Biomimetic synthesis of enamel-like hydroxyapatite on self-assembled monolayers

Hydroxyapatite (HAp) crystals mimicking tooth enamel in chemical composition and morphology were formed on sulfonic-terminated self-assembled monolayer (SAM) in 1.5SBF with F⁻ at 50 °C for 7 days. F⁻ ions showed a marked effect on the composition and morphology of deposited HAp crystals. In the absence of F⁻ ions, HAp containing CO₃²⁻ were formed on SAM, and worm-like crystals of 200–300 nm in length aggregated to form a spherical morphology. When F⁻ was added, HAp crystals containing both CO₃²⁻ and F⁻ were formed on SAM. Needle-shaped crystals of high aspect ratio and 1–2 μm in length grew elongated along the c-axial direction. In addition, these needle-shaped crystals grew in bundles, mimicking HAp crystals in tooth enamel. After the process of ripening, the needles in bundle grew to large size of up to 10 μm in length, and still kept no crystal–crystal fusion like enamel HAp crystals. The formation of enamel-like HAp can be attributed to the substitute of F⁻ for OH⁻ by disturbing the normal progress of HAp formation on SAM. The results suggest potential applications in preparing a novel dental material by a simple method.     

Synthesis and ultrastructure of plate-like apatite single crystals as a model for tooth enamel

Hydroxyapatite (HAp) is an inorganic constituent compound of human bones and teeth, with superior biocompatibility and bioactivity characteristics. Its crystal structure is hexagonal, characterized by a(b)- and c-planes. In vertebrate long bones, HAp crystals have a c-axis orientation, while in tooth enamel, they have an a(b)-axis orientation. Many methods can be used to synthesize c-axis oriented HAp single crystals; however, to the best of our knowledge, there have been no reports on a synthesis method for a(b)-axis oriented HAp single crystals. In this study, we successfully synthesized plate-like HAp crystals at the air–liquid interface of a starting solution via an enzyme reaction of urea with urease. Crystal phase analysis and ultrastructure observations were carried out, and the results indicated that the particles were single crystals, with almost the same a(b)-axis orientation as tooth enamel. It is hoped that by utilizing their unique surface charge and atomic arrangement, the resulting particles can be used as a high-performance biomaterial, capable of adsorbing bio-related substances and a model for tooth enamel.     

氨基酸/羟基磷灰石复合材料应用于酸蚀牛牙釉质体外再矿化

早期牙釉质龋病的再矿化在龋齿的预防和修复中起着至关重要的作用。基于牙釉质基质中所含的主要氨基酸, 在10 mmol/L甘氨酸、10 mmol/L L-丝氨酸和5 mmol/L L-天冬氨酸的存在下制备了氨基酸/羟基磷灰石(AA/HAP)复合材料, 表征了其物理、化学和生物学性质, 并评估了其对酸蚀牛牙釉质的再矿化作用。与不含氨基酸的羟基磷灰石(HAP)相比, 在氨基酸的抑制作用下, AA/HAP复合材料具有更低的结晶度和更高的生物相容性。在人工唾液中用AA/HAP对酸蚀牛牙釉质进行体外再矿化。再矿化后, 分别表征了牛牙釉质样品的表面和横截面形态、成分和力学性能。结果表明, AA/HAP可以诱导表面和深层牙釉质病变的修复。复合材料中释放的氨基酸可以吸附在有机基质残基上并诱导平行排列的HAP晶体的形成, 从而使牙釉质表面显微硬度(SMH)得到显著恢复。最后, 讨论了AA/HAP复合材料对酸蚀牛牙釉质的再矿化机制。     

Unidirectionally oriented hydroxyapatite/collagen composite fabricated by using a high magnetic field

A unidirectionally oriented hydroxyapatite (HAp)/collagen composite was prepared by using a high magnetic field. In the present study, a calcium-containing collagen solution was mixed with a phosphate-containing solution and kept at a temperature of 37 °C. During this process, the heat-induced self-assembling of collagen molecules and precipitation of HAp crystals took place simultaneously. Both collagen fibrils and HAp crystals have magnetic susceptibility anisotropy, which makes it possible for them to be oriented by using a high magnetic field. In order to prepare a unidirectionally oriented structure, samples were rotated when the high magnetic field was imposed perpendicularly to the rotation axis. The results indicated that collagen fibrils have been unidirectionally oriented with weakly crystallized HAp nanocrystals closely attached along the surfaces of the collagen fibrils. A biocomposite with similar structural and composition to natural bones has been prepared.     

