、脱细胞基质之间,以及透明质酸与脱细胞基质之间相互紧密结合,在不影响注射性能的情况下延缓透明质酸和脱细胞基质的降解速率;此外当制剂注射入关节腔内后,随着关节的运动,所述靶向高分子会动态暴露,从而与关节软骨组织或/和其中的透明质酸成分动态结合,使得制剂在关节软骨部位富集,在关节软骨表面形成水凝胶包覆层,起到更好的保护和润滑作用,该制剂在骨关节炎治疗中具有良好的应用前景。
Description
技术领域
本发明属于生物医用材料领域,特别涉及到一种透明质酸-脱细胞基质制剂及其制备方法与应用。
背景技术
骨关节炎是一种以疼痛、软骨丧失和关节炎症为特征的慢性疾病,其发病率随着年龄的增长而增加。在骨关节炎中,软骨基质的组成和结构发生了显著变化。最初,出现表面纤颤,随着病理过程的继续,与软骨碎片剥落相关的深裂缝发展,最终导致底层钙化软骨和骨骼的分层和暴露。
透明质酸(HA)是许多软连接组织细胞外基质(ECM)中的一种非硫酸糖胺聚糖(GAG),高分子量HA(500-5000kDa),具有吸湿性和粘弹性,调节组织水化和渗透平衡。细胞外基质中的高分子量HA具有抗血管生成和抑制内皮细胞生长的作用。现阶段,高分子量HA仍多直接用于注射液缓解关节疼痛,然而其仍然会因降解较快,只能因其润滑功能缓解骨关节炎,不能阻止骨关节炎的进程。
发明内容
针对现有技术中存在的上述问题,本发明提供了一种透明质酸-脱细胞基质制剂及其制备方法与应用,将透明质酸与软骨脱细胞基质通过靶向高分子交联形成复合制剂,能够有效增强透明质酸的流变性能和降解性能,同时软骨脱细胞基质中天然成分和生物活性因子的存在为软骨细胞发挥功能和产生新基质提供了适宜的微环境,该复合制剂是有效缓解骨关节炎的潜在手段。
本发明通过以下技术方案来实现:
一种透明质酸-脱细胞基质制剂,包括透明质酸、软骨脱细胞基质和靶向高分子,所述靶向高分子为两端修饰多肽靶向因子的聚乙二醇。所述透明质酸可以保持关节部位的润滑状态,脱细胞基质在消化后以液体形式与透明质酸结合,可以为关节软骨修复提供较好的微环境;所述靶向高分子能够在透明质酸之间、脱细胞基质之间,以及透明质酸与脱细胞基质之间相互紧密结合,从而增强透明质酸的流变性能,延缓透明质酸和脱细胞基质的降解速率;此外当制剂注射入关节腔内后,随着关节的运动,所述靶向高分子会动态暴露,从而与关节软骨组织或/和其中的透明质酸成分动态结合,使得制剂在关节软骨部位富集,在关节软骨表面形成水凝胶包覆层,起到更好的保护和润滑作用。
作为可选方式,在上述透明质酸-脱细胞基质制剂中,所述软骨脱细胞基质在消化后以液体形式与所述透明质酸通过所述靶向高分子相互结合。
作为可选方式,在上述透明质酸-脱细胞基质制剂中,所述多肽靶向因子为透明质酸靶向肽(HABP,如RRDDGAHWQFNALTVR,STMMSRSHKTRSHHV或GAHWQFNALTVR等)和/或软骨基质靶向肽(CartBP,如WYRGRL,DPHFHL或RVMLVR等),所述靶向高分子为HABP-PEG-HABP或CartBP-PEG-CartBP或HABP-PEG-CartBP。在所述制剂中多肽靶向因子中的透明质酸靶向肽与透明质酸相结合,所述软骨基质靶向肽与脱细胞基质相结合,从而实现制剂成分之间的交联稳定。
作为可选方式,在上述透明质酸-脱细胞基质制剂中,所述多肽靶向因子在制剂中的终浓度为2-30mg/mL,优选为5-20mg/mL,进一步优选为10mg/mL或20mg/mL。
作为可选方式,在上述透明质酸-脱细胞基质制剂中,所述透明质酸的分子量范围在500-5000kDa,优选为1000-2000kDa。
作为可选方式,在上述透明质酸-脱细胞基质制剂中,所述软骨脱细胞基质为动物关节软骨脱细胞基质,所述动物包括但不限于猪、牛、羊、兔、大鼠。
作为可选方式,在上述透明质酸-脱细胞基质制剂中,所述透明质酸-脱细胞基质制剂为可注射制剂。
本发明还提供了一种所述透明质酸-脱细胞基质的制备方法,其特征在于,包括以下步骤:
(1)制备脱细胞基质;
(2)制备靶向高分子;
(3)制备透明质酸-脱细胞基质制剂。
作为可选方式,在上述制备方法中,所述步骤(1)具体为:将软管组织剪成碎片,置于PBS溶液中超声,真空冷冻干燥后,加入研磨珠研磨,将研磨得到的颗粒依次在低渗溶液和高渗溶液中清洗震荡,然后在进行冻融循环,转移至CHAPS溶液中震荡清洗,然后加入到DNA酶溶液中震荡消化,用蒸馏水清洗后离心,弃上清,将沉淀冻干后加入含胃蛋白酶的醋酸溶液中,得到脱细胞基质消化液。
作为可选方式,在上述制备方法中,所述步骤(2)具体为:将两端修饰马来酰胺基团的聚乙二醇分子溶于PBS溶液中获得溶液A,将含巯基的多肽靶向因子溶于TCEP溶液中获得溶液B,将溶液A和溶液B以物质的量比1:1进行共混后,震荡反应3-5h后转移至透析袋,用PBS透析24h后冷冻干燥保存。
