Calcium phosphate and fluorinated calcium phosphate coatings on titanium deposited by Nd:YAG laser at a high fluence

Calcium phosphate coatings are known to enhance long-term fixation, reliability and promote osteointegration of cementless titanium-based implant devices. This study was aimed at the pulsed laser deposition of calcium phosphate coatings onto titanium using hydroxyapatite and hydroxyapatite-fluorapatite targets. The deposition was carried out at the high laser beam fluence conditions, about 12 J/cm(2). The coatings were characterized with respect to their morphology, phase composition and hardness. X-ray energy dispersive analysis revealed the coatings retain their elemental composition, and fluoride content within the film is the same as in the initial target. However, unlike sintered targets, the deposited films contain no apatite-like phases. The hardness of the films, about 18 GPa, is surprisingly high compared to that of hydroxyapatite and hydroxyapatite-fluorapatite ceramic targets. The deposited coatings of 2.7-2.9 microm thickness have uniform and dense microstructure, containing the solidified droplets of the expulsed from the target phase. The uncommon structure and hardness of the films can be attributed to the melting and phase decomposition of the initial material in the laser plasma.     

Mechanical properties of calcium phosphate coatings deposited by laser ablation

Amorphous calcium phosphate and crystalline hydroxyapatite coatings with different morphologies were deposited onto Ti-6Al-4V substrates by means of the laser ablation technique. The strength of adhesion of the coatings to the substrate and their mode of fracture were evaluated through the scratch test technique and scanning electron microscopy. The effect of wet immersion on the adhesion was also assessed. The mechanisms of failure and the critical load of delamination differ significantly depending on the phase and structure of the coatings. The HA coatings with granular morphology have higher resistance to delamination as compared to HA coatings with columnar morphology. This fact has been related to the absence of stresses for the granular morphology.     

Calcium phosphate coatings obtained by Nd:YAG laser cladding: physicochemical and biologic properties

The plasma spray (PS) technique is the most popular method commercially in use to produce calcium phosphate (CaP) coatings to promote fixation and osteointegration of the cementless prosthesis. Nevertheless, PS has some disadvantages, such as the poor coating-to-substrate adhesion, low mechanical strength, and brittleness of the coating. In order to overcome the drawbacks of plasma spraying, we introduce in this work a new method to apply a CaP coating on a Ti alloy using a well-known technique in the metallurgical field: laser surface cladding. The physicochemical characterization of the coatings has been carried out by means of X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy dispersive X-ray analysis (EDX). The biologic properties of the coatings have been assessed in vitro with human osteoblast-like MG-63 cells. The overall results of this study affirm that the Nd:YAG laser cladding technique is a promising method in the biomedical field.     

Human osteoblast response to pulsed laser deposited calcium phosphate coatings

Octacalcium phosphate (OCP) and Mn(2+)-doped carbonate hydroxyapatite (Mn-CHA) thin films were deposited on pure, highly polished and chemically etched Ti substrates with pulsed laser deposition. The coatings exhibit different composition, crystallinity and morphology that might affect their osteoconductivity. Human osteoblasts were cultured on the surfaces of OCP and Mn-CHA thin films, and the cell attachment, proliferation and differentiation were evaluated up to 21 days. The cells showed a normal morphology and a very good rate of proliferation and viability in every experimental time. Alkaline phosphatase activity was always higher than the control and Ti groups. From days 7 to 21 collagen type I production was higher in comparison with control and Ti groups. The level of transforming growth factor beta 1 (TGF-beta1) was lower at 3 and 7 days, but reached the highest values during following experimental times (14 and 21 days). Our data demonstrate that both calcium phosphate coatings favour osteoblasts proliferation, activation of their metabolism and differentiation.     

Influence of thickness on the properties of hydroxyapatite coatings deposited by KrF laser ablation.

The growth of hydroxyapatite coatings obtained by KrF excimer laser ablation and their adhesion to a titanium alloy substrate were studied by producing coatings with thicknesses ranging from 170 nm up to 1.5 microm, as a result of different deposition times. The morphology of the coatings consists of grain-like particles and also droplets. During growth the grain-like particles grow in size, partially masking the droplets, and a columnar structure is developed. The thinnest film is mainly composed of amorphous calcium phosphate. The coating 350nm thick already contains hydroxyapatite, whereas thicker coatings present some alpha tricalcium phosphate in addition to hydroxyapatite. The resulting coating to substrate adhesion was evaluated through the scratch test technique. Coatings fail under the scratch test by spallating laterally from the diamond tip and the failure load increases as thickness decreases, until not adhesive but cohesive failure for the thinnest coating is observed.     

