The nature of aspidin and the evolutionary origin of bone

Bone is the key innovation underpinning the evolution of the vertebrate skeleton, yet its origin is mired by debate over interpretation of the most primitive bone-like tissue, aspidin. This has variously been interpreted as cellular bone, acellular bone, dentine or as an intermediate of dentine and bone. The crux of the controversy is the nature of unmineralised spaces pervading the aspidin matrix, which have alternatively been interpreted as having housed cells, cell processes, or Sharpey’s Fibres. Discriminating between these hypotheses has been hindered by the limits of traditional histological methods. Here we use Synchrotron X-ray Tomographic Microscopy (srXTM) to reveal the nature of aspidin. We show the spaces exhibit a linear morphology, incompatible with interpretations that they represent voids left by cells or cell processes. Instead, these spaces represent intrinsic collagen fibre bundles that form a scaffold, about which mineral was deposited. Aspidin is thus acellular dermal bone. We reject hypotheses that it is a type of dentine, cellular bone, or transitional tissue. Our study suggests the full repertoire of skeletal tissue types was established prior to the divergence of the earliest known skeletonising vertebrates, indicating that the corresponding cell types evolved rapidly following the divergence of cyclostomes and gnathostomes.