Small Charged Molecule-Mediated Fibrillar Mineralization: Implications for Ectopic Calcification.

Numerous small biomolecules exist in the human body and play roles in various biological and pathological processes. Small molecules are believed not to induce intrafibrillar mineralization alone. They are required to work in synergy with noncollagenous proteins (NCPs) and their analogs, e.g. polyelectrolytes, for inducing intrafibrillar mineralization, as the polymer-induced liquid-like precursor (PILP) process has been well-documented. In this study, we demonstrate that small charged molecules alone, such as sodium tripolyphosphate, sodium citrate, and (3-aminopropyl) triethoxysilane, could directly mediate fibrillar mineralization. We propose that small charged molecules might be immobilized in collagen fibrils to form the polyelectrolyte-like collagen complex (PLCC) via hydrogen bonds. The PLCC could attract CaP precursors along with calcium and phosphate ions for inducing mineralization without any polyelectrolyte additives. The small charged molecule-mediated mineralization process was evidenced by Cryo-TEM, AFM, SEM, FTIR, ICP-OES, etc., as the PLCC exhibited both characteristic features of collagen fibrils and polyelectrolyte with increased charges, hydrophilicity, and density. This might hint at one mechanism of pathological biomineralization, especially for understanding the ectopic calcification process.

In situ co-deposition synthesis for collagen-Astragalus polysaccharide composite with intrafibrillar mineralization as potential biomimetic-bone repair materials.

A hybrid material possessing both componential and structural imitation of bone tissue is the preferable composites for bone defect repair. Inspired by the microarchitecture of native bone, this work synthesized in vitro a functional mineralized collagen fibril (MCF) material by utilizing the method of in situ co-precipitation, which was designed to proceed in the presence of Astragalus polysaccharide (APS), thus achieving APS load within the biomineralized collagen-Astragalus polysaccharide (MCAPS) fibrils. Transmission electron microscope (TEM), selected area electron diffraction (SAED) and scanning electronic microscopy (SEM) identified the details of the intrafibrillar mineralization of the MCAPS fibrils, almost mimicking the secondary level of bone tissue microstructure. A relatively uniform and continuous mineral layer formed on and within all collagen fibrils and the mineral phase was identified as typical weak-crystalline hydroxyapatite (HA) with a Ca/P ratio of about 1.53. The proliferation of bone marrow-derived mesenchymal stem cells (BMSC) and mouse embryo osteoblast precursor cells (MC3T3-E1) obtained a significant promotion by MCAPS. As for the osteogenic properties of MCAPS, a distinct increase in the alkaline phosphatase (ALP) activity and the number of calcium nodules (CN) in BMSC and MC3T3-E1 was detected. The up-regulation of three osteogenic-related genes of RUNX-2, BMP-2 and OCN were confirmed via reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to further verify the osteogenic performance promotion of MCAPS. A period of 14 days of culture demonstrated that MCAPS-L exhibited a preferable efficacy in enhancing ALP activity and CN quantity, as well as in promoting the expression of osteogenic-related genes over MCAPS-M and MCAPS-H, indicating that a lower dose of APS within the material of MCAPS is more appropriate for its osteogenesis promotion properties.

STMP and PVPA as Templating Analogs of Noncollagenous Proteins Induce Intrafibrillar Mineralization of Type I Collagen via PCCP Process.

The phosphorylated noncollagenous proteins (NCPs) play a vital role in manipulating biomineralization, while the mechanism of phosphorylation of NCPs in intrafibrillar mineralization of collagen fibril has not been completely deciphered. Poly(vinylphosphonic acid) (PVPA) and sodium trimetaphosphate (STMP) as templating analogs of NCPs induce hierarchical mineralization in cooperation with indispensable sequestration analogs such as polyacrylic acid (PAA) via polymer-induced liquid-like precursor (PILP) process. Herein, STMP-Ca and PVPA-Ca complexes are proposed to achieve rapid intrafibrillar mineralization through polyelectrolyte-Ca complexes pre-precursor (PCCP) process. This strategy is further verified effectively for remineralization of demineralized dentin matrix both in vitro and in vivo. Although STMP micromolecule fails to stabilize amorphous calcium phosphate (ACP) precursor, STMP-Ca complexes facilely permeate into intrafibrillar interstices and trigger phase transition of ACP to hydroxyapatite within collagen. In contrast, PVPA-stabilized ACP precursors lack liquid-like characteristic and crystallize outside collagen due to rigid conformation of PVPA macromolecule, while PVPA-Ca complexes infiltrate into partial intrafibrillar intervals under electrostatic attraction and osmotic pressure as evidenced by intuitionistic 3D stochastic optical reconstruction microscopy (3D-STORM). The study not only extends the variety and size range of polyelectrolyte for PCCP process but also sheds light on the role of phosphorylation for NCPs in biomineralization.

