US 12,453,801 B2
Nucleic acid-calcium phosphate nanoparticle complexes and application thereof in biomineralization
Lina Niu, Xi'an (CN); Jihua Chen, Xi'an (CN); Kia Jiao, Xi'an (CN); Minjuan Shen, Xi' an (CN); Yuxuan Ma, Xi'an (CN); Jianfei Yan, Xi'an (CN); Qianqian Wan, Xi'an (CN); and Jing Li, Xi'an (CN)
Assigned to The Fourth Military Medical University of Chinese People's Liberation Army, Xi' (CN)
Appl. No. 17/632,064
Filed by The Fourth Military Medical University of Chinese People's Liberation Army, Xi'an (CN)
PCT Filed Aug. 7, 2020, PCT No. PCT/CN2020/108009
§ 371(c)(1), (2) Date Feb. 1, 2022,
PCT Pub. No. WO2021/196490, PCT Pub. Date Oct. 7, 2021.
Claims priority of application No. 202010246314.5 (CN), filed on Mar. 31, 2020.
Prior Publication US 2023/0372585 A1, Nov. 23, 2023
Int. Cl. A61L 27/24 (2006.01); A61L 27/12 (2006.01); A61L 27/46 (2006.01); A61L 27/54 (2006.01); C12N 15/87 (2006.01)
CPC A61L 27/24 (2013.01) [A61L 27/12 (2013.01); A61L 27/46 (2013.01); A61L 27/54 (2013.01); C12N 15/87 (2013.01); A61L 2300/258 (2013.01); A61L 2300/412 (2013.01); A61L 2300/624 (2013.01); A61L 2400/12 (2013.01); A61L 2430/02 (2013.01); A61L 2430/12 (2013.01)] 19 Claims
 
1. A method producing a biological mineralizer for the mineralization of collagen fiber products and for preparing hydroxyapatite-collagen organic-inorganic composite materials,
the method comprising the forming of a complex by the electrostatic adsorption of nucleic acid and amorphous calcium phosphate nanoparticles in solution comprising the steps in the following sequential order of:
1) mixing a solution containing nucleic acids with a solution of calcium ions so that there is electrostatic adsorption between the phosphate groups of the nucleic acid and the calcium ions and
2) adding to the solution formed in step 1) a solution of potassium ions so that there is electrostatic adsorption of the free phosphate ions with the calcium ions adsorbed on the nucleic acid, thereby forming the complex of nucleic acid and amorphous calcium phosphate nanoparticles in solution,
wherein the concentration of calcium ions in the biological mineralizer is 2-5 mM, and the concentration of phosphate ions in the biological mineralizer is 1-5 mM,
wherein the ratio of the calcium ions to the phosphate ions in the biological mineralizer is from 2:1 to 1:1,
wherein the nucleic acid is total DNA or total RNA isolated from mammalian cells,
wherein the final working concentration of DNA is 70-250 μg/ml, or the final working concentration of RNA is 150-250 μg/ml, and
wherein the particle size of the biological mineralizer complex is 40-100 nm.