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CN1241879C - Method for forming osteolith class layer on surface of calcium phosphate ceramics - Google Patents

Method for forming osteolith class layer on surface of calcium phosphate ceramics Download PDF

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Publication number
CN1241879C
CN1241879C CN 200410033613 CN200410033613A CN1241879C CN 1241879 C CN1241879 C CN 1241879C CN 200410033613 CN200410033613 CN 200410033613 CN 200410033613 A CN200410033613 A CN 200410033613A CN 1241879 C CN1241879 C CN 1241879C
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calcium phosphate
phosphate ceramic
gram
body fluid
simulated body
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CN 200410033613
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CN1562894A (en
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田杰谟
王晨
董利民
李兆新
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Tsinghua University
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Tsinghua University
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Abstract

The present invention relates to a method for forming a bone-like apatite layer on the surface of tricalcium phosphate ceramics, which belongs to the technical field of processing activity on the surface of the tricalcium phosphate ceramics. The method comprises the following steps: soaking the tricalcium phosphate ceramics with 1 to 12 mol sodium hydroxide solutions; soaking with simulated body fluid with different concentration; forming the bone-like apatite layer on the surface. The present invention has the advantages of low technology cost, good bioactivity on the surface of the tricalcium phosphate ceramics, and can be used as support materials of artificial bones and bone tissue engineering in clinical medicine.

