CN115970056A - A kind of 3D printing material for bone repair, bone repair material and its preparation method and application - Google Patents
A kind of 3D printing material for bone repair, bone repair material and its preparation method and applicationDownload PDFInfo
- Publication number
- CN115970056A CN115970056ACN202310070891.7ACN202310070891ACN115970056ACN 115970056 ACN115970056 ACN 115970056ACN 202310070891 ACN202310070891 ACN 202310070891ACN 115970056 ACN115970056 ACN 115970056A
- Authority
- CN
- China
- Prior art keywords
- bone repair
- printing
- repair material
- bone
- printed
- 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.)
- Granted
Links
- 210000000988bone and boneAnatomy0.000titleclaimsabstractdescription128
- 239000000463materialSubstances0.000titleclaimsabstractdescription120
- 230000008439repair processEffects0.000titleclaimsabstractdescription114
- 2380000101463D printingMethods0.000titleclaimsabstractdescription70
- 238000002360preparation methodMethods0.000titledescription2
- 238000000034methodMethods0.000claimsabstractdescription14
- 239000003638chemical reducing agentSubstances0.000claimsabstractdescription9
- 102000004169proteins and genesHuman genes0.000claimsabstractdescription8
- 108090000623proteins and genesProteins0.000claimsabstractdescription8
- 239000003431cross linking reagentSubstances0.000claimsabstractdescription6
- 238000007639printingMethods0.000claimsdescription24
- 102000016943MuramidaseHuman genes0.000claimsdescription19
- 108010014251MuramidaseProteins0.000claimsdescription19
- 108010062010N-Acetylmuramoyl-L-alanine AmidaseProteins0.000claimsdescription19
- 229960000274lysozymeDrugs0.000claimsdescription19
- 235000010335lysozymeNutrition0.000claimsdescription19
- 239000004325lysozymeSubstances0.000claimsdescription19
- SXRSQZLOMIGNAQ-UHFFFAOYSA-NGlutaraldehydeChemical compoundO=CCCCC=OSXRSQZLOMIGNAQ-UHFFFAOYSA-N0.000claimsdescription9
- RWSXRVCMGQZWBV-WDSKDSINSA-NglutathioneChemical compoundOC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=ORWSXRVCMGQZWBV-WDSKDSINSA-N0.000claimsdescription8
- UXVMQQNJUSDDNG-UHFFFAOYSA-LCalcium chlorideChemical compound[Cl-].[Cl-].[Ca+2]UXVMQQNJUSDDNG-UHFFFAOYSA-L0.000claimsdescription7
- 235000018102proteinsNutrition0.000claimsdescription6
- AZKVWQKMDGGDSV-BCMRRPTOSA-NGenipinChemical compoundCOC(=O)C1=CO[C@@H](O)[C@@H]2C(CO)=CC[C@H]12AZKVWQKMDGGDSV-BCMRRPTOSA-N0.000claimsdescription4
- 108010024636GlutathioneProteins0.000claimsdescription4
- XUJNEKJLAYXESH-UHFFFAOYSA-NcysteineNatural productsSCC(N)C(O)=OXUJNEKJLAYXESH-UHFFFAOYSA-N0.000claimsdescription4
- 235000018417cysteineNutrition0.000claimsdescription4
- AZKVWQKMDGGDSV-UHFFFAOYSA-NgenipinNatural productsCOC(=O)C1=COC(O)C2C(CO)=CCC12AZKVWQKMDGGDSV-UHFFFAOYSA-N0.000claimsdescription4
- 229960003180glutathioneDrugs0.000claimsdescription4
- 108091003079Bovine Serum AlbuminProteins0.000claimsdescription3
- 229940098773bovine serum albuminDrugs0.000claimsdescription3
- 238000002791soakingMethods0.000claimsdescription3
- XUJNEKJLAYXESH-REOHCLBHSA-NL-CysteineChemical compoundSC[C@H](N)C(O)=OXUJNEKJLAYXESH-REOHCLBHSA-N0.000claimsdescription2
- NQTADLQHYWFPDB-UHFFFAOYSA-NN-HydroxysuccinimideChemical compoundON1C(=O)CCC1=ONQTADLQHYWFPDB-UHFFFAOYSA-N0.000claimsdescription2
- 229910019142PO4Inorganic materials0.000claimsdescription2
- NBIIXXVUZAFLBC-UHFFFAOYSA-KphosphateChemical compound[O-]P([O-])([O-])=ONBIIXXVUZAFLBC-UHFFFAOYSA-K0.000claimsdescription2
- 239000010452phosphateSubstances0.000claimsdescription2
- SRSAMLMFVUJPSR-UHFFFAOYSA-N2-carboxyethylphosphanium;chlorideChemical compoundCl.OC(=O)CCPSRSAMLMFVUJPSR-UHFFFAOYSA-N0.000claims1
- 229940050560calcium chloride anhydrousDrugs0.000claims1
- BXGTVNLGPMZLAZ-UHFFFAOYSA-Nn'-ethylmethanediimine;hydrochlorideChemical compoundCl.CCN=C=NBXGTVNLGPMZLAZ-UHFFFAOYSA-N0.000claims1
- 102000009091Amyloidogenic ProteinsHuman genes0.000abstractdescription4
- 108010048112Amyloidogenic ProteinsProteins0.000abstractdescription4
- 239000007864aqueous solutionSubstances0.000description40
- 239000000243solutionSubstances0.000description21
- 230000000694effectsEffects0.000description17
- PBVAJRFEEOIAGW-UHFFFAOYSA-N3-[bis(2-carboxyethyl)phosphanyl]propanoic acid;hydrochlorideChemical compoundCl.OC(=O)CCP(CCC(O)=O)CCC(O)=OPBVAJRFEEOIAGW-UHFFFAOYSA-N0.000description15
- 230000007547defectEffects0.000description15
- 241001465754MetazoaSpecies0.000description12
- 241000700159RattusSpecies0.000description12
- 238000002474experimental methodMethods0.000description12
- 210000001519tissueAnatomy0.000description7
- KCXVZYZYPLLWCC-UHFFFAOYSA-NEDTAChemical compoundOC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=OKCXVZYZYPLLWCC-UHFFFAOYSA-N0.000description6
- 229930040373ParaformaldehydeNatural products0.000description6
- 230000018044dehydrationEffects0.000description6
- 238000006297dehydration reactionMethods0.000description6
- 239000012188paraffin waxSubstances0.000description6
- 229920002866paraformaldehydePolymers0.000description6
- 238000004445quantitative analysisMethods0.000description6
- 210000003625skullAnatomy0.000description6
- 238000010186stainingMethods0.000description6
- 208000014674injuryDiseases0.000description3
- 230000011164ossificationEffects0.000description3
- 239000008055phosphate buffer solutionSubstances0.000description3
- 238000012360testing methodMethods0.000description3
- 238000004132cross linkingMethods0.000description2
- 230000006378damageEffects0.000description2
- 230000004048modificationEffects0.000description2
- 238000012986modificationMethods0.000description2
- 230000008733traumaEffects0.000description2
- FPQQSJJWHUJYPU-UHFFFAOYSA-N3-(dimethylamino)propyliminomethylidene-ethylazanium;chlorideChemical compoundCl.CCN=C=NCCCN(C)CFPQQSJJWHUJYPU-UHFFFAOYSA-N0.000description1
- 206010065687Bone lossDiseases0.000description1
- 206010028980NeoplasmDiseases0.000description1
- 208000027418Wounds and injuryDiseases0.000description1
- 239000002390adhesive tapeSubstances0.000description1
- 230000032683agingEffects0.000description1
- 230000009286beneficial effectEffects0.000description1
- 239000012620biological materialSubstances0.000description1
- 239000000872bufferSubstances0.000description1
- 239000007853buffer solutionSubstances0.000description1
- 239000003153chemical reaction reagentSubstances0.000description1
- 230000003111delayed effectEffects0.000description1
- 238000010586diagramMethods0.000description1
- 201000010099diseaseDiseases0.000description1
- 208000037265diseases, disorders, signs and symptomsDiseases0.000description1
- 238000005516engineering processMethods0.000description1
- 230000028993immune responseEffects0.000description1
- 238000002513implantationMethods0.000description1
- 208000015181infectious diseaseDiseases0.000description1
- 230000028709inflammatory responseEffects0.000description1
- 230000000399orthopedic effectEffects0.000description1
- 230000008506pathogenesisEffects0.000description1
- 230000010412perfusionEffects0.000description1
- 239000002994raw materialSubstances0.000description1
- 238000001356surgical procedureMethods0.000description1
- 230000001225therapeutic effectEffects0.000description1
- 230000017423tissue regenerationEffects0.000description1
Images
Classifications
- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P10/00—Technologies related to metal processing
- Y02P10/25—Process efficiency
Landscapes
- Materials For Medical Uses (AREA)
Abstract
Translated fromChineseDescription
Translated fromChinese技术领域technical field
本发明涉及3D打印领域，特别是一种骨修复的3D打印材料、骨修复材料及其制备方法和应用。The invention relates to the field of 3D printing, in particular to a 3D printing material for bone repair, a bone repair material and a preparation method and application thereof.