作为可选方式,在上述制备方法中,所述步骤(3)具体为:将透明质酸溶于PBS中,加入步骤(1)中制备的脱细胞基质消化液,搅拌均匀得到透明质酸/脱细胞基质混合液;将步骤(2)中制备的靶向高分子溶于PBS后再滴加到上述透明质酸/脱细胞基质混合液中混匀即到所述透明质酸-脱细胞基质制剂。
本发明还提供了一种上述透明质酸-脱细胞基质制剂的应用,其特征在于,将其用于制备骨关节炎治疗药物或材料。
本说明书中公开的所有特征,或公开的所有方法或过程中的步骤,除了互相排斥的特征和/或步骤以外,均可以以任何方式组合。
本发明的有益效果:
(1)本发明所述制剂中所述透明质酸可以保持关节部位的润滑状态,脱细胞基质以消化液形式与透明质酸结合,提供了二型胶原、糖胺聚糖等天然软骨组织成分,可以为关节软骨修复提供较好的微环境。
(2)本发明所述制剂中所述靶向高分子能够在透明质酸之间、脱细胞基质之间,以及透明质酸与脱细胞基质之间相互紧密结合,在不影响注射性能的情况下延缓透明质酸和脱细胞基质的降解速率。
(3)当本发明所述制剂注射入关节腔内后,随着关节的运动,所述靶向高分子会动态暴露,从而与关节软骨组织或/和其中的透明质酸成分动态结合,使得制剂在关节软骨部位富集,在关节软骨表面形成水凝胶包覆层,起到更好的保护和润滑作用,在骨关节炎治疗中具有良好的应用前景。
附图说明:
图1为本发明所述制剂的制备示意图。
图2为注射后制剂动态变化示意图。
图3为复合不同量高分子的制剂倒置示意图(HD未额外添加多肽高分子,HD-5中多肽含量5mg/L,HD-10多肽含量10mg/L)。
图4为本发明所述制剂的SEM观察图(HD未额外添加多肽高分子,HD-5中多肽含量5mg/L,HD-10多肽含量10mg/L)
图5为本发明所述制剂可注射示意图。
图6为本发明所述制剂的挤推力测试结果。(HD未额外添加多肽高分子,HD-5中多肽含量5mg/L,HD-10多肽含量10mg/L)
图7为本发明实施例6中所述样品在两种不同溶液中的降解曲线。
图8为本发明实施例7中所述样品在关节部位留存效果的照片。
具体实施方式:
为了使本发明的目的、技术方案及优点更加清楚明白,下面结合具体实施方式对本发明作进一步的详细描述,但不应将此理解为本发明上述主题的范围仅限于下述实施例。
实施例1
(1)制备脱细胞基质
首先,从新鲜的猪关节软骨上刮下透明软骨组织,将其剪成小片,置于PBS溶液中超声5min,真空冷冻干燥后置于EP管中,加入研磨珠,在低温研磨机中研磨5~10min。其中,研磨温度为-20℃,频率为60Hz,机器转速为3000r/min。将预研磨得到的颗粒置于2mLEP管中,依次在低渗溶液和高渗溶液中清洗震荡3min,重复3次,然后再进行5次冻融循环(液氮和37℃水浴)。再将颗粒移入到1%CHAPS溶液中清洗3次,在800r/min的转速下震荡6h;然后加入到100U/mL的DNA酶溶液中,在37℃恒温摇床中震荡9h,每3h换一次新鲜DNA酶溶液。最后用蒸馏水多次清洗基质,去除所有残留的洗涤剂。12000rpm离心收集脱细胞基质,冻干后存储在-20℃备用。将脱细胞基质加入含胃蛋白酶(胃蛋白酶:脱细胞基质=1:10)的0.1M醋酸溶液,在室温条件下消化48h,得到dECM溶液。
(2)制备靶向高分子
将两端修饰的马来酰胺基团的聚乙二醇(MW:2000)分子溶于PBS溶液中,将含巯基的多肽靶向因子(软骨基质靶向肽及透明质酸靶向肽)溶于TCEP溶液中用以还原二硫键。将两溶液以物质的量比1:1进行共混后,震荡反应约3-5h后转移至透析袋(MW:2000-3000),用PBS透析24h后冷冻干燥保存,得到HABP-PEG-HABP、HABP-PEG-CartBP及CartBP-PEG-CartBP。
(3)制剂的制备
将高分子量的透明质酸溶于PBS中,加入脱细胞基质消化液,以质量比1:1搅拌均匀,制剂的有效成分均为。将步骤(2)得到的靶向高分子溶于PBS后再滴加至透明质酸/脱细胞基质消化液,分别制备成靶向高分子终浓度为2mg/mL(HD-2)、5mg/mL(HD-5)、10mg/mL(HD-10)和20mg/mL(HD-20)的制剂。混合均匀后即可注入关节处成为新型透明质酸制剂。
实施例2倒置试验
取实施例1制备透明质酸-脱细胞基质制剂置于Ep管中,倒置隔一段时间观察制剂的流动性。
结果如图3所示:通过倒置实验可以观察到添加了多肽高分子的制剂随着添加量增加,材料的交联度增加,流动性变差,可以在注射后实现较好的留存效果。
实施例3微观形貌观察
取实施例1制备透明质酸-脱细胞基质制剂,分别注入模具后冻干处理在SEM下观察微观形态变化。
结果如图4所示:随着多肽的添加,制剂逐渐出现交联的微观形貌,当添加量为10mg/mL时,交联明显,整体出现了多孔结构的效果。
实施例4可注射性能测试
取实施例1制备的透明质酸-脱细胞基质制剂,用1mL的注射器(针头规格:26G,套管规格5mm*75mm)分别吸取不同组别的制剂,装上针头后进行注射。
结果如图5所示:各组均可以实现注射,添加了多肽的制剂仍可以保留可注射性能。
实施例5挤推力测试