Polyelectrolyte-Cation Complexes Using PAsp-Sr Complexes Induce Biomimetic Mineralization with Antibacterial Ability.

Remineralized dentin with an antibacterial ability is still a significant challenge in dentistry. Previously, a polyelectrolyte-calcium complexes pre-precursor (PCCP) process is proposed for rapid collagen mineralization. In the present study, the expansion concept of the PCCP process is explored by replacing the calcium with other cations, such as strontium. The results of transmission electron microscopy (TEM), 3D stochastic optical reconstruction microscopy, energy-dispersive X-ray analysis, Fourier transform infrared spectroscopy, and high-resolution TEM with selected area electron diffraction demonstrate that biomimetic mineralization of collagen fibrils and demineralized dentin could be fulfilled with Sr&F-codoped hydroxyapatite (HAp) after they are treated with poly-aspartic acid-strontium (PAsp-Sr) suspension followed by a phosphate&fluoride solution. Moreover, dentin remineralized with Sr&F-codoped HAp exhibits in vitro and in vivo antibacterial ability against Streptococcus mutans. The cytotoxicity and oral mucosa irritation tests reveal excellent biocompatibility of mineralization mediums (PAsp-Sr suspension and phosphate&fluoride solution). The demineralized dentin's mechanical properties (elastic modulus and microhardness) could be restored almost to that of the intact dentin. Hence, the expansion concept of the PCCP process that replaces calcium ions with some cationic ions along with fluorine opens up new horizons for generating antibacterial remineralized dentin containing ions-doped HAp with excellent biocompatibility via biomimetic mineralization technology.

Biomimetic Intrafibrillar Mineralization of Native Tendon for Soft-Hard Interface Integration by Infiltration of Amorphous Calcium Phosphate Precursors.

Soft and hard tissues possess distinct biological properties. Integrating the soft-hard interface is difficult due to the inherent non-osteogenesis of soft tissue, especially of anterior cruciate ligament and rotator cuff reconstruction. This property makes it difficult for tendons to be mineralized and integrated with bone in vivo. To overcome this challenge, a biomimetic mineralization strategy is employed to engineer mineralized tendons. The strategy involved infiltrating amorphous calcium phosphate precursors into collagen fibrils, resulting in hydroxyapatite deposition along the c-axis. The mineralized tendon presented characteristics similar to bone tissue and induced osteogenic differentiation of mesenchymal stem cells. Additionally, the interface between the newly formed bone and tendon is serrated, suggesting a superb integration between the two tissues. This strategy allows for biomineralization of tendon collagen and replicating the hallmarks of the bone matrix and extracellular niche, including nanostructure and inherent osteoinductive properties, ultimately facilitating the integration of soft and hard tissues.

Hyaluronic acid-mediated collagen intrafibrillar mineralization and enhancement of dentin remineralization.

Non-collagenous proteins (NCPs) in the extracellular matrix (ECM) of bone and dentin are known to play a critical regulatory role in the induction of collagen fibril mineralization and are embedded in hyaluronic acid (HA), which acts as a water-retaining glycosaminoglycan and provides necessary biochemical and biomechanical cues. Our previous study demonstrated that HA could regulate the mineralization degree and mechanical properties of collagen fibrils, yet its kinetics dynamic mechanism on mineralization is under debate. Here, we further investigated the role of HA on collagen fibril mineralization and the possible mechanism. The HA modification can significantly promote intrafibrillar collagen mineralization by reducing the electronegativity of the collagen surface to enhance calcium ions (Ca2+) binding capacity to create a local higher supersaturation. In addition, the HA also provides additional nucleation sites and shortens the induction time of amorphous calcium phosphate (ACP)-mediated hydroxyapatite (HAP) crystallization, which benefits mineralization. The acceleration effect of HA on intrafibrillar collagen mineralization is also confirmed in collagen hydrogel and in vitro dentin remineralization. These findings offer a physicochemical view of the regulation effect of carbohydrate polymers in the body on biomineralization, the fine prospect for an ideal biomaterial to repair collagen-mineralized tissues.

