J. Waletzko-Hellwig (Rostock, DE)
Biomechanics and Implant Technology Research LaboratoryPresenter Of 1 Presentation
P053 - Differentiation Capacity of Rabbit Derived Chondrocytes Using Different FBS Concentration and Biophysical Stimulation
Abstract
Purpose
A widely used technique for the treatment of cartilage lesions is matrix-induced autologous chondrocyte implantation. However, in some cases the defect area is not sufficiently filled with hyaline cartilage matrix, so strategies need to be improved by tissue engineering approaches. One possibility represents the additional biophysical stimulation during pre-cultivation to promote differentiation of the cartilaginous cells. In order to investigate such an approach in an animal study, a prior analysis and parameter verification with suitable animal cells is necessary.
Methods and Materials
Chondrocytes derived from articular cartilage of New Zealand White rabbits were isolated and the optimal FBS concentration was verified for further biophysical stimulation experiments. Therefore, cells were cultivated with 1 %, 5 % or 10 % FBS in monolayers or on a collagenous scaffold over five days at 37 °C and 5% CO2. The most promising FBS concentration was used for further electrostimulation experiments. For this purpose, cells were seeded on collagenous scaffolds and exposed to alternating electrical fields ranging from 0.01 to 0.6 V/m for seven days according to Krueger et al. (2021). Afterwards, metabolic activity and gene expression levels of chondrogenic markers were analysed.
Results
While the metabolic activity of cells increased with higher FBS concentration, a simultaneous reduction in differentiation capacity (Col2: p=0.0024 1% vs. 10% FBS; p=0.0004 1% vs. 20% FBS) in 2D and 3D culture was visible. Electrical stimulation with very low levels of alternating electric fields (0.01 V/m) maintained chondrogenic differentiation capacity, with no significant changes compared to the control.
Conclusion
The results indicate that the FBS concentration directly affects cartilaginous cell metabolism and differentiation whereby 1% FBS showed the most promising results. Cell experiments with application of alternating electrical fields revealed less influence, however further experiments with a longer stimulation period as well as combination with mechanical stimuli are planned to increase chondrogenic differentiation.
Presenter Of 1 Presentation
P053 - Differentiation Capacity of Rabbit Derived Chondrocytes Using Different FBS Concentration and Biophysical Stimulation
Abstract
Purpose
A widely used technique for the treatment of cartilage lesions is matrix-induced autologous chondrocyte implantation. However, in some cases the defect area is not sufficiently filled with hyaline cartilage matrix, so strategies need to be improved by tissue engineering approaches. One possibility represents the additional biophysical stimulation during pre-cultivation to promote differentiation of the cartilaginous cells. In order to investigate such an approach in an animal study, a prior analysis and parameter verification with suitable animal cells is necessary.
Methods and Materials
Chondrocytes derived from articular cartilage of New Zealand White rabbits were isolated and the optimal FBS concentration was verified for further biophysical stimulation experiments. Therefore, cells were cultivated with 1 %, 5 % or 10 % FBS in monolayers or on a collagenous scaffold over five days at 37 °C and 5% CO2. The most promising FBS concentration was used for further electrostimulation experiments. For this purpose, cells were seeded on collagenous scaffolds and exposed to alternating electrical fields ranging from 0.01 to 0.6 V/m for seven days according to Krueger et al. (2021). Afterwards, metabolic activity and gene expression levels of chondrogenic markers were analysed.
Results
While the metabolic activity of cells increased with higher FBS concentration, a simultaneous reduction in differentiation capacity (Col2: p=0.0024 1% vs. 10% FBS; p=0.0004 1% vs. 20% FBS) in 2D and 3D culture was visible. Electrical stimulation with very low levels of alternating electric fields (0.01 V/m) maintained chondrogenic differentiation capacity, with no significant changes compared to the control.
Conclusion
The results indicate that the FBS concentration directly affects cartilaginous cell metabolism and differentiation whereby 1% FBS showed the most promising results. Cell experiments with application of alternating electrical fields revealed less influence, however further experiments with a longer stimulation period as well as combination with mechanical stimuli are planned to increase chondrogenic differentiation.