A biomaterial with bioactive glass nanoparticles (nBG) and Ca2+ incorporated into alginate matrix was developed. Films characterization was carried out by SEM, IR, tensile strength measurements, bioactivity assay, degradation and swelling studies. Ca2+ release from films was analyzed. Freeze-dried-scaffolds were also fabricated. Films showed the development of a homogeneous matrix and the mechanical properties were improved when nBG were incorporated. The bioactive nature of nBG containing films was confirmed by studies in simulated body fluid. Degradation was negligible and a good swelling capacity was observed. Moreover Ca2+ was released in a controlled manner. In scaffolds fabricated by freeze-drying, pores were seen to be uniform and well distributed. According to the characterization results, these composite biomaterials are attractive candidates for the fabrication of bone tissue engineering scaffolds.Fil: Cattalini, Juan Pablo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Garcia, J,. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Boccaccini, A. R.. Universitat Erlangen-Nuremberg; Alemania;Fil: Lucangioli, Silvia Edith. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Analítica y Fisicoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Mouriño, Viviana Silvia Lourdes. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Tecnología Farmacéutica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentin

Boccaccini, A. R.

Cattalini, Juan Pablo

Garcia, J,

Lucangioli, Silvia Edith

Mouriño, Viviana Silvia Lourdes

Procedia Engineering

J.P. Cattalini

J. García

A.R. Boccaccini

S. Lucangioli

V. Mouriño

Crossref

A New Calcium Releasing Nano-composite Biomaterial for Bone Tissue Engineering Scaffolds

LAReferencia - Red Federada de Repositorios Institucionales de Publicaciones Científicas Latinoamericanas

