本文選題：三維打印技術(shù)　切入點：納米羥基磷灰石　出處：《吉林大學(xué)》2017年碩士論文

【摘要】：目的:因意外傷害、炎癥、腫瘤及先天畸形所致骨缺損在臨床上十分常見,這種缺損常常會影響患者的正常生理形態(tài)和生理功能,從而嚴(yán)重影響患者的生活[1]。目前臨床上用于治療這類骨缺損的方式有自體骨移植,異體骨移植以及人工骨移植等傳統(tǒng)治療方法。但是這些傳統(tǒng)的治療方法都存在著各自的局限性[2]。隨著材料科學(xué)、生命科學(xué)和工程技術(shù)的發(fā)展,組織工程技術(shù)在應(yīng)用于骨缺損的修復(fù)方面越來越受到人們的重視。組織工程包括三個基本要素,分別為種子細(xì)胞、支架、生物因子。其中支架作為聯(lián)系細(xì)胞與周圍環(huán)境的的橋梁,對于組織工程的成敗有重要的影響。因而,支架的選擇十分重要。理想的骨組織工程支架材料的研究范圍十分廣泛,它應(yīng)具有良好的生物相容性、骨誘導(dǎo)性、骨傳導(dǎo)性,與人骨相適宜的生物力學(xué)性能,合理的生物降解性以及足夠的孔隙率和孔隙尺寸結(jié)構(gòu)。同時在臨床應(yīng)用方面,支架材料又應(yīng)具備較高的可塑性,來源充足,加工簡單,價格低廉等優(yōu)點。在本實驗中,采用三維打印技術(shù)制備的羥基磷灰石(HA)/β-磷酸三鈣(β-TCP)復(fù)合雙相磷酸鈣陶瓷(BCP)支架,探究其在骨組織工程中的應(yīng)用潛力。方法:通過三維打印技術(shù)制備HA/β-TCP復(fù)合納米雙相磷酸鈣陶瓷支架,其中HA與β-TCP以質(zhì)量比為HA/β-TCP:3/7的比例混合。并以相同材料制備的壓模片。使用掃描電鏡(SEM)觀察支架形態(tài),X線衍射(XRD)檢測支架物相。分離培養(yǎng)兔骨髓間充質(zhì)干細(xì)胞(BMSC),將支架與兔BMSC共培養(yǎng);通過CCK-8實驗、堿性磷酸酶實驗檢測支架的生物相容性以及種子細(xì)胞在支架上的增殖分化。結(jié)果:三維打印技術(shù)制備的BCP支架高3mm、直徑分別3mm和8mm,圓柱形。壓模片的形狀與BCP支架相同。三維打印支架的孔隙結(jié)構(gòu)大小均勻,互相連通,孔隙的孔徑大小在350-450μm之間�？紫侗诒砻婵梢娫S多微孔結(jié)構(gòu)。CCK-8結(jié)果顯示:在體外培養(yǎng)1d、3d、5d后,空白組的細(xì)胞數(shù)量明顯高于實驗組及對照組(P0.05)。培養(yǎng)1d后,對照組細(xì)胞數(shù)量高于實驗組(P0.05),而3d、5d后,對照組與實驗組細(xì)胞數(shù)量已無明顯差異(P0.05)。培養(yǎng)7d后,對照組的細(xì)胞數(shù)量低于實驗組及空白組(P0.05),實驗組與空白組的細(xì)胞數(shù)量無明顯差異。ALP活性檢測試驗顯示:在體外培養(yǎng)4、7、14d后,實驗組的ALP活性均明顯高于對照組(P0.05)。培養(yǎng)4d后對照組的ALP活性則高于空白組(P0.05)。而培養(yǎng)7、14d后,對照組與空白組的ALP活性無明顯差異。結(jié)論:通過上述實驗,結(jié)果表明:通過三維打印技術(shù)制備的HA/β-TCP復(fù)合雙相磷酸鈣陶瓷支架,具有良好的三維孔隙結(jié)構(gòu)且孔隙間互相交通。支架的生物相容性良好,BMSC能夠在支架上很好的粘附、增殖及分化。相較于平面結(jié)構(gòu)的支架,BCP支架的多孔結(jié)構(gòu)以及微孔形成了更大的表面積,有利于細(xì)胞粘附增殖分化。通過本實驗,證明了三維打印技術(shù)制備的雙相磷酸鈣陶瓷支架在骨組織工程中的應(yīng)用潛力,并為后續(xù)的實驗及臨床應(yīng)用奠定了前期基礎(chǔ)。
[Abstract]:Objective: bone defects caused by accidental injury, inflammation, tumor and congenital malformation are very common in clinical practice. These defects often affect the normal physiological morphology and function of patients. This seriously affected the life of the patients [1] .At present, autogenous bone transplantation is used to treat this kind of bone defect in clinic. Traditional treatments such as allogeneic bone transplantation and artificial bone transplantation have their own limitations. [2] with the development of material science, life science and engineering technology, Tissue engineering technology has attracted more and more attention in the application of bone defect repair. Tissue engineering includes three basic elements: seed cells and scaffolds. Biological factors. Scaffolds, as a bridge between cells and surrounding environment, play an important role in the success or failure of tissue engineering. Therefore, the selection of scaffolds is very important. It should have good biocompatibility, bone inductivity, bone conductivity, biomechanical properties suitable for human bone, reasonable biodegradability and adequate porosity and pore size structure. In this experiment, hydroxyapatite / 尾 -tricalcium phosphate (尾 -TCP) composite biphasic calcium