Patnaik Rajashree, Jannati Shirin, Sivani Bala Mohan, Rizzo Manfredi, Naidoo Nerissa, Banerjee Yajnavalka
Mohammed Bin Rashid University of Medicine and Health Sciences, Dubai, United Arab Emirates.
Department of Molecular Biology, Lund University, Lund, Lund, Sweden.
JMIR Res Protoc. 2023 Jul 28;12:e42964. doi: 10.2196/42964.
Chondrocytes are the primary cells 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 to assess their response to different bioactive compounds can significantly advance our knowledge of cartilage biology and contribute to the discovery of novel therapeutic approaches.
We developed a novel, streamlined protocol for generating chondrocytes from bone marrow-derived mesenchymal stem cells (BMSCs) in a 3D culture system that offers significant implications for the study of cartilage biology and the discovery of potential therapeutic interventions for cartilage-related and associated disorders.
We developed a streamlined protocol for generating chondrocytes from 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 with 2D monolayer cultures. A proinflammatory chondrocyte model was created by treating the chondrocytes with lipopolysaccharide and was subsequently used to evaluate the anti-inflammatory effects of vitamin D, curcumin, and resveratrol.
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 the decreased tumor necrosis factor-α levels, in response to vitamin D, curcumin, and resveratrol treatment.
Our novel protocol offers an accessible and cost-effective approach for generating chondrocytes from BMSCs and for evaluating potential therapeutic leads in the context of inflammatory chondrocyte-related diseases. Although our approach has several advantages, further investigation is required to address its 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.
软骨细胞是维持软骨完整性和功能的主要细胞。它们在软骨内环境稳定及对炎症反应中的作用，对于理解各种软骨相关疾病的进展及潜在治疗干预措施至关重要。开发一种可获取且经济高效的模型来生成有活力的软骨细胞，并评估它们对不同生物活性化合物的反应，能够显著推进我们对软骨生物学的认识，并有助于发现新的治疗方法。
我们开发了一种新颖、简化的方案，用于在三维培养系统中从骨髓间充质干细胞（BMSC）生成软骨细胞，这对软骨生物学研究以及发现软骨相关和伴发疾病的潜在治疗干预措施具有重要意义。
我们开发了一种简化方案，使用“管内”培养方法在三维培养系统中从BMSC生成软骨细胞。这种基于简单微球的三维培养系统允许细胞聚集和球体形成，促进细胞 - 细胞及细胞 - 细胞外基质相互作用，与二维单层培养相比，能更好地模拟体内细胞环境。通过用脂多糖处理软骨细胞创建了促炎软骨细胞模型，随后用于评估维生素D、姜黄素和白藜芦醇的抗炎作用。
所建立的方案成功生成了大量有活力的软骨细胞，通过阿尔新蓝和甲苯胺蓝染色得以表征，并展示了在评估各种生物活性化合物抗炎作用方面的通用性。经维生素D、姜黄素和白藜芦醇处理后，软骨细胞炎症减轻，表现为肿瘤坏死因子 -α水平降低。
我们的新方案为从BMSC生成软骨细胞以及在炎症性软骨细胞相关疾病背景下评估潜在治疗先导物提供了一种可获取且经济高效 的方法。尽管我们的方法有几个优点，但仍需要进一步研究来解决其局限性，例如使用我们的方案生成的软骨细胞与其他既定方法衍生的软骨细胞之间的潜在差异，并完善该模型以实现更广泛的适用性和临床转化。