背景技术Background technique
随着人口老龄化的加重，因创伤、肿瘤、感染等造成的骨缺损患者逐年增加。超过临界标准的骨缺损通常难以修复，很容易导致骨修复延迟甚至骨不愈合。临界骨缺损的临床治疗是整形外科中最具挑战性的课题之一，需要植入生物材料进行修复。然而受伤、疾病或外伤引起的大规模骨损伤往往缺乏自我修复能力，尽管自体移植和同种异体移植已在临床上得到批准并显示令人满意的治疗效果，但在临床上也往往会出现供体部位的发病、免疫以及炎症反应。3D打印组织工程骨支架具有高保真度、复杂多孔结构、优秀的生物相容性，可以快速精准的满足患者需求，越来越多的研究者开始使用3D打印技术进行骨组织修复。With the aggravation of population aging, the number of patients with bone defects caused by trauma, tumor, infection, etc. is increasing year by year. Bone defects exceeding the critical standard are usually difficult to repair, which can easily lead to delayed bone repair or even bone nonunion. The clinical treatment of critical bone defects is one of the most challenging topics in orthopedic surgery, requiring the implantation of biomaterials for repair. However, large-scale bone damage caused by injury, disease, or trauma often lacks the ability to repair itself. Although autografts and allografts have been clinically approved and have shown satisfactory therapeutic effects, there are often cases of donor bone loss in clinical practice. pathogenesis, immune and inflammatory responses of body parts. 3D printed tissue engineering bone scaffolds have high fidelity, complex porous structure, and excellent biocompatibility, and can quickly and accurately meet the needs of patients. More and more researchers have begun to use 3D printing technology for bone tissue repair.
因鉴于此，特提出此发明。Because of this, propose this invention especially.
发明内容Contents of the invention
为了解决现有的技术问题，本发明提供了一种基于类淀粉样蛋白质的3D打印材料，通过该种打印材料配合特定的3D打印方法可按需制备不同形状且生物相容性优秀的骨修复材料，该种骨修复材料具备良好的机械性能，同时具备优异的骨修复能力。In order to solve the existing technical problems, the present invention provides a 3D printing material based on amyloid protein, through which a specific 3D printing method can be used to prepare bone repairs with different shapes and excellent biocompatibility This kind of bone repair material has good mechanical properties and excellent bone repair ability.
为实现上述目的，本发明采用的技术方案如下：To achieve the above object, the technical scheme adopted in the present invention is as follows:
首先，本发明提供了一种用于骨修复的3D打印材料，所述3D打印材料包括蛋白质原料和还原剂，且所述蛋白质原料的浓度为30-250mg/mL，所述还原剂的浓度为1-100mg/mL。First, the present invention provides a 3D printing material for bone repair, the 3D printing material includes a protein material and a reducing agent, and the concentration of the protein material is 30-250 mg/mL, and the concentration of the reducing agent is 1-100mg/mL.
优选或可选地，所述蛋白质原料为溶菌酶或牛血清白蛋白。Preferably or alternatively, the protein material is lysozyme or bovine serum albumin.
优选或可选地，所述还原剂为三(2-羧乙基)膦盐酸盐、半胱氨酸、谷胱甘肽中的任意一种。Preferably or alternatively, the reducing agent is any one of tris(2-carboxyethyl)phosphine hydrochloride, cysteine, and glutathione.
优选或可选地，所述3D打印材料的pH为1-4。Preferably or optionally, the pH of the 3D printing material is 1-4.
优选或可选地，所述3D打印材料的使用温度为37-100℃。Preferably or optionally, the use temperature of the 3D printing material is 37-100°C.
另一方面，本发明还提供了一种采用上述的用于骨修复的3D打印材料制备骨修复材料的方法，按下述步骤依次进行：On the other hand, the present invention also provides a method for preparing a bone repair material using the above-mentioned 3D printing material for bone repair, followed by the following steps:
(1)将3D打印材料预热至使用温度后，灌注3D打印机的挤压器内；(1) After preheating the 3D printing material to the operating temperature, pour it into the extruder of the 3D printer;
(2)将3D打印材料通过3D打印机打印为3D打印模型(2) Print the 3D printing material into a 3D printing model through a 3D printer
(3)将3D打印模型置于交联剂内浸泡，取出即得骨修复材料。(3) Soak the 3D printed model in the cross-linking agent and take it out to obtain the bone repair material.
优选或可选地，步骤(2)中，打印的气压为0.1-1.0MPa,打印速度为0.1-100mm/s，打印使用的针头直径为0.2-0.5mm。Preferably or optionally, in step (2), the printing air pressure is 0.1-1.0 MPa, the printing speed is 0.1-100 mm/s, and the diameter of the needle used for printing is 0.2-0.5 mm.
优选或可选地，步骤(3)中使用的角连接为磷酸盐、无水氯化钙、1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐/N-羟基琥珀酰亚胺、戊二醛或京尼平中的任意一种。Preferably or alternatively, the corner link used in step (3) is phosphate, anhydrous calcium chloride, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride/N - Any of hydroxysuccinimide, glutaraldehyde, or genipin.
优选或可选地，所述交联剂的浓度为0.1-10％(w/w)。Preferably or alternatively, the concentration of the cross-linking agent is 0.1-10% (w/w).