Description

The calcium phosphate ceramic surface forms the method for bone like apatite layer
Technical field
The invention belongs to calcium phosphate ceramic surface-active-treatment technical field, particularly a kind of method that makes the calcium phosphate ceramic surface form bone like apatite layer.
Background technology
The inorganic components of the chemical constitution of calcium phosphate ceramic and human body bone is close, has excellent biological compatibility, is the very important bone impairment renovation material of a class.The preparation method of calcium phosphate ceramic mainly contains synthetic method, coral conversion method and nature bone calcination method etc., all to handle in the method for preparing process through the high temperature sintering about 1000 ℃, thereby the biological activity of calcium phosphate ceramic is reduced, implant as medical material, influenced cell and adhesion, propagation and the growth organized thereon.Activity is carried out on the calcium phosphate biological ceramic surface handle, make its surface form the bone like apatite layer close with inorganic components with the human body bone structure, the application in clinical medicine has important practical value for calcium phosphate ceramic.At present, at " biomedical engineering magazine " 2002 the 19th volumes the 2nd phase 186-190 page or leaf, Duan Yourong etc. have delivered exercise question and have been " the influence research that fine and close CaP ceramic surface pattern forms bone like apatite layer in dynamic simulation body fluid ", in this article, reported calcium phosphate ceramic has been soaked, formed the method for bone like apatite layer on the calcium phosphate ceramic surface with flow simulating body fluid.The device that this method is used is comparatively complicated, technology cost height; The speed that forms osteoid apatite on the calcium phosphate ceramic surface is slow, and quantity is few, and osteoid apatite is inhomogeneous in the calcium phosphate ceramic surface arrangement, has influenced the practical application of calcium phosphate ceramic in clinical medicine.
Summary of the invention
The objective of the invention is in order to overcome the deficiency that above-mentioned calcium phosphate ceramic surface forms the osteoid apatite layer method, provide a kind of calcium phosphate ceramic to form the method for osteoid apatite, before calcium phosphate ceramic is soaked with simulated body fluid, earlier with the aqueous sodium hydroxide solution of 1~12 mole of different concns to the calcium phosphate ceramic surface preparation, and then, make the calcium phosphate ceramic surface form bone like apatite layer through the simulated body fluid immersion treatment.
Technology of the present invention may further comprise the steps:
(1) calcium phosphate ceramic is cleaned up in deionized water or distilled water, in 80~100 ℃ of oven dry;
(2) use analytical pure sodium hydroxide reagent and deionized water or distilled water to prepare the aqueous sodium hydroxide solution of 1~12 volumetric molar concentration;
(3) in 1000 ml deionized water or distilled water, add sodium-chlor (NaCl) 8.00~24.00 grams, sodium bicarbonate (NaHCO 3) 0.35~1.01 gram, Repone K (KCl) 0.23~0.69 gram, dipotassium hydrogen phosphate (K 2HPO 43H 2O) 0.23~0.69 gram, magnesium chloride (MgCl 26H 2O) 0.31~0.93 gram, calcium chloride (CaCl 22H 2O) 0.28g~0.84 gram, sodium sulfate (Na 2SO 4) 0.07~0.21 gram, Tutofusin tris ((CH 2OH) 3CNH 2) 6.06~18.18 grams, prepare simulated body fluid, and the pH value of simulated body fluid is transferred to 7.2~7.4 with the hydrochloric acid soln of 1 volumetric molar concentration;
(4) the cleaned calcium phosphate ceramic of step (1) is placed Glass Containers, add the aqueous sodium hydroxide solution of 1~12 volumetric molar concentration of step (2) preparation, and calcium phosphate ceramic is fully immersed in the aqueous sodium hydroxide solution, normal temperature was placed 2~6 days down;
(5) with the calcium phosphate ceramic after step (4) immersion treatment, clean repeatedly, after the pH value of scavenging solution is 6.5~7.5, in 80~100 ℃ of oven dry with deionized water or distilled water;
(6) calcium phosphate ceramic after step (5) cleaning, the oven dry is placed Glass Containers, the simulated body fluid that adds step (3) preparation, and calcium phosphate ceramic is fully immersed in the simulated body fluid, 36~38 ℃ of temperature, under the condition of humidity 60~90%, immersion treatment 2~20 days, during every 24~48 hours, remove old liquid, change the fresh simulated body fluid of same concentrations;
(7) calcium phosphate ceramic after step (6) processing is taken out, clean up with deionized water or distilled water, in 80~100 ℃ of oven dry.
Characteristics of the present invention:
Treatment process is simple, and cost is low.
Adopt aqueous sodium hydroxide solution that calcium phosphate ceramic is soaked pre-treatment, and then use the simulated body fluid immersion treatment, because the calcium phosphate ceramic surface has great amount of hydroxy group, make calcium ion, phosphate anions etc. are adsorbed on the calcium phosphate ceramic surface easily, thereby make osteoid apatite fast in its surperficial sedimentation velocity, quantity is many, be evenly distributed, the surfactivity height of calcium phosphate ceramic implants this calcium phosphate ceramic, is convenient to osteocyte and new organization in the calcium phosphate ceramic surface adhesion, propagation and growth, also help calcium phosphate ceramic implant back and the combining of body osseous tissue, the growth of promotion new bone tissue and from the reconstruction of body bone.
Calcium phosphate ceramic after the present invention handles is mainly used in damaged reparation of bone and bone tissue engineering stent material, also can use as the tooth dental repair material.
Embodiment
Embodiment 1
(1) porous calcium phosphate ceramic with the preparation of synthetic method cleans up in deionized water or distilled water, in 80 ℃ of oven dry;
(2) prepare the aqueous sodium hydroxide solution of 12 volumetric molar concentrations with analytical pure sodium hydroxide reagent and deionized water or distilled water;
(3) in 1000 ml deionized water or distilled water, add sodium-chlor (NaCl) 24.00 grams, sodium bicarbonate (NaHCO 3) 1.01 grams, Repone K (KCl) 0.69 gram, dipotassium hydrogen phosphate (K 2HPO 43H 2O) 0.69 gram, magnesium chloride (MgCl 26H 2O) 0.93 gram, calcium chloride (CaCl 22H 2O) 0.84 gram, sodium sulfate (Na 2SO 4) 0.21 gram, Tutofusin tris ((CH 2OH) 3CNH 2) 18.18 grams, prepare simulated body fluid, and the pH value of simulated body fluid is transferred to 7.2~7.4, the preparation simulated body fluid with the hydrochloric acid soln of 1 volumetric molar concentration;