phosphate ceramic scaffold was prepared by three-dimensional printing technique. Methods: HA/ 尾 -TCP composite nano-biphasic calcium phosphate ceramic scaffolds were prepared by three-dimensional printing technique. The HA and 尾 -TCP were mixed in the ratio of HA/ 尾 -TCPW / 3 / 7 and the compression tablets were prepared from the same material. The morphology of the scaffold was observed by scanning electron microscopy (SEM) and X-ray diffraction (XRD) was used to detect the phase of the scaffold. The rabbit bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured. The scaffold was co-cultured with rabbit BMSC. Through the CCK-8 experiment, Alkaline phosphatase assay was used to detect the biocompatibility of the scaffold and the proliferation and differentiation of seed cells on the scaffold. Results: the BCP scaffold prepared by 3D printing technique was 3 mm high, with diameter of 8 mm and 3mm diameter of 8 mm. The same rack. The pore structure of the three-dimensional printing bracket is uniform, The pore size ranged from 350 渭 m to 450 渭 m. The results of CCK-8 showed that the number of cells in the blank group was significantly higher than that in the experimental group and the control group after 1 d of culture for 3 d or 5 d. After 1 day of culture, the number of cells in the blank group was significantly higher than that in the experimental group and the control group. The number of cells in the control group was higher than that in the experimental group, but there was no significant difference in the number of cells between the control group and the experimental group after 3 days. The number of cells in the control group was lower than that in the experimental group and the blank group, and there was no significant difference in the number of cells between the experimental group and the blank group. The activity of ALP in the experimental group was significantly higher than that in the control group (P 0.05). The activity of ALP in the control group was higher than that in the blank group after 4 days of culture. However, there was no significant difference in the activity of ALP between the control group and the blank group after 7d of culture. The results show that the HA/ 尾 -TCP composite biphasic calcium phosphate ceramic scaffold prepared by 3D printing technique has good three-dimensional pore structure and good biocompatibility. Proliferation and differentiation. Compared with planar scaffold BCP scaffolds, the porous structure and micropores form a larger surface area, which is conducive to cell adhesion, proliferation and differentiation. The application potential of the biphasic calcium phosphate ceramic scaffold prepared by 3D printing technology in bone tissue engineering is proved, and the preliminary foundation for subsequent experiment and clinical application is established.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R318.08;TP391.73

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