优选或可选地，步骤(3)中浸泡的时间为0-12小时Preferably or alternatively, the soaking time in step (3) is 0-12 hours
第三方面，本发明还提供了一种骨修复材料，采用上述的方法制备而成。In the third aspect, the present invention also provides a bone repair material prepared by the above-mentioned method.
以及上述的骨修复材料在骨修复领域的应用。And the application of the above-mentioned bone repair material in the field of bone repair.
有益效果Beneficial effect
本发明提供了一种基于类淀粉样蛋白质的3D打印材料，该种打印材料在3D打印并浸泡于交联剂之后可制得机械性能良好的骨修复材料，上述方法制得的骨修复材料具有良好的生物相容性和骨修复能力，同时因其采用3D打印的方法制备，形状可以按需定制。The present invention provides a 3D printing material based on amyloid protein, which can produce a bone repair material with good mechanical properties after 3D printing and soaking in a cross-linking agent. The bone repair material prepared by the above method has Good biocompatibility and bone repair ability, and because it is prepared by 3D printing, the shape can be customized on demand.
附图说明Description of drawings
图1为本发明效果实施例1提供的未交联的立方体网格图片；Fig. 1 is the uncrosslinked cube mesh picture provided by effect example 1 of the present invention;
图2为本发明效果实施例2提供的立方体网格图片；Fig. 2 is the cubic grid picture provided by effect embodiment 2 of the present invention;
图3为本发明效果实施例3提供的骨修复材料的交联前后的力学性能测试结果图；Fig. 3 is a graph showing the mechanical property test results before and after cross-linking of the bone repair material provided by Effect Example 3 of the present invention;
图4为效果实施例4-9提供的骨修复材料在不同时间的修复效果CT图；Fig. 4 is the repair effect CT diagram of the bone repair material provided by effect embodiment 4-9 at different times;
图5为效果实施例4-9提供的骨修复材料在不同时间的骨修复体积结果图。Fig. 5 is a graph showing the bone repair volume results at different times of the bone repair materials provided by effect examples 4-9.
具体实施方式Detailed ways
为了便于理解本发明，下面将结合说明书附图和较佳实验例对本发明作更全面、细致地描述，但本发明的保护范围并不限于以下具体的实施例。In order to facilitate the understanding of the present invention, the present invention will be described more comprehensively and in detail below in conjunction with the accompanying drawings and preferred experimental examples, but the protection scope of the present invention is not limited to the following specific examples.
除非另有定义，下文中所使用的所有专业术语与本领域技术人员通常理解的含义相同。本文中所使用的专业术语只是为了描述具体实施例的目的，并不是旨在限制本发明的保护范围。Unless otherwise defined, all technical terms used hereinafter have the same meanings as commonly understood by those skilled in the art. The terminology used herein is only for the purpose of describing specific embodiments, and is not intended to limit the protection scope of the present invention.
除非另有特别说明，本发明中用到的各种原材料、试剂、仪器和设备等均可通过市场购买得到或者可通过现有方法制备得到。Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.
实施例1Example 1
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为80mg/mL的溶菌酶水溶液和50Mm pH＝1的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温3小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 80mg/mL and 50Mm tris(2-carboxyethyl)phosphine hydrochloride aqueous solution with pH=1, keep it warm at 50°C for 3 hours, and pour it into the extruder of the 3D printer ;
选择直径0.25mm的枪头，以0.12MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为长宽均为3cm，厚度为0.5cm，格数为12×12的网格状结构。Choose a gun tip with a diameter of 0.25mm, and print the 3D printing model at a pressure of 0.12MPa and a speed of 12mm/s. In this example, the 3D printing model has a length and width of 3cm, a thickness of 0.5cm, and a grid of 12×12 grid structure.
打印完成后，将打印出的3D打印模型置于10mg/mL的氯化钙溶液中浸泡2h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 10 mg/mL calcium chloride solution for 2 hours, and the bone repair material product is obtained after taking it out.
实施例2Example 2
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为100mg/mL的溶菌酶水溶液和50mM pH＝3的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温2小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 100 mg/mL and 50 mM tris(2-carboxyethyl)phosphine hydrochloride aqueous solution at pH=3, keep it warm at 50°C for 2 hours, and pour it into the extruder of the 3D printer ;
选择直径0.25mm的枪头，以0.12MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为长宽均为4cm，厚度为0.5cm，格数为12×12的网格状结构。Select a gun tip with a diameter of 0.25mm, and print the 3D printing model at a pressure of 0.12MPa and a speed of 12mm/s. In this example, the 3D printing model has a length and width of 4cm, a thickness of 0.5cm, and a grid of 12×12 grid structure.
打印完成后，将打印出的3D打印模型置于30mg/mL的氯化钙溶液中浸泡2h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 30 mg/mL calcium chloride solution for 2 hours, and the bone repair material product is obtained after taking it out.
实施例3Example 3
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为120mg/mL的溶菌酶水溶液和50mM pH＝2的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温2小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 120 mg/mL and 50 mM tris(2-carboxyethyl)phosphine hydrochloride aqueous solution at pH=2, keep it warm at 50°C for 2 hours, and pour it into the extruder of the 3D printer ;
选择直径0.25mm的枪头，以0.12MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为直径3cm，厚度为0.5cm的圆柱体网格状结构。Select a gun tip with a diameter of 0.25 mm, and print a 3D printing model at a pressure of 0.12 MPa and a speed of 12 mm/s. In this embodiment, the 3D printing model is a cylindrical grid structure with a diameter of 3 cm and a thickness of 0.5 cm.
打印完成后，将打印出的3D打印模型置于10×PBS缓冲液中浸泡2h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 10×PBS buffer solution for 2 hours, and the bone repair material product is obtained after taking it out.
实施例4Example 4
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为120mg/mL的溶菌酶水溶液和40mM pH＝1的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温2小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 120 mg/mL and 40 mM tris(2-carboxyethyl)phosphine hydrochloride aqueous solution at pH=1, keep it warm at 50°C for 2 hours, and pour it into the extruder of the 3D printer ;
选择直径0.41mm的枪头，以0.15MPa的压力、13mm/s的速度打印3D打印模型，本实施例中，3D打印模型为直径4cm，厚度为0.5cm的圆柱体网格状结构。Select a gun tip with a diameter of 0.41 mm, and print a 3D printing model at a pressure of 0.15 MPa and a speed of 13 mm/s. In this embodiment, the 3D printing model is a cylindrical grid structure with a diameter of 4 cm and a thickness of 0.5 cm.
打印完成后，将打印出的3D打印模型置于1％京尼平水溶液中浸泡2h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 1% genipin aqueous solution for 2 hours, and the bone repair material product is obtained after taking it out.
实施例5Example 5
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为120mg/mL的溶菌酶水溶液和40mg/mL pH＝4的谷胱甘肽水溶液等体积混合后，在60℃下保温5小时，灌注3D打印机的挤压器中；After mixing equal volumes of 120mg/mL lysozyme aqueous solution and 40mg/mL pH=4 glutathione aqueous solution, keep it warm at 60°C for 5 hours, and pour it into the extruder of the 3D printer;
选择直径0.41mm的枪头，以0.15MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为直径4cm，厚度为0.5cm的圆柱体网格状结构。Select a gun tip with a diameter of 0.41 mm, and print a 3D printing model at a pressure of 0.15 MPa and a speed of 12 mm/s. In this embodiment, the 3D printing model is a cylindrical grid structure with a diameter of 4 cm and a thickness of 0.5 cm.
