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Histone Extraction from Human Articular Cartilage for the Study of Epigenetic Regulation in Osteoarthritis
Authors:Carmen Nú  ñ  ez-Carro,Margarita Blanco-Blanco,Tatiana Montoya,Karla M. Villagrá  n-Andrade,Tamara Hermida-Gó  mez,Francisco J. Blanco,Marí  a C. de André  s
Abstract:Osteoarthritis (OA) is a chronic disease that affects articular cartilage, causing its degeneration. Although OA is one of the most prevalent pathologies globally, there are no definitive treatments available. Recently, research has focused on elucidating the complex interplay that takes place between inflammatory processes and epigenetic regulation, showing that histone post-translational modifications (PTMs) can exert a pronounced effect on the expression of OA-related genes. OA chondrocytes enhance the production of interleukin 1β (IL-1β) and interleukin 8 (IL-8), which are epigenetically regulated. These cytokines upregulate the synthesis of matrix metalloproteinases (MMPs) and aggrecanases, which promote the extracellular matrix (ECM) destruction. This motivates the study of histone PTMs to investigate the epigenetic regulation of proinflammatory molecules, but the absence of specific protocols to extract histones from human articular cartilage has complicated this task. The lack of effective methods can be explained by the structural complexity and low cellularity of this tissue, which are responsible for the biomechanical properties that allow the movement of the joint but also complicate histone isolation. Here, we provide a histone extraction procedure specifically adapted for cryopreserved human articular cartilage that can be useful to understand epigenetic regulation in OA and accelerate the search for novel strategies.
Keywords:histone   chondrocytes   cartilage   methods   epigenetics   osteoarthritis   inflammation
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