Name: synthesis of graphene-hydroxyapatite nanocomposites for potential use in bone tissue engineering
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Sougata Ghosh博士感谢印度政府科学技术部科学和技术部（DST）和印度贾瓦哈拉尔·尼赫鲁高级科学研究中心在纳米科学技术博士后海外奖学金（参考JNC / AO / A.0610.1（4）2019年8月19日的2019-2260）下获得资助。Sougata Ghosh博士感谢泰国曼谷泰国农业大学获得博士后奖学金，并资助重塑大学计划（参考号：6501.0207/10870，日期为2021年11月9日）。作者要感谢Kostas高级纳米表征设施（KANCF）对表征实验的帮助。KANCF是东北大学科斯塔斯研究所（KRI）内的共享多学科研究和教育设施。

1. 沉淀合成nHAP
<ol><li>使用含有1 M Ca（NO3）2∙4H 2 O和0.67 M（NH 4）H2PO 4的反应混合物的50 mL反应混合物合成原始nHAP，然后滴加NH4OH（25%）以保持pH值约为1018。</li>
	<li>此后，通过超声照射（UI）搅拌反应混合物30分钟（500 W功率和20 kHz超声频率）。</li>
	<li>让所得溶液在室温下成熟120小时，直到nHAP的白色沉淀沉淀...

<ol>
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虽然已经研究了各种金属，聚合物，陶瓷及其组合作为骨科植入物和固定配件，但HAP被认为是最优选的材料之一，因为它与骨骼本身具有化学相似性，因此细胞相容性高20，21，22。本研究的HAP取向不同，对其独特性质有显著影响，如促骨生成、骨整合、骨导等。此外，改变HAP的取向可以影响纳米复合材料的力学性能，以进一步...

石墨烯具有由sp杂交碳组成的片状二维结构。其他几种同素异形体可归因于石墨烯的扩展蜂窝网络（例如，石墨烯片的堆叠形成3D石墨，而从相同的材料上滚落导致1D纳米管1的形成）。同样，0D富勒烯是由于包裹2而形成的。石墨烯具有有吸引力的物理化学和光电特性，包括室温3，4下的双极场效应和量子霍尔效应。检测单分子吸附事件和极高的载流子迁移率增加了石墨烯5，6的吸引力。此外，石墨烯纳米带（GNRs）具有窄宽度和大平均自由程，低电阻率和高电流密度和高电子迁移率被认为是有前途的互连材料7。因此，正在探索GNR在无数设备中的应用，最近在纳米医学中的应用，特别是组织工程和药物递送8。
在各种创伤性疾病中，骨损伤被认为是最具挑战性的疾病之一，因为在稳定骨折，再生和用新骨替换，抵抗感染以及重新对齐骨不结合方面存在困难9，10。外科手术仍然是股骨干骨折的唯一替代方案。应该指出的是，中美洲和欧洲每年花费近5200万美元用于治疗骨损伤11.
用于骨组织工程应用的生物活性支架可以通过掺入纳米羟基磷灰石（nHAP）而更有效，因为它们类似于骨骼本身的微和纳米结构特性12。HAP，化学表示为Ca10（PO4）6（OH）2 ，Ca / P摩尔比为1.67，是生物医学应用的最优选，特别是用于治疗牙周缺损，硬组织的替代以及制造用于骨科手术的植入物13，14。因此，用GNRs增强的基于nHAP的生物材料的制备可以具有优异的生物相容性，并且由于其促进骨整合和骨传导的能力而可能是有利的15，16。这种杂交复合支架可以保持生物学特性，例如细胞粘附，扩散，增殖和分化17。在本文中，我们通过合理地改变nHAP和GNRs的空间排列，报告了两种用于骨组织工程的新型纳米复合材料的制备，如图 1所示。本文评估了两种不同nHAP-GNRs排列的化学和结构特性。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Ammonium phosphate monobasic</td>
			<td>Sigma-Aldrich</td>
			<td>216003-100G</td>
			<td>Synthesis</td>
		</tr>
		<tr>
			<td>Calcium nitrate tetrahydrate</td>
			<td>Sigma-Aldrich</td>
			<td>237124</td>
			<td>Synthesis</td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Hettich</td>
			<td>EBA 200S</td>
			<td>Recovery</td>
		</tr>
		<tr>
			<td>Fourier transform infrared spectrometer</td>
			<td>Brucker</td>
			<td>Vertex 70</td>
			<td>Characterization</td>
		</tr>
		<tr>
			<td>Graphene nanoribbon</td>
			<td>Sigma-Aldrich</td>
			<td>922714</td>
			<td>Synthesis</td>
		</tr>
		<tr>
			<td>High resolution transmission electron microscope</td>
			<td>Thermo Fisher Scientific</td>
			<td>Themis Titan 300</td>
			<td>Characterization</td>
		</tr>
		<tr>
			<td>Magnetic stirrer</td>
			<td>IKA</td>
			<td>C-MAG HS7 S68</td>
			<td>Functionalization</td>
		</tr>
		<tr>
			<td>Micropipettes</td>
			<td>TreffLab</td>
			<td>06H35687</td>
			<td>Reagent preparation</td>
		</tr>
		<tr>
			<td>pH meter</td>
			<td>Eutech pH5+</td>
			<td>ECPH503PLUSK</td>
			<td>Reagent preparation</td>
		</tr>
		<tr>
			<td>Thermogravimetric analyzer</td>
			<td>TA Instruments</td>
			<td>SDT Q600</td>
			<td>Characterization</td>
		</tr>
		<tr>
			<td>Ultrasonic bath</td>
			<td>Bandelin</td>
			<td>DT100</td>
			<td>Functionalization</td>
		</tr>
		<tr>
			<td>Universal Oven</td>
			<td>Memmert</td>
			<td>UF55</td>
			<td>Functionalization</td>
		</tr>
		<tr>
			<td>Weighing balance</td>
			<td>Precisa</td>
			<td>XB220A</td>
			<td>Reagent preparation</td>
		</tr>
		<tr>
			<td>X-ray diffractometer</td>
			<td>Brucker</td>
			<td>D8-Advanced</td>
			<td>Characterization</td>
		</tr>
	</tbody>
</table>

