Influence of implant material and surface on differentiation and proliferation of human adipose derived stromal cells”

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Abstract

Introduction For guided regeneration of periimplant hard and soft tissues it is of crucial importance to evaluate the interaction of different cofactors and to know the decisive activating cascades. The classic triad of tissue engineering and (personalised) tissue regeneration is the interplay of cells, biologically active proteins and a carrier matrix (scaffold). Regarding the typical periimplant defect it does not seem sensible to support the physiological course of regeneration by replacing the lacking tissue by extra corporally designed and fabricated constructs; the vascular coupling and therefore the integration of the implanted cells often remains difficult. It is much more promising to try to stimulate the body's own, intrinsic regenerative capacity and hence to significantly increase the tissue restoration in situ. In the context of personalized regenerative concepts, the principle of an autologous approach combining individual cells, a (resorbable) matrix and bioactive mediators seems to provide the most reliable results. We used human adipose derived stromal cells (hADSC) isolated from abdominal fat tissue as a promising source for mesenchymal stromal cells. These multipotent autologous cells showed the best properties. Results were compared to edaphic cells as human mandibular osteoblasts (hOB) and human gingival fibroblasts (HGF). Methods In this study the characteristics' of hADSC cultivated on two different dental implant materials (titanium and zirconium) was analysed. Furthermore, the influences of different surface textures (rough and polished / machined) were analysed (titanium SLA- and PT-surface, zirconium ZLA- and M-surface). Cells were seeded onto the dental material (test bodies) and analysed at three different time points (1d-7d-14d). Cell proliferation and viability (MTT / living cell count) was analysed. Further, cytotoxic effects (LDH) and stem cell activity / osteogenic activity (ALP) was analysed. Alteration in cell morphology (SEM) and direct morphological information of cytoskeleton alteration (f-actin staining) was analysed. Alterations of hADSC concerning their pluripotency and their potential to differentiate into osteogenic linage expression of different markers were analysed at gene (RT-qPCR) and at protein (ELISA) level. Statistical analysis was carried out by one way ANOVA and p<0.05 (SPSS; version 24). Results As postulated, the cultivation on zirconium and here on the ZLA-surface showed a significant better proliferation rate for hADSC and also for hOB and HGF cells (p<0.05). Cytotoxic effects became apparent during the first days for all tested materials, but were negligible. At time point 14d a cell layer was observable on all test bodies. Compared to polished / machined surface showed the rough surfaces a significant better stimulation of gene and of protein expression during time (p<0.05). Stem cell potential and osteogenic differentiation was more stimulated by the rough surface (p<0.05).

Speakers from the University of Münster

Jung, Susanne	Clinic for Cranio-Maxillofacial Surgery
Kleinheinz, Johannes	Clinic for Cranio-Maxillofacial Surgery
Sielker, Sonja	Clinic for Cranio-Maxillofacial Surgery