Hydroxyapatite and chloroapatite derived from sardine by-products

doi:10.1016/j.ceramint.2014.05.030

Ceramics International

Volume 40, Issue 8, Part B, September 2014, Pages 13231-13240

https://doi.org/10.1016/j.ceramint.2014.05.030 Get rights and content

Abstract

In this paper, phosphate-based compounds used in biomedicine were extracted from bones and scales of European sardines (Sardina pilchardus); this is the first time that different parts of the same fish are used for the extraction of these kinds of materials. The bones and scales behave very differently with processing, producing different materials when annealed between 600 and 1000 °C.

The bones formed a mixture of hydroxyapatite (Ca₁₀(PO₄)₆(OH)₂, HAp) and β-tri-calcium phosphate (β-Ca₃(PO₄)₂, β-TCP), with a higher content of β-TCP obtained with increasing temperature. This bi-phasic material has a high added value, as it is employed as a bioceramic; in fact HAp has good biocompatibility while β-TCP has better resorbability than HAp, despite being less biocompatible.

With scales, on the other hand, either a HAp-based material or a chlorine-substitute HAp containing material (chloroapatite (Ca₁₀(PO₄)₆Cl₂, ClAp) were produced. HAp-based material was obtained with a simple annealing process; for ClAp, on the other hand, a combined washing–annealing process was used. ClAp is also used in biomedicine, due to its improved resorption, mechanical properties and bioactivity. This is the first time ClAp of marine origin was produced.

Introduction

Calcium phosphate-based compounds are used in several technological applications. Many of these compounds show very high biocompatibility; because of this, they are employed as biomaterials, in particular to fabricate bone implants, bone cements and dental pastes [1], [2]. Their use in this field is mainly due to the compositional similarities that these compounds have with human bones and teeth.

Hydroxyapatite (Ca₁₀(PO₄)₆(OH)₂, HAp) is probably the most used compound in this field. It is the main component of human bones and, therefore, it is the species which best mimics its mineral composition and behaviour. Tri-calcium phosphate (Ca₃(PO₄)₂, TCP) is also employed in this field. It can exist in two possible forms, α and β, and normally the β form gets converted into the α with annealing at temperatures higher than 1250–1300 °C [3]. For many biomaterial applications, a mixture of HAp and TCP is often considered [4]; this is because TCP has better resorbability than HAp, despite being less biocompatible.

Apart from these applications in biomedicine, HAp has other uses too, especially for environmental remediation. HAp can be employed to remove heavy metals from contaminated waters and/or soils [5], [6]; moreover, some forms of HAp show photocatalytic activity and can be used to decompose organic contaminants [7], [8].

Chlorine-substituted HAp is also a compound with interesting potential. Chloroapatite (Ca₁₀(PO₄)₆Cl₂, ClAp), where hydroxyl groups are completely substituted by chlorine, can be used in electronics as a phosphor material, due to its fluorescence [9]. Compounds with only partial substitution, however, have been investigated for possible applications in biomedicine; the presence of some chlorine in the HAp lattice can actually improve its resorption, as well as mechanical properties [10]. Literature data also show that, in some cases, HAp with high chlorine substitution showed greater bioactivity than non-substituted HAp [11].

The majority of HAp used today is synthetic, prepared with various techniques [1]. However, HAp and other calcium phosphate materials can also be prepared from natural sources and/or wastes and by-products. The use of agri-food by-products, in particular, has attracted more and more interest in recent years. This is due to the production of increasing amounts of waste and/or by-products, which have to be disposed of with environmental impact. Extracting and/or obtaining compounds which have high value is, therefore, a way of addressing this problem while valorising such waste.

HAp can be obtained from by-products of the food industry, such as animal or fish bones; in fact, HAp is the main component of the bones themselves, with the remaining part being made of organic matter (i.e. collagen). Several studies have been published on this topic: bovine and pig bones, for instance, were used to extract HAp [12], [13], and several kinds of fish bones have also been used, including cod fish, swordfish and tuna [14], [15], [16], [17].

Fish scales, made of HAp and collagen, can also be employed to extract HAp. Literature reports exist of HAp extracted from fish scales for applications in both biomedical and environmental fields [18], [19], [20], [21], [22].

In this paper, we report a study on the use of wastes from European sardines (Sardina pilchardus) to extract phosphate-based compounds; both bones and scales were considered. It is the first time that different parts of the same fish were used for the extraction of this kind of compound. The materials obtained were characterised by several techniques (X-ray diffraction, thermogravimetry, IR spectroscopy and SEM micrography) to determine their characteristics, and address the possible differences according to the sources.

Section snippets

Sample storage and pre-annealing treatment

Sardine bones and scales were provided by A Poveira (Povoa de Varzim, Portugal). After collection, they were stored at −15 °C. Before treatment, the bones were defrosted and cleaned manually with hot water to remove impurities (i.e. fragments of meat, skin, etc.). They were then dried at 40 °C overnight. The scales were defrosted and dried at 40 °C overnight. For selected experiments, scales were soaked in water prior to, or after, the annealing, to remove sodium chloride. The process was

Annealing and characterisation of the bones

Fig. 1 shows the XRD patterns of the bones annealed at different temperatures between 600 and 1000 °C. It can be seen that a second phase as well as HAp is detected in all samples, which is β-TCP—the peaks corresponding to β-TCP are marked with a β in Fig. 1. The presence of this compound was previously reported in HAp samples of marine origin [15], [17].

