明胶溶胀行为对自固化多孔骨水泥性能和结构的影响

目的 研究明胶溶胀行为对多孔骨水泥性能和结构的影响。方法 在α-磷酸钙骨水泥体系中加入生物明胶，研究明胶对骨水泥水化产物、抗压强度和产物微结构所产生的影响。结果利用明胶的溶胀行为与水化过程中体系pH值变化的相关性，可制备具有大孔和微孔结构的骨水泥。结论加入明胶促进羟基磷灰石的成核，提高骨水泥的抗压强度。     

磷酸钙骨水泥复合骨形态发生蛋白6和血管内皮生长因子修复骨缺损

背景：单独将骨形态发生蛋白或血管内皮生长因子植入体内易被血液冲刷掉而不能最大限度发挥诱导成骨和血管生成作用,同时缺少载体的支撑作用。 目的：观察骨形态发生蛋白6、血管内皮生长因子及磷酸钙骨水泥联合应用在骨缺损修复过程中的作用。 方法：制作新西兰兔双侧股骨内侧髁骨缺损模型,左侧分别植入磷酸钙骨水泥/骨形态发生蛋白6/血管内皮生长因子、磷酸钙骨水泥/骨形态发生蛋白6及磷酸钙骨水泥,右侧不植入任何物质作为空白对照。植入8,16周通过硬组织切片组织学观察、电镜扫描等手段观察新骨形成情况。 结果与结论：各组材料的组织相容性良好,未见明显炎症组织反应。植入8周时,磷酸钙骨水泥/骨形态发生蛋白6/血管内皮生长因子组骨水泥-骨组织交界处基本上被新生骨小梁包绕,材料进一步降解,新生骨小梁表面可见大量活跃的成骨细胞;16周时,新生骨小梁继续长入,进一步增长、增粗、增多,有大量新生编织骨成网格状长入材料中,骨水泥材料降解明显,与周围组织结合紧密,降解与骨长入同步,此组不同时间点成骨速度及成骨效果均明显优于其他两组材料(P < 0.05)。表明3种材料联合应用可协同促进骨缺损修复。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程全文链接：     

磷酸钙骨水泥的改性研究进展

磷酸钙骨水泥作为一种新型人工骨替代材料 ,以其良好的生物相容性和骨传导性被广泛应用于临床骨缺损修复。但其存在固化时间较长、机械性能不足及降解缓慢等缺点 ,使其应用受到一定限制 ,故需要对其进行改性研究 ,本文就此做一综述如下     

磷酸钙骨水泥的研究和临床进展

磷酸钙骨水泥是一种极好生物相容性和生物可降解的非陶瓷型羟基磷灰石类生物材料,以被广泛应用到骨科、外科、口腔科等医学领域.本文主要介绍该材料在生物性能、操作性能及临床应用方面的研究进展.     

磷酸钙骨水泥的生物学研究进展

本综述了磷酸钙骨水泥作为一种新型人工骨替代材料近年来的生物学基础及提高其生物学性能的研究,阐明了磷酸钙骨水泥以其良好的生物学特性,使其在骨缺损修复领域具有广阔的应用前景。     

磷灰石骨水泥材料的物性研究

为探讨自行研制的磷灰石骨水材料材料作为生物材料应用的可能性,采用磷酸钙、碳酸钙和磷酸氢钙为起始原料,经高温固相反应合成制得仅含有羧磷灰石和α－磷酸三钙两晶相组成的粉剂,以柠檬酸水溶液为基本组成配制若干种固化液。结果显示,当固化液采用单一柠檬酸时,３０％（ｗ／ｗ）浓度水溶液的压缩强度为２４．９６ＭＰａ,凝固时间为６分钟,水中溶解率为１．４０％;当液剂中添加钾和钠盐时,可使固化体的压缩强度增强;当添加     

