Biphasic Calcium Phosphate Block Research Articles

Dual-phase blocks for regeneration of critical-sized bone defects

Repair of critical-size bone defects (CBD) remains a challenge in orthopedic surgery due to the restricted regenerative capacity of bone tissue. Herein, multi-functional biphasic calcium phosphate (BCP) blocks decorated with gelatin packs were developed for bone regeneration of CBDs. The gelatin packs consist of gelatin microspheres (GMSs) and an enzymatic crosslinked gelatin layer for bone morphogenetic protein-2 (BMP-2) and antibiotics delivery. The porous interior of the BCP block was filled with GMSs containing BMP-2 and tightly packed with an enzymatic crosslinked gelatin layer. The outermost part of the block was then covered with antibiotics-loaded GMSs. The BCP platform elaborately decorated for each zone improved the adhesion of osteoblasts and enabled the sequential release of antibiotics and BMP-2 and sustained release of BMP-2, realizing on-demand drug release according to the treatment stage. The programmed drug release of this platform significantly improved the regeneration of CBDs. In vivo experiments using dog's mandible defect models clearly demonstrated the clinical application potential of this BCP platform.

Efficacy of Application of Periodontal Ligament Stem Cells in Bone Regeneration: A Systematic Review of Animal Studies

Introduction: We aimed to systematically review the animal studies that have investigated the efficacy of periodontal ligament stem cells (PDLSCs) in bone regeneration. Methods: Web of science, Scopus, PubMed, Embase, Cochrane Library, and ProQuest were searched to September 2022 for animal studies investigating bone regeneration using PDLSCs. Results: Twenty studies were included. Calvaria and alveolar defects were treated with stem cells. The cells were mainly carried with hydrogel, hydroxyapatite, and collagen scaffolds. PDLSCs promoted osteogenesis compared with cell-free scaffolds, except in one study where biphasic calcium phosphate block scaffolds alone or with Bone morphogenic protein 2 had superior outcomes in calvaria defects. Controversial results were reported when comparing the osteogenic potential of PDLSCs and bone marrow stem cells. Two studies reported higher potential in BMSCs, and two others reported higher bone formation and more bone quality in PDLSCs. Conclusion: With the limitations of this study, PDLSCs might have promising potential to accelerate bone regeneration in artificial defects; however, due to high heterogeneity in the outcomes of the present studies, before moving forward to human experiments, further preclinical in vivo studies are needed.

Scanning Electron Microscopic Evaluation of the Internal Fit Accuracy of 3D-Printed Biphasic Calcium Phosphate Block: An Ex Vivo Pilot Study.

The aim of this study was to assess the internal fit accuracy of a three-dimensional (3D)-printed biphasic calcium phosphate (BCP) block compared with a 3D-milled poly methyl methacrylate (PMMA) block by scanning electron microscope (SEM) analysis. In a total of 20 porcine rib bones, two different types of defects having two adjacent walls and a floor were produced: a defect with a flat floor (flat defect; N = 10) and a defect with a concave floor (curved defect; N = 10). Each defect was grafted with either the 3D-printed BCP block or the 3D-milled PMMA block fabricated following the computer aided design. The defects were then cut cross-sectionally and evaluated under the SEM. The extents of internal contact and gap were measured and statistically analyzed (p < 0.05). All blocks in both BCP and PMMA groups were successfully fit to the flat and curved defects. The internal contact ratio was significantly higher in the BCP group (flat defect: 0.47 ± 0.10; curved defect: 0.29 ± 0.05) compared with the PMMA group (flat defect: 0.21 ± 0.13; curved defect: 0.17 ± 0.04; p < 0.05). The internal gap area was similar between the two groups regardless of the defect types (p > 0.05). The internal fit accuracy of the 3D-printed BCP block was reliable in both the flat and curved defects when compared with the accuracy of the 3D-milled PMMA block.