Behavior in simulated body fluid of calcium phosphate coatings obtained by laser ablation

Three types of calcium phosphate coatings onto titanium alloy substrates, deposited by the laser ablation technique, were immersed in a simulated body fluid in order to determine their behavior in conditions similar to the human blood plasma. Neither the hydroxyapatite coating nor the amorphous calcium phosphate coating do dissolve and the alpha-tricalcium phosphate phase of the coating of beta-tricalcium phosphate with minor alpha phase slightly dissolves. Precipitation of an apatitic phase is favored onto the hydroxyapatite coating and onto the coating of beta-tricalcium phosphate with minor alpha phase. Onto the titanium alloy substrate reference there is also precipitation but at larger induction times. However, onto the amorphous calcium phosphate coating no precipitate is formed.     

Structural and morphological study of pulsed laser deposited calcium phosphate bioceramic coatings: influence of deposition conditions, laser parameters, and target properties

Calcium phosphate (CaP) bioceramics, especially hydroxyapatite (HA), have been used as coatings on implants owing to their biocompatible properties. The commercial practice for applying HA coating, plasma spraying, has some disadvantages which limit the long-term stability of the implants. Pulsed laser deposition (PLD) is being investigated as an alternative technique. The purpose of this research was to systematically study the effect of various parameters of the PLD process on the properties of CaP coatings. In this study, three types of HA targets and two laser wavelengths were used to make six categories of coatings. Predominantly crystalline HA coatings were produced under all six categories at optimum conditions, although small amounts of minor phases sometimes were found. Sufficient coating/substrate bond strength was also obtained. A wide variety of coating morphologies was obtained, from rather dense and uniform to rough and porous. The important factors that affected the morphology included target properties, vacuum level, deposition temperature, and laser wavelength and energy density. PLD's ability to produce both amorphous and crystalline, and both smooth/dense and rough/porous coatings may be a unique advantage.     

Properties of heat-treated calcium phosphate coatings deposited by high-velocity oxy-fuel (HVOF) spray

The influence of crystallization, upon heat treatment, on the properties of high-velocity oxy-fuel (HVOF) sprayed hydroxyapatite (HA) coatings was investigated. The characterization of the HA coating was performed by X-ray diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR). Differential Scanning Calorimeter (DSC) was employed to determine the crystallization temperature of the amorphous phase in an as-sprayed HA coating. The study demonstrated the effect of crystallization on the coating properties by considering the changes in materials chemistry, crystallinity level, and mechanical performance. Results showed that complete crystallization of the amorphous phase occurred at approximately 700 degrees C and the crystallization temperature was dependent on sample heating rate in the DSC test. The changes of ion groups were detected by FTIR, before and after the phase transformation. The crystallization of the coating after annealing at 750 degrees C resulted in a significant increase of the coatings' adhesive strength and shear strength, which attained maximum values 34 +/- 3 and 14.1 -/+ 0.8 MPa, respectively. Young's modulus increased from 21 +/- 1 to 25 +/- 2 GPa. Microhardness measurements confirmed the changes in coating properties. It is also found that the transformation from the amorphous phase has crystalline HA as the only resultant phase detected by XRD.     

