微孔发泡注射成型制备三维多孔聚乳酸骨组织工程支架研究
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摘要
微孔发泡注射成型采用超临界CO_2/N_2做为物理发泡剂，具有可显著减轻制品重量、缩短成型周期和极大改善制品翘曲变形和尺寸稳定性等特点，将微孔发泡注射成型技术应用于制备三维多孔连通结构的生物可降解骨组织工程支架，不涉及任何有机溶剂且批量生产显著降低成本，对人类骨组织重建和再生医学的发展具有重要的现实意义和科学研究价值。
聚乳酸强度高、成本低廉、生物相容性好且可降解，目前在组织工程领域已得到广泛应用。然而将聚乳酸这种半结晶性聚合物应用微孔发泡注射成型制备真三维连通结构且具有高孔隙率的支架仍存在较多加工难题，因此研究聚乳酸的微孔发泡注射成型成核机理和加工特性，对于制备理想结构的生物骨组织工程支架具有重要的指导意义。据此，本文从结晶、流变特性和异相成核等角度着手，做了以下几方面的工作：
首先，介绍了微孔发泡注射成型过程泡孔成核和长大的机理，研究了超临界流体作用下结晶对聚合物流变特性和发泡形态结构的影响。针对半结晶型聚合物气体溶解和扩散相对困难、溶解度低，发泡温度窗口窄，熔体强度对温度敏感等特点，以及超临界流体注入会降低聚合物熔体粘度的客观实际情况，研究了结晶对半结晶型聚合物泡孔成核和长大的影响机理。研究发现，结晶有助于提高聚合物熔体的储能模量和熔体弹性，对制备良好形态结构的发泡制品具有积极作用。结晶不仅可以增加成核点数目，而且作为一种异相成核剂存在能够促进泡孔成核使得泡孔密度增大，同时还可以有效阻止泡孔长大从而减小泡孔的平均尺寸。
其次，为了克服气熔混合不足和纳米粒子在复合材料中分散不均带来的制品“皮芯”结构和力学性能下降的问题，将预发泡技术引入微孔发泡注射成型工艺中，开发了一种气熔混合效果好、可有效改善发泡结构和性能的方法。采用挤出发泡制备的含气发泡粒料，使得纳米复合材料在进入微孔发泡注射成型机之前已经基本充分形成了分散均匀的微纳结构，而且提高了超临界流体与聚合物熔体的配混效果。实验结果表明，采用预发泡颗粒制备的微孔注射成型纳米复合材料，皮芯层的均匀度明显改善，泡孔平均尺寸减小，泡孔密度增大，提高了发泡制品的力学性能。
然后，根据理想骨组织工程支架的结构特征，利用共混法调控了聚乳酸共混体系的降解速率和力学性能，研究了它们的热性能、动态热机械性能和发泡形态结构。通过调整加工工艺参数，制备了具有双尺度模式泡孔结构的高孔隙率发泡制品。人类骨髓间质干细胞的培养实验结果表明，所制备的支架酸碱度适当，无毒性，细胞附着和生长繁殖良好，在PLA/PHBV(55:45)支架上培育14天的细胞呈现骨髓前体细胞典型树突结构。MTS分析测定结果表明，细胞在所制备的支架上繁殖速度较快，数目多，其中PLA/PHBV(55:45)试样尤为突出，验证了该种配方和工艺条件下制备的双尺度连通发泡结构的支架适合细胞培养，具有良好的载体作用。
最后，从熔体粘弹特性、结晶和异相成核机理等三重角度研究了纳米粒子改善发泡制品泡孔形态结构的机理和方法，讨论了纳米粒子对泡孔成核和结晶成核的双重作用，分析了纳米粒子增强复合材料的流变特性的响应规律。对制备真三维多孔连通、可调控的力学性能和降解速率的纳米填充复合高分子骨组织工程支架具有重要的研究价值。实验结果表明：聚乳酸纳米复合材料在整个频率扫描范围内呈现非常明显的剪切变稀现象，这种pseudo solid like行为充分说明了纳米粘土与高分子之间形成很强的界面作用，这种类似于物理交联点的作用限制了高分子链的松弛和滑移，增强了熔体粘度。
Microcellular injection molding produces components with excellent dimensionalstability while using lower injection pressures, shorter cycle time, and less material. Duringthis process, supercritical fluid (SCF) CO_2or N_2was used as the physical blowing agent.The microcellular injection molding technology is being applied to the fabrication of the3Dinterconnected biodegradable bone tissue engineering scaffold, taking advantage of thecontrolled weight saving and significant reduction s in cycle time without chemical solventinvolved. Most notably, the process is being applied in the bone regeneration in thebio-medical area.