磷酸钙骨水泥/纤维蛋白胶复合材料填充桡骨缺损的生物相容性

BACKGROUND: The chemical compositions and structure of calcium phosphate bone cement are similar to those of human bone, which can fill the bone collapse caused by fracture and induce osteogenesis, but its degradation rate is slow. OBJECTIVE: To evaluate the biocompatibility of the calcium phosphate cement/fibrin glue and the feasibility of repairing radius defects. METHODS: In vitro cytotoxicity experiment: Mouse fibroblasts were cultured in the calcium phosphate bone cement/fibrin glue extracts, phenol solution, and RPMI 1640 culture medium containing 10% fetal bovine serum, respectively, to detect the cytotoxicity grade. Hemocompatibility experiment: Calcium phosphate bone cement/fibrin glue extracts, normal saline and distilled water were respectively added into the rabbit anticoagulation, to detect the hemolytic rate. Forty-five New Zealand white rabbits were enrolled and modeled into bilateral radius defects, followed by randomly allotted into three groups: blank control group without any intervention, experimental and control groups were given the implantation with calcium phosphate bone cement/fibrin glue and autologous radius, respectively. X-ray, histology, bone mineral density and biomechanical test were performed at postoperative 4, 8 and 16 weeks. RESULTS AND CONCLUSION: The toxicity grade of the calcium phosphate cement/fibrin glue was 0 to 1. The hemolytic rate of the calcium phosphate cement/fibrin glue was 3.15%. At 16 weeks postoperatively, X-ray showed that in the experimental and control groups, the fracture line disappeared completely, pulp cavity was recanalized, and in plastic completely. Histology showed that the reconstructed bone trabecular was obvious, plate layer of bone was mature, and medullary cavity recanalization appeared in the control group; there were a large number of new grid-shaped woven bone tissues growing into the material in the experimental group, with overt degradation, and degradation rate was in parallel to bone ingrowth. The bone density, the maximum load, maximum stress and failure energy in the experimental and control groups were significantly higher than those in the blank control group (P < 0.05), and all above indicators showed no significant differences between the experimental and control groups. These results manifest that the calcium phosphate bone cement/fiber protein glue composite material holding a good biocompatibility can promote bone tissue regeneration for bone defect repair, achieving similar curative effect with autologous bone transplantation.     

磷酸钙骨水泥的生物学研究进展

本文综述了磷酸钙骨水泥作为一种新型人工骨替代材料近年来的生物学基础及提高其生物学性能的研究 ,阐明了磷酸钙骨水泥以其良好的生物学特性 ,使其在骨缺损修复领域具有广阔的应用前景。     

磷酸钙的固有骨诱导性及其应用

BACKGROUND: With good biocompatibility, osteoconduction and biodegradability, calcium phosphate ceramics is considered as a substitute of autologous bone; furthermore, it also has the potentiel of osteoinduction after structure optimization.     

磷酸钙骨水泥的改性研究进展

磷酸钙骨水泥作为一种新型人工骨替代材料,以其良好的生物相容性和骨传导性被广泛应用于临床骨缺损修复。但其存在固化时间较长、机械性能不足及降解缓慢等缺点,使其应用受到一定限制,故需要对其进行改性研究,本就此做一综述如下。     

Proliferation and differentiation of osteoblast-like MC3T3-E1 cells on biomimetically and electrolytically deposited calcium phosphate coatings

Biomimetic and electrolytic deposition are versatile methods to prepare calcium phosphate coatings. In this article, we compared the effects of biomimetically deposited octacalcium phosphate and carbonate apatite coatings as well as electrolytically deposited carbonate apatite coating on the proliferation and differentiation of mouse osteoblast-like MC3T3-E1 cells. It was found that MC3T3-E1 cells cultured on the biomimetically deposited carbonate apatite coating demonstrated the greatest proliferation rate and the highest differentiation potential. Cells on the biomimetically deposited octacalcium phosphate coating had lower proliferation rate before day 7, but higher after that, than those on the electrolytically deposited carbonate apatite coating. There was no difference on the expression of early differentiation markers, that is, alkaline phosphatase activity and collagen content, between biomimetically deposited octacalcium phosphate and electrolytically deposited carbonate apatite coatings. However, higher expression of late differentiation markers, that is, osteocalcin and bone sialoprotein mRNA, was found on the biomimetically deposited octacalcium phosphate coating on day 14. These results suggest that the difference in in vitro osteoblast cell performance of calcium phosphate coatings might relate to their physicochemical properties. Biomimetic carbonate apatite coating is the most favorable surface for the proliferation and differentiation of MC3T3-E1 cells.     

