用于牙髓損傷修復的新型微納米生物活性玻璃的仿生制備及性能研究
發(fā)布時間：2018-10-05 07:38
【摘要】：當牙髓受到齲病和外傷的侵襲，容易引發(fā)牙髓損傷，從而影響牙齒的健康與功能，因此保存活髓或誘導牙髓牙本質(zhì)再生是牙髓病學領(lǐng)域的研究熱點。生物活性玻璃作為第三代生物活性材料具有生物相容性、骨傳導和骨誘導性，是一類重要的骨修復材料，而牙本質(zhì)的組成和形成機制與骨類似，提示它用于牙髓牙本質(zhì)修復再生治療的可能性。然而，傳統(tǒng)生物活性玻璃易團聚、形貌不規(guī)則、尺寸結(jié)構(gòu)不可控等問題影響了其活性離子的釋放，限制了其在組織缺損修復中的應用。人體天然的組織因其具有獨特的微納米結(jié)構(gòu)而展現(xiàn)出傳統(tǒng)人工合成材料無法比擬的生物功能優(yōu)勢。因此從仿生角度出發(fā)，本文采用溶膠-凝膠技術(shù)結(jié)合有機模板自組裝技術(shù)，設計制備了具有形貌、結(jié)構(gòu)可控，尺寸、組分可調(diào)的新型微納米生物活性玻璃，并研究了微納米生物活性玻璃形貌、尺寸、結(jié)構(gòu)的控制機理、物理化學性質(zhì)以及體外磷灰石形成能力，研究了不同形貌、不同尺寸、不同組成的微納米生物活性玻璃對人牙髓細胞增殖、分化行為的影響，并通過裸鼠皮下埋植生物活性玻璃與牙髓組織的復合體，考察生物活性玻璃在體內(nèi)誘導牙髓細胞礦化的能力。主要研究工作和結(jié)論如下： （1）采用堿催化溶膠-凝膠技術(shù)結(jié)合模板自組裝技術(shù)，，以溴化十六烷基三甲銨（CTAB）作為模板劑能夠有效的控制生物活性玻璃顆粒的形態(tài)從球形（SBG）到短棒形（SRBG）再到長棒形（LRBG），孔結(jié)構(gòu)從無序到有序，CTAB的濃度決定了顆粒的形態(tài)，催化劑氨水的濃度決定了顆粒的直徑大小。三種形貌的生物活性玻璃均具有良好的體外磷灰石形成能力，且相比球形和短棒形顆粒，長棒形生物活性玻璃具有更強的體外磷灰石形成能力。生物活性玻璃顆粒對人牙髓細胞增殖與顆粒濃度有關(guān)，高濃度（200μg/mL）時對細胞增殖有抑制作用，低濃度（100μg/mL）時，能夠促進細胞增殖。且在低濃度條件下，從促進細胞增殖和ALP蛋白表達的效果來看，SBG SRBG LRBG。 （2）以十二胺（DDA）作為水解催化劑和結(jié)構(gòu)形成模板劑，采用堿催化溶膠-凝膠技術(shù)結(jié)合模板協(xié)同組裝技術(shù)能夠有效制備出單分散微納米生物活性玻璃球形顆粒（MBGS）。調(diào)節(jié)DDA加入量可以在微納米尺寸范圍內(nèi)調(diào)控生物玻璃顆粒大小。體外磷灰石形成能力以及離子釋放研究表明生物活性玻璃尺寸越小，離子釋放速率更快，其體外磷灰石形成能力越強。在生物活性玻璃濃度為100μg/mL條件下，生物活性玻璃顆粒的尺寸大小對細胞增殖和ALP蛋白表達的影響有明顯區(qū)別，更小尺寸的生物活性玻璃MBGS-1顯示出更好的細胞相容性。 （3）在第三章制備工藝基礎(chǔ)之上，摻入一定量的Sr以取代Ca，制備摻Sr微納米生物活性玻璃（BG-Sr），生物活性玻璃依然呈現(xiàn)單分散的規(guī)則微納米球形形態(tài)，且對顆粒大小、晶相結(jié)構(gòu)以及化學組成沒有明顯的影響，但隨著Sr摻入量的增多，生物活性玻璃微球的比表面積增大，Si離子釋放速度增大，體外磷灰石形成能力降低。Sr離子的濃度對細胞增殖和分化有一定影響，摻入適量的Sr（6mol%）呈現(xiàn)較好的細胞增殖能力、ALP分化能力以及細胞礦化能力，而當Sr摻入量達到15mol%時，其細胞增殖和分化能力都受到抑制。 （4）采用溶膠-凝膠技術(shù)結(jié)合模板自組裝技術(shù)，選用CTAB作為模板劑，氨水為催化劑，成功制備空腔大小可控的微納米中空介孔生物活性玻璃球（HMBGS）。且平均粒徑以及內(nèi)部空腔隨著醇水比的增大而增大，隨CTAB濃度的增大而減小。同時HMBGS均具有較高的比表面積（444.003~609.512m2/g）和良好的體外磷灰石形成活性。 （5）進一步考察生物活性玻璃體內(nèi)誘導牙髓細胞礦化的能力，將45S5生物玻璃、58S生物玻璃以及單分散微納米生物活性玻璃球MBGS分別與剪碎的裸鼠牙髓組織混合，進行皮下埋植6周后，通過HE染色和Masson染色觀察發(fā)現(xiàn)，不會出現(xiàn)明顯的炎癥細胞，提示生物活性玻璃具有良好的生物相容性。三組生物活性玻璃均能夠促進牙髓細胞外基質(zhì)的分泌和礦化，且相比45S5組和58S組，MBGS組新生的礦化基質(zhì)更多，提示MBGS促進礦化作用更強。
[Abstract]:When endodontic pulp is affected by caries and trauma, dental pulp injury can be easily induced, thus affecting the health and function of teeth. Bioactive glass, as the third generation bioactive material, has biocompatibility, bone conduction and osteoinducibility, is a kind of important bone repair material, and the composition and formation mechanism of dentin is similar to bone, suggesting it can be used in the treatment of dental pulp dentin repair and regeneration. However, the problems of easy agglomeration, irregular morphology and uncontrollable size structure of traditional bioactive glass affect the release of active ions and limit their application in tissue defect repair. the natural tissue of the human body exhibits the biological functional advantage of traditional artificial synthetic materials due to its unique micro-nano structure. Therefore, the micro-nano biological active glass with morphology, controllable structure, size and composition can be prepared by combining sol-gel technology and organic template self-assembly technology, and the morphology and size of micro-nano biological active glass are studied. The mechanism, physical and chemical properties of the structure and the ability of apatite formation in vitro were studied. The effects of different morphology, size and composition of micro-nano bioactive glass on the proliferation and differentiation of human dental pulp cells were studied. and the ability of bioactive glass to induce the mineralization of dental pulp cells in vivo is investigated through the composite of the biological active glass and the dental pulp tissue under the skin of the naked mouse. The main research work and conclusions are as follows: (1) adopting alkali catalysis sol-gel technology to combine template self-assembly According to the technology, the morphology of bioactive glass particles can be effectively controlled from spherical (SBG) to short rod shape (SRBG) to long rod shape (LRBG) as a template by using bromohexadecyltrimethylamine (CTAB) as a template agent, the concentration of ammonia in the catalyst determines the diameter of the particles, The bioactive glass with three kinds of morphology has good in vitro apatite forming ability, and has stronger in vitro apatite formation compared with spherical and short rod shaped particles. Ability. Bioactive glass particles have an inhibitory effect on