多方式引入硅元素改性磷酸钙骨水泥及其性能研究
发布时间：2018-10-13 18:22
【摘要】：磷酸钙骨水泥(CPC)具有良好的可塑型性、生物相容性和生物活性以及自固化特性,被广泛用于骨修复材料领域。但由于CPC存在固化时间过长、力学强度不高、抗溃散性差以及骨诱导性不足等缺点,限制了其在临床上的应用。硅(Si)元素对维持机体的正常生长和骨骼的形成极其重要,它参与骨的钙化过程,尤其在新骨的形成过程中发挥着重要作用。为了克服CPC的这些缺点,本研究将Si元素以不同形式引入CPC中,制备综合性能优异的含Si骨水泥,为解决CPC存在的问题探讨新的途径和方法。将无定形硅酸钙(ACS)添加到CPC中,制备出ACS/CPC复合骨水泥。ACS的加入能够促进CPC的水化反应,使CPC的凝结时间缩短、抗压强度提高。当ACS的添加量为5 wt%时,ACS/CPC的凝结时间比纯CPC的凝结时间(40 min)缩短了11分钟,抗压强度(35.7±3.6 MPa)提高了35%,可注射性仍然保持在95%以上,满足临床操作的要求。ACS/CPC在模拟体液(SBF)中沉积羟基磷灰石(HA)的速度比纯CPC更快,使得ACS/CPC在浸泡后期的失重率减小。细胞生物学研究表明,相比于纯CPC,ACS/CPC能显著促进小鼠骨髓间质干细胞(mBMSCs)和人脐静脉内皮细胞(HUVECs)的粘附、增殖,以及促进mBMSCs向成骨细胞分化。在CPC中添加硅灰石(WS),对WS粉末粒径以及含量对CPC理化性能的影响进行了系统的研究。在WS颗粒尺寸大于106μm时,WS/CPC的凝结时间随着WS粒径的增大而延长,抗压强度下降。WS粒径在53-106μm的WS/CPC具有更好的理化性能。因此,选择WS的最佳粒径范围为53-106μm。WS能够促进CPC的水化反应,但不影响其水化产物的物相。WS的含量超过20 wt%时,WS/CPC的可注射性急剧下降。在CPC中添加WS可以缩短CPC的凝结时间、提高抗压强度,同时其孔隙率和可降解性有所降低。较之纯CPC,含有适量WS的WS/CPC对mBMSCs和HUVECs表现出更好的反应性。采用化学沉淀法在部分结晶磷酸钙(PCCP)中掺杂Si元素,经分析可知Si能够进入PCCP中的结晶相HA晶格,使晶胞参数发生改变,晶胞体积增大。少量掺杂Si使其中的HA相结晶变得较完善,晶粒主要为针状晶体,但过量掺杂Si又会使HA的结晶性变差。将掺Si的PCCP(Si-PCCP)与DCPA混合得到Si掺杂的磷酸钙骨水泥(Si-CPC)。相比纯CPC,Si-CPC的凝结时间有所缩短,抗压强度有所提高,并且Si-CPC比纯CPC具有更高的生物活性。细胞学实验表明,相比纯CPC,Si-CPC能够显著促进mBMSCs和HUVECs的粘附、增殖以及mBMSCs的分化。根据上述三种含硅组分(ACS、WS和Si-PCCP)溶解度的不同,通过将三者组合来调控Si的释放速率,制备出可控Si释放的磷酸钙骨水泥。同时添加两种或者三种含Si组分的骨水泥比单一含Si组分的骨水泥具有更优越的理化性能和细胞生物学性能,三种含Si组分共掺的骨水泥(3SiCPC)性能最优。3SiCPC在SBF中降解14天后达到平衡,溶液中的Si浓度变化较小,在28天内基本保持稳定在较高的浓度值。培养基中Si浓度在适当范围内时,多种含Si组分共掺的CPC具有更好的细胞反应性。通过在上述可控Si释放的磷酸钙骨水泥3SiCPC中添加槐豆胶(LBG)和多巴胺(DA)作为抗溃散剂,制备了抗溃散性良好的可注射型磷酸钙骨水泥。这两种抗溃散剂的添加均能明显改善3SiCPC的抗溃散性。在LBG含量不超过1.0 wt%时,3SiCPC的凝结时间稍有延长、抗压强度和可注射性略有降低。并且,含有1.0 wt%LBG的3SiCPC对mBMSCs表现出良好的细胞相容性。当DA的添加量不超过0.4 wt%时,3SiCPC的凝结时间稍有延长,抗压强度略有提高。相比纯3SiCPC,含有0.4 wt%DA的3SiCPC更有利于蛋白的吸附,对mBMSCs和HUVECs具有更好的细胞响应。
[Abstract]:Calcium phosphate cement (CPC) has good formability, biocompatibility and bioactivity, as well as self-curing properties, and is widely used in the field of bone repair materials. However, due to the long curing time of CPC, the mechanical strength is not high, the fracture resistance is poor and the osteoinducibility is insufficient and so on, the application of the CPC in clinic is limited. Silicon (Si) element plays an important role in maintaining the normal growth of the body and the formation of bone, it participates in the calcification process of bone, especially plays an important role in the formation of new bone. In order to overcome these disadvantages of CPC, this study introduces Si elements into CPC in different forms, prepares Si-containing cement with excellent comprehensive properties, and discusses new ways and methods for solving the problems of CPC. An ACS/ CPC composite bone cement was prepared by adding amorphous calcium silicate (ACS) into CPC. The addition of ACS can promote the hydration reaction of CPC, shorten the coagulation time of CPC and improve the compressive strength. When the addition amount of ACS is 5 wt%, the coagulation time of ACS/ CPC is shortened by 11 minutes compared with the coagulation time (40 min) of pure CPC, the compressive strength (35. 7 MPa 3. 6 MPa) is improved by 35%, the injectability is still kept above 95%, and the requirement of clinical operation is met. ACS/ CPC deposited hydroxyapatite (HA) in simulated body fluid (SBF) faster than pure CPC, so that the weight loss rate of ACS/ CPC in late infusion was decreased. Cell biological studies have shown that ACS/ CPC can significantly promote the adhesion and proliferation of bone marrow mesenchymal stem cells (mBMSCs) and human umbilical vein endothelial cells (HUVECs) in mice compared with pure CPC, and promote the differentiation of mBMSCs into osteoblasts. The influence of particle size and content of WS powder on the physical and chemical properties of CPC was studied by adding wollastonite (WS) in CPC. When the size of WS particles is larger than 106. m u.m, the coagulation time of WS/ CPC is extended with the increase of WS particle diameter and the compressive strength decreases. WS particle size is 53-106. m u.m and WS/ CPC has better physical and chemical properties. Therefore, the optimum particle size range of the selected WS is 53-106. m u.m. WS can promote the hydration reaction of CPC, but does not affect the product phase of its hydration product. When the content of WS exceeds 20 wt%, the injection properties of WS/ CPC are drastically reduced. Adding WS to CPC can shorten the coagulation time of CPC, improve the compressive strength, and decrease its porosity and degradability. Compared with pure CPC, WS/ CPC containing a proper amount of WS showed better reactivity to mBMSCs and HUVECs. Si element is doped in partial crystalline calcium phosphate (PCCP) by chemical precipitation method. Through analysis, Si can enter crystalline phase HA crystal lattice in PCCP, change cell parameters and increase unit cell volume. With a small amount of Si doped with Si, the crystal of HA phase becomes more perfect, and grains are mainly needle-like crystals, but excessive doping of Si can lead to poor crystallinity of HA. Si-doped calcium phosphate cement (Si-CPC) was obtained by mixing Si-PCCP and DCPA. Compared with pure CPC, the condensation time of Si-CPC is shortened, the compressive strength is improved, and Si-CPC has higher bioactivity than pure CPC. The cytology test showed that compared with pure CPC, Si-CPC could significantly promote the adhesion, proliferation and differentiation of mBMSCs and HUVECs. According to the different solubility of the three silicon-containing components (ACS, WS and Si-PCCP), calcium phosphate cement with controllable Si release was prepared by combining the three combinations to control the release rate of Si. At the same time, the bone cement with two or three Si-containing components has better physical and chemical properties and cell biological properties than bone cement with a single Si-containing component, and the three-Si-containing co-doped bone cement (3SiCPC) performance is optimal. 3SiCPC is equilibrated in SBF for 14 days, the concentration of si in the solution is small and remains substantially stable over 28 days at higher concentration values. When the concentration of Si in the culture medium is within proper range, a variety of CPC containing Si-containing components has better cell reactivity. The injectable calcium phosphate cement was prepared by adding locust bean gum (LBG) and dopamine (DA) into the calcium phosphate cement 3SiCPC released by the controllable Si as an anti-collapse agent. The addition of these two anti-crush agents can obviously improve the anti-sibility of 3SiCPC. When the LBG content does not exceed 1. 0% by weight, the coagulation time of 3SiCPC is slightly prolonged, and the compressive strength and injectability are slightly reduced. Also, 3SiCPC containing 1.0 wt% LBG exhibited good cell compatibility to mBMSCs. When the addition amount of DA is not more than 0.4 wt%, the coagulation time of 3SiCPC is slightly prolonged, and the compressive strength is slightly improved. Compared with pure 3SiCPC, 3SiCPC containing 0.4 wt% DA is more favorable for protein adsorption, and has better cell response to mBMSCs and HUVECs.