A seminal perspective on the role of chondroitin sulfate in biomineralization.

Chondroitin sulfate (CS) is an important extracellular matrix component of mineralized tissues. It participates in biomineralization, osteoblast differentiation and promotes bone tissue repair in vitro. However, the mechanism in which CS functions is unclear. Accordingly, an in-depth investigation of how CS participates in mineralization was conducted in the present study. Chondroitin sulfate was found to directly induce intrafibrillar mineralization of the collagen matrix. The mineralization outcome was dependent on whether CS remained free in the extracellular matrix or bound to core proteins; mineralization only occurred when CS existed in a free state. The efficacy of mineralization appeared to increase with ascending CS concentration. This discovery spurred the authors to identify the cause of heterotopic ossification in the Achilles tendon. Chondroitin sulfate appeared to be a therapeutic target for the management of diseases associated with heterotopic calcification. A broader perspective was presented on the applications of CS in tissue engineering.

Intrafibrillar Mineralization and Immunomodulatory for Synergetic Enhancement of Bone Regeneration via Calcium Phosphate Nanocluster Scaffold.

Inspired by the bionic mineralization theory, organic-inorganic composites with hydroxyapatite nanorods orderly arranged along collagen fibrils have attracted extensive attention. Planted with an ideal bone scaffold will contribute greatly to the osteogenic microenvironment; however, it remains challenging to develop a biomimetic scaffold with the ability to promote intrafibrillar mineralization and simultaneous regulation of immune microenvironment in situ. To overcome these challenges, a scaffold containing ultra-small particle size calcium phosphate nanocluster (UsCCP) is prepared, which can enhance bone regeneration through the synergetic effect of intrafibrillar mineralization and immunomodulatory. By efficient infiltration into collagen fibrils, the UsCCP released from the scaffold achieves intrafibrillar mineralization. It also promotes the M2-type polarization of macrophages, leading to an immune microenvironment with both osteogenic and angiogenic potential. The results confirm that the UsCCP scaffold has both intrafibrillar mineralization and immunomodulatory effects, making it a promising candidate for bone regeneration.

Calcium Phosphate Delivery Systems for Regeneration and Biomineralization of Mineralized Tissues of the Craniofacial Complex.

Calcium phosphate (CaP)-based materials have been extensively used for mineralized tissues in the craniofacial complex. Owing to their excellent biocompatibility, biodegradability, and inherent osteoconductive nature, their use as delivery systems for drugs and bioactive factors has several advantages. Of the three mineralized tissues in the craniofacial complex (bone, dentin, and enamel), only bone and dentin have some regenerative properties that can diminish due to disease and severe injuries. Therefore, targeting these regenerative tissues with CaP delivery systems carrying relevant drugs, morphogenic factors, and ions is imperative to improve tissue health in the mineralized tissue engineering field. In this review, the use of CaP-based microparticles, nanoparticles, and polymer-induced liquid precursor (PILPs) amorphous CaP nanodroplets for delivery to craniofacial bone and dentin are discussed. The use of these various form factors to obtain either a high local concentration of cargo at the macroscale and/or to deliver cargos precisely to nanoscale structures is also described. Finally, perspectives on the field using these CaP materials and next steps for the future delivery to the craniofacial complex are presented.

Promotion Effect of Carboxymethyl Chitosan on Dental Caries via Intrafibrillar Mineralization of Collagen and Dentin Remineralization.

OBJECTIVE: To observe ultrastructural changes during the process of carboxymethyl chitosan (CMC)-mediated intrafibrillar mineralization, we evaluated the biomimetic remineralization potential of CMC in type-I collagen fibrils and membranes, and further explored the bond strength as well as the bond interfacial integrity of the biomimetic remineralized artificial caries-affected dentin (ACAD). METHODS: A mineralized solution containing 200 µg/mL CMC was used to induce type-I collagen biomimetic remineralization in ACAD, while traditional mineralization without CMC was used as a control. The process and pattern of mineralization were investigated by transmission electron microscopy (TEM), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS) as well as structured illumination microscopy (SIM). The Vickers hardness test was used to quantify the dentin hardness, while the microtensile bond strength (µTBS) test was used to assess the bond strength and durability. The bond interfacial integrity was evaluated by a confocal laser scanning microscope (CLSM). RESULTS: TEM, SEM, and SIM images showed that CMC had a positive effect on stabilizing amorphous calcium phosphate (ACP) and promoting intrafibrillar mineralization, while extrafibrillar mineralization was formed without CMC. Furthermore, hardness evaluation and µTBS proved that CMC significantly increased dentin hardness and bond strength. CLSM indicated that CMC could create a significantly better bond interfacial integrity with less of a micro-gap in ACAD. SIGNIFICANCE: CMC possessed the ability to promote intrafibrillar mineralization and remineralization in demineralized caries dentin lesions, as well as improve bond performance, which implied its potential in carious dentin demineralization or dentin hypersensitivity and possibly even as a possible material for indirect pulp-capping, to deal with deep caries. HIGHLIGHTS: CMC possessed the ability to induce intrafibrillar mineralization effectively; the bond strength and bond durability of demineralized caries dentin were improved via CMC-induced remineralization; the CMC-induced remineralization complex is a potential material for indirect pulp-capping, to deal with deep caries.