cell proliferation at a high concentration (200 & mu; g/ mL) with respect to human dental pulp cell proliferation and particle concentration, and can promote cells at low concentrations (100 & mu; g/ mL) Proliferation. In low concentrations, SBG SRBG LR was seen from the effect of promoting cell proliferation and ALP expression. BG. (2) adopts dodecylamine (DDA) as the hydrolysis catalyst and structure to form template agent, and adopts alkali-catalyzed sol-gel technology combined with template co-assembly technology to effectively prepare monodisperse micro-nano biological active glass spherical particles (MBGS). Adjust the amount of DDA to adjust the bio-glass within the micro-nano-size range. Particle size, apatite formation ability in vitro and ion release study show that the smaller the size of bioactive glass, the faster the ion release rate and the apatite shape in vitro The stronger the bio-active glass particle size on cell proliferation and the expression of ALP protein under the condition of the bioactive glass concentration of 100. m u.g/ mL, the biological active glass MBGS-1 with smaller size is better than that of the biological active glass MBGS-1. Cell compatibility. (3) On the basis of the preparation of the third chapter, a certain amount of Sr is added to replace Ca, so as to prepare the Sr micro-nano biological active glass (BG-Sr), and the bioactive glass still exhibits a monodisperse regular micro-nano-spherical shape. In addition, with the increase of Sr incorporation, the specific surface area of bioactive glass microspheres increases, the release rate of Si ions increases, and in vitro phosphorus The concentration of Sr ions has a certain effect on cell proliferation and differentiation, and a proper amount of Sr (6mol%) has better cell proliferation ability, ALP differentiation ability and cell mineralization ability, but when Sr incorporation reaches 15mol%, its cell proliferation and differentiation (4) adopting a sol-gel technology combined template self-assembly technology, adopting CTAB as a template agent and ammonia as a catalyst, and successfully preparing the micro-nano hollow porous biological activity with controllable cavity size; The average particle size and the internal cavity increase with the increase of the water ratio of the alcohol, At the same time, HMBGS has higher specific surface area (444. 003 ~ 609. 512m2/ g) and good (5) the ability of biological active glass to induce the mineralization of dental pulp cells is further investigated, 45S5 biological glass, 58S biological glass and monodisperse micro-nano biological active glass spheres MBGS are respectively connected with the cut bare rat dental pulp. After 6 weeks of subcutaneous injection, the tissue was mixed and observed by HE staining and Masson staining, and no obvious inflammatory cells were present, suggesting that the organism The bioactive glass has good biocompatibility, and the three groups of bioactive glass can promote the secretion and mineralization of the extracellular matrix of the pulp, and compared with 45S5 and 58S, the MBGS group has more mineralized matrix,
【學位授予單位】：華南理工大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R318.08;TQ317
本文編號：2252556
[Abstract]:When endodontic pulp is affected by caries and trauma, dental pulp injury can be easily induced, thus affecting the health and function of teeth. Bioactive glass, as the third generation bioactive material, has biocompatibility, bone conduction and osteoinducibility, is a kind of important bone repair material, and the composition and formation mechanism of dentin is similar to bone, suggesting it can be used in the treatment of dental pulp dentin repair and regeneration. However, the problems of easy agglomeration, irregular morphology and uncontrollable size structure of traditional bioactive glass affect the release of active ions and limit their application in tissue defect repair. the natural tissue of the human body exhibits the biological functional advantage of traditional artificial synthetic materials due to its unique micro-nano structure. Therefore, the micro-nano biological active glass with morphology, controllable structure, size and composition can be prepared by combining sol-gel technology and organic template self-assembly technology, and the morphology and size of micro-nano biological active glass are studied. The mechanism, physical and chemical properties of the structure and