打印完成后，将打印出的3D打印模型置于2％戊二醛水溶液中浸泡2h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 2% glutaraldehyde aqueous solution for 2 hours, and the bone repair material product is obtained after taking it out.
实施例6Example 6
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为100mg/mL的溶菌酶水溶液和30mg/mL pH＝4的谷胱甘肽水溶液等体积混合后，在60℃下保温5小时，灌注3D打印机的挤压器中；After mixing equal volumes of 100mg/mL lysozyme aqueous solution and 30mg/mL pH=4 glutathione aqueous solution, keep it warm at 60°C for 5 hours, and pour it into the extruder of the 3D printer;
选择直径0.41mm的枪头，以0.15MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为直径4cm，厚度为0.5cm的圆柱体网格状结构。Select a gun tip with a diameter of 0.41 mm, and print a 3D printing model at a pressure of 0.15 MPa and a speed of 12 mm/s. In this embodiment, the 3D printing model is a cylindrical grid structure with a diameter of 4 cm and a thickness of 0.5 cm.
打印完成后，将打印出的3D打印模型置于5％戊二醛水溶液中浸泡4h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 5% glutaraldehyde aqueous solution for 4 hours, and the bone repair material product is obtained after taking it out.
实施例7Example 7
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为100mg/mL的溶菌酶水溶液和50mg/mL pH＝4的半胱氨酸水溶液等体积混合后，在50℃下保温8小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 100 mg/mL and cysteine aqueous solution with a pH of 50 mg/mL = 4, heat it at 50°C for 8 hours, and pour it into the extruder of the 3D printer;
选择直径0.25mm的枪头，以0.2MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为长宽为5cm，厚度为0.5cm，12×12的方形网格状结构。Select a gun tip with a diameter of 0.25mm, and print the 3D printing model at a pressure of 0.2MPa and a speed of 12mm/s. In this example, the 3D printing model is a square grid with a length and width of 5cm and a thickness of 0.5cm, and 12×12 shape structure.
打印完成后，将打印出的3D打印模型置于5％戊二醛水溶液中浸泡4h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 5% glutaraldehyde aqueous solution for 4 hours, and the bone repair material product is obtained after taking it out.
实施例8Example 8
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为160mg/mL的溶菌酶水溶液和100mg/mL pH＝4的半胱氨酸水溶液等体积混合后，在50℃下保温8小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 160 mg/mL and cysteine aqueous solution with a pH of 100 mg/mL = 4, heat it at 50°C for 8 hours, and pour it into the extruder of the 3D printer;
选择直径0.25mm的枪头，以0.2MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为直径4cm，厚度为8cm的圆柱体网格状结构。Select a gun tip with a diameter of 0.25 mm, and print a 3D printing model at a pressure of 0.2 MPa and a speed of 12 mm/s. In this embodiment, the 3D printing model is a cylindrical grid structure with a diameter of 4 cm and a thickness of 8 cm.
打印完成后，将打印出的3D打印模型置于0.1％京尼平水溶液中浸泡12h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 0.1% genipin aqueous solution for 12 hours, and the bone repair material product is obtained after taking it out.
实施例9Example 9
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将牛血清白蛋白用PBS缓冲液配置成浓度为80mg/mL的溶液，并和50mM pH＝2的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在37℃下保温6小时，灌注3D打印机的挤压器中；Bovine serum albumin was prepared into a solution with a concentration of 80 mg/mL in PBS buffer, mixed with an equal volume of 50 mM tris(2-carboxyethyl)phosphine hydrochloride aqueous solution at pH=2, and incubated at 37°C for 6 hour, in the extruder of perfusion 3D printer;
选择直径0.25mm的枪头，以0.4MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为直径4cm，厚度为8cm的圆柱体网格状结构。Select a gun tip with a diameter of 0.25 mm, and print a 3D printing model at a pressure of 0.4 MPa and a speed of 12 mm/s. In this embodiment, the 3D printing model is a cylindrical grid structure with a diameter of 4 cm and a thickness of 8 cm.
打印完成后，将打印出的3D打印模型置于0.1％戊二醛溶液中浸泡6h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 0.1% glutaraldehyde solution for 6 hours, and the bone repair material product is obtained after taking it out.
效果实施例Effect example
效果实施例1Effect Example 1
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为180mg/mL的溶菌酶水溶液和50Mm pH＝1的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温3小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 180mg/mL and 50Mm tris(2-carboxyethyl)phosphine hydrochloride aqueous solution with pH=1, keep it warm at 50°C for 3 hours, and pour it into the extruder of the 3D printer ;
选择直径0.25mm的枪头，以0.12MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为长宽5cm，厚度为0.5cm，12×12的方形网格状结构。Select a gun tip with a diameter of 0.25mm, and print the 3D printing model at a pressure of 0.12MPa and a speed of 12mm/s. In this example, the 3D printing model is a square grid shape with a length and width of 5cm and a thickness of 0.5cm. structure.
打印完成后如附图1，将打印出的3D打印模型置于培养皿中，取出即得骨修复材料产品。After the printing is completed, as shown in Figure 1, place the printed 3D printed model in a petri dish, and take it out to obtain the bone repair material product.
效果实施例2Effect Example 2
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为180mg/mL的溶菌酶水溶液和50Mm pH＝1的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温3小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 180mg/mL and 50Mm tris(2-carboxyethyl)phosphine hydrochloride aqueous solution with pH=1, keep it warm at 50°C for 3 hours, and pour it into the extruder of the 3D printer ;
选择直径0.25mm的枪头，以0.12MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为长宽5cm，厚度为0.5cm，12×12的方形网格状结构。Select a gun tip with a diameter of 0.25mm, and print the 3D printing model at a pressure of 0.12MPa and a speed of 12mm/s. In this example, the 3D printing model is a square grid shape with a length and width of 5cm and a thickness of 0.5cm. structure.
打印完成后如附图2，将打印出的3D打印模型置于1％戊二醛溶液中浸泡2h，取出即得骨修复材料产品。After the printing is completed, as shown in Figure 2, the printed 3D printed model is soaked in 1% glutaraldehyde solution for 2 hours, and the bone repair material product is obtained after taking it out.
效果实施例3Effect Example 3
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为180mg/mL的溶菌酶水溶液和50Mm pH＝2的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温3小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 180 mg/mL and 50 Mm tris(2-carboxyethyl)phosphine hydrochloride aqueous solution with pH=2, incubate at 50°C for 3 hours, and pour it into the extruder of the 3D printer ;
选择直径0.25mm的枪头，以0.12MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型类似哑铃状或狗骨头状尺寸两端长宽1cm×1cm，中间长宽为1cm×0.5cm。Choose a gun tip with a diameter of 0.25mm, and print the 3D printing model with a pressure of 0.12MPa and a speed of 12mm/s. In this example, the 3D printing model is similar to a dumbbell or a dog bone. The width is 1cm×0.5cm.
