Assessing the advantages of 3D bioprinting and 3D spheroids in deciphering the osteoarthritis healing mechanism using human chondrocytes and polarized macrophages

Abstract

The molecular niche of an osteoarthritic microenvironment comprises of the native chondrocytes, the circulatory immune cells, and their respective inflammatory mediators. Although, M2 macrophages infiltrate the joint tissue during osteoarthritis (OA) to initiate cartilage repair, the mechanistic crosstalk that dwells underneath is still unknown. Our study established a co-culture system of human OA chondrocytes and M2 macrophages in 3D spheroids and 3D bioprinted silk-gelatin constructs. It is already well established that Silk fibroin-gelatin bioink support chondrogenic differentiation due to upregulation in Wnt/β-catenin pathway. Additionally, the presence of anti-inflammatory M2 macrophages significantly upregulated the expression of chondrogenic biomarkers (COL-II, ACAN) with an attenuated expression of the chondrocyte hypertrophy (COL-X), chondrocyte dedifferentiation (COL-I) and matrix catabolism (MMP-1 and MMP-13) genes even in the absence of the interleukins. Furthermore, the 3D bioprinted co-culture model displayed an upper hand in stimulating cartilage regeneration and OA inhibition than the spheroid model, underlining the role of silk fibroin-gelatin in encouraging chondrogenesis. Additionally, the 3D bioprinted silk-gelatin constructs further supported the maintenance of stable anti-inflammatory phenotype of M2 macrophage. Thus, the direct interaction between the primary OAC and M2 macrophages in 3D context along with the release of the soluble anti-inflammatory factors by the M2 cells significantly contributed to a better understanding regarding the molecular mechanisms responsible for immune cell-mediated OA healing.

Macrophages in guided bone regeneration: potential roles and future directions

“…Classically activated M1 macrophages are generally considered proinflammatory, mediating inflammatory responses and osteoclastogenesis. In contrast, selectively activated M2 macrophages, known as anti-inflammatory or proregenerative macrophages, are responsible for immunomodulation and tissue remodeling (8,12,22).…”