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Synthesis and characterization of rod-like amino acids/nanohydroxyapatite composites to inhibit osteosarcoma.

ABSTRACT: In this study, rod-like hydroxyapatite (HA) with uniform morphology and controllable particle size modified by doping with two different amino acids (alanine and threonine) was synthesized by a microwave hydrothermal method. The physical and chemical properties of the composites were tested by utilizing X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), general thermogravimetric analysis (TG) and scanning electron microscopy (SEM). The SEM and XRD results show that the presence of amino acids (especially threonine) can significantly reduce the aspect ratio and crystallinity of hydroxyapatite. Pure hydroxyapatite and modified hydroxyapatite doped with two different proportions of amino acids were cultured with mouse osteoblasts (MC3T3-E1) for 1, 3 and 5 days, respectively, nanohydroxyapatite modified by threonine has better biocompatibility compared with pure hydroxyapatite. The amino acid-modified hydroxyapatite samples were co-cultured with osteosarcoma cells (MG63) for 1, 4 and 7 days, respectively, and showed better inhibitory effects on osteosarcoma cells. The nanohydroxyapatite doped with amino acids could be used as a potential drug that promotes bone repair and inhibits the growth of osteosarcoma cells.

SUBMITTER: Chen Z

PROVIDER: S-EPMC9756758 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature

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Synthesis and characterization of rod-like amino acids/nanohydroxyapatite composites to inhibit osteosarcoma.

Chen Zhengxiong Z Wang Xinyu X Luo Jing J Zhang Bowen B Shen Fei F Li Binbin B Yang Jing J

RSC advances 20221216 55

In this study, rod-like hydroxyapatite (HA) with uniform morphology and controllable particle size modified by doping with two different amino acids (alanine and threonine) was synthesized by a microwave hydrothermal method. The physical and chemical properties of the composites were tested by utilizing X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), general thermogravimetric analysis (TG) and scanning electron microscopy (SEM). The SEM and XRD results show that the pres ...[more]

PMID: 36545101

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Project description:β-hydroxy amino acids, such as serine, threonine, and phenylserine, are important compounds for medical purposes. To date, there has been only limited exploration of thermostable serine hydroxylmethyltransferase (SHMT) for the synthesis of these amino acids, despite the great potential that thermostable enzymes may offer for commercial use due to their high stability and catalytic efficiencies. ITBSHMT_1 (ITB serine hydroxylmethyltransferase clone number 1) from thermophilic and methanol-tolerant bacteria Pseudoxanthomonas taiwanensis AL17 was successfully cloned. Biocomputational analysis revealed that ITBSHMT_1 contains Pyridoxal-3'-phosphate and tetrahydrofolatebinding residues. Structural comparisons show that ITBSHMT_1 has 5 additional residues VSRQG on loop near PLP-binding site as novel structural feature which distinguish this enzyme with other characterized SHMTs. In silico mutation revealed that the fragment might have very essential role in maintaining of PLP binding on structure of ITBSHMT_1. Recombinant protein was produced in Escherichia coli Rosetta 2(DE3) in soluble form and purified using NiNTA affinity chromatography. The purified protein demonstrated the best activity at 80 °C and pH 7.5 based on the retro aldol cleavage of phenylserine. Activity decreased significantly in the presence of 3 mM transition metal ions but increased in the presence of 30 mM β-mercaptoethanol. ITBSHMT_1 demonstrated Vmax, Km, Kcat, and Kcat/Km at 242 U/mg, 23.26 mM, 186/s, and 8/(mM.s), respectively. The aldol condensation reaction showed the enzyme's best activity at 80 °C for serine, threonine, or phenylserine, with serine synthesis showing the highest specific activity. Biocomputational analysis revealed that high intramolecular interaction within the 3D structure of ITBSHMT_1 might be correlated with the enzyme's high thermal stability. The above data suggest that ITBSHMT_1 is a potential and novel enzyme for the production of various β-hydroxy amino acids.

Project description:ObjectivesThis work aimed to design, synthesize and characterize replacement natural moisturizing factor (NMF) composed of modified hygroscopic linear amino acids to pre-empt or repair skin barrier dysfunction.MethodsFollowing synthesis and characterization, thermo-gravimetric analysis and quantum mechanics molecular modelling quantified and depicted water binding to the new compounds. Deliquescence relative humidity demonstrated the water-scavenging ability of the compounds, whereas snake skin moisturizing studies showed they increased water uptake into snake skin.ResultsFrom thermal analysis, N-hydroxyglycine showed greatest water-holding capacity followed by N-hydroxyserine, l-homoserine and α-hydroxyglycine; coupled with quantum mechanics molecular modelling, between 8 and 12 molecules of water could associate with each molecule of either N-hydroxyglycine, N-hydroxyserine or l-homoserine. All of our modified amino acids were efficacious and induced similar or greater water uptake compared with the established moisturizing compounds hyaluronic acid, glycerine and urea in snake skin. Incorporated at 10% in Oilatum, N-hydroxyserine induced >200% greater moisture uptake into dry snake skin compared to treatment with water alone, with efficacy related to the molecule structure and ability to bind to 12 water molecules. Oilatum cream spiked with all our unnatural amino acid hydrotropes increased water uptake into snake skin compared with Oilatum alone. The compound series was designed to elucidate some structure - efficacy relationships. Amino acid chirality did not affect the water-holding capacity but did affect uptake into skin. Compounds with high melting points and bond energies tended to decrease water-holding capacity. With isosteric replacement, the more electronegative atoms gave greater water-holding capacities.ConclusionsThis work demonstrates the potential of unnatural amino acid hydrotropes as skin moisturizers and has developed some predictive 'rules' for further design and refinement of chemical structures.

Dataset Information

Synthesis and characterization of rod-like amino acids/nanohydroxyapatite composites to inhibit osteosarcoma.

Publications

Synthesis and characterization of rod-like amino acids/nanohydroxyapatite composites to inhibit osteosarcoma.

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