打印完成后，将打印出的3D打印模型置于1％戊二醛溶液中浸泡2h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 1% glutaraldehyde solution for 2 hours, and the bone repair material product is obtained after taking it out.
用胶带粘住样品长度方向的两端，再夹持于试验机夹头。传感器量程为100N，设定拉升速度为5mm/min，跨距为15-20mm。此测试均在室温下进行，每组样品测试三次，取平均值，计算误差带，记录拉伸力，截面积，计算拉伸强度。Adhere both ends of the sample in the length direction with adhesive tape, and then clamp it to the chuck of the testing machine. The measuring range of the sensor is 100N, the setting pulling speed is 5mm/min, and the span is 15-20mm. This test is carried out at room temperature, each group of samples is tested three times, the average value is taken, the error band is calculated, the tensile force and cross-sectional area are recorded, and the tensile strength is calculated.
如附图3测试得出，未交联的3D打印骨修复材料的拉伸强度为0.117MPa,用1％戊二醛溶液中浸泡2h后的3D打印骨修复材料的拉伸强度可达1.02Mpa,因此通过交联可以使支架获得相较于未交联支架而言更好的拉伸强度。As shown in Figure 3, the tensile strength of the uncrosslinked 3D printed bone repair material is 0.117MPa, and the tensile strength of the 3D printed bone repair material soaked in 1% glutaraldehyde solution for 2 hours can reach 1.02Mpa , so cross-linking can make scaffolds obtain better tensile strength than uncross-linked scaffolds.
效果实施例4Effect Example 4
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为180mg/mL的溶菌酶水溶液和50Mm pH＝1的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温3小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 180mg/mL and 50Mm tris(2-carboxyethyl)phosphine hydrochloride aqueous solution with pH=1, keep it warm at 50°C for 3 hours, and pour it into the extruder of the 3D printer ;
选择直径0.25mm的枪头，以0.12MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为直径为5mm,厚度为40mm，4层圆形网格状。Select a gun tip with a diameter of 0.25mm, and print a 3D printing model with a pressure of 0.12MPa and a speed of 12mm/s. In this embodiment, the 3D printing model is 5mm in diameter, 40mm in thickness, and has a 4-layer circular grid shape.
打印完成后，将打印出的3D打印模型置于1％氯化钙溶液中浸泡6h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 1% calcium chloride solution for 6 hours, and the bone repair material product is obtained after taking it out.
所有动物实验均遵循天津市南开医院动物试验伦理委员会批准。3只雄性SD大鼠(8周龄，230-250g)，左侧为空白对照组，右侧为交联支架填充组。大鼠麻醉后制备双侧直径5mm缺损，右侧放入打印的支架材料，最后分层缝合各层组织。在1个月后分别取出完整颅骨，浸泡在4％多聚甲醛溶液中，进行固定，随后进行micro-ct扫描，使用ct-vox软件获得重建后3d图像，进一步使用ct-an软件进行定量分析成骨相关指标。随后，将扫描后样本放在EDTA溶液中进行脱钙，大约4周左右，至样本可无阻力穿透为止。后续进一步进行脱水，石蜡包埋，切片，HE及Masson三色染色，观察缺损处周围骨组织的修复情况。All animal experiments were approved by the Animal Experiment Ethics Committee of Tianjin Nankai Hospital. Three male SD rats (8 weeks old, 230-250g), the left side is the blank control group, and the right side is the cross-linked scaffold-filled group. After the rats were anesthetized, a defect with a diameter of 5 mm on both sides was prepared, and the printed scaffold material was placed on the right side, and finally each layer of tissue was sutured in layers. After 1 month, the complete skulls were taken out, soaked in 4% paraformaldehyde solution, and fixed, followed by micro-ct scanning, using ct-vox software to obtain reconstructed 3D images, and further using ct-an software for quantitative analysis Osteogenesis-related indicators. Subsequently, the scanned sample is placed in EDTA solution for decalcification, about 4 weeks, until the sample can penetrate without resistance. Further dehydration, paraffin embedding, sectioning, HE and Masson's trichrome staining were performed to observe the repair of bone tissue around the defect.
效果实施例5Effect Example 5
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为180mg/mL的溶菌酶水溶液和50Mm pH＝1的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温3小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 180mg/mL and 50Mm tris(2-carboxyethyl)phosphine hydrochloride aqueous solution with pH=1, keep it warm at 50°C for 3 hours, and pour it into the extruder of the 3D printer ;
选择直径0.25mm的枪头，以0.12MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为直径为5mm,厚度为40mm，4层圆形网格状。Select a gun tip with a diameter of 0.25mm, and print a 3D printing model with a pressure of 0.12MPa and a speed of 12mm/s. In this embodiment, the 3D printing model is 5mm in diameter, 40mm in thickness, and has a 4-layer circular grid shape.
打印完成后，将打印出的3D打印模型置于1％氯化钙溶液中浸泡6h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 1% calcium chloride solution for 6 hours, and the bone repair material product is obtained after taking it out.
所有动物实验均遵循天津市南开医院动物试验伦理委员会批准。3只雄性SD大鼠(8周龄，230-250g)，左侧为空白对照组，右侧为交联支架填充组。大鼠麻醉后制备双侧直径5mm缺损，右侧放入打印的支架材料，最后分层缝合各层组织。在3个月后分别取出完整颅骨，浸泡在4％多聚甲醛溶液中，进行固定，随后进行micro-ct扫描，使用ct-vox软件获得重建后3d图像，进一步使用ct-an软件进行定量分析成骨相关指标。随后，将扫描后样本放在EDTA溶液中进行脱钙，大约4周左右，至样本可无阻力穿透为止。后续进一步进行脱水，石蜡包埋，切片，HE及Masson三色染色，观察缺损处周围骨组织的修复情况。All animal experiments were approved by the Animal Experiment Ethics Committee of Tianjin Nankai Hospital. Three male SD rats (8 weeks old, 230-250g), the left side is the blank control group, and the right side is the cross-linked scaffold-filled group. After the rats were anesthetized, a defect with a diameter of 5 mm on both sides was prepared, and the printed scaffold material was placed on the right side, and finally each layer of tissue was sutured in layers. After 3 months, the complete skulls were taken out, soaked in 4% paraformaldehyde solution, fixed, followed by micro-ct scanning, using ct-vox software to obtain reconstructed 3D images, and further using ct-an software for quantitative analysis Osteogenesis-related indicators. Subsequently, the scanned sample is placed in EDTA solution for decalcification, about 4 weeks, until the sample can penetrate without resistance. Further dehydration, paraffin embedding, sectioning, HE and Masson's trichrome staining were performed to observe the repair of bone tissue around the defect.
效果实施例6Effect Example 6
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为180mg/mL的溶菌酶水溶液和50Mm pH＝1的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温3小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 180mg/mL and 50Mm tris(2-carboxyethyl)phosphine hydrochloride aqueous solution with pH=1, keep it warm at 50°C for 3 hours, and pour it into the extruder of the 3D printer ;
选择直径0.25mm的枪头，以0.12MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为直径为5mm,厚度为40mm，4层圆形网格状。Select a gun tip with a diameter of 0.25mm, and print a 3D printing model with a pressure of 0.12MPa and a speed of 12mm/s. In this embodiment, the 3D printing model is 5mm in diameter, 40mm in thickness, and has a 4-layer circular grid shape.
