An Ex Vivo 3 Dimensional Organotypic Culture Model: A First Attempt to Stimulate Endochondral Ossification of the Growth Plate in Vitro
Eingereicht von: Sriveena Srinivasaiah, MSc
Firma/Universität: Medical University of Graz – University Clinics for Orthopedics and Traumatology
Kooperationspartner: Helmholtz-Zentrum Geesthacht GmbH; Mayo clinic
Bone tissue engineering and reconstruction remains as a very challenging approach in the field of orthopedics because of its structural complexities that often exhibits limited reaction to the chemical and physical factors in vitro, most of these processes are curbed by lack of native extracellular matrix anchoring vital proteins that are responsible for various signaling cascades involved in the recruitment of growth and development of a normal bone. So far, years of experiment have shown that it is very difficult to maintain a functional bone tissue in vitro. On the latest trend the organotypic culture model has been one of the key inventions in various fields promoting a large demand for the various clinical investigations and therapeutic interventions. Developing an alternative tissue engineering constructs could be a beneficial measure from a biological perspective, as it enables comprehensive understanding of developmental and reparative processes influenced by a large number of physical and chemical manifestations involved in bone formation.
A functional bone organotypic model was developed for the first time using a natural bone template consisting 3D organisation of bone tissue with its heterogeneous cell population, extracellular matrix and intrinsic growth factors. The organotypic culture shows similar mechanisms involved in long bone formation there by providing a great opportunity to understand the underlying mechanisms involved in bone development. The results so far validate that the bone slices respond to osteogenic conditions up to 2 weeks promoting osteogenic differentiation of mesenchymal stem cells present in the native template there by eventually leading to endochondral and perichondral ossification of the bone slices. In order to establish this above test system bone slices were generated from a 4-day-old newborn rat femur and subjected to osteogenic stimuli. The bone slices were assessed for different parameter by histological evaluation to estimate the bone turnover. This current data shows the quantitative evaluations corresponding to in vitro endochondral bone formation. The current objective is to perform proteomic investigations by high throughput screening by mass spectrometric proteomics and RNA sequencing in collaboration with Mayo clinic, Rochester. The research team is anticipating that this approach would enable us to detect some of the key biomolecules involved in bone development, with greater chances of discovering novel biomolecules that could perhaps be very crucial in the process of bone development. The over impact of this study can be very beneficial in the field of orthopedics and biomaterials as it provides more realistic platform for preclinical investigatations.