Bone Regeneration Using a Three-Dimensional Hexahedron Channeled BCP Block Combined with Bone Morphogenic Protein-2 in Rat Calvarial Defects.

It is important to obtain sufficient bone mass before implant placement on alveolar bone, and synthetic bone such as biphasic calcium phosphate (BCP) has been studied to secure this. This study used a BCP block bone with a specific structure of the three-dimensional (3D) hexahedron channel and coating with recombinant human bone morphogenetic protein-2 (rhBMP-2) impregnated carboxymethyl cellulose (CMC) was used to examine the enhancement of bone regeneration of this biomaterial in rat calvarial defect. After the preparation of critical-size calvarial defects in fifteen rats, defects were divided into three groups and were implanted with the assigned specimen (n = 5): Boneplant (untreated 3D hexahedron channeled BCP block), Boneplant/CMC (3D hexahedron channeled BCP block coated with CMC), and Boneplant/CMC/BMP (3D hexahedron channeled BCP block coated with CMC containing rhBMP-2). After 4 weeks, the volumetric, histologic, and histometric analyses were conducted to measure the newly formed bone. Histologically, defects in the Boneplant/CMC/BMP group were almost completely filled with new bone compared to the Boneplant and Boneplant/CMC groups. The new bone volume (P < 0.05) and area (P < 0.001) in the Boneplant/CMC/BMP group (20.12% ± 2.17, 33.79% ± 3.66) were much greater than those in the Boneplant (10.77% ± 4.8, 16.48% ± 9.11) and Boneplant/CMC (10.72% ± 3.29, 16.57% ± 8.94) groups, respectively. In conclusion, the 3D hexahedron channeled BCP block adapted rhBMP-2 with carrier CMC showed high possibility as an effective bone graft material.

Bone regeneration using three-dimensional hexahedron channel structured BCP block in rabbit calvarial defects.

The purpose of this study is to evaluate the efficacy of bone regeneration and volume maintenance of the three-dimensional (3D) structured biphasic calcium phosphate (BCP) block with porous hexahedron channels in a rabbit calvarial model. In this work, four circular defects (diameter: 8 mm) in calvarium of rabbits were randomly assigned to (1) negative control (control), (2) 3D hexahedron channel structured BCP block, (3) deproteinized bovine bone mineral particle, and (4) deproteinized porcine bone mineral particle. Animals were euthanized at 2 (n = 5) and 8 weeks (n = 5). Outcome measures included micro-computed tomography (CT) and histomorphometrical analysis. Results indicated that in micro-CT, BCP group showed the highest new bone volume with significant difference compared to control (p = 0.008) at 8 weeks. Histomorphometrically, total augmented area of BCP group was the highest with significant difference compared to control (p = 0.008) at 8 weeks. BCP group also maintained total volume of the original defect without collapsing. BCP block with 3D hexahedron channel structure seems to have favorable osteogenic and volume maintaining ability and highly porous structure might attribute to new bone formation. Further studies regarding the optimal internal structure and porosity of the BCP block bone substitute are needed. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 2254-2262, 2019.

Evaluation of In Vivo Osteogenic Potential of Bone Morphogenetic Protein 2-Overexpressing Human Periodontal Ligament Stem Cells Combined with Biphasic Calcium Phosphate Block Scaffolds in a Critical-Size Bone Defect Model.

