Biomedical subjects

Paul Pilet

Publications and source records attributed to Paul Pilet.

13 recordsLinked to original sources

PubMed2007

Hybrid composites of calcium phosphate granules, fibrin glue, and bone marrow for skeletal repair.

Synthetic bone substitutes, such as calcium phosphate ceramics, give good results in clinical applications. In order to adapt to surgical sites, bioceramics come in the form of blocks or granules, and are either dense or porous. Combining these bioceramics with fibrin glue provides a mouldable and self-hardening composite biomaterial with the biochemical properties of each component. Critical-sized defects in the femoral condyle of rabbits were filled with TricOs/fibrin glue/bone marrow hybrid/composite material. The TricOs granules (1-2 mm) were composed of hydroxyapatite and beta tricalcium phosphate (60/40 in weight). The fibrin glue was composed of fibrinogen, thrombin and other biological factors and mixed with MBCP granules either simultaneously or sequentially. Bone marrow was also added to the MBCP/fibrin composite prior to filling the defects. After 3, 6, 12, and 24 weeks of implantation, the newly-formed bone was analysed with histology, histomorphometry and mechanical tests. The newly-formed bone had grown centripetally. Simultaneous application of fibrin glue showed better results for mechanical properties than sequential application after 6 weeks. Around 40% of bone had formed after 24 weeks in the three groups. Although the addition of bone marrow did not improve bone formation, the MBCP/fibrin material could be used in clinical bone filling applications.

Animals↗

PubMed2006

Physico-chemical-mechanical and in vitro biological properties of calcium phosphate cements with doped amorphous calcium phosphates.

Calcium phosphate cements (CPCs) are successfully used as bone substitutes in dentistry and orthopaedic applications. This study investigated the physico-chemical-mechanical properties of and in vitro biological properties (cell response) of CPCs prepared with amorphous calcium carbonate phosphate (ACCP) doped with magnesium (ACCP-Mg), zinc (ACCp-Zn) or fluoride (ACCP-F) ions. The experimental CPC consisted of alpha-TCP, doped ACCP, and MPCM powders as matrix and biphasic calcium phosphate (BCP) granules. X-ray diffraction analysis showed that the matrix converted to apatite with poor crystallinity (reflecting small crystal size) after setting for 24 h, while BCP remained apparently unchanged. Cements with ACCP-F (F-CPC) had shorter setting times and greater compressive strength compared to cements with ACCP-Mg (Mg-CPC) or ACCP-Zn (Zn-CPC). Scanning electron microscopy (SEM) showed that crystals set on Mg-CPC and Zn-CPC were smaller compared to those on F-CPC. The total porosity of Mg-CPC was greater compared to Zn-CPC or F-CPC. Osteoblast-like cells, MC3T3-E1, remained viable and maintained their ability to express alkaline phosphatase in contact with the CPCs with doped ACCPs.

3T3 Cells↗

PubMed2006

Injectable calcium phosphate scaffold and bone marrow graft for bone reconstruction in irradiated areas: an experimental study in rats.

The purpose of this study was to assess the possibilities for bone reconstruction of an injectable calcium phosphate scaffold (ICPS) associated with a bone marrow (BM) graft after irradiation in a rat model. External irradiation was delivered to 12 out of 27 inbred rats. Three weeks later, four osseous defects were created per animal and were kept empty or filled with either ICPS alone, BM graft alone or with a mixture of BM and ICPS. Three weeks after implantation, bone specimens were studied under light microscopy and by scanning electron microscopy. Filling irradiated defects with ICPS alone was not accompanied by the formation of new bone. The BM graft associated with ICPS significantly increased ceramic degradation (p<0.01) and bone ingrowth (p<0.01) in the irradiated areas. The results are evidence for the meaning of the BM in driving the bone repair in irradiated animals.

Animals↗

PubMed2006

The modulation of gene expression in osteoblasts by thrombin coated on biphasic calcium phosphate ceramic.

