|Title:||Intervertebral disc regeneration : small and large scale systems|
|Authors :||Bono, Epifania|
|Conference details:||Ceramics, cells and tissues - 12th annual seminar & meeting, Faenza, 2009|
|License (according to publishing contract) :||Licence according to publishing contract|
|Type of review:||Not specified|
|Subjects :||Regeneration; Disc; Intervertebal|
|Subject (DDC) :||610: Medicine and health|
|Abstract:||Objectives: The intervertebral disc (IVD) degeneration is an aberrant cell-mediated response to progressive structural failure. One of the major causes of the back pain, the IVD degeneration affects about 60% of the population, increasing the costs of the health system. Although the problem of disc degeneration has been approached from many sides, from orthopaedic surgery to molecular biology, it remains unsolved. The present study suggests a tissue engineering alternative to the current surgical therapies, focusing its attention on the regeneration of the nucleus pulposus and on the standardization of its in vitro basic methods. The first objective of this investigation is to analyze the potentiality of an injectable biomaterial combination as carrier for autologus disc cells in the IVD augmentation process. Moreover, in order to reduce the costs of tissue engineering, the second objective of this study regards the establishment of an automated system able to isolate, propagate and characterize in large scale human primary disc cells. Methods: Disc-like cells were isolated by enzymatic digestion from intervertebral disc tissue, harvested during discectomy, and expanded in monolayer cultures before seeding onto a three dimensional system. The cells were seeded onto a macroporous scaffold and subsequently encapsulated into a hydrogel. The viability of cells as well as their capability to proliferate, migrate into the biomaterial combination and synthesize the extracellular matrix was detected by cell and biochemical assays at different culture time points. Moreover, disc cells were genotypically characterized analysing by RT-PCR the expression pattern of specific genes. The disc cell isolation and the monolayer cultures procedures were furthermore performed with the automated liquid handling robot supplied from Tecan® (Tecan® freedom EVO®). Results: Disc-like cells were able to adhere and proliferate onto the three dimensional system showing a homogenous distribution into the surrounding hydrogel structure after few days of culture. Cells synthesized the required extracellular matrix and re-acquired their original genotypic features indicating their redifferentiation in vitro. The automated isolation of primary disc-like cells from human biopsies showed comparable results to the manual one. The isolated cells compared for the number, viability and morphology with the manual processing. Furthermore, cells seeded and maintained in bidimensional culture by the robot revealed the same morphology and growth rate of manually treated cells. Conclusions: This study indicates that the three dimensional developed structure can be a valid system for the treatment of early-stages IVD degeneration. Moreover, the current results obtained with the EVO® robot suggests its feasibility in the automated isolation of disc cells from human biopsy and its propagation in large scale for a future clinical application.|
|Departement:||Life Sciences und Facility Management|
|Organisational Unit:||Institute of Chemistry and Biotechnology (ICBT)|
|Publication type:||Conference Other|
|Appears in Collections:||Publikationen Life Sciences und Facility Management|
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