Repair Of Rat Calvarial Defects Research Articles

The effectiveness of hydroxyapatite-beta tricalcium phosphate incorporated into stem cells from human exfoliated deciduous teeth for reconstruction of rat calvarial bone defects.

To investigate the effects of stem cells from the pulp of human exfoliated deciduous teeth (SHED) on biphasic calcium phosphate granules (BCP) to repair rat calvarial defects as compared to autogenous bone grafting. A defect with a 6-mm diameter was produced on the calvaria of 50 rats. BCP granules were incorporated into SHED cultures grown for 7 days in conventional (CM) or osteogenic (OM) culture media. The animals were allocated into 5 groups of 10, namely: clot, autogenous bone, BCP, BCP+SHED in CM (BCP-CM), and BCP+SHED in OM (BCP-OM). The presence of newly formed bone and residual biomaterial particles was assessed by histometric analysis after 4 and 8 weeks. The autogenous group showed the largest newly formed bone area at week 8 and in the entire experimental period, with a significant difference in relation to the other groups (P<0.05). At week 8, BCP-CM and BCP-OM groups showed homogeneous new bone formation (P=0.13). When considering the entire experimental period, the BCP group had the highest percentage of residual particle area, with no significant difference from the BCP-CM group (P=0.06) and with a significant difference from the BCP-OM group (P=0.01). BCP-CM and BCP-OM groups were homogeneous throughout the experimental period (P=0.59). BCP incorporated into SHED cultures showed promising outcomes, albeit less pronounced than autogenous grafting, for the repair of rat calvarial defects. BCP incorporated into SHED cultures showed to be an alternative in view of the disadvantages to obtain autogenous bone graft.

Delivery vehicle of muscle-derived irisin based on silk/calcium silicate/sodium alginate composite scaffold for bone regeneration.

Irisin is a cytokine produced by skeletal muscle and usually plays a pivotal role in inducing fat browning and regulating energy expenditure. In recent years, it was found that irisin might be the molecular entity responsible for muscle-bone connectivity and is useful in osteogenesis induction. To study its effect on bone regeneration, we developed silk/calcium silicate/sodium alginate (SCS) composite scaffold based on an interpenetrating network hydrogel containing silk fibroin, calcium silicate, sodium alginate. Then we loaded irisin on the SCS before coating it with polyvinyl alcohol (PVA). The SCS/P scaffold was physically characterized and some in vitro and in vivo experiments were carried out to evaluate the scaffold effect on bone regeneration. The SCS/P scaffold was showed a porous sponge structure pursuant to scanning electron microscopy analysis. The release kinetics assay demonstrated that irisin was stably released from the irisin-loaded hybrid system (i/SCS/P system) to 50% within 7 days. Moreover, osteoinductive studies using bone marrow stem cells (BMSCs) in vitro exhibited the i/SCS/P system improved the activity of alkaline phosphatase (ALP) and enhanced the expression levels of a series of osteogenic markers containing Runx-2, ALP, BMP2, Osterix, OCN, and OPN. Alizarin red staining also demonstrated the promotion of osteogenesis induced by i/SCS/P scaffolds. In addition, in vivo studies showed that increased bone regeneration with better mineralization and higher quality was found during the repair of rat calvarial defects through utilizing the i/SCS/P system. These data provided strong evidence that the composite i/SCS/P would be a promising substitute for bone tissue engineering.

Osteoinductive Agents-Incorporated Three-Dimensional Biphasic Polymer Scaffold for Synergistic Bone Regeneration.

Large-scale bone defects are difficult to be regenerated entirely in the clinical practice. Bone tissue engineering has drawn more attention as an alternative to bone grafting owing to its convenience and flexibility. However, the low bioactivity of scaffolds and adverse effects of growth factors have hindered its practical application. Herein, the properties of poly(lactic-co-glycolic acid) (PLGA) scaffold, including hydrophilicity and mechanical strength, were improved by a gelatin coating incorporated with two small molecules, alendronate (ALD) and naringin (NG). Interestingly, these two drugs demonstrated a synergistic effect for the repair of rat calvarial defect, as ALD had an inhibitory impact on osteoclast activity and NG had an osteogenic effect on mesenchymal stem cells. From the results of histopathological staining and microcomputed tomography, the PLGA scaffold incorporated with gelatin, ALD, and NG (PLGA+Gelatin/ALD/NG) almost completely repaired the rat calvarial defect with physiological integrity at 16 weeks. In all, this biphasic scaffold can be a promising alternative to the conventional scaffold for clinical application.