Histological structures and acidic etching sensitivities of the enamels at the occlusal pit parts in the deciduous and permanent teeth

The purpose of this study is to compare the histological structures and acidic etching sensitivities of the enamels at the occlusal pit parts between the deciduous molars and permanent molars. They were observed by the polarizing and scanning electron microscopies. The enamel rods were less made slender by EDTA etching and the outlines of the apatite crystals, constituting the enamel rods, were clearer at the occlusal pit part of the deciduous molar than that of the permanent molar in reverse of that at the cusp part. It is thought that the enamel at the occlusal pit part of the permanent molar is more easily decayed by the dental caries than that of the deciduous molar because the former is more easily decayed by the acidic etching than the latter in reverse at the cusp part. It is considered that the thin superficialmost layer of the enamel at the occlusal pit part of the permanent molar has originally higher degree of resistance to the dental caries.     

In situ X-ray scattering evaluation of heat-induced ultrastructural changes in dental tissues and synthetic hydroxyapatite

Human dental tissues consist of inorganic constituents (mainly crystallites of hydroxyapatite, HAp) and organic matrix. In addition, synthetic HAp powders are frequently used in medical and chemical applications. Insights into the ultrastructural alterations of skeletal hard tissues exposed to thermal treatment are crucial for the estimation of temperature of exposure in forensic and archaeological studies. However, at present, only limited data exist on the heat-induced structural alterations of human dental tissues. In this paper, advanced non-destructive small- and wide angle X-ray scattering (SAXS/WAXS) synchrotron techniques were used to investigate the in situ ultrastructural alterations in thermally treated human dental tissues and synthetic HAp powders. The crystallographic properties were probed by WAXS, whereas HAp grain size distribution changes were evaluated by SAXS. The results demonstrate the important role of the organic matrix that binds together the HAp crystallites in responding to heat exposure. This is highlighted by the difference in the thermal behaviour between human dental tissues and synthetic HAp powders. The X-ray analysis results are supported by thermogravimetric analysis. The results concerning the HAp crystalline architecture in natural and synthetic HAp powders provide a reliable basis for deducing the heating history for dental tissues in the forensic and archaeological context, and the foundation for further development and optimization of biomimetic material design.     

Remineralization of initial enamel caries in vitro using a novel peptide based on amelogenin

Dental caries is the most common oral disease with high incidence, widely spread and can seriously affect the health of oral cavity and the whole body. Current caries prevention measures such as fluoride treatment, antimicrobial agents, and traditional Chinese herbal, have limitations to some extent. Here we design and synthesize a novel peptide based on the amelogenin, and assess its ability to promote the remineralization of initial enamel caries lesions. We used enamel blocks to form initial lesions, and then subjected to 12-day pH cycling in the presence of peptide, NaF and HEPES buffer. Enamel treated with peptide or NaF had shallower, narrower lesions, thicker remineralized surfaces and less mineral loss than enamel treated with HEPES. This peptide can promote the remineralization of initial enamel caries and inhibit the progress of caries. It is a promising anti-caries agent with various research prospects and practical application value.     

Long-Wave Infrared Thermophotonic Imaging of Demineralization in Dental Hard Tissue

Dental caries remains the most prevalent chronic disease in both children and adults worldwide. To address this prevalence through disease prevention and management, dentists need tools capable of detecting caries at early stages of formation. Looking into the physics of light propagation in teeth, this study presents a clinically and commercially viable platform technology for thermophotonic detection of early dental caries using an inexpensive long-wavelength infrared (LWIR; 8 \(\upmu \)m to 14 \(\upmu \)m) camera. The developed system incorporates intensity-modulated light to generate a thermal-wave field inside enamel and uses the subsequent infrared emission of the thermal-wave field to detect early caries. It was found that the greater light absorption at caries sites shifts the thermal-wave field centroid, providing contrast between early caries and intact enamel. Use of LWIR detection band in dental samples is novel and beneficial over the conventional mid-wavelength infrared band (3 \(\upmu \)m to 5 \(\upmu \)m) as it suppresses the masking effect of the instantaneous radiative emission from subsurface features due to the minimal transmittance of enamel in the LWIR band. The efficacy of the LWIR system is verified though experiments carried out on nonbiological test samples as well as on teeth with natural and artificially induced caries. The results suggest that the developed LWIR technology is an affordable early dental caries detection system suitable for commercialization/translation to Dentistry.     