the ability of apatite formation in vitro were studied. The effects of different morphology, size and composition of micro-nano bioactive glass on the proliferation and differentiation of human dental pulp cells were studied. and the ability of bioactive glass to induce the mineralization of dental pulp cells in vivo is investigated through the composite of the biological active glass and the dental pulp tissue under the skin of the naked mouse. The main research work and conclusions are as follows: (1) adopting alkali catalysis sol-gel technology to combine template self-assembly According to the technology, the morphology of bioactive glass particles can be effectively controlled from spherical (SBG) to short rod shape (SRBG) to long rod shape (LRBG) as a template by using bromohexadecyltrimethylamine (CTAB) as a template agent, the concentration of ammonia in the catalyst determines the diameter of the particles, The bioactive glass with three kinds of morphology has good in vitro apatite forming ability, and has stronger in vitro apatite formation compared with spherical and short rod shaped particles. Ability. Bioactive glass particles have an inhibitory effect on cell proliferation at a high concentration (200 & mu; g/ mL) with respect to human dental pulp cell proliferation and particle concentration, and can promote cells at low concentrations (100 & mu; g/ mL) Proliferation. In low concentrations, SBG SRBG LR was seen from the effect of promoting cell proliferation and ALP expression. BG. (2) adopts dodecylamine (DDA) as the hydrolysis catalyst and structure to form template agent, and adopts alkali-catalyzed sol-gel technology combined with template co-assembly technology to effectively prepare monodisperse micro-nano biological active glass spherical particles (MBGS). Adjust the amount of DDA to adjust the bio-glass within the micro-nano-size range. Particle size, apatite formation ability in vitro and ion release study show that the smaller the size of bioactive glass, the faster the ion release rate and the apatite shape in vitro The stronger the bio-active glass particle size on cell proliferation and the expression of ALP protein under the condition of the bioactive glass concentration of 100. m u.g/ mL, the biological active glass MBGS-1 with smaller size is better than that of the biological active glass MBGS-1. Cell compatibility. (3) On the basis of the preparation of the third chapter, a certain amount of Sr is added to replace Ca, so as to prepare the Sr micro-nano biological active glass (BG-Sr), and the bioactive glass still exhibits a monodisperse regular micro-nano-spherical shape. In addition, with the increase of Sr incorporation, the specific surface area of bioactive glass microspheres increases, the release rate of Si ions increases, and in vitro phosphorus The concentration of Sr ions has a certain effect on cell proliferation and differentiation, and a proper amount of Sr (6mol%) has better cell proliferation ability, ALP differentiation ability and cell mineralization ability, but when Sr incorporation reaches 15mol%, its cell proliferation and differentiation (4) adopting a sol-gel technology combined template self-assembly technology, adopting CTAB as a template agent and ammonia as a catalyst, and successfully preparing the micro-nano hollow porous biological activity with controllable cavity size; The average particle size and the internal cavity increase with the increase of the water ratio of the alcohol, At the same time, HMBGS has higher specific surface area (444. 003 ~ 609. 512m2/ g) and good (5) the ability of biological active glass to induce the mineralization of dental pulp cells is further investigated, 45S5 biological glass, 58S biological glass and monodisperse micro-nano biological active glass spheres MBGS are respectively connected with the cut bare rat dental pulp. After 6 weeks of subcutaneous injection, the tissue was mixed and observed by HE staining and Masson staining, and no obvious inflammatory cells were present, suggesting that the organism The bioactive glass has good biocompatibility, and the three groups of bioactive glass can promote the secretion and mineralization of the extracellular matrix of the pulp, and compared with 45S5 and 58S, the MBGS group has more mineralized matrix,
【學位授予單位】：華南理工大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R318.08;TQ317
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