(4) the cleaned calcium phosphate ceramic of step (1) is placed Glass Containers, add the aqueous sodium hydroxide solution of 12 volumetric molar concentrations of step (2) preparation, and calcium phosphate ceramic is fully immersed in the aqueous sodium hydroxide solution, normal temperature was placed 6 days down;
(5) with the calcium phosphate ceramic after step (4) immersion treatment, clean repeatedly, after the pH value of scavenging solution is 6.5~7.5, in 80 ℃ of oven dry with deionized water or distilled water;
(6) calcium phosphate ceramic after step (5) cleaning, the oven dry is placed Glass Containers, the simulated body fluid that adds step (3) preparation, and calcium phosphate ceramic is fully immersed in the simulated body fluid, be placed on 36~37 ℃ of temperature, in the thermostat container of humidity 60~70%, immersion treatment 18 days, during every 48 hours, remove old liquid, change the fresh simulated body fluid of same concentrations;
(7) set by step after handle (6), calcium phosphate ceramic is taken out, clean up,,, show on the calcium phosphate ceramic surface to have formed bone like apatite layer by scanning electron microscopic observation and Infrared spectroscopy in 80 ℃ of oven dry with deionized water or distilled water.
Embodiment 2
(1) porous calcium phosphate ceramic with the preparation of coral conversion method cleans up in deionized water or distilled water, in 90 ℃ of oven dry;
(2) prepare the aqueous sodium hydroxide solution of 8 volumetric molar concentrations with analytical pure sodium hydroxide reagent and deionized water or distilled water;
(3) in 1000 ml deionized water or distilled water, add sodium-chlor (NaCl) 12.00 grams, sodium bicarbonate (NaHCO 3) 0.525 gram, Repone K (KCl) 0.345 gram, dipotassium hydrogen phosphate (K 2HPO 43H 2O) 0.345 gram, magnesium chloride (MgCl 26H 2O) 0.465 gram, calcium chloride (CaCl 22H 2O) 0.42 gram, sodium sulfate (Na 2SO 4) 0.105 gram, Tutofusin tris ((CH 2OH) 3CNH 2) 9.09 grams, prepare simulated body fluid, and the pH value of simulated body fluid is transferred to 7.2~7.4 with the hydrochloric acid soln of 1 volumetric molar concentration; The preparation simulated body fluid;
(4) the cleaned calcium phosphate ceramic of step (1) is placed Glass Containers, add the aqueous sodium hydroxide solution of 8 volumetric molar concentrations of step (2) preparation, and calcium phosphate ceramic is fully immersed in the aqueous sodium hydroxide solution, normal temperature was placed 4 days down;
(5) with the calcium phosphate ceramic after step (4) immersion treatment, clean repeatedly, after the pH value of scavenging solution is 6.5~7.5, in 90 ℃ of oven dry with deionized water or distilled water;
(6) with the calcium phosphate ceramic after step (5) cleaning, the oven dry, place Glass Containers, the simulated body fluid that adds step (3) preparation, and calcium phosphate ceramic is fully immersed in the simulated body fluid, be placed on 36~37 ℃ of temperature, in the thermostat container of humidity 70~80%, immersion treatment 10 days, every 36 hours, remove old liquid during this time, change the fresh simulated body fluid of same concentrations;
(7) set by step after handle (6), calcium phosphate ceramic is taken out, clean up,,, show on the calcium phosphate ceramic surface to have formed bone like apatite layer by scanning electron microscopic observation and Infrared spectroscopy in 90 ℃ of oven dry with deionized water or distilled water.
Embodiment 3
(1) porous calcium phosphate ceramic with the preparation of nature bone calcination method cleans up in deionized water or distilled water, in 100 ℃ of oven dry;
(2) prepare the aqueous sodium hydroxide solution of 1 volumetric molar concentration with analytical pure sodium hydroxide reagent and deionized water or distilled water;
(3) in 1000 ml deionized water or distilled water, add sodium-chlor (NaCl) 8.00 grams, sodium bicarbonate (NaHCO 3) 0.35 gram, Repone K (KCl) 0.23 gram, dipotassium hydrogen phosphate (K 2HPO 43H 2O) 0.23 gram, magnesium chloride (MgCl 26H 2O) 0.31 gram, calcium chloride (CaCl 22H 2O) 0.28g gram, sodium sulfate (Na 2SO 4) 0.07 gram, Tutofusin tris ((CH 2OH) 3CNH 2) 6.06 grams, prepare simulated body fluid, and the pH value of simulated body fluid is transferred to 7.2~7.4, the preparation simulated body fluid with the hydrochloric acid soln of 1 volumetric molar concentration;
(4) the cleaned calcium phosphate ceramic of step (1) is placed Glass Containers, add the aqueous sodium hydroxide solution of 1 volumetric molar concentration of step (2) preparation, and calcium phosphate ceramic is fully immersed in the aqueous sodium hydroxide solution, normal temperature was placed 2 days down;
(5) with the calcium phosphate ceramic after step (4) immersion treatment, clean repeatedly, after the pH value of scavenging solution is 6.5~7.5, in 100 ℃ of oven dry with deionized water or distilled water;
(6) the calcium phosphate ceramic material after step (5) cleaning, the oven dry is placed Glass Containers, the simulated body fluid that adds step (3) preparation, and calcium phosphate ceramic is fully immersed in the simulated body fluid, be placed on 36~37 ℃ of temperature, in the thermostat container of humidity 80~90%, immersion treatment 3 days, during every 24 hours, remove old liquid, change the fresh simulated body fluid of same concentrations;
(7) set by step after handle (6), calcium phosphate ceramic is taken out, clean up,,, show on the calcium phosphate ceramic surface to have formed bone like apatite layer by scanning electron microscopic observation and Infrared spectroscopy in 100 ℃ of oven dry with deionized water or distilled water.
Scleroblast is inoculated in the calcium phosphate ceramic of resulting surface generation bone like apatite layer among above-mentioned three embodiment, vitro culture 7 days, competence for added value with the mtt assay test cell, vitro culture 14 days, measure alkaline phosphatase activities (ALP), the result is as shown in table 1, this shows, the competence for added value of cell and alkaline phosphatase activities all than under the similarity condition without the present invention handle to calcium phosphate ceramic height in the same old way.
The MTT absorbance of the compound cultivation of table 1 calcium phosphate ceramic/scleroblast and ALP value
Embodiment 1 Embodiment 2 Embodiment 3 To in the same old way
Mtt assay records absorbance 1.245 0.823 2.014 0.508
Alkaline phosphatase (ALP) (U/L) 16.451 15.523 18.264 10.825

