The idea of a tissue engineered nucleus implant is to seed cells in a three-dimensional collagen matrix. This matrix may serve as a scaffold for a tissue engineered nucleus implant. The aim of this study was to investigate whether implantation of the collagen matrix into a spinal segment after nucleotomy is able to restore disc height and flexibility. The implant basically consists of condensed collagen type-I matrix. For clinical use, this matrix will be used for reinforcing and supporting the culturing of nucleus cells. In experiments, matrixes were concentrated with barium sulfate for X-ray purposes and cell seeding was disclaimed in order to evaluate the biomechanical performance of the collagen material. Six bovine lumbar functional spinal units, aging between 5 and 6 months, were used for the biomechanical in-vitro test. In each specimen, an oblique incision was performed, the nucleus was removed and replaced by a collagen-type-I matrix. Specimens were mounted in a custom-built spine tester, and subsequently exposed to pure moments of 7.5 Nm to move within the three anatomical planes. Each tested stage (intact, nucleotomy and implanted) was evaluated for range of motion, neutral zone and change in disc height. Removal of the nucleus significantly reduced disc height by 0.84 mm in respect to the intact stage and caused an instability in the segment. Through the implantation of the tissue engineered nucleus it was possible to restore this height and stability loss, and even to increase slightly the disc height of 0.07 mm compared with the intact stage. There was no statistical difference between the stability provided by the implant and intact stage. Results of movements in lateral bending and axial rotation showed the same trend compared to flexion/extension. However, implant extrusions have been observed in three of six cases during the flexibility assessment. The results of this study directly reflect the efficacy of vital nucleus replacement to restore disc height and to provide stability to intervertebral discs. However, from a biomechanical point of view, the challenge is to employ an appropriate annulus fibrosus sealing method, which is capable to keep the nucleus implant in place over a long-time period. Securing the nucleus implant inside the disc is one of the most important biomechanical prerequisites if such a tissue engineered implant shall have a chance for clinical application.