This investigation aims to utilise the cell-cell communications between osteoblasts and osteoclasts to create a functional tissue engineered construct that is closer to physiological remodelling than current single cell tissue engineered constructs. A ratio of osteoblasts:osteoclasts was optimised as well as a culture medium that supports both cell types. Four different materials, each with excellent properties for tissue engineering including biocompatible and biodegradable, were compared for their ability to support co-cultures. These materials are; silk fibroin, from Bombyx mori (water vapour and methanol stabilised), chitosan and Poly (1-lactin acid) (PLLA). Silk fibroin and chitosan were shown to support the growth and differentiation of both osteoblasts and osteoclasts in both mono and co-cultures, PLLA did not support osteoclast growth as determined by Tartrate resistant acid phosphatase stain and DNA concentration. The 2D films showed signs of degradation after 10 days culture according to differential scanning calorimetry, fourier transform infra red and gel permeation chromatography. Silk fibroin 3D sponges were manufactured to determine if the co-cultures adhere to, proliferate and differentiate in a 3D environment. Static and dynamic (rotary bioreactor) conditions were compared to determine if the bioreactor conditions enhanced the co-culture of osteoblasts and osteoclasts. The cells were shown to adhere to and proliferate on the sponges by scanning electron microscopy and DNA analysis. The sponges also showed early signs of degradation. The use of silk fibroin and a co-culture system appear to provide excellent potential for bone tissue engineering.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:505664 |
| Date | January 2008 |
| Creators | Jones, Gemma |
| Publisher | Keele University |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
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