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

ABSTRACT: 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.

SUBMITTER: Lan X

PROVIDER: S-EPMC8602093 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:This study aimed to develop poly (vinyl alcohol) grafted glycidyl methacrylate/cellulose nanofiber (PVA-g-GMA/CNF) injectable hydrogels for meniscus tissue engineering. PVA-g-GMA is an interesting polymer for preparing cross-linking injectable hydrogels with UV radiation, but it has poor mechanical properties and low cell proliferation. In this study, CNF as a reinforcing agent was selected to improve mechanical properties and cell proliferation in PVA-g-GMA injectable hydro-gels. The effect of CNF concentration on hydrogel properties was investigated. Both PVA-g-GMA and PVA-g-GMA hydrogels incorporating 0.3, 0.5, and 0.7% (w/v) CNF can be formed by UV curing at a wavelength of 365 nm, 6 mW/cm2 for 10 min. All hydrogels showed substantial microporosity with interconnected tunnels, and a pore size diameter range of 3-68 µm. In addition, all hydrogels also showed high physicochemical properties, a gel fraction of 81-82%, porosity of 83-94%, water content of 73-87%, and water swelling of 272-652%. The water content and swelling of hydrogels were increased when CNF concentration increased. It is worth noting that the reduction of porosity in the hydrogels occurred with increasing CNF concentration. With increasing CNF concentration from 0.3% to 0.7% (w/v), the compressive strength and compressive modulus of the hydrogels significantly increased from 23 kPa to 127 kPa and 27 kPa to 130 kPa, respectively. All of the hydrogels were seeded with human cartilage stem/progenitor cells (CSPCs) and cultured for 14 days. PVA-g-GMA hydrogels incorporating 0.5% and 0.7% (w/v) CNF demonstrated a higher cell proliferation rate than PVA-g-GMA and PVA-g-GMA hydrogels incorporating 0.3% (w/v) CNF, as confirmed by MTT assay. At optimum formulation, 10%PVA-g-GMA/0.7%CNF injectable hydrogel met tissue engineering requirements, which showed excellent properties and significantly promoted cell proliferation, and has a great potential for meniscus tissue engineering application.

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

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