Comparison of Synthetic vs. Biogenic Polymeric Process-Directing Agents for Intrafibrillar Mineralization of Collagen.

With the aging population, there is a growing need for mineralized tissue restoration and synthetic bone substitutes. Previous studies have shown that a polymer-induced liquid-precursor (PILP) process can successfully mineralize collagen substrates to achieve compositions found in native bone and dentin. This process also leads to intrafibrillar apatitic crystals with their [001] axes aligned roughly parallel to the long axis of the collagen fibril, emulating the nanostructural organization found in native bone and dentin. When demineralized bovine bone was remineralized via the PILP process using osteopontin (OPN), the samples were able to activate mouse marrow-derived osteoclasts to similar levels to those of native bone, suggesting a means for fabricating bioactive bone substitutes that could trigger remodeling through the native bone multicellular unit (BMU). In order to determine if OPN derived from bovine milk could be a cost-effective process-directing agent, the mineralization of type I collagen scaffolds using this protein was compared to the benchmark polypeptide of polyaspartic acid (sodium salt; pAsp). In this set of experiments, we found that OPN led to much faster and more uniform mineralization when compared with pAsp, making it a cheaper and commercially attractive alternative for mineralized tissue restorations.

A Dopamine Acrylamide Molecule for Promoting Collagen Biomimetic Mineralization and Regulating Crystal Growth Direction.

The reconstruction of the intra/interfibrillar mineralized collagen microstructure is extremely important in biomaterial science and regeneration medicine. However, certain problems, such as low efficiency and long period of mineralization, are apparent, and the mechanism of interfibrillar mineralization is often neglected in the present literature. Thus, we propose a novel model of biomimetic collagen mineralization that uses molecules with the dual function of cross-linking collagen and regulating collagen mineralization to construct the intrafibrillar and interfibrillar collagen mineralization of the structure of mineralized collagen hard tissues. In the present study completed in vitro, N-2-(3,4-dihydroxyphenyl) acrylamide (DAA) is used to bind and cross-link collagen molecules and further stabilize the self-assembled collagen fibers. The DAA-collagen complex provides more affinity with calcium and phosphate ions, which can reduce the calcium phosphate/collagen interfacial energy to promote hydroxyapatite (HA) nucleation and accelerate the rate of collagen fiber mineralization. Besides inducing intrafibrillar mineralization, the DAA-collagen complex mineralization template can realize interfibrillar mineralization with the c-axis of the HA crystal on the surface of collagen fibers and between fibers that are parallel to the long axis of collagen fibers. The DAA-collagen complex, as a new type of mineralization template, may provide a new collagen mineralization strategy to produce a mineralized scaffold material for tissue engineering or develop bone-like materials.

Effects of bioactive glass with high phosphorus content on mineralization of type I collagen fibrils.

PURPOSE: To study effects of bioactive glass with high phosphorus content (10.8% P2O5, 54.2% SiO2, 35% CaO, mol%, named PSC) on mineralization of type I collagen fibrils. METHODS: (1) PSC, and PSC combining 0.1 mg/mL, 0.5 mg/mL, or 1.0 mg/mL polyacrylic acid (PAA), were used to induce the mineralization of self-assembled type I collagen fibrils. After 3 and 7 days of mineralization, collagen fibrils were observed by transmission electron microscopy (TEM) and selected area electron diffraction (SAED). (2) PSC suspension was dialyzed in simulated body fluid (SBF), or in SBF containing 0.1 mg/mL, 0.5 mg/mL, or 1.0 mg/mL PAA, to form amorphous calcium phosphate (ACP), then observed by TEM. RESULTS: (1) PSC and PSC combining 0.1 mg/mL or 0.5 mg/mL PAA induced mainly extrafibrillar mineralization. PSC combining 1.0 mg/mL PAA induced both extrafibrillar and intrafibrillar mineralization. (2) The ACP induced by PSC or PSC combining 0.1 mg/mL PAA partly formed lattice structure after 24 h. The particle size of the ACP induced by PSC combining 0.5 mg/mL PAA was 100-150 nm, and that induced by PSC combining 1.0 mg/mL PAA was 30-50 nm. CONCLUSION: PSC induced mainly extrafibrillar mineralization, and PSC combining an appropriate concentration (1.0 mg/mL) of PAA induced both extrafibrillar and intrafibrillar mineralization.

