Calcium Phosphate Bone Graft Research Articles

Staphylococcal biofilm formation on the surface of three different calcium phosphate bone grafts: a qualitative and quantitative in vivo analysis.

Differences in physico-chemical characteristics of bone grafts to fill bone defects have been demonstrated to influence in vitro bacterial biofilm formation. Aim of the study was to investigate in vivo staphylococcal biofilm formation on different calcium phosphate bone substitutes. A foreign-body guinea-pig infection model was used. Teflon cages prefilled with β-tricalcium phosphate, calcium-deficient hydroxyapatite, or dicalcium phosphate (DCP) scaffold were implanted subcutaneously. Scaffolds were infected with 2 × 103 colony-forming unit of Staphylococcus aureus (two strains) or S. epidermidis and explanted after 3, 24 or 72 h of biofilm formation. Quantitative and qualitative biofilm analysis was performed by sonication followed by viable counts, and microcalorimetry, respectively. Independently of the material, S. aureus formed increasing amounts of biofilm on the surface of all scaffolds over time as determined by both methods. For S. epidermidis, the biofilm amount decreased over time, and no biofilm was detected by microcalorimetry on the DCP scaffolds after 72 h of infection. However, when using a higher S. epidermidis inoculum, increasing amounts of biofilm were formed on all scaffolds as determined by microcalorimetry. No significant variation in staphylococcal in vivo biofilm formation was observed between the different materials tested. This study highlights the importance of in vivo studies, in addition to in vitro studies, when investigating biofilm formation of bone grafts.

Comparative study of bone regeneration in critical cranial bone defects using bone marrow adult stem cells and calcium phosphate

ObjectiveTo compare the use of Bone Marrow adult Stem Cells (BMSCs), differentiated in vitro into osteoblasts, associated to calcium phosphate versus autogenous bone graft, in the repair process of critical size bone defects. Materials and methodOn 36 Wistar adult rats, bilateral full-thickness defects on parietal bone were created. The defects were either repaired with calcium phosphate (group I), calcium phosphate+(BMSCs) (group II) or autogenous bone graft (group III), and the opposite side with blood clot (Control Group). In all cases a collagen membrane was used. The animals were sacrificed at 30 and 60 days, and all specimens were collected for further histological and histomorfometric study. ResultsAt 30 days, group III (autogenous bone graft) evidences a statistical difference on bone formation when compared to the experimental and control groups (p≤0.05). At 60 days group II (BS+BMSCs) and group III (autogenous bone) showed a similar bone formation and has only a statistical difference when compared to group I (BS) and control group. ConclusionThe use of calcium phosphate in conjunction with BMSCs resulted in a similar behavior in the process of bone repair in critical size defects, when compared with autogenous bone graft.

Comparative study of bone regeneration in critical cranial bone defects using bone marrow adult stem cells and calcium phosphate

ObjectiveTo compare the use of Bone Marrow adult Stem Cells (BMSCs), differentiated in vitro into osteoblasts, associated to calcium phosphate versus autogenous bone graft, in the repair process of critical size bone defects. Materials and methodOn 36 Wistar adult rats, bilateral full-thickness defects on parietal bone were created. The defects were either repaired with calcium phosphate (group I), calcium phosphate+(BMSCs) (group II) or autogenous bone graft (group III), and the opposite side with blood clot (Control Group). In all cases a collagen membrane was used. The animals were sacrificed at 30 and 60 days, and all specimens were collected for further histological and histomorfometric study. ResultsAt 30 days, group III (autogenous bone graft) evidences a statistical difference on bone formation when compared to the experimental and control groups (p≤0.05). At 60 days group II (BS+BMSCs) and group III (autogenous bone) showed a similar bone formation and has only a statistical difference when compared to group I (BS) and control group. ConclusionThe use of calcium phosphate in conjunction with BMSCs resulted in a similar behavior in the process of bone repair in critical size defects, when compared with autogenous bone graft.