The relative intensities of the peaks belonging to the two phases change depending on the annealing temperature; this indicates a change in the

Conclusions

Sardine bones and scales were successfully used to extract apatite- and calcium-phosphate-based materials; it is the first time that two parts of the same fish are used to extract compounds presenting high potential as biomaterial.

Results show that bones can be used to obtain materials in which HAp is the main component, through a calcination process. With scales, on the other hand, HAp-based material or ClAp-containing ones can be produced using a simple calcination and a combined

Acknowledgements

This work was funded through the ValorPeixe Project (QREN, AdI 13634). Authors also acknowledge PEst-C/CTM/LA0011/2013 and PEst-OE/EQB/LA0016/2013 programmes. C. Piccirillo thanks the FCT for Research Grant (SFRH/BPD/86483/2012), while R.C. Pullar wishes to thank the FCT Ciência2008 programme for supporting his work.

References (39)

S.V. Dorozhkin
Bioceramics of calcium orthophosphates
Biomaterials
(2010)
C. Piccirillo et al.
Bacteria immobilization on hydroxyapatite surface for heavy metals removal
J. Environ. Manag.
(2013)
H. Tanaka et al.
FTIR studies of adsorption and photocatalytic decomposition under UV irradiation of dimethyl sulfide on calcium hydroxyapatite
Adv. Powder Technol.
(2012)
S. Kannan et al.
Synthesis and mechanical behaviour of chloroapatite and chloroapatite/β-TCP composites
J. Eur. Ceram. Soc.
(2007)
N.A.M. Barakat et al.
Physiochemical characterization of hydroxyapatite extracted from bovine bones by three different methods: extraction of biologically desirable HAp
Mater. Sci. Eng. C
(2008)
M. Boutinguiza et al.
Biological hydroxyapatite obtained from fish bones
Mater. Sci. Eng. C
(2012)
M. Boutinguiza et al.
Hydroxyapatite nanoparticles obtained by fiber laser-induced fracture
Appl. Surf. Sci.
(2009)
C. Piccirillo et al.
Extraction and characterization of apatites-and calcium phosphate-based materials from cod fish bones
Mater. Sci. Eng. C
(2013)
S. Kongsri et al.
Nanocrystalline hydroxyapatite from fish scale waste: preparation, characterization and application for selenium adsorption in aqueous solution
Chem. Eng. J.
(2013)
Y.C. Huang et al.
Hydroxyapatite extracted from fish scale: effects on MG63 osteoblast-like cells
Ceram. Int.
(2011)

C. Piccirillo et al.

Bacteria immobilisation on hydroxyapatite surface for heavy metal removal

J. Env. Manag.

(2013)

H. Zhang et al.

Characterization and thermal behavior of calcium deficient hydroxyapatite whiskers with various Ca/P ratios

Mater. Chem. Phys.

(2011)

K. Haberko et al.

Natural hydroxyapatite – its behaviour during heat treatment

J. Eur. Ceram. Soc.

(2006)

V. Ferraro et al.

Extraction of high added value biological compounds from sardines, sardine-type fish and mackerel canning residues

Mater. Sci. Eng. C

(2013)

R. Chakraborty et al.

Application of calcined waste fish (Labeo rohita) scale as low-cost heterogenous catalyst for biodiesel synthesis

Bioresour. Technol.

(2011)

J. Oliva et al.

Biogenic hydroxyapatite (Apatite II^TM) dissolution kinetics and metal removal from acid mine drainage

J. Hazard. Mater.

(2012)

S.V. Dorozhkin

Calcium orthophosphates in dentistry

J. Mater. Sci.: Mater. Med.

(2013)

J. Peña et al.

Hydroxyapatite, tricalcium phosphate and biphasic materials prepared by a liquid mix technique

J. Eur. Ceram. Soc.

(2003)

S. Kannan et al.

Synthesis and thermal stability of hydroxyapatite-β-tricalcium phosphate composites with co-substituted sodium, magnesium and fluorine

Chem. Mater.

(2006)

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Hydroxyapatite and chloroapatite derived from sardine by-products

Abstract

Introduction

Section snippets

Sample storage and pre-annealing treatment

Annealing and characterisation of the bones

Conclusions

Acknowledgements

Biomaterials

J. Environ. Manag.

Adv. Powder Technol.

J. Eur. Ceram. Soc.

Mater. Sci. Eng. C

Mater. Sci. Eng. C

Appl. Surf. Sci.

Mater. Sci. Eng. C

Chem. Eng. J.

Ceram. Int.

J. Env. Manag.

Mater. Chem. Phys.

J. Eur. Ceram. Soc.

Mater. Sci. Eng. C

Bioresour. Technol.

J. Hazard. Mater.

Calcium orthophosphates in dentistry

J. Mater. Sci.: Mater. Med.

Hydroxyapatite, tricalcium phosphate and biphasic materials prepared by a liquid mix technique

J. Eur. Ceram. Soc.

Synthesis and thermal stability of hydroxyapatite-β-tricalcium phosphate composites with co-substituted sodium, magnesium and fluorine

Chem. Mater.