JMIR Publications

ABSTRACT

Background:

Chondrocytes are the primary cell type responsible for maintaining cartilage integrity and function. Their role in cartilage homeostasis and response to inflammation is crucial for understanding the progression and potential therapeutic interventions for various cartilage-related disorders. Developing an accessible and cost-effective model to generate viable chondrocytes and assess their response to different bioactive compounds can significantly advance our knowledge of cartilage biology and contribute to the discovery of novel therapeutic approaches.

Objective:

The development of our novel, streamlined protocol for generating chondrocytes from BMSCs in a 3D culture system offers significant implications for the study of cartilage biology and the discovery of potential therapeutic interventions for cartilage-related disorders.

Methods:

We developed a streamlined protocol for generating chondrocytes from bone marrow-derived mesenchymal stem cells (BMSCs) in a 3D culture system using an "in-tube" culture approach. This simple pellet-based 3D culture system allows for cell aggregation and spheroid formation, facilitating cell-cell and cell-extracellular matrix interactions that better mimic the in vivo cellular environment compared to 2D monolayer cultures. A pro-inflammatory chondrocyte model was created by treating the chondrocytes with lipopolysaccharide (LPS) and subsequently employed to evaluate the anti-inflammatory effects of vitamin D, curcumin, and resveratrol.

Results:

The established protocol successfully generated a large quantity of viable chondrocytes, characterized by Alcian blue and Toluidine blue staining, and demonstrated versatility in assessing the anti-inflammatory effects of various bioactive compounds. The chondrocytes exhibited reduced inflammation, as evidenced by decreased tumor necrosis factor-alpha (TNF-α) levels, in response to vitamin D, curcumin, and resveratrol treatment.

Conclusions:

Our novel protocol offers an accessible and cost-effective approach for generating chondrocytes from BMSCs and evaluating potential therapeutic leads in the context of inflammatory chondrocyte-related diseases. Although our approach has several advantages, further investigation is required to address limitations, such as the potential differences between chondrocytes generated using our protocol and those derived from other established methods, and to refine the model for broader applicability and clinical translation.