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Current Medical Imaging

Editor-in-Chief

ISSN (Print): 1573-4056
ISSN (Online): 1875-6603

Research Article

3D Printed Chitosan Composite Scaffold for Chondrocytes Differentiation

Author(s): Nitin Sahai*, Manashjit Gogoi and Ravi Prakash Tewari

Volume 17, Issue 7, 2021

Published on: 17 December, 2020

Page: [832 - 842] Pages: 11

DOI: 10.2174/1573405616666201217112939

Price: $65

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

Graphical Abstract

[1]
Mironov V, Boland T, Trusk T, Forgacs G, Markwald RR. Organ printing: computer-aided jet-based 3D tissue engineering. Trends Biotechnol 2003; 21(4): 157-61.
[http://dx.doi.org/10.1016/S0167-7799(03)00033-7] [PMID: 12679063]
[2]
European Symposium on Computer Aided Process Engineering -ESCAPE-1. Computers and Chemical Engineering 1992.
[3]
Sun W, Lal P. Recent development on computer aided tissue engineering-a review. Comput Methods Programs Biomed 2002; 67(2): 85-103.
[http://dx.doi.org/10.1016/S0169-2607(01)00116-X] [PMID: 11809316]
[4]
An J, Teoh JEM, Suntornnond R, Chua CK. Design and 3D printing of scaffolds and tissues. Engineering 2015; 1(2): 261-8.
[http://dx.doi.org/10.15302/J-ENG-2015061]
[5]
Sun W, Starly B, Nam J, Darling A. Bio-CAD modeling and its applications in computer-aided tissue engineering. CAD Comput Aided Des 2005; 37(11): 1097-114.
[http://dx.doi.org/10.1016/j.cad.2005.02.002]
[6]
Lee M, Wu BM. Recent advances in 3D printing of tissue engineering scaffolds. Methods Mol Biol 2012; 868: 257-67.
[http://dx.doi.org/10.1007/978-1-61779-764-4_15] [PMID: 22692615]
[7]
Sun W, Starly B, Darling A, Gomez C. Computer-aided tissue engineering: application to biomimetic modelling and design of tissue scaffolds. Biotechnol Appl Biochem 2004; 39(Pt 1): 49-58.http://www.ncbi.nlm.nih.gov/pubmed/14556653
[http://dx.doi.org/10.1042/BA20030109] [PMID: 14556653]
[8]
Wettergreen MA, Bucklen BS, Sun W, Liebschner MAK. Computer-aided tissue engineering of a human vertebral body. Ann Biomed Eng 2005; 33(10): 1333-43.
[http://dx.doi.org/10.1007/s10439-005-6744-1] [PMID: 16240082]
[9]
Ozbolat IT, Peng W, Ozbolat V. Application areas of 3D bioprinting. Drug Discov Today 2016; 21(8): 1257-71.
[http://dx.doi.org/10.1016/j.drudis.2016.04.006] [PMID: 27086009]
[10]
Choudhury D, Anand S, Naing MW. The arrival of commercial bioprinters - Towards 3D bioprinting revolution! Int J Bioprint 2018; 4(2): 139.
[http://dx.doi.org/10.18063/ijb.v4i2.139] [PMID: 33102917]
[11]
Ozbolat IT. Bioprinter Technologies.3D Bioprinting. Academic Press 2017.
[12]
Pereira FDAS, Parfenov V, Khesuani YD, Ovsianikov A, Mironov V. Commercial 3D Bioprinters.3D Printing and Biofabrication. Springer 2018; pp. 235-49.
[13]
Vijayavenkataraman S, Yan WC, Lu WF, Wang CH, Fuh JYH. 3D bioprinting of tissues and organs for regenerative medicine. Adv Drug Deliv Rev 2018; 132: 296-332.
[http://dx.doi.org/10.1016/j.addr.2018.07.004] [PMID: 29990578]
[14]
Cui X, Boland T, D’Lima DD, Lotz MK. Thermal inkjet printing in tissue engineering and regenerative medicine. Recent Pat Drug Deliv Formul 2012; 6(2): 149-55.
[http://dx.doi.org/10.2174/187221112800672949] [PMID: 22436025]
[15]
Zhang YS, Yue K, Aleman J, et al. 3D bioprinting for tissue and organ fabrication. Ann Biomed Eng 2017; 45(1): 148-63.
[http://dx.doi.org/10.1007/s10439-016-1612-8] [PMID: 27126775]
[16]
Chia HN, Wu BM. Recent advances in 3D printing of biomaterials. J Biol Eng 2015; 9: 4.
[http://dx.doi.org/10.1186/s13036-015-0001-4] [PMID: 25866560]
[17]
Patra S, Young V. A review of 3D printing techniques and the future in biofabrication of bioprinted tissue. Cell Biochem Biophys 2016; 74(2): 93-8.
