Concentrated growth factor (CGF) is 100% blood-derived, cross-linked fibrin glue with platelets and growth factors. Human CGF clot is transformed into membrane by a compression device, which has been widely used clinically. However, the mechanical properties of the CGF membranes have not been well characterized. The aims of this study were to measure the tensile strength of human CGF membrane and observe its behavior as a scaffold of BMP-2 in ectopic site over the skull. The tensile test of the full length was performed at the speed of 2mm/min. The CGF membrane (5 × 5 × 2 mm³) or the CGF/BMP-2 (1.0 μg) membrane was grafted onto the skull periosteum of nude mice (5-week-old, male), and harvested at 14 days after the graft. The appearance and size of the CGF membranes were almost same for 7 days by soaking at 4 °C in saline. The average values of the tensile strength at 0 day and 7 days were 0.24 MPa and 0.26 MPa, respectively. No significant differences of both the tensile strength and the elastic modulus were found among 0, 1, 3, and 7 days. Supra-periosteal bone induction was found at 14 days in the CGF/BMP-2, while the CGF alone did not induce bone. These results demonstrated that human CGF membrane could become a short-term, sticky fibrin scaffold for BMP-2, and might be preserved as auto-membranes for wound protection after the surgery.

The aim of this study was to evaluate the bio-absorption and bone regeneration of human tooth-derived dentin scaffold, entitled as perforated root-demineralized dentin matrix (PR-DDM), after in vivo implantation into the critical-size iliac defects. The dentin scaffolds were prepared from human vital, non-functional teeth. Thirty artificial macro-pores (Ø 1 mm) were added after removing the enamel portion. The modified teeth were supersonically demineralized in 0.34 N HNO₃ for 30 min. The microstructure was observed by scanning electron microscope (SEM). The 3D micro-CT and histological analysis were carried out to evaluate the bio-absorption of PR-DDM at 2 and 4 months. A smooth dentin collagen surface with symmetrical macro-pores and tube-type dentinal tubules (Ø 1–2 µm) with micro-cracks were observed on the perforated region. A significant number of custom-made macro-pores disappeared, and the size of the macro-pores became significantly wider at 4 months compared with the 2 months (p < 0.05) evaluated by 3D micro-CT. Histological images revealed the presence of multinucleated giant cells attached to the scalloped border of the PR-DDM. The morphological changes due to bio-absorption by the cellular phagocytes were comparable to the 3D micro-CT and histological images at 2 and 4 months. Altogether, the results demonstrated that the PR-DDM block was gradually absorbed by multinucleated giant cells and regenerated bone. Human PR-DDM might serve as a unique scaffold for extraoral bone regeneration.

Osteoinduction in muscles by porous ceramics has been reported to be a real phenomenon. In this study, osteoinduction in connective tissues was found in highly porous hydroxyapatite (HAp) ceramics with large specific surface areas. We have developed the combination method of the partial dissolution-precipitation (PDP) technique involving the stirring-supersonic treatment in 1.7 × 10⁻² N HNO₃ solution containing Ca²⁺ and PO₄³⁻ to improve the surface and the bulk of commercially available synthetic HAp block (82.5% in porosity, 50–300 µm in pore size). The modified HAp was named as a partially dissolved and precipitated HAp (PDP-HAp). The PDP-HAp exhibited the porosities of 85–90%, the macropore sizes of 50–200 µm, and the specific surface areas of 1.0–2.0 m²/g, with microcracks. The aim of this study was to observe bone induction by the PDP-HAp with or without BMP-2 in scalp tissues of four-week-old rats. Young rats were divided into the PDP-HAp alone group and the PDP-HAp/BMP-2 group for a long-term observation. In the PDP-HAp group, bone induction occurred inside the many pores at nine months, and the ratio of induced bone was 12.0%. In the PDP-HAp/BMP-2 group, bone induction occurred in almost all pores at three months, and compact bone was found at nine months. The ratios of induced bone were 77.0% at three months and 86.0% at nine months. We believe that osteoinduction by the PDP-HAp might be different from the process of BMP-loaded HAp-induced bone formation, because the PDP-HAp has osteogenic microporous compartments with partially absorbable HAp crystals. The PDP technique may contribute to create bioceramics with osteoinductive property for bone regenerative medicine.

Autologous blood-derived fibrin glue with platelets, called the concentrated growth factor (CGF), can be prepared immediately by only the decided centrifuge without the addition of coagulation factors. Collagen materials combined with recombinant human BMP-2 have been commercially available for clinical use. The fresh CGF is auto-clot with wettability and elasticity, while most collagen membranes are derived from the cow or pig. The fresh CGF has wettability and elasticity, while collagen membranes are dry materials without elasticity. The aim of this study was to observe the microstructures of human CGF membrane and evaluate its behavior as a delivery scaffold of rhBMP-2 in the subcutaneous tissues of nude mice. Twenty-four nude mice (5-week-old, male) were used for the assessment of in vivo ectopic bone formation. Mice were received the CGF membrane as the controls and the CGF/rhBMP-2 membrane as the experimental group in the subcutaneous tissues, and harvested at 7, 10, and 14 days after the graft. Harvested samples were evaluated for the histological examination and the histomorphometric measurement was conducted to compare the residue of the CGF, as well as the new bone. Mature fibrin fibers assembled from multiple fibrillary elements and platelets with the rhBMP-2 membrane induced several bony islands and cartilage without residues of CGF at 14 days, while the CGF membrane alone was almost absorbed at 10 days and failed to induce bone formation at 14 days. These results demonstrated that the fresh, human CGF membrane could contribute to a short-term, sticky fibrin matrix for the delivery of rhBMP-2.

A vital tooth-derived demineralized dentin matrix (DDM) is defined as acid-insoluble, cross-linked collagen with growth factors. The objective of this study is to evaluate the feasibility of the perforated-DDM for bone regeneration. Thirty perforations were added to the human vital wisdom tooth after the removal of enamel portion. The modified tooth was partially demineralized in 0.34 N HNO 3 for 30 min and grafted into the critical defect of sheep iliac to evaluate bone regeneration. Sheep were sacrificed for µ-CT and histological evaluation at 2 and 4 months after the graft. New bone was connected directly with the grafted DDM at 2 months. The volume of new bone was comparable with the µ-CT, 3D, and the histological findings at 2 and 4 months. The amount of new bone tissue in the critical defects at 4 months was higher than that at 2 months. The results revealed that the human tooth-derived block material with artificial porous structure, the perforated-DDM, successfully contributed to bone ingrowth as the unique scaffold. This novel human tooth-derived scaffold may be a promising biomaterial to induce bone regeneration in the tri-cortical critical-sized bone defects.