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Leonardo – Biomaterial for Bone Grafting PRECLINICAL And

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Leonardo – Biomaterial for Bone Grafting PRECLINICAL And PRECLINICAL and CLINICAL STUDIES on BIOACTIVE GLASS and LEONARDO – Oral surgery Leonardo – Biomaterial for bone grafting 4. Biocompatibility evaluation (A) 12 PROLIFERATION AND GROWTH OF BONE Preclinical studies on Leonardo granules 4.1 Biocompatibility tests following NF EN ISO 10993-1 standard 11 CRYSTALLIZATION OF MATRIX Biocompatibility evaluation has been conducted following the table 10 GENERATION OF MATRIX of the NF EN ISO 10993-1 standard. Since bioactive glass is a 1. General informations on bioactive glasses very low, but can vary with pH. Tricalcium phosphate ß (ß-TCP) is 100 9 DIFFERENTATION OF STEM CELLS well-known material which has been clinically used for more than much more soluble than HA and degrades rapidly in vivo. Bioactive Bioactive glasses are classified as ceramic biomaterials, and more 20 years, biocompatibility tests which have been conducted aim precisely as bioactive ceramic materials, just like hydroxyapatite glass 45S5 can be considerate as perfectly absorbable since it com- 8 ATTACHMENT OF STEM CELLS 20 at evaluating the toxicity of manufacturing residues. [1] pletely dissolves in order to form carbonated hydroxyapatite. Its and other calcium phosphate ceramics . Bioactive glasses are 7 ACTION OF MACROPHAGES mainly composed of silicon dioxide (SiO ), sodium oxide (Na O), degradation rate depends on its composition. 10 2 2 Here is the list of the tests which have been conducted and the calcium oxide (CaO) and phosphorous pentoxide (P O ). L. L. Hench 6 ADSORPTION OF BIOLOGICAL MOIETIES IN HCA LAYER 2 5 2 providers involved in the study: was the inventor of bioactive glass 45S5 in 1971, called Bioglass® LOG TIME (HOURS) 5 CRYSTALLIZATION OF HYDROXYL CARBONATE APATITE (HCA) 2. Description of mechanism of bioactivity (composed, in weigh, of 45 % SiO2, 24.5 % Na2O, 24.5 % CaO and 1 ADSORPTION OF AMORPHOUS Ca+PO +CO 6 % P O ). Since then, different types of bioactive glasses or glass- A cascade of physical and chemical reactions occurs at the periph- 4 4 3 BIOCOMPOTIBILITY TESTS PERFORMED 2 5 SURFACE REACTION STAGES ery of bioactive glasses in contact with body fluids. A hydrolytic ceramic materials (glasses incorporating hydroxyapatite crystals 3 POLYCONDENSATION OF SiOH + SiOH Si-O-Si PROLIFERATION AND GROWTH OF BONE and/or wollastonite), with different properties, have been devel- degradation of bioactive glass happens[2], followed by numerous and oped[1]. complex reactions. Figure 1 below summarizes the cascade of the 1&2 FORMATION OF SiOH BONDS IRRITATION (ISO 10993-10) 11 reactions occurring at the surface of bioactive glass during the BIOACTIVE GLASS DELAYED-TYPE HYPERSENSITIVITY (ISO 10993-10) Bioactive glasses are effective materials for bone grafting or pros- formation of a solid grip with the bone. thesis coating. They have been used in different types of surgeries: Figure 1: Sequence of interfacial reactions involved in forming a bond between ACUTE SYSTEMIC TOXICITY (ISO 10993-11) The five first reactions take place in the periphery of bioactive glass bone and a bioactive glass. L. L. Hench[1] orthopedic surgery, oral surgery, cranio-facial surgery and spinal AMES TEST (ISO 10993-3) surgery. These materials are known for their excellent biocompat- and do not depend on the presence of tissue. This reactions has been ibility. In contact with body fluids, they are able to create a solid studied and could take place in distilled water or buffer solution 3. Osteostimulation PYROGENICITY (USP 31) like TRIS or SBF[3,4]. These reactions make up a layer of crystallized Recent in vitro studies have been led to the understanding of the grip at the interface implant/host tissues. These biomaterials do not CYTOTOXICITY (ISO 10993-5) promote any fibrous encapsulation at the interface implant/tissue; carbon hydroxyapatite at the top of material. The implant fastens mechanism of action of bioactive glass[5,6]. They show that ionic the connection between the implant and the bone tissues is thus down with the tissue for the step 6 to 11 like as growth of bone. products such as silicon and calcium have an influence on the stem cells differentiation and on the growth of bone. Besides, the bone direct. The surface of bioactive glasses serves as a support for bone The set of biocompatibility tests performed show that bioactive mineralization happens thanks to phosphor transit. The dissolution regrowth. The degree of bioactivity depends on the composition of glass 45S5 products: bioactive glasses, in particular on the proportion of silica (SiO ), products of bioactive glass 45S5 develop an extracellular environ- 2 are not cytotoxic [2] ment that is capable of supporting osteoblast phenotype expression sodium oxide (Na2O), phosphorus pentoxide and calcium oxide (CaO) . and extracellular matrix deposition and mineralization in vitro[6]. do not induce irritation do not induce delayed-type hypersensitivity Absorbable biomaterials are known for their ability to gradual- 5 I. D. Xynos et