打印完成后，将打印出的3D打印模型置于1％氯化钙溶液中浸泡6h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 1% calcium chloride solution for 6 hours, and the bone repair material product is obtained after taking it out.
所有动物实验均遵循天津市南开医院动物试验伦理委员会批准。3只雄性SD大鼠(8周龄，230-250g)，左侧为空白对照组，右侧为交联支架填充组。大鼠麻醉后制备双侧直径5mm缺损，右侧放入打印的支架材料，最后分层缝合各层组织。在6个月后分别取出完整颅骨，浸泡在4％多聚甲醛溶液中，进行固定，随后进行micro-ct扫描，使用ct-vox软件获得重建后3d图像，进一步使用ct-an软件进行定量分析成骨相关指标。随后，将扫描后样本放在EDTA溶液中进行脱钙，大约4周左右，至样本可无阻力穿透为止。后续进一步进行脱水，石蜡包埋，切片，HE及Masson三色染色，观察缺损处周围骨组织的修复情况。All animal experiments were approved by the Animal Experiment Ethics Committee of Tianjin Nankai Hospital. Three male SD rats (8 weeks old, 230-250g), the left side is the blank control group, and the right side is the cross-linked scaffold-filled group. After the rats were anesthetized, a defect with a diameter of 5 mm on both sides was prepared, and the printed scaffold material was placed on the right side, and each layer of tissue was finally sutured in layers. After 6 months, the complete skulls were taken out, soaked in 4% paraformaldehyde solution, and fixed, followed by micro-ct scanning, using ct-vox software to obtain reconstructed 3D images, and further using ct-an software for quantitative analysis Osteogenesis-related indicators. Subsequently, the scanned sample is placed in EDTA solution for decalcification for about 4 weeks until the sample can penetrate without resistance. Further dehydration, paraffin embedding, sectioning, HE and Masson's trichrome staining were performed to observe the repair of bone tissue around the defect.
效果实施例7Effect Example 7
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为180mg/mL的溶菌酶水溶液和50Mm pH＝2的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温5小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 180mg/mL and 50Mm tris(2-carboxyethyl)phosphine hydrochloride aqueous solution with pH=2, incubate at 50°C for 5 hours, and pour it into the extruder of the 3D printer ;
选择直径0.25mm的枪头，以0.12MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为直径为5mm,厚度为40mm，4层圆形网格状。Select a gun tip with a diameter of 0.25mm, and print a 3D printing model with a pressure of 0.12MPa and a speed of 12mm/s. In this embodiment, the 3D printing model is 5mm in diameter, 40mm in thickness, and has a 4-layer circular grid shape.
打印完成后，将打印出的3D打印模型置于10×磷酸盐缓冲液溶液中浸泡4h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 10×phosphate buffer solution for 4 hours, and the bone repair material product is obtained after taking it out.
所有动物实验均遵循天津市南开医院动物试验伦理委员会批准。3只雄性SD大鼠(8周龄，230-250g)，左侧为空白对照组，右侧为交联支架填充组。大鼠麻醉后制备双侧直径5mm缺损，右侧放入打印的支架材料，最后分层缝合各层组织。在1个月后取出完整颅骨，浸泡在4％多聚甲醛溶液中，进行固定，随后进行micro-ct扫描，使用ct-vox软件获得重建后3d图像，进一步使用ct-an软件进行定量分析成骨相关指标。随后，将扫描后样本放在EDTA溶液中进行脱钙，大约4周左右，至样本可无阻力穿透为止。后续进一步进行脱水，石蜡包埋，切片，HE及Masson三色染色，观察缺损处周围骨组织的修复情况。All animal experiments were approved by the Animal Experiment Ethics Committee of Tianjin Nankai Hospital. Three male SD rats (8 weeks old, 230-250g), the left side is the blank control group, and the right side is the cross-linked scaffold-filled group. After the rats were anesthetized, a defect with a diameter of 5 mm on both sides was prepared, and the printed scaffold material was placed on the right side, and each layer of tissue was finally sutured in layers. After 1 month, the complete skull was taken out, soaked in 4% paraformaldehyde solution, and fixed, followed by micro-ct scanning, using ct-vox software to obtain reconstructed 3D images, and further using ct-an software for quantitative analysis Bone-related indicators. Subsequently, the scanned sample is placed in EDTA solution for decalcification for about 4 weeks until the sample can penetrate without resistance. Further dehydration, paraffin embedding, sectioning, HE and Masson's trichrome staining were performed to observe the repair of bone tissue around the defect.
效果实施例8Effect Example 8
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为180mg/mL的溶菌酶水溶液和50Mm pH＝2的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温5小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 180mg/mL and 50Mm tris(2-carboxyethyl)phosphine hydrochloride aqueous solution with pH=2, incubate at 50°C for 5 hours, and pour it into the extruder of the 3D printer ;
选择直径0.25mm的枪头，以0.12MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为直径为5mm,厚度为40mm，4层圆形网格状。Select a gun tip with a diameter of 0.25mm, and print a 3D printing model with a pressure of 0.12MPa and a speed of 12mm/s. In this embodiment, the 3D printing model is 5mm in diameter, 40mm in thickness, and has a 4-layer circular grid shape.
打印完成后，将打印出的3D打印模型置于10×磷酸盐缓冲液溶液中浸泡4h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 10×phosphate buffer solution for 4 hours, and the bone repair material product is obtained after taking it out.
所有动物实验均遵循天津市南开医院动物试验伦理委员会批准。3只雄性SD大鼠(8周龄，230-250g)，左侧为空白对照组，右侧为交联支架填充组。大鼠麻醉后制备双侧直径5mm缺损，右侧放入打印的支架材料，最后分层缝合各层组织。在3个月后取出完整颅骨，浸泡在4％多聚甲醛溶液中，进行固定，随后进行micro-ct扫描，使用ct-vox软件获得重建后3d图像，进一步使用ct-an软件进行定量分析成骨相关指标。随后，将扫描后样本放在EDTA溶液中进行脱钙，大约4周左右，至样本可无阻力穿透为止。后续进一步进行脱水，石蜡包埋，切片，HE及Masson三色染色，观察缺损处周围骨组织的修复情况。All animal experiments were approved by the Animal Experiment Ethics Committee of Tianjin Nankai Hospital. Three male SD rats (8 weeks old, 230-250g), the left side is the blank control group, and the right side is the cross-linked scaffold-filled group. After the rats were anesthetized, a defect with a diameter of 5 mm on both sides was prepared, and the printed scaffold material was placed on the right side, and finally each layer of tissue was sutured in layers. After 3 months, the complete skull was taken out, soaked in 4% paraformaldehyde solution, fixed, followed by micro-ct scanning, using ct-vox software to obtain reconstructed 3D images, and further using ct-an software for quantitative analysis Bone-related indicators. Subsequently, the scanned sample is placed in EDTA solution for decalcification, about 4 weeks, until the sample can penetrate without resistance. Further dehydration, paraffin embedding, sectioning, HE and Masson's trichrome staining were performed to observe the repair of bone tissue around the defect.
