Dataset Information

Generation of an in vitro model of the outer annulus fibrosus-cartilage interface.

ABSTRACT: Current treatments for degenerative disc disease do not restore full biological functionality of the intervertebral disc (IVD). As a result, regenerative medicine approaches are being developed to generate a biological replacement that when implanted will restore form and function of the degenerated IVD. Tissue-engineered models to date have focused on the generation of nucleus pulposus and annulus fibrosus IVD components. However, these tissues need to be integrated with a cartilage endplate in order for successful implantation to occur. The purpose of this study was to generate an in vitro annulus fibrosus-cartilage interface model which would enable us to better understand the biological and biomechanical implications of such interfaces. It was hypothesized that in vitro-formed outer annulus fibrosus (OAF) and cartilage tissues would integrate in direct-contact coculture to yield an interface containing extracellular matrix with aspects resembling the native OAF-CEP interface. In vitro-formed tissues were generated using bovine OAF cell-seeded angle-ply, multi-lamellated polycarbonate urethane scaffolds and articular chondrocytes, which were then placed in direct-contact coculture. 2-week old OAF tissues integrated with 3-day old cartilage by 1?week of coculture. Immunohistochemical staining of 2-week interfaces showed that distributions of collagen type I, collagen type II, and aggrecan were similar to the native bovine interface. The apparent tensile strength of the in vitro interface increased significantly between 2 and 4?weeks of coculture. In summary, an annulus fibrosus-cartilage interface model can be formed in vitro which will facilitate the identification of conditions required to generate an entire tissue-engineered disc replacement suitable for clinical use.

SUBMITTER: Chong JE

PROVIDER: S-EPMC7323466 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Generation of an in vitro model of the outer annulus fibrosus-cartilage interface.

Chong Jasmine E JE Santerre J Paul JP Kandel Rita A RA

JOR spine 20200506 2

Current treatments for degenerative disc disease do not restore full biological functionality of the intervertebral disc (IVD). As a result, regenerative medicine approaches are being developed to generate a biological replacement that when implanted will restore form and function of the degenerated IVD. Tissue-engineered models to date have focused on the generation of nucleus pulposus and annulus fibrosus IVD components. However, these tissues need to be integrated with a cartilage endplate in ...[more]

PMID: 32613164

Similar Datasets

Project description:Low back pain caused by degenerative disc disease affects many people worldwide and brings huge economical burden. Thus, attentions have focused on annulus fibrosus (AF) tissue engineering for treatment of intervertebral disc degeneration. To engineer a functional replacement for the AF, it is important to fabricate scaffolds that mimic the structural and mechanical properties of native tissue. AF-derived stem cells are promising seed cells for AF tissue engineering due to their tissue specificity. In the present study, decellularized AF matrix (DAFM)/chitosan hybrid hydrogels were fabricated using genipin as a crosslinker. AF stem cells were cultured on hydrogel scaffolds with or without basic fibroblast growth factor (bFGF), and cell proliferation, morphology, gene expression, and AF tissue synthesis were examined. Overall, more collagen-I, collagen-II, and aggrecan were secreted by AF stem cells grown on hydrogels with bFGF compared to those without. These results support the application of DAFM/chitosan hybrid hydrogels as an appropriate candidate for AF tissue engineering. Furthermore, incorporation of bFGF into hydrogels promoted AF-related tissue synthesis. Impact Statement The investigation of annulus fibrosus (AF)-related tissue secretion and gene expression in extracellular matrix (ECM) of AF-derived stem cells (AFSCs) provided theoretical and practical basis for the choice of scaffold materials and growth factors for AF tissue engineering. The innovations of the present work are obvious. First, AFSCs were used because they are more easily differentiated into AF cells, thereby producing more AF-related ECM. Second, the decellularized AF matrix (DAFM) was derived from native AF tissue, but had reduced immunogenicity after decellularization. Furthermore, the DAFM structure mimicked the fibrous network of actual AF tissue, which was advantageous to AFSC adhesion and growth. Third, basic fibroblast growth factor was successfully incorporated into the DAFM, showed gradual sustained release, and effectively promoted production of AF tissue ECM factors collagen-I, collagen-II, aggrecan, and glycosaminoglycan.