For many years, fibrin sealants were associated with bone substitutes to promote bone healing. However, the osteoblastic response to fibrin sealant components remains poorly documented. In this study, MC3T3-E1 osteoblastic cells were cultured on biphasic calcium phosphate ceramic (MBCP) coated with Tissucol components (thrombin and fibrinogen). Analysis of osteoblastic differentiation markers by RT-PCR revealed that MBCP coated with Tissucol stimulated mRNA levels for osteocalcin and alkaline phosphatase (ALP). Of all the components of Tissucol, thrombin has been reported to affect osteoblastic behavior. Our results demonstrated that low thrombin concentrations (0.5-5 U/ml) stimulated mRNA levels for ALP, whereas high thrombin concentrations (50-100 U/ml) decreased mRNA levels for ALP and PTH/PTHrP receptor and also increased mRNA level for the osteoclastogenesis inhibitor OPG. As thrombin stimulated angiogenesis, we then wondered whether thrombin could influence the expression of angiogenic factors. Low thrombin concentrations were shown to up-regulate mRNA levels for VEGF-B and VEGF-R1, suggesting an autocrine/paracrine role for VEGF-B. Higher thrombin concentrations also up-regulated mRNA for VEGF-A and neuropilin-1. In conclusion, the association of MBCP with thrombin and fibrinogen appears to be a convenient scaffold for bone cell differentiation. Thrombin could also acts at the cellular level by increasing the angiogenic potential of osteoblasts as well as their responsiveness to thrombin and VEGF.

Animals↗

PubMed2006

Quantitative and reliable in vitro method combining scanning electron microscopy and image analysis for the screening of osteotropic modulators.

The increased generation and up-regulated activity of bone resorbing cells (osteoclasts) play a part in the impairment of bone remodeling in many bone diseases. Numerous drugs (bisphosphonates, calcitonin, selective estrogen receptor modulators) have been proposed to inhibit this increased osteoclastic activity. In this report, we describe a pit resorption assay quantified by scanning electron microscopy coupled with image analysis. Total rabbit bone cells with large numbers of osteoclasts were cultured on dentin slices. The whole surface of the dentin slice was scanned and both the number of resorption pits and the total resorbed surface area were measured. Resorption pits appeared at 48 h and increased gradually up to 96 h. Despite the observation of a strong correlation between the total resorption area and the number of pits, we suggest that area measurement is the most relevant marker for osteoclastic activity. Osteotropic factors stimulating or inhibiting osteoclastic activity were used to test the variations in resorption activity as measured with our method. This reproducible and sensitive quantitative method is a valuable tool for screening for osteoclastic inhibitors and, more generally, for investigating bone modulators.

Animals↗

PubMed2006

Bone repair using a new injectable self-crosslinkable bone substitute.

A new injectable and self-crosslinkable bone substitute (IBS2) was developed for filling bone defects. The IBS2 consisted of a chemically modified polymer solution mixed with biphasic calcium phosphate (BCP) ceramic particles. The polymer hydroxypropylmethyl cellulose was functionalized with silanol groups (Si-HPMC) and formed a viscous solution (3 wt %) in alkaline medium. With a decrease in pH, self-hardening occurred due to the formation of intermolecular -Si-O- bonds. During setting, BCP particles, 40 to 80 microm in diameter, were added to the polymer solution at a weight ratio of 50/50. The resulting injectable material was bilaterally implanted into critically sized bone defects at the distal femoral epiphyses of nine New Zealand White rabbits. The IBS2 filled the bone defects entirely and remained in place. After 8 weeks, bone had grown centripetally and progressed towards the center of the defects. Newly formed bone, ceramic, and nonmineralized tissue ratios were 24.6% +/- 5.6%, 21.6% +/- 5.8%, and 53.7% +/- 0.1%, respectively. Mineralized and mature bone was observed between and in contact with the BCP particles. The bone/ceramic apposition was 73.4% +/- 10.6%. The yield strength for the IBS2-filled defects was 16.4 +/- 7.2 MPa, significantly higher than for the host trabecular bone tissue (2.7 +/- 0.4 MPa). This study showed that BCP particles supported the bone healing process by osteoconduction while the Si-HPMC hydrogel created intergranular space for bone ingrowth. This new injectable and self-crosslinkable bone substitute could be used conveniently in orthopedic surgery for filling critical-size bone defects.

Animals↗

PubMed2005

Micro-architecture of calcium phosphate granules and fibrin glue composites for bone tissue engineering.

Calcium phosphate ceramics are currently used as bone graft substitutes in various types of clinical applications. Fibrin glue is also used in surgery due to its haemostatic, chemotactic and mitogenic properties. By combining these two biomaterials, new composite scaffolds were prepared. In this study, we attempt to analyse whether thrombin concentration in the fibrin glue could influence the properties of the composite. The association between fibrin glue and calcium phosphate ceramic granules was characterized at the ultra structural level. Micro and macroporous biphasic calcium phosphate ceramic granules with a diameter of 1-2mm composed of hydroxyapatite and beta-tricalcium phosphate (60/40) were associated to fibrin glue. The composites were observed by scanning and transmission electron microscopy and microcomputed tomography. Fibre thickness, porosity and homogeneity of the fibrin clot were modified by increased the thrombin concentration. Mixing fibrin glue with calcium phosphate granules (1:2) did not modify the microstructure of the fibrin clot in the composite. Nevertheless, thrombin interacted with the bioceramic by inducing the nucleation of crystalline precipitate at the ceramic/fibrin glue interface. Combining fibrin sealant and calcium phosphate ceramics could lead to new scaffolds for bone tissue engineering with the synergy of the properties of the two biomaterials.

