Francoise Moreau

An injectable cellulose-based hydrogel for the transfer of autologous nasal chondrocytes in articular cartilage defects.

Articular cartilage has a low capacity for spontaneous repair. To promote the repair of this tissue, the transfer of autologous chondrocytes using a three‐dimensional matrix appears promising. In this context, the aim of the present work was to investigate the potential use of autologous rabbit nasal chondrocytes (RNC) associated with an injectable self‐setting cellulose‐based hydrogel (Si‐HPMC). Firstly, the influence of Si‐HPMC on chondrocytic phenotype was investigated by real‐time PCR for specific chondrocyte markers (type II collagen and aggrecan) and type I collagen. Thereafter, autologous RNC were amplified in vitro for 4 weeks before transplantation with Si‐HPMC into a rabbit articular cartilage defect followed by analysis 6 weeks later. Implants were histologically characterized for the presence of sulfated GAG and type II collagen. Transcripts analysis indicated that dedifferentiated RNC recovered expression of the main chondrocytic markers after in vitro three‐dimensional culture within Si‐HPMC. Histological analysis of autologous RNC transplanted in an articular cartilage defect revealed the formation of repair tissue with a histological organization similar to that of healthy articular cartilage. In addition, immunohistological analysis of type II collagen suggested that the repair tissue was a hyaline‐like cartilage. Si‐HPMC hydrogel associated with nasal chondrocytes therefore appears a promising injectable tissue engineering device for the repair of articular cartilage. Biotechnol. Bioeng. 2009;102: 1259–1267.

A comparison between bone reconstruction following the use of mesenchymal stem cells and total bone marrow in association with calcium phosphate scaffold in irradiated bone

The purpose of this study was to compare bone reconstruction using either mesenchymal stem cells (MSCs) or total bone marrow (TBM) in association with biphasic calcium phosphate (BCP) granules after irradiation in a rat model. Three weeks after an external irradiation of the hind limbs of rats, four bone defects were created per animal. The defects were filled with either BCP alone, or with a mixture of BCP and TBM, or with a mixture of BCP and MSCs (adipose-derived or bone marrow-derived MSCs). Three weeks after implantations, new-bone formation was assessed. Histological examination showed osteoconductive and osteointegrative properties of BCP in irradiated tissue. The BCP-TBM mixture significantly improved bone ingrowth (p<0.05). The BCP-MSCs mixtures did not provide new-bone formation over and above that induced by BCP alone. This gives grounds for suspecting that there is a link between this result and the cellular and vascular weakness observed in irradiated bone. The BCP-TBM mixture may have induced an increased vascularization of irradiated bone. This could be due to the presence of all components in TBM that were lacking in the BCP-MSCs mixtures. BCP associated with TBM appears to be the most efficient material for bone substitution in irradiated areas.

Mandibular Segmental Defect Regenerated With Macroporous Biphasic Calcium Phosphate, Collagen Membrane, and Bone Marrow Graft in Dogs

OBJECTIVE To reconstruct segmental mandibulectomy using calcium phosphate ceramics and collagen membrane with a delayed bone marrow grafting in experimental animals. DESIGN Defects of segmental mandibulectomy were filled with calcium phosphate granules and wrapped with a collagen membrane in 4 dogs and left empty as a control in 2 dogs. Two months later, a bone marrow graft was injected into the center of the implants. Animals were humanely killed after a 16-week delay. SUBJECTS Six adult beagles were included in this study. INTERVENTION Segmental mandibulectomy. MAIN OUTCOME MEASURE Bone ingrowth and material resorption in the reconstructed segment. RESULTS Successful osseous colonization bridged the whole length of the defects. The good new bone formation at the center and the periosteum-like formation at the periphery suggest the osteoinductive role of the bone marrow graft and the healing scaffold role of the membrane. CONCLUSIONS This model succeeded in regenerating a large segmental defect in the mandible. An investigation with a postimplantation radiation delivery schedule is required with the use of this model, which should be considered as a preclinical study for a bone tissue engineering approach in patients with cancer-related bone defects.

Reconstruction of irradiated bone segmental defects with a biomaterial associating MBCP+®, microstructured collagen membrane and total bone marrow grafting: An experimental study in rabbits

The bone tissue engineering models used today are still a long way from any oncologic application as immediate postimplantation irradiation would decrease their osteoinductive potential. The aim of this study was to reconstruct a segmental critical size defect in a weight-bearing bone irradiated after implantation. Six white New Zealand rabbits were immediately implanted with a biomaterial associating resorbable collagen membrane EZ(R) filled and micro-macroporous biphasic calcium phosphate granules (MBCP+(R)). After a daily schedule of radiation delivery, and within 4 weeks, a total autologous bone marrow (BM) graft was injected percutaneously into the center of the implant. All the animals were sacrificed at 16 weeks. Successful osseous colonization was found to have bridged the entire length of the defects. Identical distribution of bone ingrowth and residual ceramics at the different levels of the implant suggests that the BM graft plays an osteoinductive role in the center of the defect. Periosteum-like formation was observed at the periphery, with the collagen membrane most likely playing a role. This model succeeded in bridging a large segmental defect in weight-bearing bone with immediate postimplantation fractionated radiation delivery. This has significant implications for the bone tissue engineering approach to patients with cancer-related bone defects.