效果实施例9Effect Example 9
本实施例提供了一种骨修复材料。This embodiment provides a bone repair material.
该骨修复材料按下述方法制备而成：The bone repair material is prepared as follows:
将浓度为180mg/mL的溶菌酶水溶液和50Mm pH＝2的三(2-羧乙基)膦盐酸盐水溶液等体积混合后，在50℃下保温5小时，灌注3D打印机的挤压器中；After mixing equal volumes of lysozyme aqueous solution with a concentration of 180mg/mL and 50Mm tris(2-carboxyethyl)phosphine hydrochloride aqueous solution with pH=2, incubate at 50°C for 5 hours, and pour it into the extruder of the 3D printer ;
选择直径0.25mm的枪头，以0.12MPa的压力、12mm/s的速度打印3D打印模型，本实施例中，3D打印模型为直径为5mm,厚度为40mm，4层圆形网格状。Select a gun tip with a diameter of 0.25mm, and print a 3D printing model with a pressure of 0.12MPa and a speed of 12mm/s. In this embodiment, the 3D printing model is 5mm in diameter, 40mm in thickness, and has a 4-layer circular grid shape.
打印完成后，将打印出的3D打印模型置于10×磷酸盐缓冲液溶液中浸泡4h，取出即得骨修复材料产品。After the printing is completed, the printed 3D printed model is soaked in 10×phosphate buffer solution for 4 hours, and the bone repair material product is obtained after taking it out.
所有动物实验均遵循天津市南开医院动物试验伦理委员会批准。3只雄性SD大鼠(8周龄，230-250g)，左侧为空白对照组，右侧为交联支架填充组。大鼠麻醉后制备双侧直径5mm缺损，右侧放入打印的支架材料，最后分层缝合各层组织。在6个月后取出完整颅骨，浸泡在4％多聚甲醛溶液中，进行固定，随后进行micro-ct扫描，使用ct-vox软件获得重建后3d图像，进一步使用ct-an软件进行定量分析成骨相关指标。随后，将扫描后样本放在EDTA溶液中进行脱钙，大约4周左右，至样本可无阻力穿透为止。后续进一步进行脱水，石蜡包埋，切片，HE及Masson三色染色，观察缺损处周围骨组织的修复情况。All animal experiments were approved by the Animal Experiment Ethics Committee of Tianjin Nankai Hospital. Three male SD rats (8 weeks old, 230-250g), the left side is the blank control group, and the right side is the cross-linked scaffold-filled group. After the rats were anesthetized, a defect with a diameter of 5 mm on both sides was prepared, and the printed scaffold material was placed on the right side, and each layer of tissue was finally sutured in layers. After 6 months, the complete skull was taken out, soaked in 4% paraformaldehyde solution, fixed, and then scanned by micro-ct, using ct-vox software to obtain the reconstructed 3D image, and further using ct-an software for quantitative analysis Bone-related indicators. Subsequently, the scanned sample is placed in EDTA solution for decalcification for about 4 weeks until the sample can penetrate without resistance. Further dehydration, paraffin embedding, sectioning, HE and Masson's trichrome staining were performed to observe the repair of bone tissue around the defect.
通过附图4和5中的CT和骨体积统计结果表明，1个月后Blank组骨体积分数为9％，CaCl2组骨体积分数为17％，PBS组骨体积分数为19％；3个月后Blank组骨体积分数为16％，CaCl2组骨体积分数为40％，PBS组骨体积分数为46％；6个月后Blank组骨体积分数为34％，CaCl2组骨体积分数为55％，PBS组骨体积分数为70％。The results of CT and bone volume statistics in accompanying drawings 4 and 5 showed that the bone volume fraction of the Blank group was 9%, the bone volume fraction of the CaCl2 group was 17%, and the bone volume fraction of the PBS group was 19% after 1 month; One month later, the bone volume fraction of the Blank group was 16%, that of the CaCl2 group was 40%, and that of the PBS group was 46%. After 6 months, the bone volume fraction of the Blank group was 34%, and that of the CaCl2 group was 55%, and the bone volume fraction in the PBS group was 70%.
综上所述，在1、3和6个月的CT结果表明，PBS组和CaCl2组的骨修复能力均要原高于Blank组，其中PBS组的骨修复效果更优，其骨修复体积是Blank组的2倍以上。In summary, the CT results at 1, 3 and 6 months showed that the bone repair ability of the PBS group and the CaCl2 group were higher than that of the Blank group, and the bone repair effect of the PBS group was better, and the bone repair volume It is more than 2 times that of the Blank group.
由此可见，本发明提供的基于类淀粉样蛋白质的3D打印材料，具有良好的生物相容性和骨修复能力。It can be seen that the 3D printing material based on amyloid protein provided by the present invention has good biocompatibility and bone repair ability.
最后应说明的是：以上实施例仅用以说明本发明的技术方案，而非对其限制；尽管参照前述实施例对本发明进行了详细的说明，本领域的普通技术人员应当理解：其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分技术特征进行等同替换；而这些修改或者替换，并不使相应技术方案的本质脱离本发明各实施例技术方案的精神和范围。Finally, it should be noted that: the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: it can still be Modifications are made to the technical solutions described in the foregoing embodiments, or equivalent replacements are made to some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the various embodiments of the present invention.