紫外激光辐射区肝组织β连环蛋白及过氧化物酶体增殖物激活受体γ蛋白的表达

背景：研究表明经典的wnt信号途径可通过抑制成脂分化关键因子过氧化物酶体增殖物激活受体γ而抑制细胞向脂肪细胞的分化。 目的：观察紫外光辐射肝组织致创后自然恢复期不同阶段β连环蛋白及过氧化物酶体增殖物激活受体γ的表达。 方法：将雄性SD大鼠随机分4组：激光致创组大鼠是以355 nm紫外激光辐射肝叶形成损伤区,器械致创对照组则以手术剪肝叶致创,假手术组仅做腹部切开及缝合,正常组大鼠仅实施正常喂养。 结果与结论：紫外激光可在肝组织形成边缘整齐的消融区并在周围形成可自然修复的有限致损区域。免疫组织化学图像平面测量表明紫外激光辐射后自然恢复1周大鼠肝组织中β连环蛋白表达低于各对照组(P < 0.01),而自然恢复表达β连环蛋白 辐射区周围组织表达高于正常(P < 0.01)。蛋白印迹结果显示辐射致创组辐射后自然同时过氧化物酶体增殖物激活受体γ蛋白2周 恢复高于1周恢复(P < 0.01),且恢复期肝组织中可同时表达过氧化物酶体增殖物激活受体γ 1及2。结果证实,辐射区旁组织自然恢复各阶段存在不同的β连环蛋白及过氧化物酶体增殖物激活受体γ表达调控,2周自然恢复后辐射区旁组织成脂进程得到加强。     

Micro- and nano-testing of calcium phosphate coatings produced by pulsed laser deposition

Micro- and nano-testing methods have been explored to study the thin calcium phosphate coatings with high adhesive strength. The pulsed laser deposition (PLD) technique was utilised to produce calcium phosphate coatings on metal substrates, because this type of coatings exhibit much higher adhesive strength with substrates than conventional plasma-sprayed coatings. Due to the limitations of the conventional techniques to evaluate the mechanical properties of these thin coatings (1 microm thick), micro-scratch testing has been applied to evaluate the coating-to-substrate adhesion, and nano-indentation to determine the coating hardness and elastic modulus. The test results showed that the PLD produced amorphous and crystalline HA coatings are more ductile than titanium substrates, and the PLD coatings are not delaminated from the substrates by scratch. Also, the results showed that the crystalline HA coating is superior in internal cohesion to the amorphous one, even though the lower elastic modulus of amorphous coating could be more mechanically compatible with natural bone.     

XPS, EDX and FTIR analysis of pulsed laser deposited calcium phosphate bioceramic coatings: the effects of various process parameters

Many techniques have been used to produce calcium phosphate, especially hydroxyapatite (HA), coatings on metallic implant surfaces for improved biocompatibility. Although some techniques have produced coatings used clinically, the long-term stability of the coating/implant is still questionable. As a new technique for making HA coatings, pulsed laser deposition (PLD) shows some advantages in controlling the coatings' crystal structure and composition. In this study, three types of HA target and two wavelengths of laser were used to produce calcium phosphate coatings. Despite PLDs ability to improve the crystal structure by incorporating water vapor into the deposition process, the characterization with EDX and XPS showed that coatings had different Ca/P ratios from that of the pure HA targets, which almost assured the presence of non-HA phases. FTIR spectra also showed differences in phosphate bands of coatings and targets although the difference in data collecting modes might have been a factor. The observed differences might be related to the differences between the surface and bulk chemistries of the coatings. Nevertheless, when evaluating the suitability of the PLD technique for making HA coatings, the possibility of the formation of non-HA phases cannot be excluded, although it may not necessarily be a negative factor.     

Bone growth on and resorption of calcium phosphate coatings obtained by pulsed laser deposition

Three different calcium phosphate coatings of crystalline hydroxyapatite (HA), alpha- and beta-tricalcium phosphate (alpha+beta-TCP), or amorphous calcium phosphate (ACP) obtained by pulsed laser deposition on Ti-6Al-4V were incubated in a potentially osteogenic primary cell culture (rat bone marrow) in order to evaluate the amount and mode of mineralized bone matrix formation after 2 weeks with special emphasis on the type of interfacial structure that was created. Evaluation techniques included fluorescence labeling and scanning electron microscopy. The resistance to cellular resorption by osteoclasts was also studied. Bone matrix delaminated from the ACP coatings, while it remained on the HA and the alpha+beta-TCP coatings even after fracturing. A cementlike line was seen as the immediate contiguous interface with the nondegrading dense HA surface and with the surface of the remaining porous beta-TCP coating. Highly dense and crystalline HA coatings do not dissolve but are capable of establishing a strong bond with the bone matrix grown on top. Chemical and mechanical bonding were considered in this case. Cellular resorption was practically not observed on the HA coatings, but it was observed on the alpha+beta-TCP coatings. Resorption took place as dissolution that was due to the acidic microenvironment.     