Poly lactic acid is a compostable polymer that can be derived completely fromrenewable resources. It has been used for the medical and biodegradability. One of thecritical steps in the production of microcellular polymers is the dissolution of gas into apolymer matrix. For gas and semi-crystalline thermoplastic systems, the solution formationprocess is notably more complex. The crystallization results in a solution that is relativelydifficult to microcellular process requiring relatively high temperature compared toamorphous polymer/gas solutions. In addition, the crystallization of the solution results in alower solubility, increased matrix stiffness, and a lower diffusivity. This paper is to present asystematic study of the gas dissolution process in semi-crystalline polymers in an attempt toprovide further insight and engineering analysis into the roles of gas dissolution withinduced crystallization, viscoelastic behavior, and crystallniity in microcellular processing.
Firstly, the mechanism of the cell nucleation and growth theories were introduced tofurther investigate the effect of the crystallinity and rheological properties on the cellmorphology. To study the solution’s gas dissolution and crystallization characteristics, ouranalysis include an experimental characterization of the carbon dioxide-inducedcrystallization occurring during microcellular polymer processing indicating a critical gasconcentration is required for crystallization and an experimental estimation of theviscoelastic behavior of semi-crystalline solution, and an experimental investigation of the effects of crystallinity on microcellular processing and the resulting cell morphology.
Secondly, the dispersion of the nano-particle in the nanocomposites is a knownproblem either in the conventional or microcellular injection molding process. To improvingthe morphology and structure of the nanocomposites, the pre-foam technology wasintroduced to the microcellular injection molding to produce a single polymer-gas solutionand uniform foaming parts with uniform properties. This method involves producing thepellets with gas laden by extrusion foaming to improve the dispersion of the nano-particle inthe microcellular injection molding specimens and also decrease the skin-core structureunder the shear flow.
Thirdly, based on the requirements of3D interconnected bone tissue engineeringscaffold, the thermal, dynamic thermo-mechanical property, and the cell morphology of thePLA blend were studied to tune the mechanical properties and biodegradable rate to accessthe optimal processing condition. One kind of bio-model cell structure was produced withhigh porosity. The cell culture experiments were conducted on the PLA/PHBV scaffold, itwas found that the cell can attach on the scaffold very well, and the cell proliferation wassignificant, indicating the scaffold was suitable for the cell attachment and proliferation.
Finally, the effect of the nanoclay on the nucleation of crystallinity and cell was studied.Based on the viscoelastic behavior characterization of polymer-gas solutions, this study hasfound that solutions of crystalline polymer and gas tend to have higher storage modulicompared with their amorphous counterparts. The crystallization and the resulting change inviscoelastic behavior tend to play a major role in microcellular processing. It was also foundthat crystallization influences microcellular processing through its effects on cell nucleationmechanisms resulting in larger cell densities and cell growth mechanisms resulting insmaller cell sizes.
聚乳酸强度高、成本低廉、生物相容性好且可降解，目前在组织工程领域已得到广泛应用。然而将聚乳酸这种半结晶性聚合物应用微孔发泡注射成型制备真三维连通结构且具有高孔隙率的支架仍存在较多加工难题，因此研究聚乳酸的微孔发泡注射成型成核机理和加工特性，对于制备理想结构的生物骨组织工程支架具有重要的指导意义。据此，本文从结晶、流变特性和异相成核等角度着手，做了以下几方面的工作：
首先，介绍了微孔发泡注射成型过程泡孔成核和长大的机理，研究了超临界流体作用下结晶对聚合物流变特性和发泡形态结构的影响。针对半结晶型聚合物气体溶解和扩散相对困难、溶解度低，发泡温度窗口窄，熔体强度对温度敏感等特点，以及超临界流体注入会降低聚合物熔体粘度的客观实际情况，研究了结晶对半结晶型聚合物泡孔成核和长大的影响机理。研究发现，结晶有助于提高聚合物熔体的储能模量和熔体弹性，对制备良好形态结构的发泡制品具有积极作用。结晶不仅可以增加成核点数目，而且作为一种异相成核剂存在能够促进泡孔成核使得泡孔密度增大，同时还可以有效阻止泡孔长大从而减小泡孔的平均尺寸。
其次，为了克服气熔混合不足和纳米粒子在复合材料中分散不均带来的制品“皮芯”结构和力学性能下降的问题，将预发泡技术引入微孔发泡注射成型工艺中，开发了一种气熔混合效果好、可有效改善发泡结构和性能的方法。采用挤出发泡制备的含气发泡粒料，使得纳米复合材料在进入微孔发泡注射成型机之前已经基本充分形成了分散均匀的微纳结构，而且提高了超临界流体与聚合物熔体的配混效果。实验结果表明，采用预发泡颗粒制备的微孔注射成型纳米复合材料，皮芯层的均匀度明显改善，泡孔平均尺寸减小，泡孔密度增大，提高了发泡制品的力学性能。
然后，根据理想骨组织工程支架的结构特征，利用共混法调控了聚乳酸共混体系的降解速率和力学性能，研究了它们的热性能、动态热机械性能和发泡形态结构。通过调整加工工艺参数，制备了具有双尺度模式泡孔结构的高孔隙率发泡制品。人类骨髓间质干细胞的培养实验结果表明，所制备的支架酸碱度适当，无毒性，细胞附着和生长繁殖良好，在PLA/PHBV(55:45)支架上培育14天的细胞呈现骨髓前体细胞典型树突结构。MTS分析测定结果表明，细胞在所制备的支架上繁殖速度较快，数目多，其中PLA/PHBV(55:45)试样尤为突出，验证了该种配方和工艺条件下制备的双尺度连通发泡结构的支架适合细胞培养，具有良好的载体作用。
