CN112479735A - CaSO-containing food4beta-TCP composite ceramic material, preparation method and application - Google Patents
CaSO-containing food4beta-TCP composite ceramic material, preparation method and application Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title claims abstract description 13
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- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 claims abstract description 65
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- AMVQGJHFDJVOOB-UHFFFAOYSA-H aluminium sulfate octadecahydrate Chemical compound O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.O.[Al+3].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O AMVQGJHFDJVOOB-UHFFFAOYSA-H 0.000 claims abstract description 16
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Abstract
The invention discloses a catalyst containing CaSO4beta-TCP composite ceramic material, its preparation method and application, said material is formed from beta-TCP ceramic and CaSO4·2H2The mass ratio of the O is 1: 5-11, wherein the mass ratio of the beta-tricalcium phosphate to the hydroxyapatite in the beta-TCP ceramic is 7: 3. The method comprises grinding and sieving aluminum sulfate octadecahydrate powder, mixing with calcium sulfate dihydrate, placing in a pressure cooker, heating, pressurizing, maintaining pressure, and drying to obtain alpha-calcium sulfate hemihydrate, and mixing calcium sulfate dihydrate with doping material to obtain CaSO4·2H2The composite ceramic material powder consists of O and beta-TCP ceramic. The application comprises preparing the powder of the composite ceramic material into a porous calcium sulfate/beta-TCP biological scaffold; the biological stent is implanted into a human body. The bone repair material prepared by the invention has good biocompatibility and biodegradability, and has good application prospects in the fields of local administration, tooth repair and the like.
Description
Technical Field
The invention relates to the technical field of preparation of medical materials, in particular to a medical material containing CaSO4And beta-TCP, a preparation method and application thereof.
Background
Calcium sulfate has been used as a filler material in bone defect repair for centuries, but since previous methods of preparation have been limited by a number of factors, calcium sulfate has not been sufficiently pure and has been limited in medical use.
Due to scientific progress and development of medical conditions, calcium sulfate is increasingly widely used in medical treatment. Calcium sulfate is used in various medical treatments at present, and good achievement is achieved.
In general, the mechanism of calcium sulfate osteogenesis can be divided into two areas:
(1) calcium sulfate is a bone conduction substance, has a plurality of functions, and can be filled in the defect position firstly; secondly, the shape of the damaged bone can be restored; finally, the soft tissue can be prevented from growing up and entering new bones. The degradation and the supporting effect can be simultaneously carried out in the degradation process of the calcium sulfate.
(2) The calcium sulfate has bone induction activity, and the mechanism is that a slightly acidic environment is locally formed in the degradation and absorption process of the calcium sulfate to cause local decalcification and release of bone matrix osteogenesis induction factors. Calcium sulfate is continuously studied because of its irreplaceable mechanical properties and biological activity to form various medical materials based on calcium sulfate, and these composite materials also have a surprising effect in treating bone defects.
Calcium sulfate particles are widely applied to orthopedics in medical treatment, and mainly because the medical calcium sulfate particles have bone induction activity, a slightly acidic environment is locally formed in the degradation and absorption process of the medical calcium sulfate particles, so that local decalcification and release of bone matrix osteogenesis induction factors are caused, and further osteoblasts are promoted to proliferate and differentiate. Calcium sulfate has these advantages, calcium phosphate is incomparable, so calcium sulfate is immaterial as an important material for the treatment of bone defects. Research finds that the degradation and absorption curve of calcium sulfate is close to the bone growth curve, and moreover, the effect of the calcium sulfate on repairing bone defects by self bones is greatly different. The calcium sulfate can be used alone, can be compounded with autologous bone to enhance the osteogenesis performance when repairing massive defects, and can be compounded with antibiotics to treat infectious bone defects. The calcium sulfate can be made into a bracket, and can also be made into a bracket together with some materials with good bioactivity and good compatibility with bones to repair bone defects. In the research of bone tissue engineering, calcium-based inorganic materials such as phosphorus, sulfur, calcium silicate and the like are main scaffolds or basic scaffolds for bone tissue engineering. Many materials, which are not known to exist toxicity and harm to human body because of the lack of knowledge, calcium sulfate is a material which is not toxic to human body. In vitro cell culture studies prove that calcium sulfate has no toxic effect on osteoblasts, is beneficial to osteoblast adhesion, and can stimulate the proliferation and differentiation of osteoblasts. Calcium sulfate is so suitable for bone defect treatment and these features are not separable per se. At present, the application of calcium sulfate is very wide, and in the front of various technologies, people are exploring the improvement of calcium sulfate in various fields again and again, continuously modifying and researching the calcium sulfate, and achieving surprising results.
