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3D 打印壳聚糖复合支架促进软骨细胞分化。

3D Printed Chitosan Composite Scaffold for Chondrocytes Differentiation.

机构信息

Department of Biomedical Engineering, North-Eastern Hill University, Shillong-793022, Meghalaya, India.

Department of Applied Mechanics, Motilal Nehru National Institute of Technology, Allahabad, Prayagraj-211004, Uttar Pradesh, India.

出版信息

Curr Med Imaging. 2021;17(7):832-842. doi: 10.2174/1573405616666201217112939.

Abstract

AIMS

Our aim is to develop 3D printed chitosan-gelatin-alginate scaffolds using a costeffective in house designed 3D printer followed by its characterization. To observe chondrocyte differentiation on 3D printed scaffolds as part of scaffold application.

BACKGROUND

Cartilage is considered to be a significant tissue in humans. It is present in between the rib cage, the lobe of the ear, nasal septum in the form of hyaline cartilage, in between ribs costal cartilage, intervertebral discs in the form of fibrocartilage, meniscus, larynx, epiglottis and between various joints of bones. To replace or repair damaged tissues due to disorders or trauma, thousands of surgical procedures are performed daily. 3D printing plays a crucial role in the development of controlled porous architectures of scaffolds for cartilage tissue regeneration. Advancement in 3D printing technology like inkjet, micro- extrusion in 3D bioprinting, Laser-assisted 3D Bioprinting (LAB), stereolithography combination with biomaterials plays a crucial role in the quick development of patient-specific articulating cartilage when need in a short period frame.

OBJECTIVE

Our objective is to develop different compositions of chitosan-gelatin-alginate composite hydrogel scaffolds with controlled porosity and architectures with the application of 3D printing and observe the growth of cartilage on it. To achieve as proposed, an in-house 3D paste extruder printer was developed, which is capable of printing porous composite chitosan hydrogel scaffolds of desired architecture layer by layer. After the characterization of 3D printed chitosan composite scaffolds, the differentiation of chondrocyte was observed using hMSC.

METHODS

In present paper process for the development of chitosan-alginate-gelatin composite hydrogel, 3D printing, morphological characterization, and observation for differentiation of chondrocytes cells on 3D printed chitosan composite hydrogels is presented. The present study is divided into three parts: in first part development of composite chitosan-alginate-gelatin hydrogel with the utilization of in house customized assembled paste extruder based 3D printer, which is capable of printing chitosan composite hydrogels. In the second part, the characterization of 3D printed chitosan composite scaffolds hydrogel is performed for evaluating the morphological, mechanical, and physical properties. The prepared composite scaffolds were characterized by Fourier Transform Infrared Spectroscopy (FTIR), X-Ray Diffraction(XRD), Scanning Electron Microscopy SEM, swelling property, mechanical testing, porosity, etc. In the last part of the study, the differentiation of chondrocytes cells was observed with human Mesenchymal Stem Cells (hMSC) on 3D printed scaffolds and showed positive results for the same.

RESULTS

Stereolithography (STL) files of 3D models for porous chitosan composite were developed using Computer-Aided Design (CAD) and printed with a hydrogel flow rate within the range of 0.2-0.25 ml/min. The prepared scaffolds are highly porous, having optimum porosity, optimal mechanical strength to sustain the cartilage formation. The 3D printed chitosan composite scaffolds show supports for the differentiation of chondrocytes. The above study is helpful for in-vivo regeneration of cartilage for patients having related cartilage disorders.

CONCLUSION

This method helps in regeneration of degenerated cartilage for patient-specific and form above experiment we also concluded that 3D printed chitosan scaffold is best suited for the regeneration of chondrocyte cells.

摘要

目的

我们旨在使用成本效益高的内部设计 3D 打印机开发壳聚糖-明胶-海藻酸盐 3D 打印支架,并对其进行特性描述。观察 3D 打印支架上软骨细胞的分化,作为支架应用的一部分。

背景

软骨被认为是人体中的一种重要组织。它存在于肋骨之间、耳朵的叶状部分、鼻中隔的透明软骨、肋骨的肋软骨、椎间盘的纤维软骨、半月板、会厌和骨骼的各个关节之间。为了替代或修复因疾病或创伤而受损的组织,每天都要进行数千次手术。3D 打印在软骨组织再生的可控多孔支架结构的开发中起着至关重要的作用。3D 生物打印中喷墨、微挤出等 3D 打印技术的进步,以及激光辅助 3D 生物打印 (LAB)、立体光刻与生物材料的结合,在短时间内快速开发出特定于患者的关节软骨方面发挥了关键作用。

目的

我们的目标是开发具有不同组成的壳聚糖-明胶-海藻酸盐复合水凝胶支架,具有可控的孔隙率和结构,应用 3D 打印,并观察其软骨的生长。为了实现这一目标,我们开发了一种内部 3D 糊挤出打印机,它能够逐层打印所需结构的多孔复合壳聚糖水凝胶支架。在对 3D 打印壳聚糖复合支架进行特性描述后,使用 hMSC 观察软骨细胞的分化。

方法

在本文中,介绍了壳聚糖-藻酸盐-明胶复合水凝胶的开发、3D 打印、形态学特性描述以及 3D 打印壳聚糖复合水凝胶上软骨细胞分化的观察过程。本研究分为三个部分:第一部分是利用内部定制组装的基于 3D 打印机的糊挤出机开发壳聚糖-藻酸盐-明胶复合水凝胶,该打印机能够打印壳聚糖复合水凝胶。第二部分,对 3D 打印壳聚糖复合支架水凝胶进行特性描述,以评估其形态、机械和物理性能。对制备的复合支架进行傅里叶变换红外光谱 (FTIR)、X 射线衍射 (XRD)、扫描电子显微镜 (SEM)、溶胀性能、力学性能测试、孔隙率等表征。在研究的最后一部分,观察 3D 打印支架上人类间充质干细胞 (hMSC) 的软骨细胞分化,并取得了积极的结果。

结果

使用计算机辅助设计 (CAD) 开发了用于多孔壳聚糖复合的 3D 模型的立体光刻 (STL) 文件,并使用水凝胶流速在 0.2-0.25ml/min 的范围内进行打印。所制备的支架具有高度多孔性,具有最佳的孔隙率和最佳的机械强度,以维持软骨形成。3D 打印壳聚糖复合支架支持软骨细胞的分化。上述研究有助于为患有相关软骨疾病的患者进行软骨的体内再生。

结论

该方法有助于为特定患者的退化软骨进行再生,通过上述实验,我们还得出结论,3D 打印壳聚糖支架最适合软骨细胞的再生。

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