Penetration depth into dental casting alloys by Nd:YAG laser

This study investigated the effect of laser-beam welding conditions [voltage (V) and spot diameter (mm)] on the penetration depth into dental casting alloys. Castings (3.0 mm x 8.0 mm x 50 mm) were prepared from the metals used in this study: commercially pure titanium (CP Ti), Ti-6Al-4V, Ti-6Al-7Nb, cobalt-chromium alloy (Co-Cr) and Type IV gold alloy. Two cast blocks of each metal were butted against one another at the 8.0 x 50-mm surfaces. They were then welded at their interface under the following conditions: voltage of 160-340 V, spot diameter of 0.4-1.6 mm, and pulse duration of 10 ms. After laser welding, the blocks were separated, and the penetration depth into each alloy was measured. The data were analyzed with the use of ANOVA at the p < 0.05 level of significance. The penetration depths were as follows: CP Ti (0.29-6.45 mm), Ti-6Al-4V (0.32-5.24 mm), Ti-6Al-7Nb (0.34-5.65 mm), Co-Cr (0.24-6.15 mm), and Type IV gold alloy (0.12-5.22 mm). The voltage and spot diameter affected the penetration depth into the metals tested. When the voltage increased and the spot diameter decreased, the penetration depth increased for each metal. Selecting suitable conditions for laser welding to obtain sufficient penetration depth for the optimal thickness of the metal is important when welding prostheses.     

Characterization of calcium phosphate cements modified by addition of amorphous calcium phosphate

In this study the influence of amorphous calcium phosphate (ACP) on the setting of, and the formed apatite crystallite size in, a calcium phosphate cement (CPC) based on α-tricalcium phosphate (α-TCP) or tetracalcium phosphate (TTCP)/monocalcium phosphate monohydrate (MCPM) was investigated. Setting times at 22 °C were measured in air atmosphere; those at 37 °C were measured at 100% relative humidity. The phase composition of the set cements was investigated after 1 week using X-ray diffractometry and infrared spectroscopy and the morphology was investigated using scanning electron microscopy. The compressive strength (CS) of the set CPCs was measured after 1 day. Viability of MC3T3-E1 cells on the CPCs was analyzed after 7, 14 and 21 days of incubation using the CellTiter 96^® Aqueous Non-Radioactive Cell Proliferation Assay. The α-TCP-based cement exhibited long setting times, a high CS and was converted to a calcium-deficient hydroxyapatite (CDHAp). The TTCP/MCPM-based CPC was only partly converted to CDHAp, produced acceptable setting times and had a low CS. Addition of ACP to these two CPCs resulted in cements that exhibited good setting times, CS suitable for non-load-bearing applications and a full conversion to nanocrystalline CDHAp. Moreover, the ACP containing CPCs demonstrated good cell viability, making them suitable candidates for bone substitute materials.     

Plume dynamics and shielding by the ablation plume during Er:YAG laser ablation

Free-running Er:YAG lasers are used for precise tissue ablation in various clinical applications. The ablated material is ejected into the direction perpendicular to the tissue surface. We investigated the influence of shielding by the ablation plume on the energy deposition into an irradiated sample because it influences the ablation dynamics and the amount of material ablated. The investigations were performed using an Er:YAG laser with a pulse duration of 200 micros for the ablation of gelatin with different water contents, skin, and water. Laser flash photography combined with a dark field Schlieren technique was used to visualize gaseous and particulate ablation products, and to measure the distance traveled by the ablating laser beam through the ablation plume at various times after the beginning of the laser pulse. The temporal evolution of the transmission through the ablation plume was probed using a second free running Er:YAG laser beam directed parallel to the sample's surface. The ablation dynamics was found to consist of a vaporization phase followed by material ejection. The observation of droplet ejection during water ablation provided evidence that a phase explosion is the driving mechanism for material ejection. The laser light transmission was only slightly reduced by the vapor plume, but decreased by 25%-50% when the ejected material passed the probe beam. At radiant exposures approximately 10 times above the ablation threshold, the laser energy deposited into the sample amounted to only 61% of the incident energy for gelatin samples with 90% water content and to 86% for skin samples. For free-running Er:YAG laser pulses shielding must therefore be considered in modeling the ablation dynamics and determining the dosage for clinical applications.     

The rat esophagus: ultrastructure and radiological aspects of tissue response after 1320 nm Nd:YAG laser irradiation.

Morphological tissue response towards laser treatment was investigated in the esophagi of adult Wistar rats by light- and transmission electron microscopy. The specimens were fixed by perfusion immediately, 2 days and 14 days after laser treatment in order to assess different stages of the healing process. The epithelium of the lasercentre was completely destroyed in the immediate group. The connective tissue showed damaged cells, fused collagenous fibres and occluded blood vessels. Smooth muscle cells presented a vacuolated sarcoplasm and pycnotic nuclei. The cross striation of skeletal muscle cells had disappeared and their nuclei were karyolytic. In a distance of 4 mm from the lasercentre all wall tissues had an almost normal appearance. After 2 days the morphological feature of the lasercentre was the same as in the immediate group. In a distance of 2 mm some layers of flat and intact epithelial cells were observed below the necrotic epithelium. The adjacent connective tissue was infiltrated by inflammatory cells. After 14 days the formation of granulation tissue had caused an occlusion of the lumen in the lasercentre. In a distance of 2 mm the lumen was patent and the wall tissues had been partly restored. As the rat esophagus serves as a model for esophagotracheal fistulae in newborn children we assume the 1320 nm Nd:YAG laser to be a possible application in occlusion of these fistulae.     

Effect of Nd: YAG laser on titanium dental implants studied by AFM

Bacterial contamination of dental implants is considered the main cause of implant failure. Recently, the laser treatment of the implant surface has been proposed as an useful method for decontamination. In such a view, the present study was conducted to investigate the effects of a Nd:YAG laser on the surface morphology of a titanium dental implant by means of an atomic force microscope. We demonstrated that, when the pulse energy of the laser was kept below 30 mJ, independently from the pulse rate, the laser-treated specimens exhibited a qualitatively similar surface morphology when compared to the untreated titanium implants, suggesting that the implant surface was unaffected by the treatment, in these particular conditions. We also found that, by cooling the implant surface with an air flow? during laser irradiation, the mean temperature of the implant was maintained under 37 degrees C. All these data taken together suggest the possibility to use Nd:YAG laser for the treatment of failing dental implants.     

Reaction of myocardial mast cells during neoangiogenesis induced by YAG:Nd laser

Time course of morphofunctional changes in myocardial mast cells of Wistar rats irradiated with a YAG:Nd laser is studied. Mast cells were detected on serial sections by toluidine blue staining. A drastic increase in mast cell degranulation around the laser channel was observed on days 1–20 of experiment. Mast cell degranulation may be the first stage in neoangiogenesis in the myocardium in response to laser injury. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 126, No. 7, pp. 116–118, July, 1998     

Nd:YAG激光治疗儿童后发障的临床观察

目的探讨Nd:YAG激光治疗儿童后发障的时机、效果及并发症。方法用Nd:YAG激光对65例87眼儿童后发障患者进行治疗。结果术后视力提高1行者7眼,占8%;提高2行者20眼,占22.9%;提高3行及以上者54眼,占62%;术后视力未提高者6眼,占6.8%。结论Nd:YAG激光治疗儿童后发障是一种安全有效的方法,并应尽早进行。     

Effect of Nd:YAG laser irradiation on human dental enamel

The crystallographic alterations of the hydroxyapatite under the laser irradiation were evaluated by the x-ray diffraction pattern analysis and scanning electron microscopic observation. Recrystallization may occur in fused and resolidified dental enamel by the pulsed Nd: YAG laser irradiation. Two different grades of energy densities such as 400 pulses and 800 pulses were given to the powdered enamel. The irradiation was performed at a peak power of 500W with a pulse width of 10 msec.; the average output of 10W, spot size of 3mm and two pulses were given in every second. After the 800 pulses of Nd:YAG laser irradiation to the human dental enamel the x-ray diffraction pattern demonstrated both alpha-tricalcium phosphate and hydroxyapatite peaks. With the lower level of energy at 400 pulses, no significant differences were seen in the diffraction patterns between lased and unlased enamel. In scanning electron microscopic findings, there were no significant changes between lased and unlased enamel. When the unlased and lased enamel were exposed to acid solution, unlased enamel showed a honeycomb pattern, while the lased enamel showed preferentially removed prism core material.