取实施例1制备的透明质酸-脱细胞基质制剂,用1mL的注射器(针头规格:26G,套管规格5mm*75mm)分别吸取不同组别0.8mL左右的制剂,由日本万能试验机测定不同组制剂的注射力。
结果如图6所示:添加了多肽的制剂仍可以保留可注射性能,当多肽的添加量提高时,注射力略有增加。
实施例6降解实验
取实施例1制备的透明质酸-脱细胞基质制剂(多肽添加量分别为0,10mg/mL和20mg/mL)以及10mg/mL的HA溶液。将4种不同制剂分别浸于PBS和4U/mL的透明质酸酶溶液中,置于37℃,90rpm的摇床中,在不同时间点离心观测制剂体积变化。
结果如图7所示:加入多肽的制剂相比于未加多肽及HA溶液,在PBS和透明质酸酶中都有更好的抗降解性能。
实施例7关节部位留存效果
分别取实施例1制备的0.2mL的透明质酸-脱细胞基质制剂(多肽添加量分别为0,10mg/mL和20mg/mL)以及10mg/mL的HA溶液。为方便观察,首先用阿利新蓝溶液分别将4种制剂染色,随后将其注射至表面制造了一定的缺损(8mm*3mm*0.5mm)的兔关节滑车部位,注射后静置3min后,置于PBS溶液在500rpm转速下震荡5min后观察不同材料的留存效果。
结果如图8所示:HA溶液在震荡过程中极易脱离关节表面,加入脱细胞基质后留存效果有所加强,但也随震荡损失较多。加入多肽后,制剂在关节软骨表面留存效果大幅度提高,随震荡铺展覆盖在关节软骨表面,有较好的留存效果。
最后说明的是,以上实施例仅用以说明本发明的技术方案而非限制,尽管参照较佳实施例对本发明进行了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的宗旨和范围,其均应涵盖在本发明的权利要求范围当中。
Claims (10)
1.一种透明质酸-脱细胞基质制剂,其特征在于,包括透明质酸、软骨脱细胞基质和靶向高分子,所述靶向高分子为两端修饰多肽靶向因子的聚乙二醇。
2.根据权利要求1所述的透明质酸-脱细胞基质制剂,其特征在于,所述软骨脱细胞基质在消化后以液体形式与所述透明质酸通过所述靶向高分子相互结合。
3.根据权利要求1所述的透明质酸-脱细胞基质制剂,其特征在于,所述多肽靶向因子为软骨基质靶向肽和/或透明质酸靶向肽,所述靶向高分子为HABP-PEG-HABP或CartBP-PEG-CartBP或HABP-PEG-CartBP。
4.根据权利要求1所述的透明质酸-脱细胞基质制剂,其特征在于,所述透明质酸的分子量范围在500-5000kDa,优选为1000-2000kDa。
5.根据权利要求1所述的透明质酸-脱细胞基质制剂,其特征在于,所述软骨脱细胞基质为动物关节软骨脱细胞基质。
6.根据权利要求1所述的透明质酸-脱细胞基质制剂,其特征在于,所述透明质酸-脱细胞基质制剂为可注射制剂。
7.一种如权利要求1所述的透明质酸-脱细胞基质的制备方法,其特征在于,包括以下步骤:
(1)制备脱细胞基质;
(2)制备靶向高分子;
(3)制备透明质酸-脱细胞基质制剂。
8.根据权利要求7所述的制备方法,其特征在于,所述步骤(2)具体为:将两端修饰马来酰胺基团的聚乙二醇分子溶于PBS溶液中获得溶液A,将含巯基的多肽靶向因子溶于TCEP溶液中获得溶液B,将溶液A和溶液B以物质的量比1:1进行共混后,震荡反应3-5h后转移至透析袋,用PBS透析24h后冷冻干燥保存。
9.根据权利要求7所述的制备方法,其特征在于,所述步骤(3)具体为:将透明质酸溶于PBS中,加入步骤(1)中制备的脱细胞基质消化液,搅拌均匀得到透明质酸/脱细胞基质混合液;将步骤(2)中制备的靶向高分子溶于PBS后再滴加到上述透明质酸/脱细胞基质混合液中混匀即到所述透明质酸-脱细胞基质制剂。
10.一种如权利要求1所述的透明质酸-脱细胞基质制剂的应用,其特征在于,将其用于制备骨关节炎治疗药物或材料。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2022116491169 | 2022-12-21 | ||
CN202211649116 | 2022-12-21 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN116196329A true CN116196329A (zh) | 2023-06-02 |
Family
ID=86510986
