Establishment of cocultures of osteoblasts, Schwann cells, and neurons towards a tissue-engineered approach for orofacial reconstruction.

In orofacial reconstruction not only the osseous structures themselves but also neighboring cranial nerves need to be regenerated. To replace autologous bone implants, biocompatible tissue-engineered scaffolds are under investigation at least for bone replacement but until now these studies have not focused on parallel reconstruction of injured cranial nerves. The present study contributes to the development of optimized tissue-engineered products that will enable regeneration of both bone and nervous tissue. For the first time, cocultures of primary osteoblasts (rat or human) and primary Schwann cells (rat or human) were established. The suitability of monocultures of osteoblasts and cocultures of osteoblasts plus Schwann cells as substrate for sensory neurons as well as motoneurons was tested here. The results suggest that whereas osteoblasts provide a good substrate for sensory neurons, motoneurons depend on the presence of Schwann cells for survival and neurite outgrowth. For prolonged availability of regeneration-promoting growth factors at the site of the graft, those proteins should be delivered by the transplanted cells themselves. To enable this, we established electroporation-based nonviral transfection of osteoblasts as well as Schwann cells. Our new cell culture system will enable investigations of the effect of graft-derived growth factors on osteoblasts and Schwann cells as well as on neurite outgrowth from cocultured neurons of the sensory and motor system.