To quantify length and tortuosity, total (curved) and theoretical (chord) lengths were measured in Matlab for the whole system length and for the corresponding main branch. The curved length was defined as a cumulative sum of 3D Euclidean distances between neighboring graph points (i.e. links forming points) multiplied by voxel size. The chord length was defined as cumulative sum of 3D Euclidean distances between neighboring nodes multiplied by voxel size. The chord length therefore reflects system length where any nodes are connected by links with the shortest possible length. To analyze the relationship between BD and PV a length of the BD was divided by a PV length (curved or chord). Tortuosity was calculated as curved length divided by chord length of the same system and distributed into regions based on the generation number as previously described. Tortuosity was assessed in %, as BD and PV are not straight lines the actual tortuosity measurements were subtracted by 100% (perfectly straight line).

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