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310213104.XA Pending CN116196329A (zh) | 2022-12-21 | 2023-03-06 | 一种透明质酸-脱细胞基质制剂及其制备方法与应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN116196329A (zh) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20140369975A1 (en) * | 2012-01-19 | 2014-12-18 | The Johns Hopkins University | Biomaterials comprising hyaluronic acid binding peptides and bifunctional biopolymer molecules for hyaluronic acid retention and tissue engineering applications |
US20160158270A1 (en) * | 2013-07-19 | 2016-06-09 | The Johns Hopkins University | Biomaterials comprising hyaluronic acid binding peptides and extracellular matrix binding peptides for hyaluronic acid retention and tissue engineering applications |
CN111701073A (zh) * | 2020-06-01 | 2020-09-25 | 天津大学 | 一种基于胶原蛋白模拟肽改性透明质酸的关节注射制剂及其制备方法和应用 |
-
2023
- 2023-03-06 CN CN202310213104.XA patent/CN116196329A/zh active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20140369975A1 (en) * | 2012-01-19 | 2014-12-18 | The Johns Hopkins University | Biomaterials comprising hyaluronic acid binding peptides and bifunctional biopolymer molecules for hyaluronic acid retention and tissue engineering applications |
US20160158270A1 (en) * | 2013-07-19 | 2016-06-09 | The Johns Hopkins University | Biomaterials comprising hyaluronic acid binding peptides and extracellular matrix binding peptides for hyaluronic acid retention and tissue engineering applications |
CN111701073A (zh) * | 2020-06-01 | 2020-09-25 | 天津大学 | 一种基于胶原蛋白模拟肽改性透明质酸的关节注射制剂及其制备方法和应用 |
Non-Patent Citations (4)
Title |
---|
HEATHER J. FAUST等: "A hyaluronic acid binding peptide-polymer system for treating osteoarthritis", 《BIOMATERIALS》, pages 93 - 101 * |
LEENA-STIINA KONTTURI等: "An injectable, in situ forming type Ⅱ collagen/hyaluronic acid hydrogel vehicle for chondrocyte delivery in cartilage tissue engineering", 《DRUG DELIV. AND TRANSL. RES.》, pages 1 - 10 * |
张亚鑫: "软骨脱细胞基质治疗兔膝关节骨性关节炎的研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》, pages 066 - 192 * |
蒋婷等: "Ⅱ型胶原-透明质酸-软骨脱细胞基质制备组织工程软骨支架的实验研究", 《西部医学》, vol. 25, no. 8, pages 1132 - 1135 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Zhang et al. | Injectable Mussel‐Inspired highly adhesive hydrogel with exosomes for endogenous cell recruitment and cartilage defect regeneration | |