Comparison of three calcium phosphate bone graft substitutes from biomechanical, histological, and crystallographic perspectives using a rat posterolateral lumbar fusion model

This study evaluated the effectiveness of three calcium phosphate bone graft substitutes with different chemical compositions on spinal fusion using a rat posterolateral lumbar fusion model. Specifically, two recently developed non-dispersive tetracalcium phosphate/dicalcium phosphate anhydrous-based calcium phosphate cements (CPCs), namely a CPC consisting of equimolar amounts of the two compounds (nd-CPC) and a CPC consisting of a two-fold greater amount of dicalcium phosphate anhydrous (DCP-rich CPC), were compared with a commercial calcium phosphate bone graft (c-CPG) consisting of hydroxyapatite (60%) and β-tricalcium phosphate (40%). Single-level posterolateral lumbar fusion was performed at the L4–L5 vertebrae in fifteen adult rats (n=5 for each group). Spinal fusion was evaluated with radiographs, manual palpation, mechanical testing, micro-CT, and histology 8weeks post-surgery. In particular, the crystallographic phases in the three substitutes were identified before and 8weeks after their implantation. Manual palpation revealed stable constructs in nearly all of the spine specimens. The stiffness and bending load of fused spines in the two CPC groups were comparable to those in the c-CPG group. The radiographs specifically revealed implant resorption and bone remodeling in the DCP-rich CPC group. Analysis of 3D micro-CT images revealed that the bone volume ratio in the DCP-rich CPC group was significantly greater than those in the nd-CPC and c-CPG groups. Histology showed that the DCP-rich CPC group exhibited the highest degree of bone regeneration and osseointegration. Notably, DCP-rich CPC led to a pronounced phase transformation, generating the greatest amount of poorly crystalline apatite among the three groups, which together with adequate resorption may explain the aforementioned positive findings. We therefore conclude that of the bone graft substitutes considered, DCP-rich CPC has the greatest potential to be used in spinal fusion.

Prospective, Randomized, Controlled Trial of Silicate-Substituted Calcium Phosphate Versus rhBMP-2 in a Minimally Invasive Transforaminal Lumbar Interbody Fusion

Prospective, randomized, controlled trial. To compare arthrodesis rates between patients undergoing a primary single-level minimally invasive transforaminal lumbar interbody fusion (MIS TLIF) with either Actifuse or recombinant human bone morphogenetic protein-2 (rhBMP-2). Preclinical animal studies suggest that silicate-substituted calcium phosphate (Actifuse) bone graft substitute offers equivalent or an increased fusion rate compared with other graft enhancers/extenders and rhBMP-2. Fifty-two patients undergoing a single-level unilateral MIS TLIF were evenly randomized into 2 cohorts as follows: the Actifuse cohort received Actifuse combined with 5 mL of bone marrow aspirate (n = 26; 50%), whereas the rhBMP cohort received 4.2 mg of rhBMP-2 (n = 26; 50%). A pre hoc G*Power analysis yielded a sample size of n = 26 that was determined through a 2-tailed distribution calculation. Computed tomographic analysis was performed at 6 months and 1 year postoperatively. Pre- and postoperative visual analogue scale scores were obtained to assess the clinical outcomes. Arthrodesis was determined by 2 separate, blinded orthopedic surgeons and a board certified radiologist. At 1-year follow-up, 65% (17/26) of the Actifuse cohort and 92% (24/26) of the rhBMP-2 cohort demonstrated a radiographical arthrodesis (P = 0.01). In both study cohorts, the 1-year postoperative visual analogue scale scores significantly improved (P < 0.001). Pseudarthrosis rates at 1 year were 35.0% (9/26) and 7.7% (2/26) for the Actifuse and rhBMP-2 groups, respectively (P = 0.01, OR = 6.35, 95% CI = 1.22-33.1). A greater reoperation rate was noted in the Actifuse cohort (35.0%, 9/26) compared with the BMP-2 cohort (7.7%, 2/26; P = 0.01). One patient with BMP-2 also experienced symptomatic neuroforaminal bone growth (3.8%, n = 1/26). Silicate-substituted calcium phosphate was associated with a significantly lower rate of arthrodesis than rhBMP-2 in a MIS TLIF. The patients with pseudarthrosis in both cohorts were all clinically symptomatic with an unimproved visual analogue scale score. Additional analysis of Actifuse and other graft enhancers/extenders are needed prior to the utilization for an MIS TLIF. 2.