The most commonly used biphase calcium phosphate at present is a beta-TCP/HA biphase composite material with different proportions, which is taken as an important bone tissue engineering scaffold material and HAs better controllable degradation performance and osteogenic performance than a single calcium phosphate ceramic material, thereby receiving wide attention of people. The composite material combines the advantages of the beta-TCP and the HA of the beta-TCP, so that the easy absorbability of the beta-TCP and the bracket effect of the HA generate a synergistic effect, and simultaneously, the crystal microstructure of the material can be regulated and controlled by changing the Ca/P ratio of the beta-TCP/HA two-phase composite material, so that the osteogenic property of the material is greatly improved. Researches show that the biphasic calcium phosphate ceramic scaffold with a three-dimensional porous structure can enable the scaffold material to have osteoinductivity under certain conditions by adjusting the proportion of HAP/beta-TCP.
Wepen et al use the electrostatic spinning technology to prepare n-HAP/beta-TCP stent material, through scanning results analysis, the composite material has a net structure, through finding with osteoblast coculture, the material can promote cell adhesion and proliferation, has good biocompatibility. Reddy and the like directly mix n-HAP and beta-TCP in polyvinyl alcohol (PVA) according to different proportions to prepare an n-HAP/beta-TCP composite material, screen a Ca/P ratio with the best bone forming effect, and verify that the composite material has good bone forming performance through an in vivo implantation experiment. Ninie Wei and He Wei prepare the nano HAP/beta-TCP dual-phase ceramic biological scaffold material, implant the nano HAP/beta-TCP dual-phase ceramic biological scaffold material into a rabbit left-side radius defect model, and compare the nano HAP/beta-TCP dual-phase ceramic biological scaffold material with a single nano HAP artificial bone, and find that the nano HAP/beta-TCP dual-phase ceramic biological scaffold material has better osteogenesis performance and biodegradability.
Studies have shown that bone formation in the pores is favored only when the intercommunicating pore size is greater than 50um, and that as the intercommunicating pore size increases, the conductive properties of the material increase. However, mechanical property tests of the material show that the mechanical strength of the porous material is reduced by increasing the porosity and the pore diameter, and the mechanical strength, the pore diameter and the porosity of the porous material are greatly influenced by the preparation method. Therefore, extensive research has been conducted to develop a series of methods for preparing porous materials, and the pore-forming principle can be classified into pore-forming agent method, organic foam impregnation method, chemical foaming method, etc. Wherein, the porosity of the porous material product prepared by the pore-forming agent method is not high, generally lower than 50%, and the pore distribution is not uniform. The material prepared by the organic foam impregnation method has high porosity (70-95%), has a three-dimensional open-cell structure, is convenient to operate, has a simple preparation process, and is widely applied to preparation of porous materials. However, the porous material prepared by the foam impregnation method has low mechanical strength due to too high porosity. The chemical foaming method can prepare porous ceramics with different shapes and different pore diameters by controlling the type and the dosage of the foaming agent, but most of the pores of the porous material prepared by the method are closed, and the pore penetration rate is poor. The freeze-drying method is simple and convenient to operate, the equipment is simple, the porosity is up to more than 90%, the influence of high temperature on the activity of the biological material is effectively avoided in the preparation process, and the introduction of bioactive factors can be facilitated. However, an organic solvent is often required to be added in the freeze drying process, the preparation period of the method is long, the energy consumption is high, and meanwhile, the low temperature needs to be strictly controlled to avoid the generation of a dense layer on the surface of the material and even cause the collapse of the porous scaffold.
Disclosure of Invention
In view of the deficiencies of the prior art, one aspect of the present invention provides a CaSO-containing composition4And beta-TCP, and aims to provide an osteoinductive material which has good biological property and mechanical property and adjustable degradation rate. The material takes an organism as a reactor, does not add cell nucleus growth factors, constructs a bone graft with a certain volume in a bone or non-bone environment without influencing the function of the organism, and repairs the autologous bone defect technology. The method specifically comprises the following steps:
CaSO-containing food4And beta-TCP, which is made of CaSO4·2H2O and BCP (β -TCP ceramic, i.e. β -tricalcium phosphate and hydroxyapatite), wherein: beta-TCP ceramics and CaSO4·2H2The mass ratio of the O is 1: 5-11, and the mass ratio of the beta-tricalcium phosphate to the hydroxyapatite in the BCP is 7: 3.