Involvement of prenucleation clusters in calcium phosphate mineralization of collagen.

Involvement of thermodynamically-stable prenucleation clusters (PNCs) in the biomineralization of collagen has been speculated since their existence was reported in mineralization systems. It has been hypothesized that intrafibrillar mineralization proceeds via nucleation of inhibitor-stabilized intermediates produced by liquid-liquid separation (aka. polymer-induced liquid precursors; PILPs). Here, the contribution of PNCs and PILPs to calcium phosphate intrafibrillar mineralization of collagen was examined in a model with a semipermeable membrane that excludes nucleation inhibitor-stabilized PILPs from reaching the collagen fibrils, using cryogenic electron microscopy of reconstituted fibrils and conventional transmission electron microscopy of collagen sponges. Molecular dynamics simulation with the Interface force field (IFF) was used to confirm the existence of PILPs with amorphous calcium phosphate and elucidate details of the dynamics. Furthermore, intrafibrillar mineralization of single collagen fibrils was experimentally observed with unstabilized PNCs when anionic/cationic polyelectrolytes were used to establish Donnan equilibrium across the semipermeable membrane. Molecular dynamics simulation verified PNC formation within the collagen intrafibrillar gap zones at the atomic scale and explained the role of external PILPs. The PILPs decrease the interfibrillar water content and increase the interfibrillar ionic concentration. Nevertheless, intrafibrillar mineralization of collagen sponges with PNCs alone was inefficacious, being constrained by competition from extrafibrillar mineral precipitation. STATEMENT OF SIGNIFICANCE: Compared with conventional PILP-based intrafibrillar mineralization, mineralization of collagen fibrils using unstabilized PNCs is constrained by competition from extrafibrillar mineral deposition. The narrow window of opportunity for PNCs to produce intrafibrillar mineralization provides a plausible explanation for the feasibility of nucleation inhibitor-free intrafibrillar apatite assembly during reconstitution of type I collagen.

Three-dimensional structural interrelations between cells, extracellular matrix, and mineral in normally mineralizing avian leg tendon.

The spatial-temporal relationship between cells, extracellular matrices, and mineral deposits is fundamental for an improved understanding of mineralization mechanisms in vertebrate tissues. By utilizing focused ion beam-scanning electron microscopy with serial surface imaging, normally mineralizing avian tendons have been studied with nanometer resolution in three dimensions with volumes exceeding tens of micrometers in range. These parameters are necessary to yield sufficiently fine ultrastructural details while providing a comprehensive overview of the interrelationships between the tissue structural constituents. Investigation reveals a complex lacuno-canalicular network in highly mineralized tendon regions, where ∼100 nm diameter canaliculi emanating from cell (tenocyte) lacunae surround extracellular collagen fibril bundles. Canaliculi are linked to smaller channels of ∼40 nm diameter, occupying spaces between fibrils. Close to the tendon mineralization front, calcium-rich deposits appear between the fibrils and, with time, mineral propagates along and within them. These close associations between tenocytes, tenocyte lacunae, canaliculi, small channels, collagen, and mineral suggest a concept for the mineralization process, where ions and/or mineral precursors may be transported through spaces between fibrils before they crystallize along the surface of and within the fibrils.

Apatite minerals derived from collagen phosphorylation modification induce the hierarchical intrafibrillar mineralization of collagen fibers.