【学位授予单位】：华南理工大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R318.08
，
本文编号：2269520
[Abstract]:Calcium phosphate cement (CPC) has good formability, biocompatibility and bioactivity, as well as self-curing properties, and is widely used in the field of bone repair materials. However, due to the long curing time of CPC, the mechanical strength is not high, the fracture resistance is poor and the osteoinducibility is insufficient and so on, the application of the CPC in clinic is limited. Silicon (Si) element plays an important role in maintaining the normal growth of the body and the formation of bone, it participates in the calcification process of bone, especially plays an important role in the formation of new bone. In order to overcome these disadvantages of CPC, this study introduces Si elements into CPC in different forms, prepares Si-containing cement with excellent comprehensive properties, and discusses new ways and methods for solving the problems of CPC. An ACS/ CPC composite bone cement was prepared by adding amorphous calcium silicate (ACS) into CPC. The addition of ACS can promote the hydration reaction of CPC, shorten the coagulation time of CPC and improve the compressive strength. When the addition amount of ACS is 5 wt%, the coagulation time of ACS/ CPC is shortened by 11 minutes compared with the coagulation time (40 min) of pure CPC, the compressive strength (35. 7 MPa 3. 6 MPa) is improved by 35%, the injectability is still kept above 95%, and the requirement of clinical operation is met. ACS/ CPC deposited hydroxyapatite (HA) in simulated body fluid (SBF) faster than pure CPC, so that the weight loss rate of ACS/ CPC in late infusion was decreased. Cell biological studies have shown that ACS/ CPC can significantly promote the adhesion and proliferation of bone marrow mesenchymal stem cells (mBMSCs) and human umbilical vein endothelial cells (HUVECs) in mice compared with pure CPC, and promote the differentiation of mBMSCs into osteoblasts. The influence of particle size and content of WS powder on the physical and chemical properties of CPC was studied by adding wollastonite (WS) in CPC. When the size of WS particles is larger than 106. m u.m, the coagulation time of WS/ CPC is extended with the increase of WS particle diameter and the compressive strength decreases. WS particle size is 53-106. m u.m and WS/ CPC has better physical and chemical properties. Therefore, the optimum particle size range of the selected WS is 53-106. m u.m. WS can promote the hydration reaction of CPC, but does not affect the product phase of its hydration product. When the content of WS exceeds 20 wt%, the injection properties of WS/ CPC are drastically reduced. Adding WS to CPC can shorten the coagulation time of CPC, improve the compressive strength, and decrease its porosity and degradability. Compared with pure CPC, WS/ CPC containing a proper amount of WS showed better reactivity to mBMSCs and HUVECs. Si element is doped in partial crystalline calcium phosphate (PCCP) by chemical precipitation method. Through analysis, Si can enter crystalline phase HA crystal lattice in PCCP, change cell parameters and increase unit cell volume. With a small amount of Si doped with Si, the crystal of HA phase becomes more perfect, and grains are mainly needle-like crystals, but excessive doping of Si can lead to poor crystallinity of HA. Si-doped calcium phosphate cement (Si-CPC) was obtained by mixing Si-PCCP and DCPA. Compared with pure CPC, the condensation time of Si-CPC is shortened, the compressive strength is improved, and Si-CPC has higher bioactivity than pure CPC. The cytology test showed that compared with pure CPC, Si-CPC could significantly promote the adhesion, proliferation and differentiation of mBMSCs and HUVECs. According to the different solubility of the three silicon-containing components (ACS, WS and Si-PCCP), calcium phosphate cement with controllable Si release was prepared by combining the three combinations to control the release rate of Si. At the same time, the bone cement with two or three Si-containing components has better physical and chemical properties and cell biological properties than bone cement with a single Si-containing component, and the three-Si-containing co-doped bone cement (3SiCPC) performance is optimal. 3SiCPC is equilibrated in SBF for 14 days, the concentration of si in the solution is small and remains substantially stable over 28 days at higher concentration values. When the concentration of Si in the culture medium is within proper range, a variety of CPC containing Si-containing components has better cell reactivity. The injectable calcium phosphate cement was prepared by adding locust bean gum (LBG) and dopamine (DA) into the calcium phosphate cement 3SiCPC released by the controllable Si as an anti-collapse agent. The addition of these two anti-crush agents can obviously improve the anti-sibility of 3SiCPC. When the LBG content does not exceed 1. 0% by weight, the coagulation time of 3SiCPC is slightly prolonged, and the compressive strength and injectability are slightly reduced. Also, 3SiCPC containing 1.0 wt% LBG exhibited good cell compatibility to mBMSCs. When the addition amount of DA is not more than 0.4 wt%, the coagulation time of 3SiCPC is slightly prolonged, and the compressive strength is slightly improved. Compared with pure 3SiCPC, 3SiCPC containing 0.4 wt% DA is more favorable for protein adsorption, and has better cell response to mBMSCs and HUVECs.
【学位授予单位】：华南理工大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R318.08
，
本文编号：2269520
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