最后，从熔体粘弹特性、结晶和异相成核机理等三重角度研究了纳米粒子改善发泡制品泡孔形态结构的机理和方法，讨论了纳米粒子对泡孔成核和结晶成核的双重作用，分析了纳米粒子增强复合材料的流变特性的响应规律。对制备真三维多孔连通、可调控的力学性能和降解速率的纳米填充复合高分子骨组织工程支架具有重要的研究价值。实验结果表明：聚乳酸纳米复合材料在整个频率扫描范围内呈现非常明显的剪切变稀现象，这种pseudo solid like行为充分说明了纳米粘土与高分子之间形成很强的界面作用，这种类似于物理交联点的作用限制了高分子链的松弛和滑移，增强了熔体粘度。
Microcellular injection molding produces components with excellent dimensionalstability while using lower injection pressures, shorter cycle time, and less material. Duringthis process, supercritical fluid (SCF) CO_2or N_2was used as the physical blowing agent.The microcellular injection molding technology is being applied to the fabrication of the3Dinterconnected biodegradable bone tissue engineering scaffold, taking advantage of thecontrolled weight saving and significant reduction s in cycle time without chemical solventinvolved. Most notably, the process is being applied in the bone regeneration in thebio-medical area.
Poly lactic acid is a compostable polymer that can be derived completely fromrenewable resources. It has been used for the medical and biodegradability. One of thecritical steps in the production of microcellular polymers is the dissolution of gas into apolymer matrix. For gas and semi-crystalline thermoplastic systems, the solution formationprocess is notably more complex. The crystallization results in a solution that is relativelydifficult to microcellular process requiring relatively high temperature compared toamorphous polymer/gas solutions. In addition, the crystallization of the solution results in alower solubility, increased matrix stiffness, and a lower diffusivity. This paper is to present asystematic study of the gas dissolution process in semi-crystalline polymers in an attempt toprovide further insight and engineering analysis into the roles of gas dissolution withinduced crystallization, viscoelastic behavior, and crystallniity in microcellular processing.
Firstly, the mechanism of the cell nucleation and growth theories were introduced tofurther investigate the effect of the crystallinity and rheological properties on the cellmorphology. To study the solution’s gas dissolution and crystallization characteristics, ouranalysis include an experimental characterization of the carbon dioxide-inducedcrystallization occurring during microcellular polymer processing indicating a critical gasconcentration is required for crystallization and an experimental estimation of theviscoelastic behavior of semi-crystalline solution, and an experimental investigation of the effects of crystallinity on microcellular processing and the resulting cell morphology.
Secondly, the dispersion of the nano-particle in the nanocomposites is a knownproblem either in the conventional or microcellular injection molding process. To improvingthe morphology and structure of the nanocomposites, the pre-foam technology wasintroduced to the microcellular injection molding to produce a single polymer-gas solutionand uniform foaming parts with uniform properties. This method involves producing thepellets with gas laden by extrusion foaming to improve the dispersion of the nano-particle inthe microcellular injection molding specimens and also decrease the skin-core structureunder the shear flow.
Thirdly, based on the requirements of3D interconnected bone tissue engineeringscaffold, the thermal, dynamic thermo-mechanical property, and the cell morphology of thePLA blend were studied to tune the mechanical properties and biodegradable rate to accessthe optimal processing condition. One kind of bio-model cell structure was produced withhigh porosity. The cell culture experiments were conducted on the PLA/PHBV scaffold, itwas found that the cell can attach on the scaffold very well, and the cell proliferation wassignificant, indicating the scaffold was suitable for the cell attachment and proliferation.
Finally, the effect of the nanoclay on the nucleation of crystallinity and cell was studied.Based on the viscoelastic behavior characterization of polymer-gas solutions, this study hasfound that solutions of crystalline polymer and gas tend to have higher storage modulicompared with their amorphous counterparts. The crystallization and the resulting change inviscoelastic behavior tend to play a major role in microcellular processing. It was also foundthat crystallization influences microcellular processing through its effects on cell nucleationmechanisms resulting in larger cell densities and cell growth mechanisms resulting insmaller cell sizes.
引文
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