人体牙齿的显微组织及纳米力学性能

采用光学显微镜、能谱扫描电镜、高分辨透射电镜以及纳米力学探针等设备对人体牙齿的显微组织及纳米力学性能进行了研究。结果表明: 牙本质由规则排列的牙本质小管和基质组成,牙釉质由不同方向的釉柱及柱间质组成,牙本质与牙釉质之间有明显的界面;牙釉质中的Ca和P含量明显高于牙本质中的含量,而C含量明显低于牙本质中的含量。牙釉质中发现有大量的磷灰石晶体结构,而牙本质中则明显缺少。力学性能结果显示,牙釉质的纳米硬度平均为4.4 GPa,弹性模量为81 GPa,而牙本质的纳米硬度平均为1.0 GPa,弹性模量为26 GPa。牙釉质高的硬度和弹性模量与含有大量的羟基磷灰石晶体组织有关,Ca和P促进了该晶体组织形成。      

Microstructure and nanomechanical properties of enamel remineralized with asparagine–serine–serine peptide

A highly biocompatible peptide, triplet repeats of asparagine–serine–serine (3NSS) was designed to regulate mineral deposition from aqueous ions in saliva for the reconstruction of enamel lesions. Healthy human enamel was sectioned and acid demineralized to create lesions, then exposed to the 3NSS peptide solution, and finally immersed in artificial saliva for 24 h. The surface morphology and roughness were examined using scanning electron microscopy (SEM) and atomic force microscopy (AFM), respectively. X-ray diffraction (XRD) was used to identify the phases and crystallinity of the deposited minerals observed on the enamel surface. Attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) was used to quantitatively analyze the mineral variation by calculating the relative integrated-area of characteristic bands. Nanohardness and elastic modulus measured by nanoindentation at various treatment stages were utilized to evaluate the degree of recovery. Biomimetic effects were accessed according to the degree of nanohardness recovery and the amount of hydroxyapatite deposition. The charged segments in the 3NSS peptide greatly attracted aqueous ions from artificial saliva to form hydroxyapatite crystals to fill enamel caries, in particular the interrod areas, resulting in a slight reduction in overall surface roughness. Additionally, the deposited hydroxyapatites were of a small crystalline size in the presence of the 3NSS peptide, which effectively restrained the plastic deformations and thus resulted in greater improvements in nanohardness and elastic modulus. The degree of nanohardness recovery was 5 times greater for remineralized enamel samples treated with the 3NSS peptide compared to samples without peptide treatment.     

Transmission electron microscopic observation of a metastable phase on the thermal decomposition process of Ca-deficient hydroxyapatite

Calcium-deficient hydroxyapatite (Ca-def HAp) decomposes to stoichiometric hydroxyapatite (HAp) and β-tricalcium phosphate (β-TCP) at high temperature. In a previous study, we reported that a metastable phase with a high Ca/P molar ratio appeared in the temperature range from 700 to 800°C. In the present study, the formation process of a metastable phase and the crystallographic relationship between the Ca-rich metastable phase and HAp matrix were investigated by high-resolution transmission electron microscopy (HRTEM). Ca-def HAp was annealed at 600–850°C for 2 or 6 h in air. TEM observations were performed before and after annealing Ca-def HAp. Based on analysis of image of Ca-def HAp before annealing, several HAp crystals with different aspect ratios agglomerated. The metastable phases grew thicker by long-term annealing. HRTEM image suggested that the Ca-rich metastable phase was formed by migration to the interface and continuous accumulation of calcium ions from HAp crystals with a small aspect ratio. From HRTEM images and results of the analysis of selected area electron diffraction patterns along the [010], [110] and [001] zone axes, lattice constants of the metastable phases were determined to be a = 2.86 nm, b = 0.94 nm, and c = 0.69 nm with orthorhombic crystals system.     

