Connective Tissue Research

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Biomineralization: An Overview

Adele L. Boskey

To cite this article: Adele L. Boskey (2003) Biomineralization: An Overview, Connective Tissue
Research, 44:1, 5-9, DOI: 10.1080/03008200390152007
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Connective Tissue Research, 44(Suppl. 1): 5–9, 2003
Copyright °
c 2003 Taylor & Francis
0300-8207/03 $12.00 + .00
DOI: 10.1080/03008200390152007

Biomineralization: An Overview

Adele L. Boskey
Hospital for Special Surgery and Affiliated with Weill Medical College of Cornell University, New York,
New York, USA

COMMON CHARACTERISTICS
Biomineralization describes the deposition of mineral within or OF BIOMINERALIZATION IN DIFFERENT TAXA
outside the cells of living organisms. Examples include iron and The minerals depositing within the cells and upon the ex-
gold deposits in bacteria and other unicellular organisms, silicates
tracellular matrices in various organisms share certain common
in algae and diatoms, carbonates in diatoms and nonvertebrates,
and calcium phosphates and carbonates in vertebrates. The for- features, but serve a variety of functions including those related
mation of these deposits involves similar mechanisms, but there to locomotion, protection (shielding internal cell components or
are distinctions in each case, and those similarities and distinctions internal organs), sound reception, toxic waste disposal, tempo-
are the subject of this overview. A comparison of the apatitic min- rary ion storage, and magnetic orientation [3]. The first mineral
eral in bones and teeth is presented as a model for understanding
to deposit may be noncrystalline, and this amorphous mineral
these factors.
may then transform to more crystalline forms [2, 10]. The crys-
talline mineral phases are relatively insoluble and are generally
Keywords Biomineralization, Infrared Imaging, Mineralization
Mechanisms, Nonvertebrate Mineral. highly oriented.
Organic constituents within the cell or within the extracellu-
lar matrix regulate biomineralization. Some matrix constituents
protect the organism from being “choked” by excessive miner-
INTRODUCTION
alization (e.g., the mucins and soluble matrix proteins in echin-
Biomineralization is the process by which mineral deposits
oderms [11], the large aggregating proteoglycans in vertebrate
within or outside the cells of a variety of organisms. As reviewed
cartilage [12], and matrix gla protein in blood vessels and tra-
in detail elsewhere [1–3] these minerals include the magnetites
cheal cartilage [13]). In contrast, other matrix constituents pro-
and other iron containing deposits [4, 5], gold deposits within
mote mineralization. Two generalized types of structures seem
unicellular organisms [6], silicates in diatoms [7], and both cal-
to be most relevant: lipid membranes that provide protected en-
cium carbonates and calcium phosphates in nonvertebrate shells
vironments in which initial mineral forms [14–16] and proteins
[8–10]. It also includes the magnetities, calcium carbonates, and,
that serve as nucleators and regulators of crystal growth and ori-
more commonly, the calcium phosphate phases found in verte-
entation. Examples of these biomineral-associated proteins are
brate tissues. In all cases of biomineralization, the cell directs
found in Table 1.
the mineral formation process: by expression of proteins that act
Proteins associated with biomineralization have common fea-
as nucleators in the cell or in the extracellular matrix or which
tures. Their structures, as predicted from gene and amino acid
prevent mineral formation in unwanted sites: by production of
sequences, NMR, IR, and X-ray crystallography, are reviewed
enzymes that modify the functions of these proteins; and by the
elsewhere [17]. Most are anionic. These charged groups en-
regulation of ion transport. The proteins’ properties determine
able them to interact with highly charged mineral crystal sur-
their abilities to affect biomineralization and to interact with
faces [18]. Where secondary and tertiary structure information
other proteins, with cells, and with the biomineral.
is available for these proteins, many have been shown to have
structures that “match” one or more crystal face of the phase to
which they bind. The complementarity between crystal struc-
Received 9 November 2001; revised 1 February 2002; accepted 7 ture and protein structure is believed to result in regulation of
February 2002.
Address correspondence to Adele L. Boskey, PhD, Hospital for crystal growth and morphology as well as induction of crys-
Special Surgery, 535 E. 70th Street, New York, NY 10021, USA. E-mail: tal nucleation [17, 18]. Most of the proteins associated with
[email protected] biomineralization attain their anionic character by

5
6 A. L. BOSKEY

TABLE 1
Examples of proteins associated with biominerals.