Bone Substitutes↗

PubMed2005

MBCP biphasic calcium phosphate granules and tissucol fibrin sealant in rabbit femoral defects: the effect of fibrin on bone ingrowth.

An ageing population implies an increase in bone and dental diseases, which are in turn a source of numerous handicaps. These pathologies are an expensive burden for the European health system. As no specific bioactive materials are efficient enough to cope with this burden, we have to develop an injectable, mouldable, self-hardening bone substitute to support bone tissue reconstruction and augmentation. New, highly bioactive and suitable biomaterials have been developed to replace bone grafts in orthopedic revision and maxillofacial surgery for bone augmentation. These mouldable, self-hardening materials are based on the association of MBCP Biphasic Calcium Phosphate Granules and Tissucol Fibrin Sealant. The in vivo evaluation of ingrowth in relation to the composite was made in an experiment on rabbits. The results indicate that in the presence of fibrin sealant, newly-formed bone developed at a small distance from the surface of the calcium phosphate ceramic. Two different bone apposition processes were identified. Without the fibrin component (MBCP group), bone rested directly on the surface of the granules. This observation is commonly described as osteoconduction in calcium phosphate materials. On the contrary, the presence of the fibrinogen component seemed to modify this standard osteoconduction phenomenon: the newly-formed bone essentially grew at a distance from the surface of the granules, on the fibrillar network, and could be considered as an inductive phenomenon for osteogenic cell differentiation from mesenchymal stem cells.

Animals↗

PubMed2005

In vitro biological effects of titanium rough surface obtained by calcium phosphate grid blasting.

Surface roughness modulates the osseointegration of orthopaedic and dental titanium implants. High surface roughness are currently obtained by blasting of titanium implants with silica or aluminium oxide abrasive particles. This process may cause the release of cytotoxic silicium or aluminium ions in the peri-implant tissue. To generate a biocompatible roughened titanium surface, we currently develop an innovative grid-blasting process using biphasic calcium phosphate (BCP) particles. Titanium alloy (Ti6Al4V) discs were either polished, BCP grid-blasted or left as-machined. BCP grid-blasting created an average surface roughness of 1.57 +/- 0.07 microm compared to the original machined surface of 0.58 +/- 0.05 microm. X-ray photoelectron spectroscopy indicated traces of calcium and phosphorus and relatively less aluminium on the BCP grid-blasted surface than on the initial titanium specimen. Scanning electronic microscopy observations and measurement of mitochondrial activity (MTS assay) showed that osteoblastic MC3T3-E1 cells were viable in contact with the BCP grid-blasted titanium surface. In addition, our results indicate that MC3T3-E1 cells expressed ALP activity and conserved their responsiveness to bone morphogenetic protein BMP-2. The overall results clearly indicate that this calcium phosphate grid-blasting technique increases the roughness of titanium implants and provides a non-cytotoxic surface with regard to mouse osteoblasts.

3T3 Cells↗

PubMed2005

Novel biomaterials for bisphosphonate delivery.

One type of gem-bisphosphonate (Zoledronate) has been chemically associated onto calcium phosphate (CaP) compounds of various compositions. For that purpose, CaP powders of controlled granulometry have been suspended in aqueous Zoledronate solutions of variable concentrations. Using mainly (31)P NMR spectroscopy, two different association modes have been observed, according to the nature of the CaP support and/or the initial concentration of the Zoledronate solution. beta-tricalcium phosphate (beta-TCP) and mixtures of hydroxyapatite and beta-TCP (BCPs) appear to promote Zoledronate-containing crystals formation. On the other hand, at concentrations <0.05 mol l(-1) CDAs (calcium deficients apatites) seem to undergo chemisorption of the drug through a surface adsorption process, due to PO(3) for PO(4) exchange, that is well described by Freundlich equations. At concentrations >0.05 mol l(-1), crystalline needles of a Zoledronate complex form onto the CDAs surface. The ability of such materials to release Zoledronate, resulting in the inhibition of osteoclastic activity, was shown using a specific in vitro bone resorption model.