Human periodontal ligament stem cells (hPDLSCs) are considered potential cellular carriers for gene delivery in the field of tissue regeneration. This study tested the osseoregenerative potential of hPDLSCs transduced with replication-deficient recombinant adenovirus (rAd) containing the gene encoding bone morphogenetic protein-2 (BMP2; hPDLSCs/rAd-BMP2) in both in vivo and in vitro osteogenic environments. After the optimal condition for rAd-mediated transduction was determined, hPDLSCs were transduced to express BMP2. In vivo bone formation was evaluated in a critical-size rat calvarial bone defect model that more closely mimics the harsher in vivo milieu for bone regeneration than subcutaneous transplantation model. As support materials for bone regeneration, block-type biphasic calcium phosphate (BCP) scaffolds were combined with hPDLSCs and/or BMP2 and transplanted into critical-size bone defects in rats. Experimental groups were as follows: BCP scaffold control (group 1 [Gr1]), scaffold containing recombinant human BMP2 (rhBMP2; group 2 [Gr2]), scaffold loaded with normal hPDLSCs (group 3 [Gr3]), scaffold combined with both normal hPDLSCs and rhBMP2 (group 4 [Gr4]), and scaffold loaded with hPDLSCs transduced with rAd-BMP2 (hPDLSCs/rAd-BMP2; group 5 [Gr5]). Our data showed that new bone formation was highest in Gr2. Less mineralization was observed in Gr3, Gr4, and Gr5 in which hPDLSCs were transplanted. In vitro transwell assay demonstrated that hPDLSCs exert an inhibitory activity on BMP2-induced osteogenic differentiation. Our findings suggest that the in vivo bone regenerative potential of BMP2-overexpressing hPDLSCs could be compromised in a critical-size rat calvarial bone defect model. Thus, further investigations are required to elucidate the underlying mechanisms and to develop efficient techniques for improved tissue regeneration.

Comparative analysis of carrier systems for delivering bone morphogenetic proteins.

The objective of this study was to comparatively assess the bone regenerative capacity of absorbable collagen sponge (ACS), biphasic calcium phosphate block (BCP) and collagenated biphasic calcium phosphate (CBCP) loaded with a low dose of recombinant human bone morphogenetic protein-2 (rhBMP-2). The CBCP was characterized by X-ray diffraction and scanning electron microscopy. In rabbit calvaria, four circular 8-mm-diameter defects were created and assigned to one of four groups: (1) blood-filled group (control), (2) rhBMP-2-soaked absorbable collagen sponge (0.05 mg/mL, 0.1 mL; CS group), (3) rhBMP-2-loaded BCP (BCP group), or (4) rhBMP-2-loaded CBCP (CBCP group). The animals were sacrificed either 2 weeks or 8 weeks postoperatively. Histological and histomorphometric analyses were performed. The CBCP showed web-like collagen fibrils on and between particles. Greater dimensional stability was observed in the BCP and CBCP groups than in the control and the CS groups at 2 and 8 weeks. The new bone formation was significantly greater in the BCP and CBCP groups than in the control and CS groups at 2 weeks, but did not significantly differ among the four groups at 8 week. The CBCP group exhibited more new bone formation in the intergranular space and in the center of the defect compared to the BCP group at 2 weeks, but a similar histologic appearance was observed in both groups at 8 weeks. The dose of rhBMP-2 in the present study enhanced bone regeneration in the early healing period when loaded on BCP and CBCP in rabbit calvarial defects.

Effectiveness of biphasic calcium phosphate block bone substitutes processed using a modified extrusion method in rabbit calvarial defects

This study evaluated the mechanical and structural properties of biphasic calcium phosphate (BCP) blocks processed using a modified extrusion method, and assessed their in vivo effectiveness using a rabbit calvarial defect model. BCP blocks with three distinct ratios of hydroxyapatite (HA):tricalcium phosphate (TCP) were produced using a modified extrusion method:HA8 (8%:92%), HA48 (48%:52%), and HA80 (80%:20%). The blocks were examined using scanning electron microscopy, X-ray diffractometry, and a universal test machine. Four circular defects 8 mm in diameter were made in 12 rabbits. One defect in each animal served as a control, and the other three defects received the BCP blocks. The rabbits were sacrificed at either two weeks (n=6) or eight weeks (n=6) postoperatively. The pore size, porosity, and compressive strength of the three types of bone block were 140-170 µm, >70%, and 4-9 MPa, respectively. Histologic and histomorphometric observations revealed that the augmented space was well maintained, but limited bone formation was observed around the defect base and defect margins. No significant differences were found in the amount of new bone formation, graft material resorption, or bone infiltration among the three types of BCP block at either of the postoperative healing points. Block bone substitutes with three distinct compositions (i.e., HA:TCP ratios) processed by a modified extrusion method exhibited limited osteoconductive potency, but excellent space-maintaining capability. Further investigations are required to improve the processing method.