synthesis of graphene-hydroxyapatite nanocomposites for potential use in bone tissue engineering

开发用于骨组织工程的新型材料是纳米医学最重要的推动领域之一。已经用羟基磷灰石制造了几种纳米复合材料，以促进细胞粘附，增殖和成骨。在这项研究中，使用石墨烯纳米带（GNRs）和羟基磷灰石纳米颗粒（nHAPs）成功开发了杂化纳米复合材料，当用于生物活性支架时，可以潜在地改善骨组织再生。这些纳米结构可以是生物相容性的。在这里，使用了两种方法来制备新材料。在一种方法中，使用了一种共功能化策略，其中nHAP被合成并同时与GNR共轭，从而在GNR表面上产生nHAP的纳米杂化（表示为nHAP / GNR）。高分辨率透射电子显微镜（HRTEM）证实，nHAP/GNR复合材料由细长薄的GNRs结构（最大长度为1.8μm）和针状nHAP（长度为40-50nm）的离散斑块（150-250 nm）组成。在另一种方法中，市售的nHAP与GNRs共轭，形成GNR涂层的nHAP（表示为GNR / nHAP）（即相对于nHAP / GNR纳米杂缰相反的方向）。采用后一种方法形成的纳米杂化物表现出直径范围为50 nm至70 nm的nHAP纳米球，表面覆盖着GNRs网络。能量色散光谱、元素映射和傅里叶变换红外（FTIR）光谱证实了nHAP和GNR在两种纳米杂联中的成功整合。热重分析（TGA）表明，GNR/nHAP和gnH/GNR在加热温度升高时的损耗分别为0.5%和0.98%。具有相反方向的nHAP-GNR纳米杂交代表了用于生物活性支架的重要材料，可以潜在地促进细胞功能以改善骨组织工程应用。

断续器分析 单独来看，GNR是细长的竹状结构，在一定距离处有一些弯曲，如图 2所示。最长的GNR为1.841μm，而最小的弯曲GNR为497nm。纳米带通常显示出明显的宽度变化，这可能归因于在许多地方扭曲形成螺旋构型。这种GNRs的单向对准可能有助于获得有吸引力的特征，例如磁性、电导率或传热性7。
在室温下使用硝酸钙四水...

Watch this Scientific Journal Video about Synthesis of Graphene-Hydroxyapatite Nanocomposites for Potential Use in Bone Tissue Engineering at JoVE.com

Synthesis of Graphene-Hydroxyapatite Nanocomposites for Potential Use in Bone Tissue Engineering

采用溶液相合成制备了石墨烯纳米带和羟基磷灰石纳米颗粒的新型纳米复合材料.当这些杂交种用于生物活性支架时，可以在组织工程和骨再生中表现出潜在的应用。

石墨烯-羟基磷灰石纳米复合材料的合成在骨组织工程中的潜在应用

Developing novel materials for bone tissue engineering is one of the most important thrust areas of nanomedicine. Several nanocomposites have been ...