 Procedia Engineering  59 ( 2013 )  78 – 84 1877-7058 © 2013 The Authors. Published by Elsevier Ltd.Selection and peer-review under responsibility of the Centre for Rapid and Sustainable Product Development, Polytechnic Institute of Leiria, Centro Empresarial da Marinha Grandedoi: 10.1016/j.proeng.2013.05.096 3rd International Conference on Tissue Engineering, ICTE2013 A new calcium releasing nano-composite biomaterial for bone tissue engineering scaffolds Cattalini J.P.a, García J.a, Boccaccini A.R.b, Lucangioli S.a,c, Mouriño V.a,c,* a Faculty of Pharmacy and Biochemistry, University of Buenos Aires, 956 Junin 6th floor, Buenos Aires, CP1113, Argentina b Institute of Biomaterials,University of Erlangen-Nuremberg, 91058 Erlangen, Germany c National Research Council (CONICET), Buenos Aires, Argentina Abstract A biomaterial with bioactive glass nanoparticles (nBG) and Ca2+ incorporated into alginate matrix was developed. Films characterization was carried out by SEM, IR, tensile strength measurements, bioactivity assay, degradation and swelling studies. Ca2+ release from films was analyzed. Freeze-dried-scaffolds were also fabricated. Films showed the development of a homogeneous matrix and the mechanical properties were improved when nBG were incorporated. The bioactive nature of nBG containing films was confirmed by studies in simulated body fluid. Degradation was negligible and a good swelling capacity was observed. Moreover Ca2+ was released in a controlled manner. In scaffolds fabricated by freeze-drying, pores were seen to be uniform and well distributed. According to the characterization results, these composite biomaterials are attractive candidates for the fabrication of bone tissue engineering scaffolds.  © 2013 The Authors. Published by Elsevier Ltd. Selection and peer-review under responsibility of the Centre for Rapid and Sustainable Product Development, Polytechnic Institute of Leiria, Centro Empresarial da Marinha Grande. Keywords: Composites; Biomaterials; Scaffolds; Films; Calcium.      * Corresponding author. Tel.: 0054-11-4961-8371; fax: 0054-11-4961-8271. E-mail address: vmourino@ffyb.uba.ar Available online at www.sciencedirect.com79 J.P. Cattalini et al. /  Procedia Engineering  59 ( 2013 )  78 – 84 1. Introduction The new generation of biomaterials to develop scaffolds for bone tissue engineering (BTE) includes the elaboration of nano-composites (n-c) in order to resemble de bone extracellular matrix, with the possibility of using the scaffold itself as a drug delivery system [1]. In this context, inorganic biomaterials such as ceramics and glasses, as well as organic systems such as biodegradable polymers, are being used to fabricate composite scaffolds [2]. Among inorganic systems, bioactive glasses are a special class of highly bioactive materials exhibiting interesting properties such osteoconductive and osteocompatible characteristics, as well as the promotion of osteogenesis and angiogenesis [3]. Further, studies confirmed that the addition of ceramic nanoparticles into a polymer matrix usually improves the mechanical properties of the matrix [4]. The possibility to deliver different biological factors to the surrounding tissue to stimulate new bone formation, as well as to release different drugs intended for the treatment of bone diseases is a strategy that makes the scaffold a multifunctional matrix for BTE [5,6]. Lately, metallic ions as therapeutics agents (MITA) have been considered for new biomaterials development [7,8]. In fact, a large amount of biomaterials for BTE contain calcium ions, Ca2+ [9], which stimulate the differentiation of bone cells, osteoblasts proliferation, bone metabolism and bone mineralization [7]. According to the above considerations, this work presents the development of a new n-c biomaterial which combines alginate with nBG and incorporates Ca2+ for BTE applications. 2. Materials and Methods A mixture of sodium alginate with nBG (nominal composition of Bioglass® 45S5, size: 35-40 nm), loaded with Ca2+ ions (by using CaCl2.2H2O, which was purchased from MERCK, Germany), was casted into molds to obtain n-c films. The morphology and homogeneity of n-c films were evaluated by scanning electron microscopy (SEM). n-c films were also analyzed by Infrared (IR) spectroscopy. Mechanical properties of n-c films in compare with films without nBG were assessed by tensile strength measurements, where ten samples of each type of film were used. The bioactive nature of n-c films was tested in Simulated Body Fluid (SBF) at 37°C for 7 days and the growth of hydroxyapatite (HA) crystals on the films surface was determined by SEM and confirmed by X-ray diffraction (XRD). Degradation and swelling studies of n-c films were made at 37 °C in phosphate buffer at pH 7.4. Ca2+ release from n-c films was studied at 37 °C in phosphate buffer at pH 7.4 during 60 days and the quantification was made by a UV-direct capillary electrophoresis method previously developed [10]. Scaffolds of the same composition as n-c films were fabricated by freeze-drying. Morphology and porosity of n-c scaffolds were studied by SEM. 3. Results and Discussion 3.1. Characterization of films Figure 1 shows the surface of n-c films; a uniform and homogeneous matrix with nBG dispersed in it is observed.  Figure 1. SEM image of the surface of a n-c film before bioactivity studies. 