Protein Mineral phase(s) Protein characteristics Function

Echinoderm spicules SM30 Calcium carbonate One of 40 distinct proteins Appears late during
(amorphous and in the spicule: lectin- spicule formation—
calcite) like and acidic [33]. Size regulation
SM50 Basic, nonglycosylated Appears early—Protects
[33, 34] space for spicule
formation
Mollusks Lustrin A Calcium carbonate Basic protein with Interacts with aragonite
(aragonite) modular structure [35] binding proteins—
Adhesive
Nacrein — Carbonic anhydrase Ion transport
activity [32]
Pearlins — Family of proteins which Size regulation
influence in vitro
crystallization [36]
N16 — Acidic domains, high Inhibits in vitro aragonite
proportion of glycine, formation in absence of
tyrosine, aspartate [37] water soluble matrix;
but promotes in its
presence
Vertebrate (shell) Ovocalyxin-32 Calcite Binds to calcite faces [38] Regulator
Vertebrate (dentin) Phosphophoryn Apatite Rich in polyasp and Nucleator and inhibitor
phosphoserine [25]
Vertebrate (bone) Bone sialoprotein Apatite Rich in polyglu [27, 39] Nucleator and inhibitor
Biglycan Apatite Low MW proteoglycan Nucleator
[40]

post-translational modifications, and their compositions are mod- COMPARATIVE PROPERTIES OF THE MINERAL
ulated by enzymes (kinases, phosphatases, proteinases) that exist OF VERTEBRATE BONES AND TEETH
within the cells or tissues in which they are expressed. This sug- The bulk of the mineral phase in bones and teeth is a poorly
gests that there may be some tissue specificity that determines crystalline carbonate-rich analogue of the naturally occurring
where and when nucleators and/or regulators of crystal growth mineral hydroxyapatite. However, the properties of the mineral
will appear. in bones and teeth are different. Even excluding enamel and
The proteins associated with biomineralization may have acellular cementum, which contain much larger apatite crystals
both direct and indirect effects on mineral deposition. Indirect with relatively higher amounts of A-type carbonate relative to
activities include the transport and/or sequestering of ions; inter- dentin, dentin mineral crystals are larger/more perfect than bone
action with other proteins leading to changes in the conforma- mineral crystals and have different stoichiometries and compo-
tion of the nucleating protein or the organization of the tissue; sitions [19]. This size difference between dentin and bone min-
and recruiting of cells that modulate the mineralization process. eral crystals was reported based on X-ray diffraction data of
The proteins directly influence biomineralization by a variety homogenized tissues in the 1960s [20] and recently has been
of mechanisms. They may bind ions involved in crystal for- demonstrated at ∼7 µm spatial resolution using infrared imag-
mation, providing a unique surface structure upon which addi- ing (Figure 1). Four factors must be examined to understand why
tional mineral is deposited; they may act directly as nucleators, these small but significant differences in mineral properties may
and/or they may bind to surfaces of the crystals determining exist: 1. Nature of the cells producing the mineralized matrix.
crystal proliferation and growth [17, 18]. The distribution of 2. Rate at which the matrix is formed. 3. Extent to which the tis-
these proteins, and the time of their expression, appears to be sue is remodeled after mineral is first deposited. 4. Nature and
critical in determining the overall structure of the mineral phases relative amounts of the mineral-associated proteins within each
deposited, as exemplified by the patterns in echinoderm shells tissue.
[9, 10, 13], and the mineral characteristics of bones and teeth A recent review examined the relative expression and distri-
[19]. bution of noncollagenous proteins in different types of bone and
 BIOMINERALIZATION: AN OVERVIEW 7