Animals↗

PubMed2005

Calcium phosphate scaffold and bone marrow for bone reconstruction in irradiated area: a dog study.

Squamous cell carcinomas (SCC) of the upper aero-digestive tract are characterized by a high incidence of bone invasion; their treatment often requires large and damaging surgical resections and radiotherapy. Surgical and radiotherapeutic procedures generate irreversible effects on normal tissues, involving injuries on their reparation properties, especially on bone. The quality of life of patients undergoing major surgery and radiotherapy in maxillary and mandible areas is often reduced but could be improved by bone reconstructions. Bone reconstructions are rarely performed because surgery is complex and unsafe in irradiated bone. The aim of the study was to evaluate the bone reconstruction possibilities of macroporous biphasic calcium phosphate (MBCP) associated to autologous bone marrow (BM) graft injected after irradiation. MBCP hollowed blocks were specially designed and implanted in tibia and femur bone before irradiation in a dog model. Implants were removed after 18 weeks. This is the first report of experiments performed after radiation delivery using high fractionated doses approximating usual treatment of SCC in human. The quality of the bone adjacent to implanted MBCP and the bone ingrowth's rates were evaluated. The qualitative and quantitative role of BM grafts associated with the MBCP implants was determined, using scanning electron microscopy linked to quantitative image analysis. A direct contact between newly formed bone and MBCP implants associated to BM graft was observed, without fibrous interposition. The new-bone formation was statistically increased inside the MBCP (P=0.0126) by BM grafts. This study demonstrates that BM graft added to MBCP constitute an appropriate material to be considered in case of bone defect occurring in irradiated tissue, and could be foreseen for use after bone removal for oncologic obligations.

Animals↗

PubMed2005

Ectopic bone formation by microporous calcium phosphate ceramic particles in sheep muscles.

Calcium phosphate ceramics are widely used in bone reconstructive surgery because of their osteconductive properties. However, these materials generally lack osteoinductive properties required to support bone healing in large defects. In this article, we study the osteoinductive potential of calcium phosphate ceramic particles implanted for 6 months into the dorsal muscles of eight adult female sheep. Microporous biphasic calcium phosphate (MBCP) granules of 1-2 mm composed of hydroxyapatite and beta-tricalcium phosphate (60/40) had macropores of 450 microm, micropores of 0.43 microm, and a specific surface area of 1.8 m(2)/g. After 6 months in the back muscles of sheep, the explants composed of MBCP granules were hard and encapsulated by normal muscle tissue. Ectopic bone formation with Haversian structures was observed in close contact with the MBCP granules in histological sections. Back-scattered electron microscopy and micro-computed tomography indicated that approximately 10% of well-mineralized bone with mature osteocytes had formed between or upon the granules. The ectopic bone showed trabeculae bridging the MBCP granules. Both the number and thickness of the trabeculae formed between the MBCP particles were comparable to those measured in spongious bone. The overall results therefore confirmed the presence of mature bone after intramuscular implantation of MBCP granules. The different hypotheses explaining ectopic bone formation induced by MBCP granules are discussed. Synthetic bone substitutes with osteoinductive properties could be used in bone reconstructive surgery.

Animals↗

PubMed2004

Alveolar bone regeneration for immediate implant placement using an injectable bone substitute: an experimental study in dogs.

BACKGROUND: The aim of the present study was to assess the efficacy of a ready-to-use injectable bone substitute for bone regeneration around dental implants placed into fresh extraction sockets. METHODS: Third and fourth mandibular premolars were extracted from three beagle dogs and the interradicular septa were surgically reduced to induce a mesial bone defect. Thereafter, titanium implants were immediately placed. On the left side of the jaw, mesial bone defects were filled with an injectable bone substitute (IBS), obtained by combining a polymer and biphasic calcium phosphate ceramic granules. The right defects were left unfilled as controls. After 3 months of healing, specimens were prepared for histological and histomorphometric evaluations. RESULTS: No post-surgical complications were observed during the healing period. In all experimental conditions, histological observations revealed a lamellar bone formation in contact with the implant. Histomorphometric analysis showed that IBS triggers a significant (P<0.05) increase in terms of the number of threads in contact with bone, bone-to-implant contact, and peri-implant bone density of approximately 8.6%, 11.0%, and 14.7%, respectively. In addition, no significant difference was observed when number of threads, bone-to-implant contact, and bone density in the filled defects were compared to the no-defect sites. CONCLUSION: It is concluded that an injectable bone substitute composed of a polymeric carrier and calcium phosphate significantly increases bone regeneration around immediately placed implants.

Alveolar Process↗