Comparative study of biphasic calcium phosphate with beta-tricalcium phosphate in rat cranial defects—A molecular-biological and histological study

The aim of this study was to evaluate the in vivo biocompatibility of a biphasic calcium phosphate (BCP) bone graft substitute consisting of 60% hydroxyapatite and 40% β-tricalcium phosphate (β-TCP) in comparison to a pure β-TCP of identical shape and porosity. The materials were evaluated using an established rat cranial defect model in 24 animals. One bone defect with a diameter of 5mm was created per animal. The defects were filled with either BCP or β-TCP and left to heal for 4 weeks. Twelve samples (6 per material) were processed for histological evaluation and immunohistochemistry. The remaining 12 samples were processed for mRNA expression analysis. No signs of inflammation or adverse material reactions were detected. New bone formation in the former defect site did not differ between the two groups (BCP: 49.2%; β-TCP: 52.4%). Osteoblast-like and TRAP-positive osteoclast-like cells were found at the surface of the bone graft substitute granules. The β-TCP group showed significantly higher mRNA levels for the bone resorption marker Acp5 and osteogenic differentiation marker Runx2. The expression of IGF1, IGF2, VEGF, Phex, Alpl, Col1, Col2, Bglap and MMP8 did not differ between the groups. The in vivo biocompatibility of BCP is to a large part identical to those of TCP. Within the limitation of the animal model, the implantation study shows that BCP can be used as bone graft substitute, due to the fact that the material integrates into tissue, remains stable in the implantation bed and serves as an osteoconductive scaffold.

The role of the chemical composition of monetite on the synthesis and properties of α-tricalcium phosphate

There has been a resurgence of interest in alpha-tricalcium phosphate (α-TCP), with use in cements, polymer composites and in bi- and tri-phasic calcium phosphate bone grafts. The simplest and most established method for preparing α-TCP is the solid state reaction of monetite (CaHPO4) and calcium carbonate at high temperatures, followed by quenching.In this study, the effect of the chemical composition of reagents used in the synthesis of α-TCP on the local structure of the final product is reported and findings previously reported pertaining to the phase composition and stability are also corroborated. Chemical impurities in the monetite reagents were identified and could be correlated to the calcium phosphate products formed; magnesium impurities favoured the formation of β-TCP, whereas single phase α-TCP was favoured when magnesium levels were low. Monetite synthesised in-house exhibited a high level of chemical purity; when this source was used to produce an α-TCP sample, the α-polymorph could be obtained by both quenching and by cooling to room temperature in the furnace at rates between 1 and 10°C/min, thereby simplifying the synthesis process. It was only when impurities were minimised that the 12 phosphorus environments in the α-TCP structure could be resolved by 31P nuclear magnetic resonance; samples containing chemical impurity showed differing degrees of line-broadening. Reagent purity should therefore be considered a priority when synthesising/characterising the α-polymorph of TCP.

In vitro characterization of a synthetic calcium phosphate bone graft on periodontal ligament cell and osteoblast behavior and its combination with an enamel matrix derivative

