Abstract
Previous investigators have reported that transplanted demineralised dentin matrix (DDM) influences bone formation in vivo. However, the specific mechanism of how dentinal tubules contribute to bone formation has not been determined with regard to DDM transplantation therapy. In this study, we ultrastructurally investigated how DDM contacted the surrounding newly formed bone using a scanning electron microscopy (SEM) three-dimensional reconstruction method that is based on focused ion beam slicing and SEM (FIB/SEM). A pulverised and processed DDM derived from human teeth was implanted into rat calvarial bone defects, and a series of X-ray computed tomographic images were obtained over 12 weeks. Implants with surrounding new bone were removed and histologically examined using FIB/SEM. After obtaining objective block-face images, the target boundary face was reconstructed three-dimensionally. The osteocytes of the new bone tissue surrounding the DDM formed a network connected by their cellular processes and formed bone tissue. It is also interesting that the cellular processes of the osteocytes extended into the dentinal tubules, and that bone tissue with canaliculi had formed and filled the DDM surface.
Highlights
Demineralised dentin matrix (DDM), a collagen material with less antigenicity to release growth factors such as bone morphogenic proteins (BMPs), is clinically applied in several domestic and overseas facilities as a bone-filling agent in the maxillofacial field[1,2,3,4,5,6,7,8]
We successfully reconstructed the ultrastructural interface between the dentinal tubules and new bone tissue using FIB/scanning electron microscopy (SEM) tomography
We found that the osteocytes of the new bone tissue surrounding the demineralised dentin matrix (DDM) formed a network connected by the cellular processes and formed bone tissue and that the cellular processes of the osteocytes extended into the dentinal tubules
Summary
Demineralised dentin matrix (DDM), a collagen material with less antigenicity to release growth factors such as bone morphogenic proteins (BMPs), is clinically applied in several domestic and overseas facilities as a bone-filling agent in the maxillofacial field[1,2,3,4,5,6,7,8]. DDM transplantation therapy was developed after several studies by Urist and other investigators demonstrated that demineralised bone and dentine transplantation induced bone formation[9,10,11]. The BMPs in dentine have been investigated, and in recent years DDM studies have focused on particle size, production, and processing methods that result in more efficient bone formation[14,15]. Dentine includes type I collagen and growth factors, such as BMPs. Type I collagen includes non-collagenous proteins (NCPs), such as phosphophoryn and sialoprotein, which trigger bone resorption and generation processes. Stronger fixation may be obtained by the invasion of the new bone tissue into a porous body material with an irregular surface and highly communicable pores[20]. The relationship between dentinal tubules and new bone formation and their respective influences has not been determined
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