钛表面微弧氧化涂层的细胞生物活性

背景：微弧氧化技术可改善钛或钛合金的表面特征。 目的：研究纯钛表面微弧氧化涂层的表面性能及其对MC3T3-E1细胞早期黏附、增殖及成骨能力的影响。 方法：将46个直径10 mm、厚度2 mm圆盘状纯钛试件分为实验组和对照组。实验组置于含0.02 mol/Lβ-甘油磷酸二钠盐及0.2 mol/L乙酸钙的电解液中进行微弧氧化处理,对照组对试件进行机械抛光。扫描电子显微镜观察试件表面形貌,X射线能谱分析检测涂层表面钙磷比,X射线衍射分析检测涂层晶相构成。将MC3T3-E1细胞接种在两组试件表面,1,2,4 h电镜下观察细胞形态,在2,4,7 d通过CCK-8方法检测细胞增殖,并于7,14 d检测碱性磷酸酶活性。 结果与结论：经微弧氧化处理后,钛表面形成粗糙多孔的钙磷涂层,微弧氧化涂层主要元素为Ca、P、O及Ti,微弧氧化膜层主要由氧化钛、钛酸钙、磷酸钙及偏磷酸钙构成。电镜观察显示1 h 微弧氧化涂层表面细胞已伸出伪足,4 h呈现较典型的细胞形态。细胞在微弧氧化处理钛表面4,7 d的细胞增殖和7,14 d的碱性磷酸酶活性高于对照组。表明微弧氧化技术生成的粗糙多孔钙磷涂层能显著促进MC3T3-E1细胞的早期黏附、增殖及成骨活性。     

In vitro response of osteoblast-like and odontoblast-like cells to unsubstituted and substituted apatites

Different types of calcium phosphate compounds [calcium-deficient apatite (CDA); beta-tricalcium phosphate (beta-TCP); biphasic calcium phosphate (BCP)] are commercially available for medical and dental applications as bone substitute materials. Most of the reported in vitro studies on cell-material interactions have used osteoblast-like cells. The purpose of this study was to investigate the in vitro response of osteoblast-like (MC3T3-E1) and odontoblast-like (MDPC23) cells on unsubstituted (HA) and substituted (F-substituted) apatites. MC3T3-E1 and MDPC23 were cultured in alpha-modified medium containing 10% fetal bovine serum, ascorbic acid (50 microg/mL) and beta-glycerophosphate (2 mM). The cells were seeded on pellets made from HA, and FAp (with low, medium, and high F concentrations). Cell morphology was observed after 7 and 14 days using scanning electron microscopy (SEM). Cell attachment and differentiation were determined from the DNA content, alkaline phosphatase (ALP) activity, and total collagen content. Pellet surface composition was characterized by using Fourier Transform infrared spectroscopy. MC3T3-E1 and MDPC23 cells on HA were normal in shape and in fusion but not on FAp. Results of this study showed that the pattern of cell proliferation of osteoblast-like cells was different from that of the odontoblast-like cells. This study suggests that cell morphology, fusion, and proliferation on biomaterial surfaces depend on cell type (osteoblast-like vs odontoblast-like cell) and biomaterial composition (unsubstituted vs substituted F-apatites).     

Enhancing effect of poly(L-lactide) on the differentiation of mouse osteoblast-like MC3T3-E1 cells

Poly(L-lactide) (PLLA) has bioabsorbability and biocompatibility, and it is used as biodegradable screws, pins and plates for internal bone fixation. The purpose of this study was to clarify the effects of low molecular weight (Mw) PLLA on the proliferation and differentiation of mouse osteoblast-like MC3T3-E1 cells. MC3T3-E1 cells were cultured with the concentration of 5-50 microg/ml of PLLA with weight average Mw of 5000 (PLLA-5k) and 10,000 (PLLA-10k) for 2 weeks using the micromass culture. Both PLLAs did not affect the proliferation of MC3T3-E1 cells. However, the calcifications of MC3T3-E1 cells were stimulated with increasing the concentration of the PLLAs. Then PLLA-5k increased the calcification of MC3T3-E1 cells more than PLLA-10k. Additionally, both PLLAs increased the alkaline phosphatase (ALP) activity and calcium content of MC3T3-E1 cells up to the similar level to the calcification. These results indicated that low Mw PLLA enhanced the differentiation of MC3T3-E1 cells with no effect on the proliferation. Moreover, it was suggested that the increase of the ALP activity was a key step to stimulate the calcification of MC3T3-E1 cells. The osteoconductivity of implanted PLLA would be based on the enhancing effect of low Mw PLLA on the differentiation of the osteoblasts.     

Osteoblast response to zirconia-hybridized pyrophosphate-stabilized amorphous calcium phosphate

Calcium phosphate bioceramics, such as hydroxyapatite, have long been used as bone substitutes because of their proven biocompatibility and bone binding properties in vivo. Recently, a zirconia-hybridized pyrophosphate-stabilized amorphous calcium phosphate (Zr-ACP) has been synthesized, which is more soluble than hydroxyapatite and allows for controlled release of calcium and phosphate ions. These ions have been postulated to increase osteoblast differentiation and mineralization in vitro. The focus of this work is to elucidate the physicochemical properties of Zr-ACP and to measure cell response to Zr-ACP in vitro using a MC3T3-E1 mouse calvarial-derived osteoprogenitor cell line. Cells were cultured in osteogenic medium and mineral was added to culture at different stages in cell maturation. Culture in the presence of Zr-ACP showed significant increases in cell proliferation, alkaline phosphatase activity (ALP), and osteopontin (OPN) synthesis, whereas collagen synthesis was unaffected. In addition, calcium and phosphate ion concentrations and medium pH were found to transiently increase with the addition of Zr-ACP, and are hypothesized to be responsible for the osteogenic effect of Zr-ACP.     

