Guided bone and tissue regeneration

Guided bone regeneration (GBR) and guided tissue regeneration (GTR) are dental surgical procedures that use barrier membranes to direct the growth of new bone and gingival tissue at sites with insufficient volumes or dimensions of bone or gingiva for proper function, esthetics or prosthetic restoration. Guided bone regeneration typically refers to ridge augmentation or bone regenerative procedures; guided tissue regeneration typically refers to regeneration of periodontal attachment. Guided bone regeneration is similar to guided tissue regeneration, but is focused on development of hard tissues in addition to the soft tissues of the periodontal attachment. At present, guided bone regeneration is predominantly applied in the oral cavity to support new hard tissue growth on an alveolar ridge to allow stable placement of dental implants. When bone grafting is used in conjunction with sound surgical technique, guided bone regeneration is a reliable and validated procedure. History

Poroelastic modelling of peri-implnat gap

Evert Robberecht

Micromotion affects the tissue outcome at the peri-implant site. Interstitial fluid flow, induced by micromotion, may play a curtail part in the mesenchymal stem cell differentiation pathway necessary for bone regeneration. The aim of this project is to model the fluid flow at the peri-implant site composed of a soft tissue, for a glenoid implant for different gap sizes at physiological loading.

2008

Study of loading and delivery of drugs from various hydrogel matrices for orthopaedic applications

Tamsine Wills

Nowadays, there are still several unmet medical needs in the field of orthopaedic and trauma surgery leading to the incentive of finding alternative and innovative clinical therapeutics for the treatment of bone regeneration. For example, the entrapment of osteoinductive growth factors in a polymeric matrix with subsequent gradual release and local delivery to the site of trauma. This new technology inspired the activities relating to my project in which I investigated the potential use of synthetic hydrogels for the sustained in situ release of bioactives. The tasks relating to my project were divided into two main experiments; release assays and bioactivity assays. During the release assays, the in vitro release of three osteoinductive growth factors from five different synthetic hydrogels was investigated. The goal was to characterise the release of the protein drugs from the different polymers, selected on the basis of their swelling and degradation behaviour. The quantification of the released bioactives was assessed by performing an enzyme-linked immunosorbent assay (ELISA), for which different parameters were optimised. As a result, release profiles of the different protein drugs from the different hydrogel formulations were obtained. Then, it was investigated if the growth factors were still biologically active or if incubation in synthetic matrices influenced their properties. This was assessed by performing bioassays during which the alkaline phosphatase (ALP) activity of a mouse myoblast cell line (C2C12 cells) was evaluated. Indeed, ALP is a marker of osteoblastic activity which indicates increased levels of osteogenesis, implying biological activity of the tested osteoinductive bioactives. These results gave a good representation of the effect of the synthetic hydrogels and their incubation in release buffer on the bioactivity of the incorporated protein drugs

2009

Study of Cell Adhesion and Tracking Methods for in Vivo Cell Localisation in a Tissue-Engineering Scaffold

Thaïs Cléa Laura Séverine Clouet, Tanja Cloé Hausherr

Bone fracture healing problems, because of the loss of tissue regeneration, can affect many old or multi-pathologic patients around the world. Among the therapeutic strategies, bone substitutes, such as poly-(L-lactitve acid) with 5% wt. % ß-tricalcium phosphate (PLA/5%ß-TCP) scaffolds, are promising as a clinical solution. One strategy to tune their osteoinductive properties is the addition of bone progenitor cells. Therefore, cell- free and cell-seeded PLA/5% ß-TCP scaffolds were implanted into femoral condyles of rats to promote bone regeneration. The in vivo study showed that the bone growth inside a cell-seeded scaffold with human bone progenitor cells was delayed compared to cell-free scaffold. What happened to the cells? Did they trigger an immune response, migrate or die? Therefore, the goals of this project are to evaluate the cell adhesion in vitro to the scaffold and to propose a reliable cell labeling method to track them in future in vivo studies. In the first aim, the adhesion of the cells was qualitatively assessed by the cell morphology and the presence of fibronectin using fluorescence imaging. To quantify their adherence, seeded scaffolds were washed up to 3 times with a 0.9 % NaCl solution. The luminescence of the scaffolds and the washing solutions was measured using a cell viability assay. These experiments demonstrated that after 72h of culture, the measured adherent bone progenitor cells on the scaffolds decreased after 3 washes down to 50 % while it stayed stable after 14 days of culture. In the second aim, we focused on methods to label the cells either once implanted or prior to implantation. The first method was to translate the in situ hybridization protocol with the human specific Alu sequence from paraffin embedded samples to resin embedded ones. The obtained results were so far inconclusive. Concerning the labeling of the cells before implantation, the transmembrane label PKH26 was investigated. Its staining efficacy was high just after labeling, while it decreased drastically over time due to cell division.

2015