[http://dx.doi.org/10.1007/s12013-016-0730-0] [PMID: 27193609]
[18]
Kang HW, Lee SJ, Ko IK, Kengla C, Yoo JJ, Atala A. A 3D bioprinting system to produce human-scale tissue constructs with structural integrity. Nat Biotechnol 2016; 34(3): 312-9.
[http://dx.doi.org/10.1038/nbt.3413] [PMID: 26878319]
[19]
Hsieh FY, Hsu SH. 3D bioprinting: A new insight into the therapeutic strategy of neural tissue regeneration. Organogenesis 2015; 11(4): 153-8.
[http://dx.doi.org/10.1080/15476278.2015.1123360] [PMID: 26709633]
[20]
Ji S, Guvendiren M. Recent advances in bioink design for 3D bioprinting of tissues and organs. Front Bioeng Biotechnol 2017; 5: 23.
[http://dx.doi.org/10.3389/fbioe.2017.00023] [PMID: 28424770]
[21]
Markstedt K, Mantas A, Tournier I, Martínez Ávila H, Hägg D, Gatenholm P. 3D bioprinting human chondrocytes with nanocellulose-alginate bioink for cartilage tissue engineering applications. Biomacromolecules 2015; 16(5): 1489-96.
[http://dx.doi.org/10.1021/acs.biomac.5b00188] [PMID: 25806996]
[22]
Parak A, Pradeep P, du Toit LC, Kumar P, Choonara YE, Pillay V. Functionalizing bioinks for 3D bioprinting applications. Drug Discov Today 2019; 24(1): 198-205.
[http://dx.doi.org/10.1016/j.drudis.2018.09.012] [PMID: 30244080]
[23]
Chia HN, Wu BM. Recent advances in 3D printing of tissue engineering scaffolds. J Biol Eng 2015; 9(4): 2-14.
[24]
Mandrycky C, Wang Z, Kim K, Kim DH. 3D bioprinting for engineering complex tissues. Biotechnol Adv 2015; 1-13.
[http://dx.doi.org/10.1016/j.biotechadv.2015.12.011] [PMID: 26724184]
[25]
Murphy SV, Atala A. 3D bioprinting of tissues and organs. Nat Biotechnol 2014; 32(8): 773-85.http://www.nature.com/doifinder/10.1038/nbt.2958%5Cnhttp://www.ncbi.nlm.nih.gov/pubmed/25093879
[http://dx.doi.org/10.1038/nbt.2958] [PMID: 25093879]
[26]
Ashammakhi N, Ahadian S, Xu C, et al. Bioinks and bioprinting technologies to make heterogeneous and biomimetic tissue constructs. Mater Today Bio 2019; 1: 100008.
[http://dx.doi.org/10.1016/j.mtbio.2019.100008] [PMID: 32159140]
[27]
Lenda M, Skórka P, Mazur B, Ward A, Wilson K. Ivory crisis: Role of bioprinting technology. Science 2018; 360(6386): 277.
[PMID: 29674584]
[28]
Groll J, Burdick JA, Cho DW, et al. A definition of bioinks and their distinction from biomaterial inks. Biofabrication 2018; 11(1): 013001.
[http://dx.doi.org/10.1088/1758-5090/aaec52] [PMID: 30468151]
[29]
Yenilmez B, Temirel M, Knowlton S, Lepowsky E, Tasoglu S. Development and characterization of a low-cost 3D bioprinter. Bioprinting 2019; 13: e00044.
[http://dx.doi.org/10.1016/j.bprint.2019.e00044]
[30]
Kahin K, Khan Z, Albagami M, Usman S, Bahnshal S, Alwazani H, et al. Development of a robotic 3D bioprinting and microfluidic pumping system for tissue and organ engineering. 2019; XVII.
[http://dx.doi.org/10.1117/12.2507237]
[31]
Dutta PK. Chitin and chitosan for regenerative medicine. Springer 2015; pp. 1-389.
[32]
Croisier F, Jérôme C. Chitosan-based biomaterials for tissue engineering. Eur Polym J 2013; 49(4): 780-92.
[http://dx.doi.org/10.1016/j.eurpolymj.2012.12.009]
[33]
Nettles DL, Elder SH, Gilbert JA. Potential use of chitosan as a cell scaffold material for cartilage tissue engineering. Tissue Eng 2002; 8(6): 1009-16.
[http://dx.doi.org/10.1089/107632702320934100] [PMID: 12542946]
[34]
Muzzarelli RAA. Chitins and chitosans for the repair of wounded skin, nerve, cartilage and bone. Carbohydr Polym 2009; 76(2): 167-82.
[http://dx.doi.org/10.1016/j.carbpol.2008.11.002]
[35]
Zhong X, Ji C, Chan AK, Kazarian SG, Ruys A, Dehghani F. Fabrication of chitosan/poly(ε-caprolactone) composite hydrogels for tissue engineering applications. J Mater Sci Mater Med 2011; 22(2): 279-88.
[http://dx.doi.org/10.1007/s10856-010-4194-2] [PMID: 21170732]
[36]
Bergonzi C, Di Natale A, Zimetti F, et al. Study of 3D-printed chitosan scaffold features after different post-printing gelation processes. Sci Rep 2019; 9(1): 362.
[http://dx.doi.org/10.1038/s41598-018-36613-8] [PMID: 30674919]
[37]
Zhang J, Allardyce BJ, Rajkhowa R, et al. 3D printing of silk particle-reinforced chitosan hydrogel structures and their properties. ACS Biomater Sci Eng 2018; 4(8): 3036-46.
[http://dx.doi.org/10.1021/acsbiomaterials.8b00804]
[38]
Roehm KD, Madihally SV. Bioprinted chitosan-gelatin thermosensitive hydrogels using an inexpensive 3D printer. Biofabrication 2017; 10(1): 015002.
[http://dx.doi.org/10.1088/1758-5090/aa96dd] [PMID: 29083312]
[39]
Ng WL, Yeong WY, Naing MW. Polyelectrolyte gelatin-chitosan hydrogel optimized for 3D bioprinting in skin tissue engineering. Int J Bioprinting 2016; 2(1): 53-62.
[http://dx.doi.org/10.18063/IJB.2016.01.009]
[40]
Liu Q, Li Q, Xu S, Zheng Q, Cao X. Preparation and properties of 3D printed alginate-chitosan polyion complex hydrogels for tissue engineering. Polymers (Basel) 2018; 10(6): 664.
[http://dx.doi.org/10.3390/polym10060664]
[41]
Mohan N, Mohanan P V, Sabareeswaran A, Nair P. Chitosan-hyaluronic acid hydrogel for cartilage repair. Int J Biol Macromol 2017; 104(B): 1936-45.
[http://dx.doi.org/10.1016/j.ijbiomac.2017.03.142]
[42]
Akkiraju H, Nohe A. Role of chondrocytes in cartilage formation, progression of osteoarthritis and cartilage regeneration. J Dev Biol 2015; 3(4): 177-92.
[http://dx.doi.org/10.3390/jdb3040177] [PMID: 27347486]
[43]
Apelgren P, Amoroso M, Lindahl A, et al. Chondrocytes and stem cells in 3D-bioprinted structures create human cartilage in vivo. PLoS One 2017; 12(12): e0189428.
[http://dx.doi.org/10.1371/journal.pone.0189428] [PMID: 29236765]
[44]
Bae SW, Lee KW, Park JH, et al. 3D bioprinted artificial trachea with epithelial cells and chondrogenic-differentiated bone marrow-derived mesenchymal stem cells. Int J Mol Sci 2018; 19(6): 1624.
[http://dx.doi.org/10.3390/ijms19061624] [PMID: 29857483]
[45]
Pomari AA do N. Montanheiro TL do A, de Siqueira CP, Silva RS, Tada DB, Lemes AP. Chitosan hydrogels crosslinked by genipin and reinforced with cellulose nanocrystals: production and characterization. J Compos Sci 2019; 3(3): 84.
[http://dx.doi.org/10.3390/jcs3030084]
[46]
Hsieh WC, Liau JJ, Li YJ. Characterization and cell culture of a grafted chitosan scaffold for tissue engineering. Int J Polym Sci 2015; 2015: 935305.
[http://dx.doi.org/10.1155/2015/935305]
[47]
Kumar S, Koh J. Physiochemical, optical and biological activity of chitosan-chromone derivative for biomedical applications. Int J Mol Sci 2012; 13(5): 6102-16.
[http://dx.doi.org/10.3390/ijms13056102] [PMID: 22754352]
[48]
Sharma C, Dinda AK, Potdar PD, Chou CF, Mishra NC. Fabrication and characterization of novel nano-biocomposite scaffold of chitosan-gelatin-alginate-hydroxyapatite for bone tissue engineering. Mater Sci Eng C 2016; 64: 416-27.
[http://dx.doi.org/10.1016/j.msec.2016.03.060] [PMID: 27127072]
[49]
Kim JH, Sim SJ, Lee DH, et al. Preparation and properties of PHEA/chitosan composite hydrogel. Polym J 2004; 36(12): 943-8.
[http://dx.doi.org/10.1295/polymj.36.943]
[50]
Chawla S, Kumar A, Admane P, Bandyopadhyay A, Ghosh S. Elucidating role of silk-gelatin bioink to recapitulate articular cartilage differentiation in 3D bioprinted constructs. Bioprinting 2017; 7: 1-13.
[http://dx.doi.org/10.1016/j.bprint.2017.05.001]

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