Bone formation of block and particulated biphasic calcium phosphate lyophilized with Escherichia coli–derived recombinant human bone morphogenetic protein 2 in rat calvarial defects

The objective of this study was to evaluate bone formation in rat calvarial defects after surgical implantation of block or particulated biphasic calcium phosphate (BCP) lyophilized with Escherichia coli-derived recombinant human bone morphogenetic protein 2 (ErhBMP-2). Critical-size calvarial osteotomy defects were created in 5 groups of Sprague-Dawley rats. Each group received one of the following: 1) sham surgery control; 2) biphasic calcium phosphate particles (CPP); 3) biphasic calcium phosphate block (CPB); 4) ErhBMP-2-coated CPP; or 5) ErhBMP-2-coated CPB. ErhBMP was coated on BCP by a stepwise lyophilizing protocol. The new bone formation was significantly greater in ErhBMP-2-treated groups compared with the untreated group. In particular, the ErhBMP-2/CPB group showed stability of augmented areas during the period of healing, due to relevant space-providing capacity. Thus, it can be concluded that CPP and CPB lyophilized with ErhBMP-2 enhance the formation of new bone, and CPB appears to be a suitable carrier for ErhBMP-2 in which a 3-dimensional structural integrity is an important consideration factor.

RhBMP-2 using biphasic calcium phosphate block as a carrier induces new bone formation in a rat subcutaneous tissue

Purpose: The carrier for the delivery of bone morphogenetic proteins(BMPs) should also serve as a scaffold for new bone growth. In addition, predictable bone formation in terms of the volume and shape should be guaranteed. This study evaluated the ectopic bone formation of recombinant human BMP-2(rhBMP-2) using a micro macroporous biphasic calcium phosphate (MBCP: mixture of β-TCP and HA) block as a carrier in a rat subcutaneous assay model. Materials and Methods: Subcutaneous pockets were created on the back of 40 male Sprague-Dawley rats. In the pockets, rhBMP-2/MBCP and MBCP alone were implanted. The blocks were evaluated by histological and histometric parameters after a healing interval of 2 weeks (each 10 rats; MBCP and rhBMP-2/MBCP) or 8 weeks (each 10 rats; MBCP and rhBMP-2/MBCP). Results: The shape and volume of the block was maintained stable over the healing period. No histological bone forming activity was observed in the MBCP alone sites after 2 weeks and there was minimal new bone formation at 8 weeks. In the rhBMP-2/MBCP sites, new bone formation was evident in the macropores of the block. The new bone area at 8 weeks was greater than at 2 weeks. There was a further increase in the quantity of new bone with the more advanced stage of remodeling. Conclusions: A MBCP block could serve as a carrier system for predictable bone tissue engineering using rhBMPs. (J Korean Acad Periodontol 2008;38:355-362)

The Effect of Recombinant Human Bone Morphogenetic Protein-2/Macroporous Biphasic Calcium Phosphate Block system on Bone Formation in Rat Calvarial Defects

대한치주과학회지 : Vol. 37, No. 2(Suppl.), 2007 The Effect of Recombinant Human Bone Morphogenetic Protein-2/Macroporous Biphasic Calcium Phosphate Block system on Bone Formation in Rat Calvarial Defects Young-Jun Lee, Sung-Won Jung, Gyung-Joon Chae, Kyoo-Sung Cho, Chang-Sung Kim* Department of Periodontology, Research Institute for Periodontal Regeneration, College of Dentistry, Yonsei University, Seoul, Korea.