The Combination of Decellularized Cartilage and Amniotic Membrane Matrix Enhances the Production of Extracellular Matrix Elements in Human Chondrocytes

Abstract

Articular cartilage lesions are challenging to regenerate, prompting the investigation of novel biomaterial-based therapeutic approaches. Extracellular matrix (ECM)-derived biomaterials are a promising option for this purpose; however, to date, the combination of amniotic membrane (AMM) and articular cartilage (ACM) has not been tested. This study evaluated different concentrations of soluble extracts from the decellularized ECM of amniotic membrane (dAMM) and articular cartilage (dACM), both individually and in combination, to determine their ability to maintain the chondrogenic phenotype in human chondrocytes. After the decellularization process 90–99% of the cellular components were removed, it retains nearly 100% of type 2 collagen and 70% of aggrecan (ACAN) for dACM, and approximately 90% of type IV collagen and 75% of ACAN for dAMM. The biological activity of soluble extracts from dACM and dAMM were evaluated on human chondrocytes. After 72 h, 1.5 mg/mL of dACM and 6 mg/mL of dAMM significantly increased (p < 0.05) the proliferation and expression of SOX9 and ACAN. Also, the combination of both (1.5 mg/mL dACM and 6 mg/mL dAMM) showed synergistic effects, enhancing chondrocyte proliferation, maintaining chondrogenic lineage, and increasing the production of cartilage ECM components, such as COLII (1.5-fold), SOX9 (2-fold), and ACAN (2-fold). These results suggest that the combined use of dACM and dAMM has potential for cartilage regeneration.