Effect of gypsum on proliferation and differentiation of MC3T3-E1 mouse osteoblastic cells

Recently, calcium sulfate dihydrate has been demonstrated as safe biodegradable osteoconductive bone void filler. However, its exact mechanism of action on bone cells is yet unknown. In this study, the influence of gypsum on gene expression and proliferation of MC3T3-E1 mouse pre-osteoblastic cells was investigated. Cells were cultured on gypsum disc, slice, polymethylmethacrylate (PMMA), or plastic culture plate for 15 days. Cell viability, alkaline phosphatase (ALP) activity and expression profile of 15 genes involved in bone metabolism were measured in cultures. Cell proliferation on gypsum was increased by almost 2-fold, while an inhibitory effect of PMMA on proliferation rate of osteoblasts was noted. Cells cultured on gypsum disc surface exhibited an increased ALP activity and markedly different gene expression profile. Quantitative real-time PCR data indicated the expression of genes that might provide a basis for an osteoinductive potential. MC3T3-E1 cells expressed genes typical of bone fracture healing like type II collagen and fibronectin 1. These effects might be related to the calcium content of gypsum and mediated likely via SMAD3. Our results suggest that gypsum can support new bone formation by its calcium content and modulatory effect on gene expression profile of bone cells.     

IL-38 通过调控PI3K/ Akt/ GSK3β/ NFATc1 信号通路抑制骨质疏松的机制研究

目的:探讨IL-38 抑制骨质疏松的作用并研究其分子机制。方法:共纳入2014 年6 月~2016 年12 月我院收治的138 例骨质疏松患者作为研究对象。另选取120 例同期在我院骨科进行骨折手术的无骨质疏松患者作为对照。采用ELISA 法检测实验对象血清IL-38 水平。构建IL-38-C57BL/6J 转基因小鼠,建立骨质疏松小鼠模型,将野生型、IL-38 转基因小鼠分别设置为假手术组(Sham 组)与卵巢切除组(Ovariectomy,OVX 组)。术后8 周取小鼠血清,检测碱性磷酸酶(ALP)、血钙及血磷水平。另取小鼠的脊柱与双侧股骨,通过病理切片分析股骨组织形态结构,用骨密度仪检测脊柱骨密度变化。将各组小鼠的骨髓基质细胞(BMSCs)进行分离并检测其体外增殖能力,Western blot 检测各组BMSCs 的PI3K、Akt、GSK3β与NFATc1 的磷酸化水平。小鼠成骨细胞MC3T3-E1 转染IL-38 后,Western blot 检测PI3K、Akt、GSK3β与NFATc1 磷酸化水平的变化,流式细胞术检测IL-38 对细胞凋亡的影响。结果:骨质疏松组患者的血清IL-38 水平显著低于对照组(P<0.05)。野生型与IL-38 转基因OVX 小鼠的血钙、血磷水平均显著高于Sham 组(P<0.05),而ALP 水平显著低于Sham 组(P<0.05)。另外,IL-38 转基因OVX 小鼠的血钙和血磷水平均显著低于野生型OVX 小鼠(P<0.05)。股骨病理切片及脊柱骨密度分析显示,野生型与IL-38 转基因OVX 小鼠均出现骨组织形态结构破坏和骨密度下降,并且IL-38 转基因OVX 小鼠的骨组织形态结构破坏和骨密度下降情况均较野生型OVX 小鼠显著减轻(P<0.05)。IL-38 转基因OVX 小鼠BMSCs 的体外增殖能力显著高于野生型OVX 组(P<0.05)。IL-38 转基因OVX 小鼠BMSCs 的PI3K、Akt 与NFATc1 磷酸化水平均显著低于野生型OVX 组(P<0.05),GSK3β磷酸化水平显著高于野生型OVX 组(P<0.05)。MC3T3-E1 细胞转染IL-38 后PI3K、Akt 与NFATc1 的磷酸化水平均显著降低(P<0.05),GSK3β的磷酸化水平显著升高(P<0.05)。流式细胞检测显示转染IL-38 后MC3T3-E1 细胞的凋亡显著减少(P<0.05)。结论:骨质疏松患者的血清IL-38 水平显著降低,IL-38 可能通过调控PI3K/ Akt/ GSK3β/ NFATc1 信号通路促进BMSCs 增殖、抑制成骨细胞凋亡,从而抑制骨质疏松的进展。     