80   J.P. Cattalini et al. /  Procedia Engineering  59 ( 2013 )  78 – 84 The interaction between Ca2+ and the films matrix, either for films with or without nBG, is shown by IR spectra, Figure 2.   Figure 2. IR spectra of a) nBG, b) alginate, c) films without nBG, d) n-c films. The addition of nBG to the alginate matrix increased significantly (p < 0.05) the tensile strength in comparison with films not containing nBG, as shown Figure 3.   Figure 3. Results of tensile strength measurements on films with and without incorporation of nBG. SEM images of films immersed in SBF are shown in Figure 4a,b. The in vitro bioactivity was confirmed by the deposition of HA crystals on the surface of the n-c films after 3 and 7 days of immersion in SBF, as indicated in the XRD spectrum in Figure 5.  81 J.P. Cattalini et al. /  Procedia Engineering  59 ( 2013 )  78 – 84   Figure 4. SEM images of (a) n-c films after 3 days in SBF showing the growth of crystals on the surface, and (b) n-c films after 7 days in SBF.  Figure 5. XRD pattern of a composite film after 7 days of incubation in SBF showing characteristics peaks of hydroxyapatite (•) and the presence of amorphous material.  3.2. Degradation and Swelling studies of films No significant changes in weight loss were observed indicating that n-c films still remained shaped until day 60 (Figure 6a). The swelling study indicated a good swelling capacity of the films with a weight gain percentage of about 100% (Figure 6b).  (a) (b) 82   J.P. Cattalini et al. /  Procedia Engineering  59 ( 2013 )  78 – 84   Figure 6. Results of (a) weight loss as function of time, showing that there is no significant weight loss until the end of the study, and (b) swelling study showing a weight gain of 100% after 15 days, where films double their weight. 3.3. Release study of films The amount of Ca2+ released increases until day 30. After that and until day 60, the release of Ca2+ tends to remain stable. About 30-35 μg/ml of Ca2+ were released from day 30 until the end of the study. Figure 7 shows the release profile of Ca2+.   Figure 7. Ca2+ release profile in phosphate buffer at pH 7.4 during 60 days. Weightloss(%)Time (days)Weightgain(%)Time (h)Ca concentration (μg/ml)Time (days)(b) (a) 83 J.P. Cattalini et al. /  Procedia Engineering  59 ( 2013 )  78 – 84 3.4. Characterization of scaffolds Figure 8 shows the morphology and porosity of n-c scaffolds. Pores are uniform and well distributed (Figure 8a), and the presence of bioactive glass nanoparticles can be observed (Figure 8b).    Figure 8. SEM images of nBG containing composite scaffolds at (a) low and (b) high magnification. 4. Conclusions Nanocomposite, biodegradable and bioactive films and scaffolds with Ca2+ releasing ability as multifunctional substrates for BTE were developed. The incorporation of nBG into alginate films significantly improved the tensile strength. Biomineralization studies indicated the deposition of apatite on the films surface, suggesting their bioactive behavior which is a consequence of the high bioreactivity of the added nBG. Films do not completely degrade after 60 days, which indicates their stability over time. The release of Ca2+ is seen to be controlled and without an initial burst effect, which might be an advantage when cellular in vitro studies, such as citocompatibility assays, are envisaged. Future assays will be focused on a full characterization of the scaffolds, the study of the Ca2+ release in relevant biological conditions and the investigation of the capability of these scaffolds to promote the proliferation of bone cells. Acknowledgement We acknowledge Prof. W. Stark and Dr D. Mohn (ETH Zurich) for providing nBG powder. (a) (b) 84   J.P. Cattalini et al. /  Procedia Engineering  59 ( 2013 )  78 – 84 References [1] Mouriño V, Boccaccini AR. Bone tissue engineering therapeutics: controlled drug delivery in three-dimensional scaffolds. J R Soc Interface 2010;7:209-227. [2] Gardin C, Ferroni L, Favero L, Stellini E, Stomaci D, Sivolella S, Bressan E, Zavan B. Nanostructured biomaterials for tissue engineered bone tissue reconstruction. Int J Mol Sci 2012;13:737-757.  [3] Hoppe A, Güldal N, Boccaccini AR. A review of the biological response to ionic dissolution products from bioactive glasses and glass-ceramics. Biomaterials 2011;32: 2757-2774.  [4] Mouriño V, Newby P, Pishbin F, Cattalini JP, Lucangioli S, Boccaccini AR. Physicochemical, biological and drug-release properties of gallium crosslinked alginate/nanoparticulate bioactive glass composite films. Soft Matter 2011;7:6705-6712. [5] Gloria A, De Santis R, Ambrosio L. Polymer-based composite scaffolds for tissue engineering. J Appl Biomater Biomech 2010;8:57-67.  [6] Bose S, Roy M, Bandyopadhyay A. Recent advances in bone tissue engineering scaffolds. Trends in Biotechnology 2012;30:546-554. [7] Mouriño V, Cattalini JP, Boccaccini AR. Metallic ions as therapeutic agents in tissue engineering scaffolds: an overview of their biological applications and strategies for new developments. J R Soc Interface 2012;9:401-419.  [8] Habibovic P, Barralet, JE. Bioinorganics and biomaterials: Bone repair. Acta Biomater 2011;7:3013-3026. [9] Dorozhkin, S. Calcium orthophosphates as bioceramics: state of the art. J Funct Biomater 2010;1:22-107.  [10] Cattalini JP, García J, Mouriño V, Lucangioli S. Development and validation of a capillary zone electrophoresis method for the determination of calcium in composite biomaterials. Curr Anal Chem 2013, CAC-EPUB-20121024-2. 