Recent studies suggest that a combination of enamel matrix derivative (EMD) with grafting material may improve periodontal wound healing/regeneration. Newly developed calcium phosphate (CaP) ceramics have been demonstrated a viable synthetic replacement option for bone grafting filler materials. This study aims to test the ability for EMD to adsorb to the surface of CaP particles and to determine the effect of EMD on downstream cellular pathways such as adhesion, proliferation, and differentiation of primary human osteoblasts and periodontal ligament (PDL) cells. EMD was adsorbed onto CaP particles and analyzed for protein adsorption patterns via scanning electron microscopy and high-resolution immunocytochemistry with an anti-EMD antibody. Cell attachment and cell proliferation were quantified using CellTiter 96 One Solution Cell Assay (MTS). Cell differentiation was analyzed using real-time PCR for genes encoding Runx2, alkaline phosphatase, osteocalcin, and collagen1α1, and mineralization was assessed using alizarin red staining. Analysis of cell attachment revealed significantly higher number of cells attached to EMD-adsorbed CaP particles when compared to control and blood-adsorbed samples. EMD also significantly increased cell proliferation at 3 and 5 days post-seeding. Moreover, there were significantly higher mRNA levels of osteoblast differentiation markers including collagen1α1, alkaline phosphatase, and osteocalcin in osteoblasts and PDL cells cultured on EMD-adsorbed CaP particles at various time points. The present study suggests that the addition of EMD to CaP grafting particles may influence periodontal regeneration by stimulating PDL cell and osteoblast attachment, proliferation, and differentiation. Future in vivo and clinical studies are required to confirm these findings. The combination of EMD and CaP may represent an option for regenerative periodontal therapy in advanced intrabony defects.

Biofilm formation by staphylococci on fresh, fresh-frozen and processed human and bovine bone grafts

Biofilm formation is a multi-step process influenced by surface properties. We investigated early and mature biofilm of Staphylococcus aureus on 4 different biological calcium phosphate (CaP) bone grafts used for filling bone defects. We investigated standardised cylinders of fresh and fresh-frozen human bone grafts were harvested from femoral heads; processed humanand bovine bone grafts were obtained preformed. Biofilm formation was done in tryptic soy broth (TSB) using S. aureus (ATCC 29213) with static conditions. Biofilm density after 3h (early biofilm) and 24h (mature biofilm) was investigated by sonication and microcalorimetry. After 3h, bacterial density was highest on fresh-frozenandfresh bone grafts. After 24h, biofilm density was lowest on freshbone grafts (p<0.001) compared to the other 3 materials, which did not differ quantitatively (p>0.05). The lowest increase in bacterial density was detected on fresh bone grafts (p<0.001). Despite normal shaped colonies, we found additional small colonies on the surface of the fresh and fresh-frozen samples by sonication. This was also apparent in microcalorimetric heat-flow curves. The four investigated CaP bone grafts showed minor structural differences in architecture but marked differences concerning serum coverage and the content of bone marrow, fibrous tissue and bone cells. These variations resulted in a decreased biofilm density on freshand fresh-frozenbone grafts after 24h, despite an increased early biofilm formation and might also be responsible for the variations in colony morphology (small colonies). Detection of small colony variants by microcalorimetry might be a new approach to improve the understanding of biofilm formation.

Microstructure and chemistry affects apatite nucleation on calcium phosphate bone graft substitutes

The bioactivity of calcium phosphate bone grafts of varying chemistry and strut-porosity was compared by determining the rate of formation of hydroxycarbonate apatite crystals on the material surface after being soaked in simulated body fluid for up to 30 days. Three groups of silicate-substituted hydroxyapatite material were tested, with each group comprising a different quantity of strut-porosity (23, 32, and 46 % volume). A commercially available porous β-tricalcium phosphate bone graft substitute was tested for comparison. Results indicate that strut-porosity of a material affects the potential for formation of a precursor to bone-like apatite and further confirms previous findings that β-tricalcium phosphate is less bioactive than hydroxyapatite.