这项工作解释了制造具有相反方向的纳米羟基磷灰石和石墨烯纳米带的新型复合材料的策略。这些生物材料对于开发骨组织工程支架具有重要意义。这是一个简单的一锅合成。这是一种快速，有效且经济的方法。这项工作与治疗骨损伤和相关疾病的背景极其相关，其中更多的骨组织再生将促进快速愈合。首先，使用50毫升的反应混合物合成羟基磷灰石的原始纳米颗粒。滴加25%氢氧化铵以保持pH值在10左右。此后，通过超声照射搅拌反应混合物30分钟。让所得溶液在室温下成熟120小时，直到羟基磷灰石纳米颗粒的白色沉淀沉淀沉淀出来。通过在室温下以1， 398×g离心5分钟来回收羟基磷灰石的纳米颗粒。用去离子水洗涤沉淀物三次。为了制备nHAP / GNR纳米复合材料，使用共官能化策略，首先将5毫克石墨烯纳米带溶解在一摩尔硝酸钙四水合物和0.67摩尔磷酸氢铵的混合物中至50毫升最终体积。在该反应过程中，滴加25%氢氧化铵以将pH值保持在约10。通过超声波搅拌所得混合物30分钟。反应完成后，使溶液在室温下不受干扰120小时直至成熟。观察羟基磷灰石纳米颗粒的凝胶沉淀物的形成，其覆盖石墨烯纳米带，随后nHAP / GNR的白色沉淀沉淀沉降。通过在室温下以1， 398×g离心5分钟洗涤沉淀物三次，然后在去离子水中重新分散。为了合成GNR / nHAP纳米复合材料，将市售的羟基磷灰石纳米颗粒悬浮在50毫升去离子水中，浓度为每毫升5毫克，并补充5毫克的石墨烯纳米带。通过超声搅拌所得混合物30分钟，然后，使混合物在室温下不受干扰120小时。成熟后，通过在室温下以1， 398×g离心5分钟，回收所得GNR / nHAP的白色沉淀物。用去离子水洗涤样品三次。对nHAP / GNR纳米复合材料的HRTEM分析显示，nHAP斑块的长度和宽度在150至250纳米之间。元素映射证实，由于存在元素钙和磷，GNRs上的中间节点斑块确实是nHAP。在GNR/nHAP纳米复合材料中，GNR在球形nHAP纳米颗粒表面形成薄膜。EDS分析显示，由于GNRs引起的碳含量明显增加，而钙和磷特有的峰值是由于nHAP引起的。nHAP/GNR光谱中碳含量的显着增加是由于大多数GNRs仅观察到一小部分新合成的nHAP。XRD分析证实了nHAP强峰的六方晶体结构分别对应于002、102、211、300、202、310、113、222和213平面，证实了所合成的nHAP的纯度。TGA分析显示，由于nHAP的结晶，在200摄氏度至500摄氏度之后，质量稳步下降。进一步的加热导致复合物的分解。由于GNRs的存在而造成的损失被发现在GNR / nHAP和nnHAP / GNR中介于0.5%和0.98%之间。试剂的添加顺序和反应条件对于获得纳米复合材料所需的反向取向最为关键。可以进行基于细胞系的研究来检查纳米复合材料促进成骨的潜力。这将为组织工程开辟一个新的方向介导的细胞反应调节领域。

Research

JoVE Journal

Bioengineering

Elastomeric PGS Scaffolds in Arterial Tissue Engineering

弹性PGS的动脉组织工程支架

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and ...

科研

生物工程

Use of Human Perivascular Stem Cells for Bone Regeneration

人类血管周围干细胞用于骨再生

Human perivascular stem cells (PSCs) can be isolated in sufficient numbers from multiple tissues for purposes of skeletal tissue engineering1-3. PSCs are ...

Encapsulation of Cardiomyocytes in a Fibrin Hydrogel for Cardiac Tissue Engineering

心肌在心脏组织工程中的纤维蛋白凝胶的封装

Culturing cells in a three dimensional hydrogel environment is an important technique for developing constructs for tissue engineering as well as studying ...

Postproduction Processing of Electrospun Fibres for Tissue Engineering

静电纤维组织工程的后期处理

Electrospinning is a commonly used and versatile method to produce  scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo ...

Capillary Force Lithography for Cardiac Tissue Engineering

毛细力光刻技术在心脏组织工程

Cardiovascular disease remains the leading cause of death worldwide1. Cardiac tissue engineering holds much promise to deliver groundbreaking medical ...

Tissue Engineering of a Human 3D in vitro Tumor Test System

Tissue Engineering of a Human 3D in vitro Tumor Test System

一个人的3D组织工程<em&gt;在体外</em&gt;肿瘤测试系统

Cancer is one of the leading causes of death worldwide.  Current therapeutic strategies are predominantly developed in 2D culture  systems, which ...

Production, Characterization and Potential Uses of a 3D Tissue-engineered Human Esophageal Mucosal Model

三维组织工程人食管黏膜型号生产，表征和潜在用途

The incidence of both esophageal adenocarcinoma and its precursor, Barrett&#8217;s Metaplasia, are rising rapidly in the western world. Furthermore ...

Engineering 3D Cellularized Collagen Gels for Vascular Tissue Regeneration

工程三维细胞化胶原凝胶的血管组织再生

Synthetic materials are known to initiate clinical complications such as inflammation, stenosis, and infections when implanted as vascular substitutes. ...

Sandwich-like Microenvironments to Harness Cell/Material Interactions

三明治般的微环境来驾驭细胞/材料相互作用

Cell culture has been traditionally carried out on bi-dimensional (2D) substrates where cells adhere using ventral receptors to the biomaterial surface. ...

Synthesis of Keratin-based Nanofiber for Biomedical Engineering

基于角蛋白纳米纤维生物医学工程合成

Electrospinning, due to its versatility and potential for applications in various fields, is being frequently used to fabricate nanofibers. Production of ...