Abstract
Alginates are important hydrogels for meniscus tissue engineering as they support the meniscal fibrochondrocyte phenotype and proteoglycan production, the extracellular matrix (ECM) component chiefly responsible for its viscoelastic properties. Here, we systematically evaluated four biomedical- and two nonbiomedical-grade alginates for their capacity to provide the best three-dimensional (3-D) microenvironment and to support proteoglycan synthesis of encapsulated human meniscal fibrochondrocytes in vitro. Biomedical-grade, high mannuronic acid alginate spheres (BioLVM, BioMVM) were the most uniform in size, indicating an effect of the purity of alginate on the shape of the spheres. Interestingly, the purity of alginates did not affect cell viability. Of note, only fibrochondrocytes encapsulated in BioMVM alginate produced and retained significant amounts of proteoglycans. Following transplantation in an explant culture model, the alginate spheres containing fibrochondrocytes remained in close proximity with the meniscal tissue adjacent to the defect. The results reveal a promising role of BioMVM alginate to enhance the proteoglycan production of primary human meniscal fibrochondrocytes in a 3-D hydrogel microenvironment. These findings have significant implications for cell-based translational studies aiming at restoring lost meniscal tissue in regions containing high amounts of proteoglycans.
Highlights
The composition of alginate directly influences, among other factors[25], the physiochemical properties of the hydrogel meshwork[26] and the behavior of the encapsulated cells[27,28]
Human meniscal fibrochondrocytes were first encapsulated in various alginates as designed in Fig. 1 to determine the best suited compound that provides a favorable 3-D microenvironment for proteoglycan synthesis based on an evaluation of the morphology of the resulting spheres over an extended period of time
Spheres composed of BioLVM displayed the smallest diameter, significantly different than those of spheres made of BioLVG (P = 0.001), BioMVM (P < 0.001), or BioMVG (P = 0.007)
Summary
The composition of alginate directly influences, among other factors[25], the physiochemical properties of the hydrogel meshwork[26] and the behavior of the encapsulated cells[27,28]. Histological evaluation of human meniscal fibrochondrocytes embedded in the different alginate spheres on day 3 after encapsulation mainly revealed cells with an oval or round morphology and were homogenously distributed without detectable differences between groups (Fig. 5). After 21 days post-encapsulation, human meniscal fibrochondrocytes encapsulated in LVG showed the highest levels of viability (~80%), significantly higher than cells encapsulated in BioLVM (P < 0.001) or in LVM, BioLVG, BioMVG, and BioMVM (P < 0.05), with values ranging from 61.7% to 80.3%, respectively (Fig. 6).
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