Back pain is a major public health issue in our society, and is strongly correlated with the degeneration of intervertebral discs (IVDs). Current therapies are conservative or surgical, and no attempt to regenerate the IVD. The first goal of our project is to create a fully decellularized bovine caudal IVD to be used as a scaffold on which to seed adult human stem cells in an attempt to engineer a healthy, replacement IVD for patients suffering from IVD degeneration and lower back pain. The goal of decellularization is to eliminate DNA content while retaining glycosaminoglycan (GAG) content. Eliminating DNA content will prevent a foreign body response by the host's immune system once the IVD is implanted. GAG is responsible for forming interfibrillar bridges with collagen fibrils and thus assisting in resisting compressive and tensile forces. By retaining GAG content in our decellularized IVDs we will maintain structural integrity of the extracellular matrix. The IVDs closest to the base of the bovine tail were targeted, because they are similar in size and biochemistry to the human IVD. We use a mix of conventional methods including freeze thaw, sonication and agitation in a solution of sodium dodecyl sulfate (SDS) and ethylene diamine tetraacetic acid (EDTA). After decellularization, half of each IVD was placed in a tissue cassette and put formalin in preparation for histological analysis, and the other half was frozen prior to biochemical analysis (DMMB and PicoGreen assay). Our results thus far are promising in eliminating DNA content but show we have a large room for improvement in retaining GAG content.
Compton, C; Hensley, A; Lehane, A; Rames, J; Skelly, M; Fernandez, C; and Mercuri, J, "Development of a Novel Biological Intervertebral Disc Scaffold" (2015). Focus on Creative Inquiry. 140.