Figure 1–2 .
Figure 1. Comparative infrared analysis of the properties of a third trimester bovine molar. (a) Photomicrograph showing the 400 × 400 µm region analyzed
by infrared imaging indicating the regions from which the spectra in (d) were extracted. (b) Infrared image of the mineral crystallinity (as defined based on the
1030:1020 cm−1 intensity ratio [41]. For illustrative purposes, enamel (E) was assigned a value of 3 as there is no measurable 1020 cm−1 subband. (c) Infrared
image of the mineral-to-matrix ratio defined as the ratio of the integrated area of the phosphate (900–1200 cm−1 ) and amide I (1580–1720 cm−1 ) bands. All
enamel values exceeded 10. (d) Typical FTIR spectra extracted from the image data showing the difference in the shapes of the phosphate band in enamel (E),
dentin close to the predentin border (D1), and dentin close to the dentin-enamel junction (DEJ). Peaks from embedding media (PMMA), amide I, and phosphate
(PO4 ) are shown. PMMA does not penetrate into dentin. Each image is 400 × 400 µm. Images provided by Dr. Kostas Verdelis, Hospital for Special Surgery.
2. Infrared image of collagen maturity in same areas as shown in Figure 1. This parameter was calculated as the intensity ratio of bands at 1660 and 1690 cm−1
[24]. This parameter has a zero value in enamel (E). Image provided by Dr. Verdelis.
8 A. L. BOSKEY

TABLE 2
Effects of some bone and dentin phosphoproteins and proteoglycans on in vitro apatite formation and growth.

Protein In vitro effect Modification In vitro effect after modification

Phosphophoryn [25] Nucleator inhibitor Dephosphorylation cleavage No activity
DSP [28] Weak nucleators Dephosphorylation No effect
DMP-1∗ Inhibitor Dephosphorylation Nucleator
Osteopontin [26, 27] Inhibitor Dephosphorylation No activity
Increased phosphorylation Nucleator
BSP [31, 39] Nucleator inhibitor Dephosphorylation No activity
Aggrecan [12] Inhibitor Desulfation No activity
Digestion Loss of inhibition
Biglycan [40] Nucleator
Decorin [40] Inhibitor
DSP = dentin sialoprotein, BSP = bone sialoprotein, DMP-1 = dentin matrix protein,
∗
= unpublished data.

cementum [21]. The proteins within bone, cementum, and dentin mineral. Phosphophoryn [25], a major constituent of dentin, is
are similar, but distinct. Dentin has little osteopontin (OPN) and an effective nucleator, and it may be most relevant in determin-
osteocalcin (OC), but more dentin matrix protein-1 (DMP-1) ing the differences between dentin and bone mineral. Detailed
and DSSP (DMP-2) [22]. Bone contains more OPN, more bone analyses of the mineral produced when phosphophoryn is ex-
sialoprotein (BSP), more OCN, and distinct proteoglycans [23]. pressed in bone as opposed to dentin cells, and structural studies
BSP expression is highest before birth, implying a role in early of how each of these proteins interacts with apatite crystals, will
development, although the protein persists in mature bone [21]. be essential for providing these explanations.
The odontoblasts, which make the dentin matrix, are more po- It should be noted that most of these proteins are highly con-
larized than the osteoblasts, which make the osteoid of bone; served through a variety of vertebrate species, and some of the
and the collagen fibrils in predentin and newly formed dentin invertebrates have protein domains analogous to those in the ver-
have been reported to be thicker and more oriented than those tebrate mineralized tissues [32]. Thus, although the processes
in nonmineralized osteoid and newly formed bone. This is illus- of biomineralization are distinct in different species and tissue
trated in Figure 2, which shows an infrared image of the collagen types, there are clearly common threads.
maturity (related to the relative proportion of pyridinoline and
immature cross links [24]) in dentin and bone. This difference ACKNOWLEDGMENTS
in collagen orientation and maturation is probably associated Dr. Boskey’s work discussed in this overview was supported
both with the transport of collagen from the cell and with the by NIH grants DE04141, AR037661, AR41325, and AR46121
nature of the noncollagenous proteins that associate with the (Core Center for Musculoskeletal Integrity).
collagen. But which of these protein compositional differences
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