Advanced core decompression, a new treatment option of avascular necrosis of the femoral head - a first follow-up

Aseptic necrosis of the femoral head (AVN) leads to destruction of the affected hip joint, predominantly in younger patients. Advanced core decompression (ACD) is a new technique that may allow better removal of the necrotic tissue by using a new percutaneous expandable reamer. A further modification is the refilling of the drill hole and the defect with an injectable, hard-setting, composite calcium sulphate (CaSO₄)-calcium phosphate (CaPO₄) bone graft substitute. Compression tests were performed on seven pairs of femoral cadaver bones. One femur of each pair was treated with ACD, while the opposite side remained untreated. Clinically, the postoperative outcome of 27 hips in 23 patients was performed by physical examination 6 weeks after ACD and at average follow-up of 9.69 months, and compared with the preoperative results. MRI was used to assess the removal of the necrotic tissue, any possible progression of AVN and evaluation of collapse. In the biomechanical analysis, the applied maximum compression force that caused the fracture did not significantly differ from the untreated opposite side. The overall results of postoperative physical examinations were significantly better than preoperatively. Five hips (18.5%) were converted to a total hip replacement. The follow-up MRIs of the other patients showed no progression of the necrotic area. The first follow-up results of ACD have been encouraging for the early stages of aseptic necrosis of the femoral head. In our opinion, an assured advantage is the high stability of the femoral neck after ACD, which allows quick rehabilitation.

The effects of microporosity on osteoinduction of calcium phosphate bone graft substitute biomaterials

The effect of increasing strut porosity on the osteoinductive ability of silicate substituted calcium phosphate (SiCaP) biomaterials was investigated in an ectopic ovine model. Implants with strut porosities of 22.5%, 32.0% and 46.0% were inserted into the parapsinalis muscle. At 8, 12 and 24weeks histological sections were prepared. Sections were examined using backscattered scanning electron microscopy and un-decalcified histology. Bone area, implant area and bone-implant contact were quantified. At 8weeks there was no significant difference between the groups in terms of bone area and implant area. However at 12weeks, the amount of bone formation observed was significantly greater in SiCaP-46 (6.17±1.51%) when compared with SiCaP-22.5 (1.33±0.84%) p=0.035. Results also showed significantly increased amounts of bone-implant contact to the SiCaP-46 scaffold (3.30±1.17%) compared with SiCaP-22.5 (0.67±0.52%, p=0.043) at 8weeks and 12weeks; (SiCaP-46 (21.82±5.59%) vs SiCaP-22.5 (3.06±1.89%), p=0.012). At 24weeks, bone formation and graft resorption had significantly increased in all groups so that the level of bone formation in the SiCaP-46 group had increased 75-fold to 30.05±8.38%. Bone formation was observed in pores <10μm. Results suggest that bone graft substitute materials with greater strut porosity are more osteoinductive.

Total scalp reconstruction with bilateral anterolateral thigh flaps

Large scalp defects can require complicated options for reconstruction, often only achieved with free flaps. In some cases, even a single free flap may not suffice. We review the literature for options in the coverage of all reported large scalp defects, and report a unique case in which total scalp reconstruction was required. In this case, two anterolateral thigh (ALT) flaps were used to resurface a large scalp and defect, covering a total of 743 cm(2). The defect occurred after resection and radiotherapy for desmoplastic melanoma, with several failed skin grafts and local flaps and osteoradionecrosis involving both inner and outer tables of the skull. The reconstruction was achieved as a single-stage reconstruction and involved wide resection of cranium and overlying soft-tissues and reconstruction with calcium phosphate bone graft substitute, titanium mesh, and two large ALT flaps. The reconstruction was successfully achieved, with minor postoperative complications including tip necrosis of one of the flaps and wound breakdown at one of the donor sites. This is the first reported case of two large ALT flaps for scalp resurfacing and may be the largest reported scalp defect to be completely resurfaced by free flaps. The use of bilateral ALT flaps can be a viable option for the reconstruction of large and/or complicated scalp defects.