Modulation of enamel matrix proteins on the formation and nano-assembly of hydroxyapatite in vitro

Natural enamel has a hierarchically nanoassembled architecture that is regulated by enamel matrix proteins (EMPs) during the formation of enamel crystals. To understand the role of EMPs on enamel mineralization, calcium phosphate (CaP) growth experiments in both the presence and absence of native rat EMPs in a single diffusion system were conducted. The morphology and organization of formed CaP crystals were examined by X-Ray Diffraction (XRD), High-Resolution Transmission Microscopy (HRTEM) and Selected Area Electron Diffraction (SAED). In the system containing the EMPs, hydroxyapatite (HAP) with hierarchical lamellar nanostructure can be formed and the aligned HAP assembly tightly bundled by 3–4 rod-like nanocrystals like an enamel prism. However, in the absence of EMPs, only a sheet-like structure of octacalcium phosphate (OCP) phase was presented. EMPs promote HAP formation and inhibit the growth of OCP on the (010) plane. It is discussed that the organized Amelogenin/Amorphous Calcium Phosphate might be the precursor to the bundled HAP crystal prism. The study benefits the understanding of biomineralization of tooth enamel.     

Synthesis of oriented hydroxyapatite crystals: Effect of reaction conditions in the presence or absence of silk sericin

The purpose of this study was to investigate the effect of silk sericin (SS) on the morphology and crystallinity of hydroxyapatite (HAp) crystals synthesized at the pH 7.4-12 and 50 °C via a biomimetic method. Using X-ray diffraction (XRD), transmission electron microscope (TEM) and high-resolution transmission electron microscope (HRTEM), it was revealed that a homogeneous assembly could be obtained in the presence of SS, which held a highly ordered microstructure and poor crystallinity. But in the absence of SS, an oriented aggregate could be prepared at 50 °C and pH 10, which held more confused crystal lattice lines. The results proved that the biomimetic structure of HAp could be constructed under the controlled reaction conditions, which may offer a new clue on the understanding of the process of biomineralization.     

Synthesis and in-depth analysis of highly ordered yttrium doped hydroxyapatite nanorods prepared by hydrothermal method and its mechanical analysis

In this study, undoped and yttrium (Y) doped nanocrystalline hydroxyapatite crystals were synthesized by the hydrothermal method at 180 °C for 24 h. Highly ordered and oriented hydroxyapatite (HAp) nanorods were prepared by yttrium doping and their nanostructure and physical properties were compared with those of undoped HAp rods. FESEM images showed that the doping with Y ions reduced the diameter (from 25 nm to 15 nm) and increased the length (from 95 nm to 115 nm) of the synthesized rods. The aspect ratio of the undoped and Y-doped nanorods were calculated to be 4.303 (SD = 0.0959) and 7.61 (SD = 0.0355), respectively. Specific surface area (SSA) analysis showed that SSA also increased from 66.74 m²/g to 68.57 m²/g with the addition of yttrium. Y-doped HAp nanorod reinforced HMWPE composites displayed the better mechanical performance than those reinforced with pure HAp nanorods. The possible strengthening of nanorods and the increase of SSA due to the reduction in the size of nanorods in the presence of yttrium may have contributed to the strengthening of Y-doped HAp/HMWPE composites.     

Effect of supersaturation on the morphology of hydroxyapatite crystals deposited by electrochemical deposition on titanium

Cathodic deposition of calcium phosphate coating on titanium using electrochemical method was investigated in our study and well-defined hexagonal rod-like hydroxyapatite (HAp) crystals were synthesized. Scanning electron microscopic photographs of HAp coatings under different deposition durations revealed the morphology change of the HAp crystals in the coating film with the experimental process: from cone-like structure to hexagonal prism with sharp-angled tip, and finally turns to rod-like one with regular hexagonal cross section. It was suggested that both morphology and longitudinal length of the HAp crystals in the coating could be regulated by an accurate control of the degree of supersaturation of the aqueous system.     