Claims (12)
Translated fromChinesePriority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310070891.7ACN115970056B (en) | 2023-01-31 | 2023-01-31 | 3D printing material for bone repair, bone repair material, and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310070891.7ACN115970056B (en) | 2023-01-31 | 2023-01-31 | 3D printing material for bone repair, bone repair material, and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115970056Atrue CN115970056A (en) | 2023-04-18 |
| CN115970056B CN115970056B (en) | 2024-07-19 |
Family
ID=85974224
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310070891.7AActiveCN115970056B (en) | 2023-01-31 | 2023-01-31 | 3D printing material for bone repair, bone repair material, and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115970056B (en) |
Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102836461A (en)* | 2012-09-25 | 2012-12-26 | 天津工业大学 | Human hair keratin shape righting material with adjustable and controllable mechanical property as well as preparation method thereof |
| CN103756955A (en)* | 2014-01-22 | 2014-04-30 | 清华大学 | Personalized bionic composite structure as well as preparation method of composite structure and method for drug screening by using composite structure |
| WO2014124546A1 (en)* | 2013-02-12 | 2014-08-21 | Eth Zurich | Hybrid nanocomposite materials of amyloid fibrils and gold |
| US20190106673A1 (en)* | 2017-10-11 | 2019-04-11 | Wake Forest University Health Sciences | Bioink compositions and methods of preparing and using the same |
| CN110305337A (en)* | 2018-03-27 | 2019-10-08 | 天津大学 | Unwrapped albumin hydrogel and preparation method thereof |
| CN110559483A (en)* | 2018-06-06 | 2019-12-13 | 广州溯原生物科技有限公司 | Design and application of cancellous bone bionic scaffold prepared by 3d printing technology |
| CN111253862A (en)* | 2020-01-17 | 2020-06-09 | 陕西师范大学 | Amyloid protein/reduced graphene oxide composite coating and preparation method and application thereof |
| CN111295183A (en)* | 2017-07-10 | 2020-06-16 | 康特拉费克特公司 | Blood component enhancement of lytic protein antibacterial activity and methods and uses thereof |
| CN111803718A (en)* | 2020-07-02 | 2020-10-23 | 西安交通大学医学院第二附属医院 | A kind of anti-fibrotic drug sustained-release coating and preparation method thereof |
| CN112264272A (en)* | 2020-10-14 | 2021-01-26 | 陕西师范大学 | A kind of multifunctional natural superhydrophobic material and its application |
| US20210324539A1 (en)* | 2018-07-26 | 2021-10-21 | President And Fellows Of Harvard College | Alpha-keratin solutions comprising alpha-kera tin intermediate filaments in liquid crystal phase, methods of preparation, and uses thereof |
| CN114712332A (en)* | 2022-03-15 | 2022-07-08 | 陕西师范大学 | Modified water-based material and preparation method and application thereof |
| CN114805888A (en)* | 2022-03-29 | 2022-07-29 | 陕西师范大学 | Method for improving surface bioactivity and osseointegration performance of polyether-ether-ketone substrate |
- 2023
- 2023-01-31CNCN202310070891.7Apatent/CN115970056B/enactiveActive
Patent Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102836461A (en)* | 2012-09-25 | 2012-12-26 | 天津工业大学 | Human hair keratin shape righting material with adjustable and controllable mechanical property as well as preparation method thereof |
| WO2014124546A1 (en)* | 2013-02-12 | 2014-08-21 | Eth Zurich | Hybrid nanocomposite materials of amyloid fibrils and gold |
| CN103756955A (en)* | 2014-01-22 | 2014-04-30 | 清华大学 | Personalized bionic composite structure as well as preparation method of composite structure and method for drug screening by using composite structure |
| CN111295183A (en)* | 2017-07-10 | 2020-06-16 | 康特拉费克特公司 | Blood component enhancement of lytic protein antibacterial activity and methods and uses thereof |
| US20190106673A1 (en)* | 2017-10-11 | 2019-04-11 | Wake Forest University Health Sciences | Bioink compositions and methods of preparing and using the same |
| CN110305337A (en)* | 2018-03-27 | 2019-10-08 | 天津大学 | Unwrapped albumin hydrogel and preparation method thereof |
| CN110559483A (en)* | 2018-06-06 | 2019-12-13 | 广州溯原生物科技有限公司 | Design and application of cancellous bone bionic scaffold prepared by 3d printing technology |
| US20210324539A1 (en)* | 2018-07-26 | 2021-10-21 | President And Fellows Of Harvard College | Alpha-keratin solutions comprising alpha-kera tin intermediate filaments in liquid crystal phase, methods of preparation, and uses thereof |
| CN111253862A (en)* | 2020-01-17 | 2020-06-09 | 陕西师范大学 | Amyloid protein/reduced graphene oxide composite coating and preparation method and application thereof |
| CN111803718A (en)* | 2020-07-02 | 2020-10-23 | 西安交通大学医学院第二附属医院 | A kind of anti-fibrotic drug sustained-release coating and preparation method thereof |
| CN112264272A (en)* | 2020-10-14 | 2021-01-26 | 陕西师范大学 | A kind of multifunctional natural superhydrophobic material and its application |
| CN114712332A (en)* | 2022-03-15 | 2022-07-08 | 陕西师范大学 | Modified water-based material and preparation method and application thereof |
| CN114805888A (en)* | 2022-03-29 | 2022-07-29 | 陕西师范大学 | Method for improving surface bioactivity and osseointegration performance of polyether-ether-ketone substrate |
Non-Patent Citations (4)
| Title |
|---|
| GANG WEI: "Self-assembling peptide and protein amyloids: from structure to tailored function in nanotechnology", CHEMICAL SOCIETY REVIEWS, vol. 46, no. 15, 22 May 2017 (2017-05-22), pages 4661 - 4708* |
| JIN GU: "An Environmentally Benign Antimicrobial Coating Based on a Protein Supramolecular Assembly", ACS APPLIED MATERIALS & INTERFACES, vol. 9, no. 1, 16 December 2016 (2016-12-16), pages 198 - 210* |
| JINGJIE CHEN: "Converting lysozyme to hydrogel: A multifunctional wound dressing that is more than antibacterial", COLLOIDS AND SURFACES B: BIOINTERFACES, vol. 219, 16 September 2022 (2022-09-16), pages 112854, XP087222201, DOI: 10.1016/j.colsurfb.2022.112854* |
| 伏成玉: "蛋白质类淀粉样聚集及涂层应用研究", 化学学报, 10 October 2023 (2023-10-10), pages 1 - 12* |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115970056B (en) | 2024-07-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4510939B2 (en) | Integral multi-stack tissue graft construct and method for making an improved integral multi-stack tissue graft construct | |
| TW529958B (en) | Oriented biopolymeric membrane | |
| US8216602B2 (en) | Nerve guide | |
| JP4074666B2 (en) | Tubular grafts from purified submucosa | |
| US5201745A (en) | Visceral surgery patch | |
| ES2213835T3 (en) | SUBMUCOSAL INTESTINAL FABRICS GRAFT FOR THE REPAIR OF NEUROLOGICAL FABRICS. | |
| Joshi et al. | 4D Printed Programmable Shape‐Morphing Hydrogels as Intraoperative Self‐Folding Nerve Conduits for Sutureless Neurorrhaphy | |
| JP2003190271A (en) | Organic-inorganic composite biomaterial and method for producing the same | |
| KR101293209B1 (en) | structure for guided tissue regeneration | |
| CN107050520A (en) | Compound bio sticking patch and preparation method thereof | |
| US11291747B2 (en) | Cell-seeded porous lung hydrogel sealant | |
| CN102238971A (en) | Solder formulation and use in tissue welding | |
| Lee et al. | Amnion‐analogous medical device for fetal membrane healing: a preclinical long‐term study | |
| WO2012086988A9 (en) | Artificial silk membrane having excellent flexibility and suturing ability and method of manufacturing the same | |
| CN107041967A (en) | A kind of feature self-assembled nanometer polypeptide hydrogel material and its application in hemostatic material is prepared | |
| Yang et al. | A novel biopolymer device fabricated by 3D printing for simplifying procedures of pancreaticojejunostomy | |
| Sloff et al. | Novel tubular constructs for urinary diversion: a biocompatibility study in pigs | |
| CN115970056A (en) | A kind of 3D printing material for bone repair, bone repair material and its preparation method and application | |
| Jamalpour et al. | 3D-printed bi-layered polymer/hydrogel construct for interfacial tissue regeneration in a canine model | |
| AU2004255202B2 (en) | Oriented biopolymeric membrane for meningeal tissue repair | |
| US9370604B2 (en) | Planar implant | |
| Brouwer et al. | Repair of surgically created diaphragmatic defect in rat with use of a crosslinked porous collagen scaffold | |
| CN102240233B (en) | Degradable stent for sternum defect repair and manufacturing method thereof | |
| CN108324336A (en) | A kind of New cranial bottom repair materials and its preparation method and application | |
| Hoeppner et al. | Small intestinal submucosa as a bioscaffold for tissue regeneration in defects of the colonic wall |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |