TEMPO-Oxidized Cellulose Nanofiber-Alginate Hydrogel as a Bioink for Human Meniscus Tissue Engineering

Objective: The avascular inner regions of the knee menisci cannot self-heal. As a prospective treatment, functional replacements can be generated by cell-based 3D bioprinting with an appropriate cell source and biomaterial. To that end, human meniscus fibrochondrocytes (hMFC) from surgical castoffs...

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Autores principales:	Xiaoyi Lan, Zhiyao Ma, Alexander R. A. Szojka, Melanie Kunze, Aillette Mulet-Sierra, Margaret J. Vyhlidal, Yaman Boluk, Adetola B. Adesida
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	tissue engineering meniscus 3D bioprinting cellulose nanofiber hypoxia Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/c1eb2524b1bd4b0fac780747b86453a6
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spelling	oai:doaj.org-article:c1eb2524b1bd4b0fac780747b86453a62021-11-05T06:57:41ZTEMPO-Oxidized Cellulose Nanofiber-Alginate Hydrogel as a Bioink for Human Meniscus Tissue Engineering2296-418510.3389/fbioe.2021.766399https://doaj.org/article/c1eb2524b1bd4b0fac780747b86453a62021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fbioe.2021.766399/fullhttps://doaj.org/toc/2296-4185Objective: The avascular inner regions of the knee menisci cannot self-heal. As a prospective treatment, functional replacements can be generated by cell-based 3D bioprinting with an appropriate cell source and biomaterial. To that end, human meniscus fibrochondrocytes (hMFC) from surgical castoffs of partial meniscectomies as well as cellulose nanofiber-alginate based hydrogels have emerged as a promising cell source and biomaterial combination. The objectives of the study were to first find the optimal formulations of TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl)-oxidized cellulose nanofiber/alginate (TCNF/ALG) precursors for bioprinting, and then to use them to investigate redifferentiation and synthesis of functional inner meniscus-like extracellular matrix (ECM) components by expanded hMFCs.Methods: The rheological properties including shear viscosity, thixotropic behavior recovery, and loss tangent of selected TCNF/ALG precursors were measured to find the optimum formulations for 3D bioprinting. hMFCs were mixed with TCNF/ALG precursors with suitable formulations and 3D bioprinted into cylindrical disc constructs and crosslinked with CaCl2 after printing. The bioprinted constructs then underwent 6 weeks of in vitro chondrogenesis in hypoxia prior to analysis with biomechanical, biochemical, molecular, and histological assays. hMFCs mixed with a collagen I gel were used as a control.Results: The TCNF/ALG and collagen-based constructs had similar compression moduli. The expression of COL2A1 was significantly higher in TCNF/ALG. The TCNF/ALG constructs showed more of an inner meniscus-like phenotype while the collagen I-based construct was consistent with a more outer meniscus-like phenotype. The expression of COL10A1 and MMP13 were lower in the TCNF/ALG constructs. In addition, the immunofluorescence of human type I and II collagens were evident in the TCNF/ALG, while the bovine type I collagen constructs lacked type II collagen deposition but did contain newly synthesized human type I collagen.Xiaoyi LanXiaoyi LanZhiyao MaAlexander R. A. SzojkaMelanie KunzeAillette Mulet-SierraMargaret J. VyhlidalYaman BolukAdetola B. AdesidaFrontiers Media S.A.articletissue engineeringmeniscus3D bioprintingcellulose nanofiberhypoxiaBiotechnologyTP248.13-248.65ENFrontiers in Bioengineering and Biotechnology, Vol 9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	tissue engineering meniscus 3D bioprinting cellulose nanofiber hypoxia Biotechnology TP248.13-248.65
spellingShingle	tissue engineering meniscus 3D bioprinting cellulose nanofiber hypoxia Biotechnology TP248.13-248.65 Xiaoyi Lan Xiaoyi Lan Zhiyao Ma Alexander R. A. Szojka Melanie Kunze Aillette Mulet-Sierra Margaret J. Vyhlidal Yaman Boluk Adetola B. Adesida TEMPO-Oxidized Cellulose Nanofiber-Alginate Hydrogel as a Bioink for Human Meniscus Tissue Engineering
description	Objective: The avascular inner regions of the knee menisci cannot self-heal. As a prospective treatment, functional replacements can be generated by cell-based 3D bioprinting with an appropriate cell source and biomaterial. To that end, human meniscus fibrochondrocytes (hMFC) from surgical castoffs of partial meniscectomies as well as cellulose nanofiber-alginate based hydrogels have emerged as a promising cell source and biomaterial combination. The objectives of the study were to first find the optimal formulations of TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl)-oxidized cellulose nanofiber/alginate (TCNF/ALG) precursors for bioprinting, and then to use them to investigate redifferentiation and synthesis of functional inner meniscus-like extracellular matrix (ECM) components by expanded hMFCs.Methods: The rheological properties including shear viscosity, thixotropic behavior recovery, and loss tangent of selected TCNF/ALG precursors were measured to find the optimum formulations for 3D bioprinting. hMFCs were mixed with TCNF/ALG precursors with suitable formulations and 3D bioprinted into cylindrical disc constructs and crosslinked with CaCl2 after printing. The bioprinted constructs then underwent 6 weeks of in vitro chondrogenesis in hypoxia prior to analysis with biomechanical, biochemical, molecular, and histological assays. hMFCs mixed with a collagen I gel were used as a control.Results: The TCNF/ALG and collagen-based constructs had similar compression moduli. The expression of COL2A1 was significantly higher in TCNF/ALG. The TCNF/ALG constructs showed more of an inner meniscus-like phenotype while the collagen I-based construct was consistent with a more outer meniscus-like phenotype. The expression of COL10A1 and MMP13 were lower in the TCNF/ALG constructs. In addition, the immunofluorescence of human type I and II collagens were evident in the TCNF/ALG, while the bovine type I collagen constructs lacked type II collagen deposition but did contain newly synthesized human type I collagen.
format	article
author	Xiaoyi Lan Xiaoyi Lan Zhiyao Ma Alexander R. A. Szojka Melanie Kunze Aillette Mulet-Sierra Margaret J. Vyhlidal Yaman Boluk Adetola B. Adesida
author_facet	Xiaoyi Lan Xiaoyi Lan Zhiyao Ma Alexander R. A. Szojka Melanie Kunze Aillette Mulet-Sierra Margaret J. Vyhlidal Yaman Boluk Adetola B. Adesida
author_sort	Xiaoyi Lan
title	TEMPO-Oxidized Cellulose Nanofiber-Alginate Hydrogel as a Bioink for Human Meniscus Tissue Engineering
title_short	TEMPO-Oxidized Cellulose Nanofiber-Alginate Hydrogel as a Bioink for Human Meniscus Tissue Engineering
title_full	TEMPO-Oxidized Cellulose Nanofiber-Alginate Hydrogel as a Bioink for Human Meniscus Tissue Engineering
title_fullStr	TEMPO-Oxidized Cellulose Nanofiber-Alginate Hydrogel as a Bioink for Human Meniscus Tissue Engineering
title_full_unstemmed	TEMPO-Oxidized Cellulose Nanofiber-Alginate Hydrogel as a Bioink for Human Meniscus Tissue Engineering
title_sort	tempo-oxidized cellulose nanofiber-alginate hydrogel as a bioink for human meniscus tissue engineering
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/c1eb2524b1bd4b0fac780747b86453a6
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TEMPO-Oxidized Cellulose Nanofiber-Alginate Hydrogel as a Bioink for Human Meniscus Tissue Engineering

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