Synthesis and characterization of hydroxyapatite–bacterial cellulose nanocomposites

Biocomposites consisting of hydroxyapatite (HAp) and natural polymers such as collagen, chitosan, chitin, and gelatin have been extensively investigated. However, studies on the combination of HAp with bacterial cellulose (BC) have not been conducted yet. In this work, our latest results concerning the biomimetic synthesis and characterization of HAp-BC nanocomposites with a 3-dimensional (3-D) network are reported. The present research focuses on characterizing the structure of this novel class of nanocomposites. Scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD), and Fourier-transformed infrared spectroscopy (FTIR) were employed to characterize the HAp-BC nanocomposites. It is found that HAp crystals are formed when the phosphorylated and CaCl₂-treated BC fibers are soaked in a 1.5 simulated body fluid (SBF). XRD reveals that the crystallite sizes of the HAp crystals are nano-sized and their crystallinities are low. The FTIR results show that the HAp crystals are partially substituted with carbonate, resembling natural bones. The nanocomposites containing HAp with structural features close to those of biological apatites are attractive for applications as artificial bones and scaffolds for tissue engineering.     

On the initial propagation of dental caries

A multi-dimensional model for dental caries is applied to study the shape of caries lesions in a realistic tooth geometry and to examine the rate of progress of caries. An upgraded model, taking into account the outer prismless enamel layer, is derived and solved. The model demonstrates the importance of this layer in delaying the onset of caries. The conclusions are discussed in light of experimental results.     

等离子活化烧结制备石墨烯/羟基磷灰石复相生物陶瓷

以羟基磷灰石(Hydroxyapatite, HAp)为基体, 石墨烯(Graphene, rGO)作为增强相, 利用等离子活化烧结制备了石墨烯/羟基磷灰石(rGO/HAp)复相生物陶瓷。系统研究了rGO添加量对HAp陶瓷基体物相结构、生物活性及断裂韧性的影响。结果表明, rGO的加入有利于提高HAp陶瓷的生物活性。同时, 复相生物陶瓷的硬度与断裂韧性随rGO添加量的增加均表现出先升高, 后显著降低的变化趋势。当rGO添加量为2wt%时, 样品的硬度与断裂韧性分别达到6.97 GPa和0.84 MPa•m^1/2, 较纯相HAp陶瓷提高了11.5%和37.3%。研究表明rGO的拔出效应是导致复相陶瓷力学性能提高的主要原因。     

Fine structure and development of the collar enamel in gars, , Actinopterygii

The fine structure of collar enamel and the cells constituting the enamel organ during amelogenesis in Lepisosteus oculatus was observed by light, scanning electron and transmission electron microscopy. In the enamel, slender crystals were arranged perpendicular to the surface and the stripes that were parallel to the surface were observed, suggesting that the enamel in Lepisosteus shares common morphological features with that in sarcopterygian fish and amphibians. Ameloblasts containing developed Golgi apparatus, rough endoplasmic reticulum (rER) and secretory granules were found in the secretory stage. In the maturation stage, a ruffled border was not seen at the distal end of the ameloblasts, while many mitochondria and lysosome-like granules were obvious in the distal cytoplasm. The enamel organ consisted of the outer dental epithelial cells, stratum reticulum cells and ameloblasts, but there was no stratum intermedium. It is likely that the ameloblasts have less absorptive function in comparison with the inner dental epithelial cells facing cap enameloid.     

Fine structure and development of the collar enamel in gars, Lepisosteus oculatus, Actinopterygii

The fine structure of collar enamel and the cells constituting the enamel organ during amelogenesis in Lepisosteus oculatus was observed by light, scanning electron and transmission electron microscopy. In the enamel, slender crystals were arranged perpendicular to the surface and the stripes that were parallel to the surface were observed, suggesting that the enamel in Lepisosteus shares common morphological features with that in sarcopterygian fish and amphibians. Ameloblasts containing developed Golgi apparatus, rough endoplasmic reticulum (rER) and secretory granules were found in the secretory stage. In the maturation stage, a ruffled border was not seen at the distal end of the ameloblasts, while many mitochondria and lysosome-like granules were obvious in the distal cytoplasm. The enamel organ consisted of the outer dental epithelial cells, stratum reticulum cells and ameloblasts, but there was no stratum intermedium. It is likely that the ameloblasts have less absorptive function in comparison with the inner dental epithelial cells facing cap enameloid.