Controlled dual delivery of BMP-2 and dexamethasone by nanoparticle-embedded electrospun nanofibers for the efficient repair of critical-sized rat calvarial defect

There is an urgent need to develop biomimetic bone tissue engineering scaffolds for the repair of critical-sized calvarial defect. In this study, we developed a new nanoparticle-embedded electrospun nanofiber scaffold for the controlled dual delivery of BMP-2 and dexamethasone (DEX). The scaffold was achieved by (1) the encapsulation of BMP-2 into bovine serum albumin (BSA) nanoparticles to maintain the bioactivity of BMP-2 and (2) the co-electrospinning of the blending solution composed of the BSA nanoparticles, DEX and the poly(ε-caprolactone)-co-poly(ethylene glycol) (PCE) copolymer. The in vitro studies showed that the bioactivity of DEX and BMP-2 was preserved in the dual-drug-loaded nanofiber scaffold, and a sequential release pattern in which most of the DEX was released in the original eight days and the BMP-2 release lasted up to 35 days was achieved. The in vitro osteogenesis study demonstrated that the drug-loaded groups exhibited a strong ability to induce differentiation toward osteoblasts. In vivo osteogenesis studies also revealed that the degrees of repair of rat calvarial defect achieved with the drug-loaded nanofiber scaffolds were significantly better than those obtained with the blank materials; in particular, the dual-drug-loaded nanofiber scaffold manifested the best repair efficacy due to a synergistic effect of BMP-2 and DEX. Therefore, the dual-drug-loaded nanofiber scaffold is deemed a strong potential candidate for the repair of bone defects in bone tissue engineering.

Bone regeneration of calvarial defect using marine calcareous-derived beta-tricalcium phosphate macrospheres

The aim of this study was to examine the bone regeneration properties of beta-tricalcium phosphate hydrothermally converted from foraminifera carbonate exoskeleton in the repair of rat calvarial defect. These natural materials possess unique interconnected porous network with uniform pore size distribution, which can be potentially advantageous. In total, 20 adult male Wistar rats received full-thickness calvarial defect with a diameter of 5 mm. The rate of newly formed bone was measured radiologically by X-ray and micro-computed tomography and by histologic examination. After 2 weeks, the beta-tricalcium phosphate group exhibited full closure of the defect site, while control group remained unrestored at the end of the 6-week experimentation. It was observed that the newly regenerated bone thickened over the course of the experiment in the beta-tricalcium phosphate group. No soft tissue reaction was observed around the beta-tricalcium phosphate implant and the rats remained healthy. These results showed that repair of the calvarial defect can be achieved by biomimetic beta-tricalcium phosphate macrospheres, which hold potential for application as bone grafts for bone augmentation surgeries.

Influence of the association between simvastatin and demineralized bovine bone matrix on bone repair in rats

Simvastatin, a drug used to lower blood cholesterol, has been reported to have an anabolic effect on bone. The purpose of this study was to evaluate the influence of simvastatin and demineralized bovine bone matrix (DBBM) on the repair of rat calvarial defects. Defects of 5 mm were created in 64 rats, divided into four groups: no local treatment (control); treatment with DBBM (DBBM); treatment with a combination of simvastatin solution (2.2 mg/50 μl) and DBBM (DBBMSIM-1); and treatment with simvastatin solution (0.5 mg/50 μl) and DBBM (DBBMSIM-2). Animals were sacrificed on postoperative day 30 or 60, after which the calvariae were X-rayed and prepared for histomorphometric evaluation. The data were submitted to statistical analysis (p < 0.05). X-rays revealed that, on postoperative day 30, animals treated with a lower dose of simvastatin presented the lowest bone density, whereas on postoperative day 60 the use of simvastatin, regardless of the dose, resulted in lower density than that observed in control and DBBM group samples. Histomorphometric analysis revealed that, on postoperative day 30, both DBBM and DBBMSIM-1 had a negative impact on bone formation. On postoperative day 60, none of the combinations tested impaired bone repair. These results showed that the association between DBBM and simvastatin had a negative impact on bone repair.

Role of fibrillar structure of collagenous carrier in bone sialoprotein-mediated matrix mineralization and osteoblast differentiation

To investigate the effects of the microstructure of collagenous carriers on the in vivo function of bone sialoprotein (BSP) in mineralization and osteoblast differentiation, we examined the ultrastructure of reconstituted type I collagen (collagen) and heat-denatured collagen (gelatin) and the in vivo responses to purified bone-derived BSP that was implanted with collagen or gelatin into surgically created 8-mm rat calvarial bone defects. Scanning and transmission electron microscopies revealed that the collagen displayed a fine fibrillar structure with interconnecting spaces between the fibrils/fibers, while the gelatin completely lost this unique three-dimensional structure after denaturation. The rates of in vivo release of BSP from the collagen scaffold were significantly lower than those from the gelatin. Collagen-BSP, but not gelatin-BSP, induced early mineral deposition in the matrix of proliferating repair cells in the calvarial defects at ∼4–7 days after implantation. Expression levels of osteoblast markers, alkaline phosphatase activity and amounts of new bone synthesized in the collagen-BSP treated defects were significantly greater than that in the gelatin-BSP treated defects ( p<0.001). The data suggest that the fibrillar microstructure of reconstituted collagen is essential for retaining BSP at a higher concentration within the defects, which enhances BSP-mediated matrix mineralization and osteoblast differentiation during the repair of rat calvarial defects.