al. Bioglass 45S5 Stimulates Osteoblast Turnover and Enhances Bone do not induce acute systemic toxicity ly degrade while being replaced by the natural host bone tissue. Formation In Vitro: Implications and Applications for Bone Tissue Engineering. are neither toxic nor mutagen Hydroxyapatite is hardly soluble and its in vivo degradation rate is 3 L. L. Hench, G. P. Latorre. Reaction kinetics of bioactive ceramics. PartIV: Calcif Tissue Int (2000) 67:321 effect of glass and solution composition. Bioceramics (1992) 5: p. 67-74 6 Olga Tsigkou et al. Differentiation of fetal osteoblasts and formation of mineralized are not pyrogen 1 L. L. Hench. The Story of Bioglass. J. Mater. Sci: Mater Med (2006) 17: p. 967-978 4 A. E. Clark, C. G. Pantano, L. L. Hench. Auger spectroscopy analysis of bioglass bone nodules by 45S5 Bioglass conditioned medium in the absence of osteogenic 2 L. L. Hench. Bioceramics. J. Am. Ceram. Soc. (1998) 81: p. 1705-28 corrosion films. J. Am. Ceram. Soc. (1976) 59: p. 37-39 supplements. Biomaterials 30 (2009) 3542–3550 | 2 3 | 4.2 Implantation of different bone grafting materials on rabbits: global evaluation of NORAKER products Clinical Data An implantation study has been conducted on rabbits in order to evaluate the biological response of tissues to different bone graft- ing materials, including bioactive glass 45S5. This study has been 1. Literature review conducted in adaptation to the ISO 10993-6 norm. Bioactive glass A literature review including products similar to Leonardo has been 45S5 product is compared to a ß-TCP control material. Granules are realised using PubMed database. implanted in 5 animals for each time, in the area of the femoral epiphysis (either distal or proximal). Observation times are 4, 12 The following products, composed of bioactive glass 45S5, are and 24 weeks. Figure 3: Figure 4: marketed: PerioGlas® (oral surgery), NovaBone® (orthopaedic sur- Histological section in the Histological section in the gery) and NovaBone-C/M® (Skull-maxillo-facial surgery) marketed implantation area of implantation area of bioactive glass 45S5 Bioactive glass 45S5 by NovaBone (developped by US biomaterials) and Biogran® (de- at 12 weeks (section by at 24 weeks (section by velopped by Orthovita). One can also find bioactive glass 53S4 or microtome/Giemsa) microtome/Giemsa) S53P4 (Vivoxid, Finlande) with a different composition to 45S5: 53 % of SiO2, 23 % of Na2O, 20 % of CaO and 4 % de P2O5. Another After 24 weeks, materials have the following biological characteristics: surface of the material, 4/ degradation of the material (with differ- product contains 60 % of SiO2 who is called 60S. A mix of bioac- ent rates), and replacement by mature bone. Materials are in particle form and damage. tive glass and wollastonite called Cerabone A/W is also available No reactions to foreign body are visible. on the market. Addition of wollastonite to the ceramic gives better The picture shows a normal marrow without any sign of residual Materials are totally inserted to the bone. mechanical properties. This product is used for hip and spine surgery. ossification. Some Bioactive glass 45S5 residues can be observed, under granuled shapes (BG) in very isolated trabeculae. Bioactive glass 45S5 and ß-TCP materials can be considered as bio- BIOAKTIVE GLASS 45S5: Figure 2: active implants which contribute bone remodeling. At first, they Histological section in the PERIOGLAS (NOVABONE) After 12 weeks, Bioactive glass 45S5 granules quickly resorbed are colonized by monocyte cells, then by a soft conjonctive tissue. implantation area of NOVABONE (NOVABONE) bioactive glass 45S5 whereas control material (ß-TCP) had a slowier degradation. Osteoblasts then differentiate at their surface and are responsible PRODUCTS BIOGRAN (ORTHOVITA/BIOMET 3i) at 4 weeks (section by for the synthesis of immature bone matrix. Finally, the immature EQUIVALENT microtome/Giemsa) Outside the cortical area, the picture shows a heap of bioactive matrix is remodelled, repaced by mature bone. During this final stage, TO LEONARDO BIOACTICE GLASS S53P4: Bioactive glass 45S5 granules (material) are of particle shapes. We glass 45S5 particulates. They are included in a soft conjonctive tis- implants are damaged with variable-speed. 12 weeks the material (MIX OF Ca, Na, Si, P) BONALIVE (VIVOXID) can see numerous monocyte cells in the existing soft connective sue which is denser at the interface with particulates. In the medul- implantation, bioactive glass 45S5 was more damaged than the BIOACTIVE GLASS AW: tissue, between the ossified areas, only at the surface of the parti- lar implantation area, some trabeculae remain. They seam to result ß-TCP control materialß-TCP. After 24 weeks, both materials were CERABONE (AAP) cles (dark blue). from the presence of the particulates. damaged and totally integrated to the bone. No foreign body was visible in the implantation sites. After 4 weeks, at histological observation, both materials (bioactive Medical device vigilance has been conducted on bioactive glass glass 45S5 and control) have a bioactive appearance. They went products available in the market.
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