Summary. In the present work, a study of H+ effects on the microstructure and morphology of hydroxyapatite synthesized by the hydrothermal method is reported. The synthesis was realized with pH values of 10, 9.6, 9, 8, and 7. To know the particle size distribution, growth habit, chemical composition, and crystalline phases present, SEM-EDS, XRD, AFM, and FTIR spectroscopy techniques were used and completed with Rietveld analysis. The obtained results showed an important effect of H+ on the morphological and crystallographic characteristics of the hydroxyapatite, demonstrating that the shape and nanoparticle size, as well as the number of crystalline phases, can change depending on the pH during the synthesis. It was observed that there is an increase in the formation of the monoclinic phase and a decrease of the hexagonal phase when the pH value diminishes from 9.6 to 7. The crystallite size also decreases from 46.69 to 19.56 nm. An explanation of the role of pH on the final characteristics of the hydroxyapatite was related to the amount of H+ and OH- ions, and it was included in this work.

Hydroxyapatite (HAp) is a natural hard tissue constituent widely used for bone and tooth replacement engineering. In the present work, synthetic HAp was obtained from calcium nitrate tetrahydrate (Ca(NO3)2·4H2O) and ammonium phosphate dibasic (NH4)2HPO4 following an optimized microwave assisted hydrothermal method. The effect of pH was evaluated by the addition of ammonium hydroxide (NH4OH). Hence, different characterization techniques were used to determine its influence on the resulted HAp powders’ size, shape, and crystallinity. By Transmission Electron Microscopy (TEM), it was observed that the reaction pH environment modifies the morphology of HAp, and a shape evolution, from sub-hedral particles at pH = 7 to rod-like nanosized HAp at pH = 10, was confirmed. Using the X-ray Diffraction (XRD) technique, the characteristic diffraction peaks of the monoclinic phase were identified. Even if the performed Rietveld analysis indicated the presence of both phases (hexagonal and monoclinic), monoclinic HAp prevails in 95% with an average crystallite size of about 23 nm. The infrared spectra (FTIR) showed absorption bands at 3468 cm−1 and 630 cm−1 associated with OH− of hydroxyapatite, and bands at 584 cm−1, 960 cm−1, and 1090 cm−1 that correspond to the PO43− and CO32− characteristic groups. In summary, this work contributes to obtaining nanosized rod-like monoclinic HAp by a simple and soft method that has not been previously reported.

En el presente trabajo se realiza la síntesis de apatitas, a través del método hidrotermal asistido por microondas, variando el pH en 5, 6, 7, 8, 9 y 10. La caracterización fisicoquímica y estructural se realizó por las técnicas:  MEB-EDS, DRX y FTIR, determinando una tamaño de cristal de 8 a 10 nm en función del pH; se observó la formación de aglomerados longitud, constituidos por fibras de entre 43.4  a 108.5 nm de diámetro y 412.4 a 1603.8 nm de longitud, así mismo se determina la presencia de planos cristalográficos característicos de HAp y ClAp . Además, se identifican los modos vibracionales pertenecientes a ambas estructuras y una elongación en la banda del grupo OH-, atribuida a la interacción entre el grupo OH- de la HAp y los iones de Cl- de la ClAp. Los resultados obtenidos contribuyen a establecer el efecto del pH en el tamaño de cristal y de partícula para la síntesis de HAp y ClAp

The present work studies the effect of Mn doping on the crystalline structure of the Hap synthesized by the hydrothermal method at 200 °C for 24 h, from Ca(OH)2 and (NH4)2HPO4, incorporating MnCl2 to 0.1, 0.5, 1.0, 1.5 and 2.0 %wt of Mn concentrations. Samples were characterized by the X-Ray Diffraction technique, which revealed the diffraction peaks that corresponded to the hexagonal and monoclinic phase of the Hap; it was observed that the average size of crystallite decreased from 23.67 to 22.69 nm as the concentration of Mn increased. TEM shows that in all samples, there are two distributions of particle sizes; one corresponds to nanorods with several tens of nanometers in length, and the other in which the diameter and length are very close. FTIR analysis revealed absorption bands corresponding to the PO4−3 and OH− groups characteristic of the Hap. It was possible to establish a substitution mechanism between the Mn and the ions of Ca+2 of the Hap. From the Alamar blue test, a cell viability of 86.88% ± 5 corresponding to the sample of Hap at 1.5 %wt Mn was obtained, considered non-cytotoxic according to ISO 10993-5. It also evaluated and demonstrated the good osteoinductive properties of the materials, which were verified by histology and immunofluorescence expression of osteogenic markers. Adhesion, viability, biocompatibility and osteoinductive properties, make these materials candidates for future applications in bone tissue engineering with likely uses in regenerative medicine. Graphical Abstract