Generation of a scaffold free cartilage-like implant from a small amount of starting material

Abstract

Introduction: An autologous cellular based treatment of a traumatic cartilage injury requires a procedure whereby a biopsy of healthy cartilage is removed from the patient and the cells isolated and expanded by monolayer passage. This increases the cell number to required levels but also leads to a de-differentiation of the cells. We aim to produce a scaffold-free, de-novo implant from a biopsy of cartilage. Methods: Bovine chondrocytes were isolated from a small biopsy and expanded. The chondrocytic phenotype of the monolayer expanded cells was recovered during a period of culture in alginate and the effect of factors such as IGF1, TFGβ1 and dexamethasone was investigated. Results: During the alginate culture period a pre-treatment with IGF1 and dexamethasone was shown to have little effect. IGF1 however increased the glycosaminoglycan/DNA (GAG/DNA) content on day 14 to 84.95±5ng/ng compared with 37.3±1.8ng/ng in the controls (P <0.001). 35S labeling demonstrated an increased GAG synthesis in the presence of IGF1 (P < 0.001). IGF1 also induced a increase of DNA content 1383±314ng/bead compared to 512±19ng/bead in the controls (P < 0.001). The cells were released from the alginate and cultured in a silicon mould for a further 14 days to obtain a three dimensional implant. Releasing the cells from the alginate and casting in a mould produced an implant of defined shape which contained no foreign material. After 31 days of culture the implants contained 152.4±13.14ng/ng GAG/DNA and 42.93±10.23ng/ng collagen II. Discussion: We believe alginate released chondrocytes provide a real alternative to artificial scaffolds.