Gan et al. | Mussel-inspired dopamine oligomer intercalated tough and resilient gelatin methacryloyl (GelMA) hydrogels for cartilage regeneration | |
Kisiel et al. | Improving the osteogenic potential of BMP-2 with hyaluronic acid hydrogel modified with integrin-specific fibronectin fragment | |
US20210038767A1 (en) | Adipose tissue products and methods of production | |
Angulo et al. | Characterization of gelatin/chitosan scaffold blended with aloe vera and snail mucus for biomedical purpose | |
US9005606B2 (en) | Silk fibroin hydrogels and uses thereof | |
US10857265B2 (en) | Injectable composition for in-situ repair and regeneration of an injured ligament or tendon and methods of use | |
CN106075584B (zh) | 可注射性软骨脱细胞外基质混合脱钙骨基质水凝胶及其制备方法 | |
Tang et al. | Stable antibacterial polysaccharide-based hydrogels as tissue adhesives for wound healing | |
CN107441556B (zh) | 一种聚氨基酸封端的组织修补材料及其制备方法 | |
CN111840642B (zh) | 一种软骨脱细胞基质复合支架的制备方法及其应用 | |
CN114085394B (zh) | 一种重组胶原蛋白双相凝胶及其制备方法和应用 | |
CN114949359B (zh) | 一种脱细胞基质微粒填充剂及其制备方法 | |
Gao et al. | Injectable and self-crosslinkable hydrogels based on collagen type II and activated chondroitin sulfate for cell delivery | |
Ratanavaraporn et al. | Osteogenic differentiation of bone-marrow-derived stem cells cultured with mixed gelatin and chitooligosaccharide scaffolds | |
CN110812529A (zh) | 一种可注射水凝胶及其制备方法 | |
CN113694254A (zh) | 骨修复材料及骨修复材料的制备方法、应用 | |
CN116196329A (zh) | 一种透明质酸-脱细胞基质制剂及其制备方法与应用 | |
KR102250064B1 (ko) | 생분해 기간 및 물성 조절 가능한 동물 연골 유래 조직 하이드로젤의 제조 방법 | |
CN110624134A (zh) | 一种负载鹿茸多肽的ⅰ型/ⅲ型胶原软骨修复支架 | |
CN115040636A (zh) | 多肽在促进软骨再生或修复中的用途 | |
Zhang et al. | Eggshell membrane powder reinforces adhesive polysaccharide hydrogels for wound repair | |
Cheng et al. | A photocrosslinked methacrylated carboxymethyl chitosan/oxidized locust bean gum double network hydrogel for cartilage repair | |
CN117582548A (zh) | 一种具有原位修复前交叉韧带功能的可注射多交联复合水凝胶支架及其制备方法 | |
Zhou et al. | Hyaluronic Acid‐Dopamine‐NCSN Hydrogel Combined With Extracellular Matrix Promotes Wound Healing |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |