Multimodal Approach to Assess Bone Regeneration and Scaffold Performance

The clinical success of an implant largely depends on its ability to promote bone repair and facilitate osteointegration. While significant advancements have been made in designing new scaffolds, a thorough evaluation of implant success at various bone length scales has not been sufficiently investigated. Bone is a hierarchically organized tissue, with an intricate structure that ranges from the organization of collagen fibrils and their mineralization at the nanometer and micrometer scales to the macroscopic arrangement of compact and cancellous bone. This hierarchical organization is crucial for ensuring bone function and mechanical performance. In this context, a comprehensive assessment of the effects of biomaterials on bone repair at multiple length scales is essential for evaluating the effectiveness and safety of implanted materials. However, preparing samples for integrated multimodal analyses poses a technical challenge. In this work, we present a protocol designed to evaluate bone regeneration across various hierarchical levels and analyze the interface between newly formed bone and the implanted scaffold. This approach provides a robust method for assessing the effectiveness of scaffolds in inducing bone repair. The combination of selected 3D and 2D imaging techniques is crucial for a proper evaluation of the complex mineralized tissue that forms during the bone healing process.