Bone Ingrowth at the Expense of a Novel Macroporous Calcium Phosphate Cement

G. Daculsi, I. Khairoun, R. LeGeros, F. Moreau , P. Pilet , X. Bourges, P. Weiss , O. Gauthier 1,4 INSERM UMR 791, Faculté de Chirurgie Dentaire, 44042 Nantes, France, Biomatlante, 5 rue Edouard Belin, Vigneux de Bretagne, 44360, France,. New York University, College of Dentistry, New York, NY 10010, Laboratoire de Chirurgie, Ecole Nationale Vétérinaire de Nantes 44307 France, Electron and Microimaging Center SC3MM, Nantes University

Repairing segmental defect with a composite associating collagen membrane and MBCP+® combined with total bone marrow graft in irradiated bone defect: an experimental study in Rabbit.

The aim of this study was to study bone marrow quality from various location and species for reconstruction of segmental critical size defect in irradiated weigh bearing bone. Sample of bone marrow aspirates from rabbits and Beagle dog were analyzed. Rabbits were implanted with a composite associating resorbable collagen membrane plus micro macroporous biphasic calcium phosphate (MBCP®) and autologous bone marrow (BM) injected after irradiation. Bone marrow samples were found to be significantly less rich in tibia than in humerus and ilium in Dog and less rich in Dog than in Rabbit (p<0,05). Successful osseous colonization bridging of the defect were obtain at 16 weeks in all animals. Identical repartition of bone ingrowth and residual ceramic at the different levels of the implant suggest an osteoinduction role of the bone marrow graft in the center of the defect. This model succeeded in reconstruct a large segmental defect in weight bearing and irradiated bone in rabbit.

PL DLLA Calcium Phosphate Composite Combined with MBCP Gel® for New Surgical Technologies: Resorbable Osteosynthesis and Bone Substitute

We have performed and evaluated a composite combining PL DLLA and PCa able to have a better control of the hydrolysis and maintain of the mechanical property on time, until wound healing was achieved. Bioabsorbable osteoconductive composite is devoted to the development of resorbable osteosynthesis for spine and osteoarticular surgeries. Such resorbable osteosynthesis will be associated to new bone substitute having high osteogenic property but without initial mechanical property. The purpose of this study was to evaluate the resorption kinetic of a composite using PL DLLA (Poly [L-Lactide-co-D,L-Lactide] acide) charged with PCa granules and the interaction with injectable bioceramic without self hardening. MBCP gel® is a composite associating a mineral phase of an intimate nanoscale melting of hydroxyapatite and beta tri calcium phosphate and an aqueous phase containing a synthetic polymer derived from cellulose HPMC (hydroxyl propyl methyl cellulose).

PL DLLA Calcium Phosphate Composite Combined with Macroporous Calcium Phosphate Cement MCPC® for New Surgical Technologies Combining Resorbable Osteosynthesis and Injectable Bone Substitute

Resorbable osteosynthesis based on PLLA and derivatives will be associated to bone substitute for bone reconstruction. We have performed rand evaluated a composite combining PL DLLA and Biphasic calcium phosphate able to have a), a better controlled hydrolysis in the purpose to preserve on time the mechanical property, and b), for long term efficiency, bone ingrowth at the expense of the osteosynthesis and the associated bone substitute. A new calcium phosphate cement MCPC® was tested with such composite. The novel macroporous calcium phosphate cement MCPC sets to poorly crystalline apatite after mixing the powder component and an aqueous solution. Interconnective macroporosity was induced on time by resorption of one part of the MCPC®. The multiphasic calcium phosphate components in the cement, are resorbed at different rates allowing the replacement by newly formed bone. This study reports the biocompatibility and the interactions of a composite using PL DLLA (Poly [L-Lactide-co-D,L-Lactide] acid) charged with biphasic calcium phosphate granules and a self setting calcium phosphate cement of new generation.

Hollow Shells Development and Characterization for Cells Carrying Purpose

Bioceramics draw attention in bone tissue engineering field since their biomimetic properties regarding bone attribute. In this context, a concept of smart bioceramics granules made of Hydroxyapatite have been set up, enhancing surface area available to body fluids containing proteins and cell adhesion for bone forming respectively thanks to microporosities and macropore concavities. New “hollow shell” granules were developed and assessed by physico-chemical characterizations, in-vitro experiments and in-vivo implantation in comparison with classical round granules. This new original galenic formulation showed promising potential in cell carrying and osteoconduction matter.

Periostal Reconstruction Using New Porcine Microstructured Collagen Membrane and Calcium Phosphate Cement: A Dog Model

The aim of this study was to evaluate the capacity of the association of calcium phosphate cement with collagen membrane for large segmental bone defect. Six adult beagle dog underwent defect creation in the ulna and was reconstructed with combination of macroporous calcium phosphate cement MCPC® and resorbable collagen membrane EZ Cure®. After 6 months of implantation, the samples were analyzed with Micro CT, light microscopy and SEM using BSE. Bridging of the defect with a lamellar and well organized bone was achieved in all animals. MCPC granules resorption was increased at the extremities of the implant. Collagen membrane at the expense of the implant was replaced by periosteum-like formation. The results demonstrate the ability of the composite to reconstruct large segmental and critical size defect in long bone.