English

A new calcium releasing nano-composite biomaterial for bone tissue engineering scaffolds

AbstractA biomaterial with bioactive glass nanoparticles (nBG) and Ca2+ incorporated into alginate matrix was developed. Films characterization was carried out by SEM, IR, tensile strength measurements, bioactivity assay, degradation and swelling studies. Ca2+ release from films was analyzed. Freeze-dried-scaffolds were also fabricated. Films showed the development of a homogeneous matrix and the mechanical properties were improved when nBG were incorporated. The bioactive nature of nBG containing films was confirmed by studies in simulated body fluid. Degradation was negligible and a good swelling capacity was observed. Moreover Ca2+ was released in a controlled manner. In scaffolds fabricated by freeze-drying, pores were seen to be uniform and well distributed. According to the characterization results, these composite biomaterials are attractive candidates for the fabrication of bone tissue engineering scaffolds

Cattalini, J.P.

García, J.

Boccaccini, A.R.

Lucangioli, S.

Mouriño, V.

Elsevier - Publisher Connector 

 Procedia Engineering  59 ( 2013 )  78 – 84 
1877-7058 © 2013 The Authors. Published by Elsevier Ltd. Open access under CC BY-NC-ND license.
Selection and peer-review under responsibility of the Centre for Rapid and Sustainable Product Development, Polytechnic Institute of Leiria, 
Centro Empresarial da Marinha Grande
doi: 10.1016/j.proeng.2013.05.096 
3rd International Conference on Tissue Engineering, ICTE2013 
A new calcium releasing nano-composite biomaterial for bone 
tissue engineering scaffolds 
Cattalini J.P.a, García J.a, Boccaccini A.R.b, Lucangioli S.a,c, Mouriño V.a,c,* 
a Faculty of Pharmacy and Biochemistry, University of Buenos Aires, 956 Junin 6th floor, Buenos Aires, CP1113, Argentina 
b Institute of Biomaterials,University of Erlangen-Nuremberg, 91058 Erlangen, Germany 
c National Research Council (CONICET), Buenos Aires, Argentina 
Abstract 
A biomaterial with bioactive glass nanoparticles (nBG) and Ca2+ incorporated into alginate matrix was developed. Films 
characterization was carried out by SEM, IR, tensile strength measurements, bioactivity assay, degradation and swelling 
studies. Ca2+ release from films was analyzed. Freeze-dried-scaffolds were also fabricated. Films showed the development of a 
homogeneous matrix and the mechanical properties were improved when nBG were incorporated. The bioactive nature of nBG 
containing films was confirmed by studies in simulated body fluid. Degradation was negligible and a good swelling capacity 
was observed. Moreover Ca2+ was released in a controlled manner. In scaffolds fabricated by freeze-drying, pores were seen to 
be uniform and well distributed. According to the characterization results, these composite biomaterials are attractive candidates 
for the fabrication of bone tissue engineering scaffolds. 
© 2013 The Authors. Published by Elsevier Ltd. Open access under CC BY-NC-ND license. 
Selection and peer-review under responsibility of the Centre for Rapid and Sustainable Product Development, Polytechnic 
Institute of Leiria, Centro Empresarial da Marinha Grande. 
Keywords: Composites; Biomaterials; Scaffolds; Films; Calcium. 
* Corresponding author. Tel.: 0054-11-4961-8371; fax: 0054-11-4961-8271. 
E-mail address: vmourino@ffyb.uba.ar 
Available online at www.sciencedirect.com
79 J.P. Cattalini et al. /  Procedia Engineering  59 ( 2013 )  78 – 84 
1. Introduction 
The new generation of biomaterials to develop scaffolds for bone tissue engineering (BTE) includes the 
elaboration of nano-composites (n-c) in order to resemble de bone extracellular matrix, with the possibility of 
using the scaffold itself as a drug delivery system [1]. In this context, inorganic biomaterials such as ceramics and 
glasses, as well as organic systems such as biodegradable polymers, are being used to fabricate composite scaffolds 
[2]. Among inorganic systems, bioactive glasses are a special class of highly bioactive materials exhibiting 
interesting properties such osteoconductive and osteocompatible characteristics, as well as the promotion of 
osteogenesis and angiogenesis [3]. Further, studies confirmed that the addition of ceramic nanoparticles into a 
polymer matrix usually improves the mechanical properties of the matrix [4]. The possibility to deliver different 
biological factors to the surrounding tissue to stimulate new bone formation, as well as to release different drugs 
intended for the treatment of bone diseases is a strategy that makes the scaffold a multifunctional matrix for BTE 
[5,6]. Lately, metallic ions as therapeutics agents (MITA) have been considered for new biomaterials development 
[7,8]. In fact, a large amount of biomaterials for BTE contain calcium ions, Ca2+ [9], which stimulate the 
differentiation of bone cells, osteoblasts proliferation, bone metabolism and bone mineralization [7]. According to 
the above considerations, this work presents the development of a new n-c biomaterial which combines alginate 
with nBG and incorporates Ca2+ for BTE applications. 
2. Materials and Methods 
A mixture of sodium alginate with nBG (nominal composition of Bioglass® 45S5, size: 35-40 nm), loaded with 
Ca2+ ions (by using CaCl2.2H2O, which was purchased from MERCK, Germany), was casted into molds to obtain 
n-c films. The morphology and homogeneity of n-c films were evaluated by scanning electron microscopy (SEM). 
n-c films were also analyzed by Infrared (IR) spectroscopy. Mechanical properties of n-c films in compare with 
films without nBG were assessed by tensile strength measurements, where ten samples of each type of film were 
used. The bioactive nature of n-c films was tested in Simulated Body Fluid (SBF) at 37°C for 7 days and the 
growth of hydroxyapatite (HA) crystals on the films surface was determined by SEM and confirmed by X-ray 
diffraction (XRD). Degradation and swelling studies of n-c films were made at 37 °C in phosphate buffer at pH 
7.4. Ca2+ release from n-c films was studied at 37 °C in phosphate buffer at pH 7.4 during 60 days and the 
quantification was made by a UV-direct capillary electrophoresis method previously developed [10]. 
Scaffolds of the same composition as n-c films were fabricated by freeze-drying. Morphology and porosity of n-
c scaffolds were studied by SEM. 
3. Results and Discussion 
3.1. Characterization of films 
Figure 1 shows the surface of n-c films; a uniform and homogeneous matrix with nBG dispersed in it is 
observed. 
 
