3D Printed Chitosan Composite Scaffold for Chondrocytes Differentiation

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.

Keywords: 3D printing; Chitosan; cell culture; chondrocytes; hydrogel; tissue engineering..