无机纳米粒子掺杂生物活性碳纳米纤维的制备及性能评价

背景：已有研究表明向生物惰性碳纳米纤维中引入具有成骨活性的β-磷酸三钙纳米粒子,可显著提高碳纳米纤维的生物活性,而某些二价离子掺杂的β-磷酸三钙也被报道能够促进新骨的生成。 目的：考察适量锌离子、镁离子的引入对β-磷酸三钙@碳纳米纤维材料形貌及成骨活性的影响。 方法：以聚丙烯腈、磷酸三乙酯、硝酸钙、硝酸锌、硝酸镁等为原料,采用溶胶-凝胶、静电纺丝与原位烧结碳化相结合的方法制备锌或镁离子掺杂的β-磷酸三钙@碳纳米纤维材料。将所得复合纳米纤维材料及未掺杂锌或镁离子掺杂的β-磷酸三钙@碳纳米纤维与成骨细胞MC3T3-E1体外共培养,观察细胞的黏附、增殖和形态变化。 结果与结论：β-磷酸三钙@碳纳米纤维形貌均匀,表面可见直径为数十纳米的无机粒子均匀分布,锌或镁离子的引入对纤维形貌无明显影响;复合纤维主要由碳元素组成,钙、锌、镁元素等均匀分布于纤维中,且各元素相对含量与投料比相符。与未掺杂锌或镁离子的β-磷酸三钙@碳纳米纤维相比,MC3T3-E1成骨细胞更易在锌或镁离子掺杂的β-磷酸三钙@碳纳米纤维材料表面黏附,细胞增殖和铺展状态也更好。表明在β-磷酸三钙@碳纳米纤维的基础上,引入锌或镁离子掺杂,能进一步提高材料的细胞相容性及生物活性。     

miR-132-3p过表达对MC3T3-E1细胞增殖分化的影响

文题释义： miRNA：是一类小的非编码RNA,长度约为22个核苷酸,其主要通过结合靶标mRNA的3'UTR区诱导靶标mRNA降解或抑制其翻译,从而在转录及转录后水平调控相关基因的表达。miRNA在细胞增殖、分化及凋亡等多种生物学活动过程中起着非常重要的调控作用,并且发现它是调控骨组织细胞增殖和分化过程的重要因子之一。 MC3T3-E1细胞：是新生C57BL/6小鼠颅顶骨中分离培养所建立的一株成骨细胞株,它能够展现骨组织中成骨细胞的各个发育阶段和各种生物学特性。作为研究成骨细胞增殖和分化的理想模型,被广泛应用于国内外各种骨组织工程学研究。 背景：机械牵引力能够影响MC3T3-E1细胞的增殖分化过程,并引起细胞内miR-132-3p的差异表达。然而,牵引力是否通过调控miR-132-3p的表达来影响成骨细胞增殖分化仍需进一步研究。 目的：明确12%牵引力作用下MC3T3-E1细胞中成骨分化标志因子及miR-132-3p表达变化,并进一步探讨miR-132-3p对细胞增殖分化的影响。 方法：MC3T3-E1细胞分别加载0%,12%牵张应力,检测应力加载后碱性磷酸酶活性、骨钙蛋白及miR-132-3p mRNA的表达水平;细胞内瞬时转染miR-132-3p模拟物及其阴性对照,qRT-PCR检测转染后碱性磷酸酶、骨钙蛋白、Runt标志转录因子2 mRNA的表达,CCK-8法检测miR-132-3p对细胞增殖能力的影响。 结果与结论：①12%牵张应力作用下,MC3T3-E1细胞中碱性磷酸酶活性、骨钙蛋白mRNA表达水平下调(P < 0.01),miR-132-3p表达水平显著升高(P < 0.05);②细胞内转染miR-132-3p后,miR-132-3p模拟物组成骨分化标志因子碱性磷酸酶、骨钙蛋白、Runt标志转录因子2 mRNA表达水平显著降低(P < 0.05);③相比于阴性对照组,miR-132-3p 模拟物转染24,48,72 h后细胞增殖能力明显降低(P < 0.001),且在转染48 h后降低最明显;④结果说明12%周期性循环牵张应力能够通过过表达miR-132-3p负向调节MC3T3-E1细胞的增殖和成骨分化。 ORCID: 0000-0003-0696-3329(孙芬) 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程