Figure 1. SEM image of the surface of a n-c film before bioactivity studies. 
80   J.P. Cattalini et al. /  Procedia Engineering  59 ( 2013 )  78 – 84 
The interaction between Ca2+ and the films matrix, either for films with or without nBG, is shown by IR spectra, 
Figure 2. 
 
 
Figure 2. IR spectra of a) nBG, b) alginate, c) films without nBG, d) n-c films. 
The addition of nBG to the alginate matrix increased significantly (p < 0.05) the tensile strength in comparison 
with films not containing nBG, as shown Figure 3. 
 
 
Figure 3. Results of tensile strength measurements on films with and without incorporation of nBG. 
SEM images of films immersed in SBF are shown in Figure 4a,b. The in vitro bioactivity was confirmed by the 
deposition of HA crystals on the surface of the n-c films after 3 and 7 days of immersion in SBF, as indicated in 
the XRD spectrum in Figure 5. 
 
81 J.P. Cattalini et al. /  Procedia Engineering  59 ( 2013 )  78 – 84 
 
 
Figure 4. SEM images of (a) n-c films after 3 days in SBF showing the growth of crystals on the surface, and (b) n-c films after 7 days in SBF. 
 
Figure 5. XRD pattern of a composite film after 7 days of incubation in SBF showing characteristics peaks of hydroxyapatite (•) and the 
presence of amorphous material.  
3.2. Degradation and Swelling studies of films 
No significant changes in weight loss were observed indicating that n-c films still remained shaped until day 60 
(Figure 6a). The swelling study indicated a good swelling capacity of the films with a weight gain percentage of 
about 100% (Figure 6b). 
 
(a) 
(b) 
82   J.P. Cattalini et al. /  Procedia Engineering  59 ( 2013 )  78 – 84 
 
 
Figure 6. Results of (a) weight loss as function of time, showing that there is no significant weight loss until the end of the study, and (b) 
swelling study showing a weight gain of 100% after 15 days, where films double their weight. 
3.3. Release study of films 
The amount of Ca2+ released increases until day 30. After that and until day 60, the release of Ca2+ tends to 
remain stable. About 30-35 μg/ml of Ca2+ were released from day 30 until the end of the study. Figure 7 shows the 
release profile of Ca2+. 
 