Effect of increased strut porosity of calcium phosphate bone graft substitute biomaterials on osteoinduction

The effect of increasing strut porosity on the osteoinductivity of porous calcium phosphate (CaP) and silicate-substituted calcium phosphate (SiCaP) bone substitute materials was investigated in an ovine ectopic model. One to two millimeter-sized granules or block implants with strut porosities of 10, 20, or 30% were inserted into the left and right paraspinalis muscle. At 12 weeks, histological sections were prepared through the center of each implant and bone contact, bone area and implant area quantified. Backscattered scanning electron microscopy (bSEM) was used to visualize bone within small pores in the struts of the scaffolds. Increased bone formation was measured in the SiCaP with 30% strut porosity (5.482% ± 1.546%) when compared with the nonsilicate CaP with the same morphology (1.160% ± 0.502%, p = 0.02), indicating that silicate substitution may increase osteoinduction. Greater bone formation was seen in scaffolds with increased strut porosity. No bone growth was found in any of the SiCaP scaffold with 10% porosity. There was no significant difference between block and granule specimens. Scanning electron microscopy and EDX in combination with histology demonstrated bone formation within pores <5 μm in size. The use of silicate-substituted CaP material with increased strut porosity may further augment repair and regeneration in bony sites.

Calcium phosphate bone graft substitutes: Failures and hopes

Despite 40 years of efforts, researchers have failed to provide calcium phosphate bone graft substitutes performing well enough to replace bone grafting procedures: their osteogenesis potential is limited, and calcium phosphates are too brittle. However, there is hope to solve the two afore-mentioned problems. First, it is now clear why nacre and bone are very tough despite a high ceramic load. Also, recent studies suggest that calcium and phosphate ions can trigger osteoinduction. The present article aims: (i) to review our current knowledge in the field of synthetic bone graft substitutes, (ii) to explain why ceramics and in particular calcium phosphates are still the most promising materials for bone graft substitution, and (iii) finally to describe the strategy to obtain osteoinductive calcium phosphate bone graft substitutes as strong as cortical bone.

Increasing strut porosity in silicate‐substituted calcium‐phosphate bone graft substitutes enhances osteogenesis

Synthetic, porous silicate-substituted calcium phosphate bone graft matrices (SiCaP; 0.8 wt % Si) with varying strut porosity were applied to ovine critical-sized defect sites as either 1-2 mm microgranules (SiCaP-23G, SiCaP-32G, and SiCaP-46G) or 1-2 mm microgranules in an aqueous poloxamer carrier (SiCaP-23P, SiCaP-32P, and SiCaP-46P). Defect sites treated with SiCaP-23G or SiCaP-23P showed evidence of bone formation at 8 and 12 weeks in central zones. More advanced neovascularization and increased bone contact was observed for graft materials with higher strut porosities. At 12 weeks, graft materials with higher strut porosities (32% and 46%) had statistically significantly higher absolute bone volumes (p < 0.05) versus those with a strut porosity of 23%. Absolute bone volume in defects treated with grafts of matched strut porosities as microgranules, or microgranules with poloxamer carrier, were similar at 12 weeks. Absolute graft volume for SiCaP-46 reduced over 12 weeks (not statistically significant). In conclusion, bone formation patterns in critically-sized defects confirm strut porosity to be a clinically relevant property of porous silicate-substituted calcium phosphate bone grafts in promoting osteogenesis. Increasing graft matrix strut porosity encouraged earlier neovascularization and increased the absolute equilibrium volume of bone growth within the graft without compromising graft stability.

Calcaneal Intraosseous Lipoma: A Case Report and Review of the Literature

Intraosseous lipomas are rare benign bone tumors. This benign neoplasm has been reported to occur in the calcaneus as well as the proximal femur. In the past, the relative absence of symptoms and radiographic similarity to a bone cyst has accounted for underdiagnosis of intraosseous lipoma. The case presented in this article was diagnosed with the help of computerized tomography and histopathologic analysis, after which the patient was treated by means of curettage and packing with calcium phosphate bone graft substitute. The purpose of this article is to increase awareness among clinicians of the existence of this unusual lesion.

