Neuronal cultures are an excellent experimental tool to study the collective behaviour of neuronal ensembles in a controlled environment. However, neurons cultured on flat surfaces present limitations in terms of their functionality, as they exhibit a synchronous dynamic behaviour that differs from the much richer repertoire of brain activity [1].In order to address this limitation, we studied the capacity to break the isotropy in connectivity and enrich dynamics by modulating the spatial arrangement of neurons in the substrate they grow. Specifically, we cultured neurons on polydimethylsiloxane (PDMS) topographical patterns with both Euclidean and fractal geometry. In addition, we explored the influence of excitatory-inhibitory neuron ratios on dynamic modulation using antagonists. Neuronal activity was recorded using calcium fluorescence imaging technique and was analysed in the context of complex networks. Networks formed on topographical patterns were characterized by rich spatiotemporal activity patterns that comprised from localized regions to the entire culture, resulting in more complex dynamics compared to standard cultures. In addition, we observed the importance of inhibition in modulating the activity pattern repertoire, either by decreasing the inhibitory synapse strength to homogenize burst events or by reducing the amount of excitatory transmission to promote segregation [2].This research demonstrates the capacity of spatial constraints to influence the activity and functional organisation of neuronal cultures, allowing an improvement in the ability to replicate brain dynamics and to provide a more realistic experimental platform.[1] J. G. Orlandi, et al., 2013, Nat. Phys.[2] M. Montalà-Flaquer, et al., 2022, iScience.