 
Figure 7. Ca2+ release profile in phosphate buffer at pH 7.4 during 60 days. 
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Time (days)
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(%
)
Time (h)
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83 J.P. Cattalini et al. /  Procedia Engineering  59 ( 2013 )  78 – 84 
3.4. Characterization of scaffolds 
Figure 8 shows the morphology and porosity of n-c scaffolds. Pores are uniform and well distributed (Figure 
8a), and the presence of bioactive glass nanoparticles can be observed (Figure 8b). 
 
 
 
Figure 8. SEM images of nBG containing composite scaffolds at (a) low and (b) high magnification. 
4. Conclusions 
Nanocomposite, biodegradable and bioactive films and scaffolds with Ca2+ releasing ability as multifunctional 
substrates for BTE were developed. The incorporation of nBG into alginate films significantly improved the tensile 
strength. Biomineralization studies indicated the deposition of apatite on the films surface, suggesting their 
bioactive behavior which is a consequence of the high bioreactivity of the added nBG. Films do not completely 
degrade after 60 days, which indicates their stability over time. The release of Ca2+ is seen to be controlled and 
without an initial burst effect, which might be an advantage when cellular in vitro studies, such as citocompatibility 
assays, are envisaged. Future assays will be focused on a full characterization of the scaffolds, the study of the Ca2+ 
release in relevant biological conditions and the investigation of the capability of these scaffolds to promote the 
proliferation of bone cells. 
Acknowledgement 
We acknowledge Prof. W. Stark and Dr D. Mohn (ETH Zurich) for providing nBG powder. 
(a) 
(b) 
84   J.P. Cattalini et al. /  Procedia Engineering  59 ( 2013 )  78 – 84 
References 
[1] Mouriño V, Boccaccini AR. Bone tissue engineering therapeutics: controlled drug delivery in three-dimensional scaffolds. J R Soc Interface 
2010;7:209-227. 
[2] Gardin C, Ferroni L, Favero L, Stellini E, Stomaci D, Sivolella S, Bressan E, Zavan B. Nanostructured biomaterials for tissue engineered 
bone tissue reconstruction. Int J Mol Sci 2012;13:737-757.  
[3] Hoppe A, Güldal N, Boccaccini AR. A review of the biological response to ionic dissolution products from bioactive glasses and glass-
ceramics. Biomaterials 2011;32: 2757-2774.  
[4] Mouriño V, Newby P, Pishbin F, Cattalini JP, Lucangioli S, Boccaccini AR. Physicochemical, biological and drug-release properties of 
gallium crosslinked alginate/nanoparticulate bioactive glass composite films. Soft Matter 2011;7:6705-6712. 
[5] Gloria A, De Santis R, Ambrosio L. Polymer-based composite scaffolds for tissue engineering. J Appl Biomater Biomech 2010;8:57-67.  
[6] Bose S, Roy M, Bandyopadhyay A. Recent advances in bone tissue engineering scaffolds. Trends in Biotechnology 2012;30:546-554. 
[7] Mouriño V, Cattalini JP, Boccaccini AR. Metallic ions as therapeutic agents in tissue engineering scaffolds: an overview of their biological 
applications and strategies for new developments. J R Soc Interface 2012;9:401-419.  
[8] Habibovic P, Barralet, JE. Bioinorganics and biomaterials: Bone repair. Acta Biomater 2011;7:3013-3026. 
[9] Dorozhkin, S. Calcium orthophosphates as bioceramics: state of the art. J Funct Biomater 2010;1:22-107.  
[10] Cattalini JP, García J, Mouriño V, Lucangioli S. Development and validation of a capillary zone electrophoresis method for the 
determination of calcium in composite biomaterials. Curr Anal Chem 2013, CAC-EPUB-20121024-2. 


A New Calcium Releasing Nano-composite Biomaterial for Bone Tissue Engineering Scaffolds 

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A new calcium releasing nano-composite biomaterial for bone tissue engineering scaffolds

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