Bone regeneration of demineralized dentin matrix with platelet-rich fibrin and recombinant human bone morphogenetic protein-2 on the bone defects in rabbit calvaria

Abstract

Background This study was to evaluate the bone formation ability of demineralized dentin matrix (DDM) combined with platelet-rich fibrinogen (PRF) and DDM combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) to improve the osteoinductive ability of DDM. Methods After four bone defects with a diameter of 8mm were created in the calvarium of each rabbit, DDM was grafted into the first defect (experimental groups 1), a combination of DDM and PRF was grafted into the second defect (experimental groups 2), and DDM with absorbed rhBMP-2 was grafted into the third defect (experimental groups 3). The fourth defect was used as the control group. Twelve healthy male rabbits (New Zealand, white rabbits) weighing around 3.0–4.0 kg were used. Among 12 rabbits, 3 rabbits were sacrificed immediately after surgery and at 2, 4, and 8 weeks after surgery, respectively. Histopathologic analysis and histomorphometric analysis were conducted to evaluate bone formation in each group. Results The PRF/DDM group did not show a significantly higher degree of new bone formation in calvarial bone defects than the DDM group at 2, 4, and 8 weeks postoperatively in histopathological findings and histomorphometric results. On the other side, the rhBMP-2/DDM group showed higher degrees of new bone formation and calcification, and the lamellae of bone matrix, which are observed in mature bone tissue, were more distinctly visible in the rhBMP-2/DDM group. Moreover, the rhBMP-2/DDM group showed a significantly higher amount of new bone formation, compared to the DDM group at 4 and 8 weeks postoperatively (P<0.05) in histomorphometric results. Conclusion The DDM has great potential as a carrier for the maintenance and sustained release of rhBMP-2, which has been recently receiving wide attention as a type of signaling molecules to promote bone formation.

Demineralized Dentin Matrix for Dental and Alveolar Bone Tissues Regeneration: An Innovative Scope Review

“… Forms and sizes The TDM/DDM were produced in different forms likely as particles, liquid extract, hydrogel, and paste, and in different shapes such as sheets, slices, chips, blocks, discs, root-shaped, barrier membranes, and press-fitted according to the defect shape. Dentin grinder and ball mill machines were used for preparing particulate dentin [ 17 , 19 , 22 , 25 , 26 , 45 , 49 – 52 , 55 , 56 – 60 , 62 , 65 , 68 , 73 , 74 , 76 , 77 , 80 ]. The diameter of the particle size was measured in either µm or mm.…”

“…The diameter of the particle size was measured in either µm or mm. The minimum diameter was less than 40 µm [ 74 ] whereas the maximum was 2 mm [ 45 , 58 , 59 ]. In certain instances, the particles were atelopeptidized [ 25 ], decorated with carboxymethyl chitosan [ 49 ], loaded with recombinant human bone morphogenic protein-2 [ 52 ], encapsulated into liposomes [ 54 ], or used as particle-based bio-ink [ 47 ] (Fig.…”

“…Correspondingly they were 10 min, 5 min, and 10 min [ 17 ], 10 min, 10 min, and 10 min [ 26 ], and 5 min, 5 min, and 10 min [ 49 ]. In addition, 0.6 N HCl [ 45 , 50 , 52 , 57 , 65 , 68 , 73 , 77 , 80 ] was used for demineralization and the exposure time was considered once and varied between 15 min [ 77 ], 30 min [ 52 , 73 ] and 12 h [ 50 ]. The particles obtained from human exfoliated deciduous teeth were demineralized once with 0.6 N HCl and exposure time was varied between 10 min, 15 min, 20 min, 25 min, 30 min, 60 min, and 90 min [ 80 ].…”

“…Sterilization was performed by maintaining the matrices in sterile phosphate-buffered saline (PBS) with 100 units/ml of penicillin and 100 μg/ml of streptomycin at 37 °C, for 72 h and then washed in sterilized deionized water for 5 min [ 17 , 21 , 22 , 24 , 28 , 30 – 36 , 38 – 44 ], socking into penicillin and streptomycin [ 72 ] only, preservation in 5% peracetic acid and 75% ethanol for 10 min [ 53 , 80 , 81 ], immersion in 5 ml alcohol/2 ml of gentamicin [ 61 , 69 – 71 ], rinsing with sterile saline for 10 min [ 75 ], washing in sterilized deionized water for 10 min [ 82 ], rinsing twice in 0.1 M Tris-HCl (pH 7.4) for 10 min [ 55 , 62 ] and using gradient ethanol concentrations [ 47 ]. Other methods of sterilization were gamma irradiation processing using Cobalt 60 radiation with a dose of 5 kGy [ 66 ], ethylene oxide gas sterilization at low temperatures [ 19 , 45 , 49 , 52 , 57 , 58 , 60 , 67 , 68 , 76 ], steam sterilization at 121°C and a pressure of 1 bar for 15 min [ 23 ], lyophilization and freeze-drying [ 46 ], or mixing was accomplished in a sterile container [ 65 ]. The matrices were preserved in α-MEM media containing 50 units/ml of penicillin and 50 μg/ml of streptomycin or in 0.1 x PBS and kept in refrigerator at 4 °C [ 18 , 20 , 24 , 27 , 28 , 30 – 32 , 34 , 36 , 39 – 42 , 44 , 47 ] cryopreserved at -196°C ...…”

Bone Regeneration of a 3D-Printed Alloplastic and Particulate Xenogenic Graft with rhBMP-2

“…However, bone substitutes cannot be entirely superior to autogenous bone because of the bone morphogenetic proteins (BMPs), which are bone-inducing substances in the natural bone matrix. The addition of growth factors, such as recombinant human bone morphogenetic protein-2 (rhBMP-2) and polydeoxyribonucleotide (PDRN), can improve the biological activity and results [11][12][13][14]. A derivative of bone and cartilage formation, rhBMP-2 was first identified in the 1960s and was approved in 2007 as an alternative to an autogenous bone graft for the maxillary sinus, closure surgery of a cleft palate, and localized alveolar ridge augmentation [12,[15][16][17].…”