Claims (1)

1, a kind of method of calcium phosphate ceramic surface formation bone like apatite layer, technology may further comprise the steps:
(1) calcium phosphate ceramic is cleaned up in deionized water or distilled water, in 80~100 ℃ of oven dry;
(2) use analytical pure sodium hydroxide reagent and deionized water or distilled water to prepare the aqueous sodium hydroxide solution of 1~12 volumetric molar concentration;
(3) in 1000 ml deionized water or distilled water, add 8.00~24.00 gram NaCl, 0.35~1.01 gram NaHCO 3, 0.23~0.69 gram KCl, 0.23~0.69 gram K 2HPO 43H 2O, 0.31~0.93 gram MgCl 26H 2O, 0.28g~0.84 gram CaCl 22H 2O, 0.07~0.21 gram Na 2SO 4, 6.06~18.18 gram (CH 2OH) 3CNH 2, prepare simulated body fluid, and the pH value of simulated body fluid be transferred to 7.2~7.4 with the hydrochloric acid soln of 1 volumetric molar concentration;
(4) the cleaned calcium phosphate ceramic of step (1) is placed Glass Containers, add the aqueous sodium hydroxide solution of 1~12 volumetric molar concentration of step (2) preparation, and calcium phosphate ceramic is fully immersed in the aqueous sodium hydroxide solution, normal temperature was placed 2~6 days down;
(5) with the calcium phosphate ceramic after step (4) immersion treatment, clean repeatedly, after the pH value of scavenging solution is 6.5~7.5, in 80~100 ℃ of oven dry with deionized water or distilled water;
(6) calcium phosphate ceramic after step (5) cleaning, the oven dry is placed Glass Containers, the simulated body fluid that adds step (3) preparation, and calcium phosphate ceramic is fully immersed in the simulated body fluid, 36~38 ℃ of temperature, under the condition of humidity 60~90%, immersion treatment 2~20 days, during every 24~48 hours, remove old liquid, change the fresh simulated body fluid of same concentrations;
(7) calcium phosphate ceramic after step (6) processing is taken out, clean up with deionized water or distillation, in 80~100 ℃ of oven dry.
CN 200410033613 2004-04-13 2004-04-13 Method for forming osteolith class layer on surface of calcium phosphate ceramics Expired - Fee Related CN1241879C (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
PT2445543E (en) * 2009-06-23 2013-04-03 Geistlich Pharma Ag Bone substitute material
CN103341206B (en) * 2013-06-09 2014-11-19 四川大学 Calcium phosphate/collagen/bone-like apatite three-level bionic bone tissue engineering scaffold and preparation method thereof
CN107050508B (en) * 2017-01-19 2020-12-18 深圳清华大学研究院 Injectable bone repair material and preparation method thereof
CN107141022B (en) * 2017-04-14 2020-08-18 华南理工大学 Whitlockite coating constructed on surface of calcium phosphate biological ceramic matrix and preparation method thereof
CN107397977B (en) * 2017-08-03 2021-01-26 广东工业大学 3D printing metal matrix surface modification method, 3D printing metal matrix biological ceramic support and preparation method thereof

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