Effect of rhTGF-β1 combined with bone grafts on human periodontal cell differentiation

Various techniques and materials have been proposed for the treatment of periodontal defects. In periodontal regeneration, periodontal ligament (PDL) cell differentiation as well as certain growth factors and their delivery system applied are critical. The purpose of this study was to evaluate the in vitro effect of recombinant human transforming growth factor-beta 1 (rhTGF-β1) combined with two different bone grafts on human PDL (hPDL) cell differentiation. The hPDL cells were treated with TGF-β1 alone or in combination with a calcified freeze-dried bone allograft (FDBA) and a porous biphasic calcium phosphate (BC) bone graft. Cell differentiation effect was estimated by measuring alkaline phosphatase (ALPase) activity and osteocalcin secretion. Results demonstrated that rhTGF-β1 alone or in combination with FDBA and BC provoked a significant (p < 0.05) increase in ALPase activity as compared with controls. The findings of this study confirmed the beneficial role of rhTGF-β1 combined with FDBA and BC as carriers in periodontal regeneration.

Back‐scattered electron imaging and elemental analysis of retrieved bone tissue following sinus augmentation with deproteinized bovine bone or biphasic calcium phosphate

To compare resorption of a synthetic biphasic calcium phosphate (BCP) bone-graft substitute with deproteinized bovine bone (DBB) used for human maxillary sinus augmentation. Eleven patients underwent bilateral maxillary sinus floor augmentation with DBB in one side and a BCP (40%beta-tricalcium phosphate (beta-TCP) and 60% hydroxyapatite) in the contralateral side. Simultaneously, with the augmentation on each side a microimplant was placed vertically from the top of the alveolar crest penetrating the residual bone and the grafting material. Eight months after initial surgery the microimplants were retrieved with a surrounding bone core. The composition of residual graft material and surrounding bone was analysed by scanning electron microscopy and energy dispersive X-ray spectroscopy. Residual graft material of both types was present as 10-500 mum particles in direct contact with, or completely surrounded by, newly formed bone; smaller particles were also present in non-mineralized tissue. In the case of BCP the bone-graft substitute interface showed evidence of superficial disintegration of particles into individual grains. Median Ca/P ratios (at.%), determined from >200 discreet sites within residual graft particles and adjacent bone, were: DBB: 1.61 (confidence interval [CI] 1.59-1.64); BCP: 1.5 (CI 1.45-1.52); DBB-augmented bone: 1.62 (CI 1.59-1.66); BCP-augmented bone: 1.52 (CI 1.47-1.55); P=0.028 for DBB vs. BCP and DBB- vs. BCP-augmented bone. The reduction in Ca/P ratio for BCP over the healing period is consistent with the dissolution of beta-TCP and reprecipitation on the surface of calcium-deficient hydroxyapatite. The beta-TCP component of BCP may be gradually substituted by calcium-deficient hydroxyapatite over the healing period. This process and superficial degranulation of BCP particles may influence the progress of resorption and healing.

Initial biocompatibility and enhanced osteoblast response of Si doping in a porous BCP bone graft substitute

Granular shape biphasic calcium phosphate (BCP) bone grafts with and without doping of silicon cations were evaluated in regards to biocompatibility and MG-63 cellular response. To do this we studied Cellular cytotoxicity, cellular adhesion and spreading behavior and cellular differentiation with alizarin red S staining. Gene expression in MG-63 cells on the implanted bone substitutes was also examined at different time points using RT-PCR. In comparison, the Si-doped BCP granule showed more cellular viability than the BCP granule without doping in MTT assay. Moreover, cell proliferation was much higher when Si doping was employed. The cells grown on the silicon-doped BCP substitutes had more active filopodial growth with cytoplasmic webbing that proceeded to the flattening stage, which was indicative of well cellular adhesion. When these cells were exposed to Si-doped BCP granules for 14 days, well differentiated MG-63 cells were observed. Osteonectin and osteopontin genes were highly expressed in the late stage of differentiation (14 days), whereas collagen type I mRNA were found to be highly expressed during the early stage (day 3). These combined results of this study demonstrate that silicon-doped BCP enhanced osteoblast attachment/spreading, proliferation, differentiation and gene expression.