Collagen is the critical organic component of bone matrix, which is the template for bone biomineralization. Phosphorylation modification of collagen plays an important role in the process of biomineralization in vivo, but its mechanism on in vitro biomimetic mineralization of bone matrix remains unclear at the molecular level. Sodium tripolyphosphate is used to phosphorylate collagen in this study and new phosphate groups appear on collagen fibrils after phosphorylation modification. The chelating amount of calcium is improved linearly with increasing the phosphorylation degree of collagen fibrils, which demonstrates that the introduced phosphate groups serve as new nucleation sites and participate in the formation of apatite minerals inside the collagen fibers. Stabilized nanosized amorphous calcium phosphate by polyacrylic acid can also permeate into collagen fibers and further transform into another layer of hydroxyapatite minerals. Both layers of apatite minerals eventually induce the formation of hierarchical intrafibrillar mineralization structure within the phosphorylated collagen fibers. The present research enriches the previous biomineralization mechanism of bone matrix, provides a facile strategy for biomimetic mineralization of collagen, and offers the basis for future investigation of the advanced bone substitute materials.

Biomineralization-Inspired Material Design for Bone Regeneration.

Synthetic substitutes of bone grafts, such as calcium phosphate-based ceramics, have shown some good clinical successes in the regeneration of large bone defects and are currently extensively used. In the past decade, the field of biomineralization has delivered important new fundamental knowledge and techniques to better understand this fascinating phenomenon. This knowledge is also applied in the field of biomaterials, with the aim of bringing the composition and structure, and hence the performance, of synthetic bone graft substitutes even closer to those of the extracellular matrix of bone. The purpose of this progress report is to critically review advances in mimicking the extracellular matrix of bone as a strategy for development of new materials for bone regeneration. Lab-made biomimicking or bioinspired materials are discussed against the background of the natural extracellular matrix, starting from basic organic and inorganic components, and progressing into the building block of bone, the mineralized collagen fibril, and finally larger, 2D and 3D constructs. Moreover, bioactivity studies on state-of-the-art biomimicking materials are discussed. By addressing these different topics, an overview is given of how far the field has advanced toward a true bone-mimicking material, and some suggestions are offered for bridging current knowledge and technical gaps.

Using biomimetically mineralized collagen membranes with different surface stiffness to guide regeneration of bone defects.

Because guided bone regeneration (GBR) process is pronouncedly affected by the micro-environment in the defect, the surface stiffness of collagen membranes as a constituent part of the micro-environment was investigated in this study. The objective of this study was to manufacture biomimetically mineralized collagen membranes with controllable surface stiffness based on biomimetic strategy and to investigate the influences of surface stiffness on GBR process. The characterization and biocompatibility of membranes were examined in vitro. The mechanical properties of membranes were evaluated on macro and micro levels using tensile test and atomic force microscope, respectively. The critical-size cranial defect model and ectopic osteogenesis were chosen to employ their performances in vivo. The results indicated that the biomimetically mineralized collagen membranes with controllable surface stiffness were manufactured based on the biomimetic theory. The in vitro experiments showed that the mineralized collagen membrane with satisfactory surface stiffness can better promote the adhesion, proliferation, and osteogenic differentiation of mesenchymal stem cells. The membranes can perform excellently in both osteoinduction and osteoconduction, which results in effective manifestations in aspects of ectopic osteogenesis and GBR in vivo. Therefore, this biomimetically mineralized collagen membrane is a promising candidate for GBR treatment in future.

Review of the mechanism of biomimetic intrafibrillar mineralization / 口腔疾病防治

@#Bone is a hierarchically structured and highly mineralized hard tissue composed of an organic phase (type I collagen and noncollagenous proteins) and an inorganic phase (nanohydroxyapatite). Intrafibrillar mineralized collagen is the basic structural unit of bone tissue and is of high significance due to its superior mechanical and biological properties. Thus, to truly understand the unique properties of bone, it is necessary to review the most basic structural level of bone. In this article, we review the recent advances in understanding the development of intrafibrillar mineralization and the prevailing theories in the formation of such intrafibrillar minerals. Understanding the mechanisms of intrafibrillar mineralization may facilitate the development of engineered bone for clinical applications and provide deeper insight into the nature of biomineralization.

Hierarchically Staggered Nanostructure of Mineralized Collagen as a Bone-Grafting Scaffold.

A hierarchical, intrafibrillarly mineralized collagen (HIMC) is achieved through a selective mineralization progress in the collagenous gap regions mediated by poly(acrylic acid) with appropriate molecular weight. The associated topographical features directly correlate with nanomechanical heterogeneities of the HIMC to accommodate a broad range of external loads. Moreover, this hierarchically staggered nanostructure provides an optimized microenvironment to improve bone regeneration by instructing host cells.