Project description:Study designOvine in vivo study.ObjectiveTo perform lateral approach lumbar surgery in an ovine model to administer an injectable riboflavin cross-linked high-density collagen (HDC) gel and to assess its ability to mitigate intervertebral disc (IVD) degeneration after induced annulus fibrosus (AF) injury.Summary of background dataBiological-based injectable gels have shown efficacy in restoring biomechanical, radiographic, and histological parameters in IVD-injured animal models. Riboflavin cross-linked HDC gel has previously demonstrated retention of nucleus pulposus (NP) tissue, reduced loss of disc height, and prevention of terminal cellular degenerative changes in rat-tail spines. However, this biological therapy has never been tested in large animal models.MethodsForty lumbar IVDs were accessed from eight sheep via lateral approach surgery. IVDs were randomly assigned to healthy control, injury and HDC treatment, or negative control with injury and no treatment. IVD injury was carried out using a drill-bit through the AF followed by needle puncture of the NP. Sheep were followed for 16 weeks and underwent qualitative/quantitative magnetic resonance imaging, x-ray, and histological analyses of collagen and proteoglycan content.ResultsThe lateral approach to the ovine lumbar spine to deliver HDC gel proved to be safe and reproducible. IVDs treated with the HDC gel revealed less degenerative changes at the microscopic level based on AF and NP histology. However, mean Pfirrmann grade, T2 relaxation time, NP voxel size, and disc height index were not significantly different between the two injury groups.ConclusionInjectable HDC gel can be administered safely via lateral approach surgery in an ovine AF injury model. IVDs treated with HDC gel demonstrated less degeneration at the microscopic level though radiographic changes were slight when comparing treated to untreated IVDs. Future studies will need to elucidate the role of injury technique and time frame for follow-up in correlating histological and radiographical outcomes.Level of evidenceN /A.

Project description:Tissue engineering repair of annulus fibrosus (AF) defects has the potential to prevent disability and pain from intervertebral disc (IVD) herniation and its progression to degeneration. Clinical translation of AF repair methods requires assessment in long-term large animal models. An ovine AF injury model was developed using cervical spinal levels and a biopsy-type AF defect to assess composite tissue engineering repair in 1-month and 12-month studies. The repair used a fibrin hydrogel crosslinked with genipin (FibGen) to seal defects, poly(trimethylene carbonate) (PTMC) scaffolds to replace lost AF tissue, and polyurethane membranes to prevent herniation. In the 1-month study, PTMC scaffolds sealed with FibGen herniated with polyurethane membranes. When applied alone, FibGen integrated with the surrounding AF tissue without herniation, showing promise for long-term studies. The 12-month long-term study used only FibGen which showed fibrous healing, biomaterial resorption and no obvious hydrogel-related complications. However, the 2?mm biopsy punch injury condition also exhibited fibrotic healing at 12?months. Both untreated and FibGen treated groups showed equivalency with no detectable differences in histological grades of proteoglycans, cellular morphology, IVD structure and blood vessel formation, biomechanical properties including torque range and axial range of motion, Pfirrmann grade, IVD height, and quantitative scores of vertebral body changes from clinical computed tomography. The biopsy-type injury caused endplate defects with a high prevalence of osteophytes in all groups and no nucleus herniation, indicating that the biopsy-type injury requires further refinement, such as reduction to a slit-type defect that could penetrate the full depth of the AF without damaging the endplate. Results demonstrate translational feasibility of FibGen for AF repair to seal AF defects, although future study with a more refined injury model is required to validate the efficacy of FibGen before translation.

Dataset Information

Generation of an in vitro model of the outer annulus fibrosus-cartilage interface.

Publications

Generation of an in vitro model of the outer annulus fibrosus-cartilage interface.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets