1.Polysaccharide isolated from wax apple suppresses ethyl carbamate-induced oxidative damage in human hepatocytes.
Tao BAO ; Naymul KARIM ; Huihui KE ; Jitbanjong TANGPONG ; Wei CHEN
Journal of Zhejiang University. Science. B 2023;24(7):574-586
Wax apple (Syzygium samarangense) has received growing research interest for its high nutritional and medicinal value due to its constituents such as polysaccharide, organic acids, flavonoids, minerals, and other substances. In this study, wax apple polysaccharide (WAP) was isolated from this plant and its protective effect against ethyl carbamate (EC)-induced oxidative damage was evaluated in human hepatocytes (L02 cells). Firstly, a series of analyses such as high-performance liquid chromatography (HPLC), high-performance gel permeation chromatography (HPGPC), Fourier transform infrared spectroscopy (FT-IR), gas chromatography/mass spectrometry (GC/MS), and 1H and 13C nuclear magnetic resonance (NMR) were conducted to identify the structure of WAP. Thereafter, in vitro cell experiments were performed to verify the protective effects of WAP against EC-induced cytotoxicity, genotoxicity, and oxidative damage in L02 cells. Our results revealed that WAP is composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, arabinose, and fucose in a molar ratio of 2.20:3.94:4.45:8.56:8.86:30.82:39.78:1.48. Using a combination of methylation and NMR spectroscopic analysis, the primary structure of WAP was identified as Araf-(1→, Glcp-(1→, →2)-Araf-(1→, →3)-Galp-(1→, →3)-Araf-(1→, and →6)-Galp-(1→. Cell experiments indicated that WAP exhibited significant protective effects on EC-treated L02 cells via suppressing cytotoxicity and genotoxicity, reducing reactive oxygen species (ROS) and O2•- formation, as well as improving mitochondrial membrane potential (MMP) and glutathione (GSH). In a nutshell, WAP has the potential as an important therapeutic agent or supplement for hepatic oxidative damage. Meanwhile, further studies are needed to prove the above effects in vivo at the biological and clinical levels.
Humans
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Syzygium/chemistry*
;
Urethane/pharmacology*
;
Spectroscopy, Fourier Transform Infrared
;
Oxidative Stress
;
Glutathione/pharmacology*
;
Hepatocytes
;
Polysaccharides/pharmacology*
2.Physiological regulation of salicylic acid on Helianthus tubeuosus upon copper stress and root FTIR analysis.
Jinxiang AI ; Jieke GE ; Ziyi ZHANG ; Wenqian CHEN ; Jiayi LIANG ; Xinyi WANG ; Qiaoyuan WU ; Jie YU ; Yitong YE ; Tianyi ZHOU ; Jinyi SU ; Wenwen LI ; Yuhuan WU ; Peng LIU
Chinese Journal of Biotechnology 2023;39(2):695-712
Phytoremediation plays an important role in the treatment of heavy metal pollution in soil. In order to elucidate the mechanism of salicylic acid (SA) on copper absorption, seedlings from Xuzhou (with strong Cu-tolerance) and Weifang Helianthus tuberosus cultivars (with weak Cu-tolerance) were selected for pot culture experiments. 1 mmol/L SA was sprayed upon 300 mg/kg soil copper stress, and the photosynthesis, leaf antioxidant system, several essential mineral nutrients and the changes of root upon copper stress were analyzed to explore the mechanism of copper resistance. The results showed that Pn, Tr, Gs and Ci upon copper stress decreased significantly compared to the control group. Meanwhile, chlorophyll a, chlorophyll b and carotenoid decreased with significant increase in initial fluorescence (F0), maximum photochemical quantum yield of PSⅡ (Fv/Fm), electron transfer rate (ETR) and photochemical quenching coefficient (qP) content all decreased. The ascorbic acid (AsA) content was decreased, the glutathione (GSH) value was increased, the superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) activity in the leaves were decreased, and the peroxidase (POD) activity was significantly increased. SA increased the Cu content in the ground and root system, and weakened the nutrient uptake capacity of K, Ca, Mg, and Zn in the root stem and leaves. Spray of exogenous SA can maintain the opening of leaf stomata, improve the adverse effect of copper on photosynthetic pigment and PSⅡ reaction center. Mediating the SOD and APX activity started the AsA-GSH cycle process, effectively regulated the antioxidant enzyme system in chrysanthemum taro, significantly reduced the copper content of all parts of the plant, and improved the ion exchange capacity in the body. External SA increased the content of the negative electric group on the root by changing the proportion of components in the root, promoted the absorption of mineral nutrient elements and the accumulation of osmoregulatory substances, strengthened the fixation effect of the root on metal copper, and avoided its massive accumulation in the H. tuberosus body, so as to alleviate the inhibitory effect of copper on plant growth. The study revealed the physiological regulation of SA upon copper stress, and provided a theoretical basis for planting H. tuberosus to repair soil copper pollution.
Antioxidants
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Copper
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Helianthus/metabolism*
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Salicylic Acid/pharmacology*
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Chlorophyll A/pharmacology*
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Spectroscopy, Fourier Transform Infrared
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Chlorophyll/pharmacology*
;
Ascorbic Acid
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Superoxide Dismutase/metabolism*
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Photosynthesis
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Glutathione
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Plant Leaves
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Stress, Physiological
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Seedlings
3.Quality evaluation methods of Poria based on yield and content of alkali extracted polysaccharide.
Yue WANG ; Xiao-Qian LIU ; Yong-Xin ZHANG ; Wei-Hong FENG ; Li-Hua YAN ; Zhi-Min WANG
China Journal of Chinese Materia Medica 2023;48(2):443-454
To improve the quality control methods of Poria and develop and utilize its resources fully, alkaline extraction was used in this study to determine the yield and content of alkali-soluble polysaccharides of Poria. The alkali-soluble extracts of Poria were obtained according to the optimum extraction conditions on the basis of single-factor test, and 30 batches of samples were determined. The structure and chemical composition of the alkali-soluble extracts was characterized by high-performance gel permeation chromatography(HPGPC), Fourier transform infrared spectrometry(FT-IR), nuclear magnetic resonance(NMR) spectroscopy and high-performance liquid chromatography(HPLC) with 1-phenyl-3-methyl-5-pyrazolone(PMP-HPLC). The results showed that the content of the alkali-soluble extracts was in the range of 46.98%-73.86%. The main component was β-(1→3)-glucan, and its molecular mass was about 1.093×10~5. Further, the content of alkali-soluble polysaccharides of Poria was measured by UV-Vis spectrophotometry and HPLC coupled with the evaporative light scattering detector(HPLC-ELSD), and 30 batches of samples were measured. The results indicated that the content of alkali-soluble polysaccharides determined by UV-Vis spectrophotometry was in the range of 73.70%-92.57%, and the content of samples from Hubei province was slightly higher than that from Yunnan province, Anhui province and Hunan province. The content of alkali-soluble polysaccharides determined by HPLC-ELSD was in the range of 51.42%-76.69%, and the samples from Hunan province had slightly higher content than that from the other three provinces. The content determined by UV-Vis spectrophotometry was higher than that by HPLC-ELSD. However, the content determined by HPLC-ELSD was close to that of alkali-soluble extract, which could accurately characterize the content of alkali-soluble polysaccharides in Poria, and the method was simple and repeatable. Therefore, it is recommended that the quantitative analysis method for alkali-soluble extract and alkali-soluble polysaccharides by HPLC-ELSD be used in the quality standards of Poria in Chinese Pharmacopeia.
Poria/chemistry*
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Spectroscopy, Fourier Transform Infrared
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China
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Polysaccharides/chemistry*
;
Reference Standards
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Chromatography, High Pressure Liquid/methods*
4.Structural characterization of PCP-Ⅰ from Poria as vaccine adjuvant and its hydrolytic oligosaccharide.
Jia-Nan GU ; Gui-Xin LIU ; Shuai LI ; Hao MA ; Jun-Jie SHAN
China Journal of Chinese Materia Medica 2023;48(16):4429-4437
Poria is an important medical herb in clinic. The authors isolated a polysaccharide(PCP-Ⅰ) from Poria in previous studies, which is composed of galactose, mannose, fucose and glucose. PCP-Ⅰ exhibited significant adjuvant effects on H1N1 influenza vaccine, hepatitis B surface antigen and anthrax protective antigen, and its adjuvant activity was stronger than aluminium adjuvant. However, little is known about the chemical structure of PCP-Ⅰ at present. In this study, weak acid hydrolysis was used to obtain the backbone oligosaccharide of PCP-Ⅰ. Then periodate oxidation, Smith degradation, methylation analysis, Fourier transform infrared spectroscopy(FT-IR), nuclear magnetic resonance(NMR) and gas chromatography-mass spectrometry(GC-MS) were performed to investigate the chemical structural features of PCP-Ⅰ and its hydrolytic oligosaccharide(PCP-Ⅰ-hy-1). These results suggested that the backbone of PCP-Ⅰ was composed of galactose with α anomeric carbon and β anomeric carbon. The linking residues of galactan are(1→),(l→6) and(1→2,6).
Adjuvants, Vaccine
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Poria
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Hydrolysis
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Spectroscopy, Fourier Transform Infrared
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Galactose
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Influenza A Virus, H1N1 Subtype
;
Polysaccharides/chemistry*
;
Oligosaccharides
;
Carbon
5.Forensic Pathological Diagnosis of Acute and Old Myocardial Infarction Using Fourier Transform Infrared Spectroscopy.
Tian TIAN ; Xin-Biao LIAO ; Fu ZHANG ; Kai-Fei DENG ; Ji ZHANG ; Ping HUANG ; Yi-Jiu CHEN ; Jian-Hua ZHANG
Journal of Forensic Medicine 2023;39(6):535-541
OBJECTIVES:
Fourier transform infrared spectroscopy (FTIR) was used to analyze myocardial infarction tissues at different stages of pathological change to achieve the forensic pathology diagnosis of acute and old myocardial infarction.
METHODS:
FTIR spectra data of early ischemic myocardium, necrotic myocardium, and myocardial fibrous tissue in the left ventricular anterior wall of the sudden death group of atherosclerotic heart disease and the myocardium of the normal control group were collected using hematoxylin-eosin (HE) and immunohistochemistry (IHC) staining as a reference, and the data were analyzed using multivariate statistical analysis.
RESULTS:
The mean normalized spectra of control myocardium, early ischemic myocardium and necrotic myocardium were relatively similar, but the mean second derivative spectra were significantly different. The peak intensity of secondary structure of proteins in early ischemic myocardium was significantly higher than in other types of myocardium, and the peak intensity of the α-helix in necrotic myocardium was the lowest. The peaks of amide Ⅰ and amide Ⅱ in the mean normalized spectra of myocardial fibrous tissue significantly shifted towards higher wave numbers, the peak intensities of amide Ⅱ and amide Ⅲ were higher than those of other types of myocardium, and the peak intensities at 1 338, 1 284, 1 238 and 1 204 cm-1 in the mean second derivative spectra were significantly enhanced. Principal component analysis (PCA) and partial least square-discriminant analysis (PLS-DA) showed that FTIR could distinguish different types of myocardium.
CONCLUSIONS
FTIR technique has the potential to diagnose acute and old myocardial infarction, and provides a new basis for the analysis of the causes of sudden cardiac death.
Humans
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Amides
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Death, Sudden, Cardiac
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Myocardial Infarction/pathology*
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Myocardium/pathology*
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Spectroscopy, Fourier Transform Infrared/methods*
;
Forensic Pathology
6.Composition and immunomodulatory activity of neutral and acidic polysaccharides isolated from Epimedii Folium.
Yan-Jun YANG ; Chang LI ; Fei-Fei CHEN ; Mao-Mao ZHU ; Xiao-Bin JIA ; Liang FENG ; Xiao-Bin TAN
China Journal of Chinese Materia Medica 2022;47(16):4358-4364
Epimedii Folium possesses many pharmacological activities including immunomodulation, anti-oxidation, and anti-tumor. Polysaccharides are the main components of Epimedii Folium, and their activities are closely related to the structure. The present study isolated a neutral polysaccharide(EPS-1-1) and an acidic polysaccharide(EPS-2-1) from the aqueous extract of Epimedii Folium through DEAE-52 cellulose anion-exchange chromatography and Sephadex G-100. The structures were characterized by chemical composition analysis, high-performance gel permeation chromatography(HPGPC), Fourier-transform infrared spectrometry(FT-IR), 1-phenyl-3-methyl-5-pyrazolone(PMP) derivatization, scanning electron microscopy(SEM), Congo red test, etc. The immunomodulatory activity of polysaccharides in vitro was determined by investigating the effects on the maturation of bone marrow-derived dendritic cells(BMDCs) and the release of inflammatory cytokines. According to the structural characterization analysis, EPS-1-1 was composed of fructose(Fuc), mannose(Man), ribose(Rib), rhamnose(Rha), glucose(Glc), galactose(Gal), xylose(Xyl), and arabinose(Ara) at 1.90∶0.67∶0.05∶0.08∶3.29∶1.51∶0.05∶0.37(molar ratio), while EPS-2-1 was mainly composed of Fuc, Man, Rha, glucuronic acid(GlcA), galacturonic acid(GalA), Glc, Gal, Xyl, and Ara at 5.25∶0.18∶0.32∶0.13∶1.14∶0.16∶0.55∶0.08∶0.2. EPS-1-1 and EPS-2-1 could promote the maturation and function of BMDCs through up-regulating the expression of MHC-Ⅱ, CD86, CD80, and CD40, and increasing the levels of inflammatory cytokines(IL-6, IL-12, and TNF-α) in vitro experiments, which suggested that EPS-1-1 and EPS-2-1 possessed good immunomodulatory activity.
Cytokines/metabolism*
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Drugs, Chinese Herbal
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Gas Chromatography-Mass Spectrometry
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Humans
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Immunomodulation
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Polysaccharides/chemistry*
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Spectroscopy, Fourier Transform Infrared
7.Co-amorphous technology to improve dissolution and physical stability of silybin.
Huan LIU ; Guo-Wei ZHAO ; Qie-Ying JIANG ; Xin-Li LIANG ; Liao-Qi OUYANG ; Hai-Bo DING ; Xu-Long CHEN ; Zheng-Gen LIAO
China Journal of Chinese Materia Medica 2022;47(1):103-110
The present study explored the effect of co-amorphous technology in improving the dissolution rate and stability of silybin based on the puerarin-silybin co-amorphous system prepared by the spray-drying method. Solid-state characterization was carried out by powder X-ray diffraction(PXRD), polarizing microscopy(PLM), Fourier transform infrared spectroscopy(FT-IR), differential scanning calorimetry(DSC), etc. Saturated powder dissolution, intrinsic dissolution rate, moisture absorption, and stability were further investigated. The results showed that puerarin and silybin formed a co-amorphous system at a single glass transition temperature which was higher than that of any crude drug. The intrinsic dissolution rate and supersaturated powder dissolution of silybin in the co-amorphous system were higher than those of the crude drug and amorphous system. The co-amorphous system kept stable for as long as three months under the condition of 40 ℃, 75% relative humidity, which was longer than that of the single amorphous silybin. Therefore, the co-amorphous technology could significantly improve the dissolution and stability of silybin.
Calorimetry, Differential Scanning
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Desiccation
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Drug Compounding/methods*
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Drug Stability
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Silybin
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Solubility
;
Spectroscopy, Fourier Transform Infrared
;
Technology
;
X-Ray Diffraction
8.Improvement in compatibility of hot melt pressure-sensitive adhesive with cinnamon volatile oil and in vitro transdermal property by physical blending.
Shuo YIN ; Tan WU ; Jian-Ying LU ; Zhen-da LIU ; Teng GUO ; Nian-Ping FENG
China Journal of Chinese Materia Medica 2021;46(21):5650-5657
Hot melt pressure-sensitive adhesive(HMPSA) has broad application potential in the field of traditional Chinese medicine(TCM) plasters due to its high drug loading, weak skin irritation, satisfactory adhesion, etc. compared with rubber plasters.However, the structure of HMPSA is prone to suffer from the damage caused by volatile oils in TCM plasters. In view of this, a kind of HMPSA with a stable structure was prepared by physical blending of DINCH, polypropylene wax and liquid rubber(LIR) in the present study, which is denoted as DPL. The dosage of cinnamon volatile oil(CVO), the model drug, was selected with viscosity, softening point and cohesion as evaluation indexes. The interaction between DPL and HMPSA was investigated by Fourier transform infrared spectroscopy(FT-IR) and differential scanning calorimetry(DSC). The compatibility of HMPSA with CVO and its transdermal ability were studied by in vitro transdermal test, adhesion, scanning electron microscopy( SEM) and rheological evaluation. The results showed that 5% CVO began to damage the structure of HMPSA. The initial adhesion and holding adhesion of DPL-modified HMPSA(DPL-HMPSA) were not significantly changed compared with those of HMPSA, whereas the 180° peel strength was decreased. FI-IR unraveled that DPL formed the n-π conjugated system with styrene-isoprene-styrene block copolymer(SIS), and there was no significant difference in the glass transition temperature according to DSC results, which indicated the good compatibility of DPL with HMPSA. With 5% CVO loaded, the drug content of DPL-HMPSA was 1. 14 times higher than that of HMPSA, and the decrease rate of drug content in DPL-HMPSA was 16% lower than that in HMPSA after 3 months. SEM demonstrated that CVO did not cause obvious structural damage to DPL-HMPSA. Rheological evaluation revealed that the storage modulus and loss factor of DPL-HMPSA were higher than those of HMPSA, and the cohesion was also stronger. The percutaneous penetration rate of cinnamaldehyde in DPL-HMPSA was 2. 25 times that of HMPSA. In conclusion, DPL-HMPSA had more stable structure, better compatibility with CVO, and higher in vitro transdermal efficiency of cinnamaldehyde than before the modification. This study can provide reference for the mitigation of the matrix structure damage caused by volatile oil components in TCM plasters and the enhancement of the content and in vitro transdermal rate of drug.
Adhesives
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Administration, Cutaneous
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Cinnamomum zeylanicum
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Oils, Volatile
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Spectroscopy, Fourier Transform Infrared
9.Relationship between Postmortem Interval and FTIR Spectroscopy Changes of the Rat Skin.
Jiao HUANG ; Yuan Yuan ZHOU ; Kai Fei DENG ; Yi Wen LUO ; Qi Ran SUN ; Zhou Ru LI ; Ping HUANG ; Jie ZHANG ; Hong Xing CAI
Journal of Forensic Medicine 2020;36(2):187-191
Objective To infer postmortem interval (PMI) based on spectral changes of the dorsal skin of rats within 15 days postmortem using Fourier transform infrared (FTIR) spectroscopy. Methods The rats were sacrificed by cervical dislocation after anesthesia, and then placed at 25 ℃ and relative humidity of 50%. The FTIR spectral data collected from the dorsal skin at PMI points were modeled with machine learning technique. Results There was no significant difference of absorption peak location among all the PMI groups but their peak intensities changed as a function of PMIs. The model for PMI estimation was constructed using partial least squares (PLS) regression, reaching a R2 of 0.92 and a root mean square error (RMSE) of 1.30 d. As shown in variable importance for projection (VIP), four spectral bands including 1 760-1 700 cm-1, 1 660-1 640 cm-1, 1 580-1 540 cm-1 and 1 460-1 420 cm-1 were determined as important contributions to model prediction. Conclusion Application of the FTIR technique to detect postmortem spectral changes of the rat skin provides a novel proposal for PMI estimation.
Animals
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Autopsy
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Postmortem Changes
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Rats
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Spectroscopy, Fourier Transform Infrared
10.Surface characterization of calcium phosphate coating formed on chitosan and alkali-treaDted titanium metal
Kyung Hee PARK ; Woon Young LEE ; Ho Ju SONG ; Yeong Joon PARK
Korean Journal of Dental Materials 2019;46(1):33-42
The calcium phosphate coating on various pretreated metals was prepared by soaking in modified simulated body fluid (m-SBF) solution. The coating structure and its surface morphologies were determined by x-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy. The results revealed significant differences in morphology and composition of the calcium phosphate coatings with and without chitosan and NaOH-pretreated commercially pure titanium (cp-Ti) substrate. The calcium phosphates formed on chitosan coated-Ti pretreated with NaOH were ~ 350 nm-sized resulting in strong bonding of the apatite layer with the substrates and a uniform gradient of stress transfer from coating materials to the Ti-substrate. After NaOH pretreatment, the hydroxyl groups bind to Ca²⁺ to attract PO₄³⁻ anions, eventually resulting in a continuous layer of calcium phosphate on chitosan coated-Ti substrate during immersion in m-SBF solution. The chitosan coated-Ti showed hydrophobic surface while NaOH pretreatment resulted in maximum hydrophilicity to the Ti substrate. Due to improved wettability of Ti by NaOH pretreatment before chitosan coating, aggregation of calcium phosphate was prevented and size-controlled composite materials were obtained.
Anions
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Body Fluids
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Calcium Phosphates
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Calcium
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Chitosan
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Clothing
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Hydrophobic and Hydrophilic Interactions
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Immersion
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Metals
;
Microscopy, Electron, Scanning
;
Spectroscopy, Fourier Transform Infrared
;
Titanium
;
Wettability
;
X-Ray Diffraction

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