Hair brushAnother aspect of the invention provides a composition comprising CaSO4And beta-TCP, comprising the following steps:
(1) taking aluminum sulfate octadecahydrate powder, fully grinding and sieving the powder, and weighing 0.45g of the powder;
(2) weighing calcium sulfate dihydrate powder, fully and uniformly mixing the calcium sulfate dihydrate powder with the weighed aluminum sulfate octadecahydrate, and filling the mixture into a 150ml beaker;
(3) buckling filter paper on a beaker filled with calcium sulfate by using weighing paper, sleeving a rubber band on the beaker, and placing the beaker in a pressure cooker;
(4) controlling the voltage of the pressure cooker to be kept between 0.14 and 0.15MPa, keeping the pressure for 10 hours and then taking out;
(5) drying at 80 deg.C for 12h to obtain alpha-calcium sulfate hemihydrate, and packaging in a sealed bag.
(6) And (3) fully and uniformly mixing the calcium sulfate dihydrate powder and the doping material according to a certain mass ratio, filling the mixture into a beaker, and repeating the steps 4 and 5 for a plurality of times.
(7) And filling the powder into a 50-hole plate, sealing the opening in a 60-DEG C oven, hydrating for 24h, and opening and drying for 24h to finally obtain the porous calcium sulfate/beta-TCP biological scaffold.
Another aspect of the present invention provides a method for forming a bone graft having mechanical properties similar to those of natural bone by in vivo bone tissue engineering, comprising4And β -TCP as a bone graft, comprising:
calcium Sulfate (CS) is an inorganic ceramic material, and has good biocompatibility and degradation properties, as well as certain osteogenic and osteoinductive activities. The osteoinduction is mainly due to the fact that mitosis of cells is promoted after calcium ions are combined with calcium sensitive receptors, and then bone marrow mesenchymal stem cells are induced to gradually differentiate to osteoblasts. Beta-tricalcium phosphate (beta-TCP) is similar to natural inorganic composition of human bone, has biocompatibility superior to other inorganic materials, is well jointed with matrix bone after being implanted, and has excellent bone conductivity; the material is gradually discharged out of the body through the process of dissolution and absorption or metabolism after being implanted into a human body, and finally, the bone defect part is replaced by new bone tissues; meanwhile, one part of the beta-tricalcium phosphate is discharged out of the body through biochemical reaction, the other part of the beta-tricalcium phosphate participates in the formation of new bones, and degraded calcium and phosphorus ions can enter a human body circulatory system and gradually form new bones, so the beta-tricalcium phosphate becomes a potential tissue engineering scaffold material with excellent comprehensive performance and is used for constructing tissue engineering artificial bones. Analysis results show that the beta-TCP porous ceramic not only can provide a three-dimensional structure for tissue growth, is beneficial to differentiation of stem cells into osteoblasts, but also can provide a good microenvironment to enable bone tissues to form.
The invention has the beneficial effects that:
(1) the bone repair material prepared by the invention adopts the material which has good biodegradability and contains CaSO4And beta-TCP;
(2) after the bone repair material is implanted into a body, active ingredients of the body can be brought in the degradation and absorption process;
(3) the composite porous bone repair scaffold material prepared from the bone repair material has the advantages of simple process, easy operation, wide raw material source, low cost, stable yield and no damage to CaSO4And the advantages of beta-TCP, and has good mechanical strength, and has good application prospect in the field of cartilage tissue engineering;
(4) the prepared porous bone repair scaffold material is beneficial to cell adhesion and proliferation, and has good biocompatibility and biodegradability.
Drawings
FIG. 1: the preparation method of the invention has a process flow chart.
FIG. 2: an SEM topography of a pure calcium sulfate sample.
FIG. 3: CaSO-containing food4And a SEM topography of the composite ceramic material of the beta-TCP.
FIG. 4: CaSO-containing food4And XRD pattern of the composite ceramic material of beta-TCP.
FIG. 5: and (5) comparing the degradation results of the composite ceramic materials with different proportions.
Detailed Description
The invention is described in more detail below with reference to the figures and examples, but the scope of the invention is not limited to the description.
Example 1
(1) Taking aluminum sulfate octadecahydrate powder, fully grinding and sieving the powder, and weighing 0.45g of the powder;
(2) weighing calcium sulfate dihydrate powder, fully and uniformly mixing the calcium sulfate dihydrate powder with the weighed aluminum sulfate octadecahydrate, and filling the mixture into a 150ml beaker;
(3) buckling filter paper on a beaker filled with calcium sulfate by using weighing paper, sleeving a rubber band on the beaker, and placing the beaker in a pressure cooker;
(4) controlling the voltage of the pressure cooker to be kept between 0.14 and 0.15MPa, keeping the pressure for 10 hours and then taking out;
(5) drying at 80 deg.C for 12h to obtain alpha-calcium sulfate hemihydrate, and packaging in a sealed bag.
The SEM topography of the calcium sulfate material prepared in this example is shown in fig. 2. As can be seen from the figure, the powder has a needle-like and scaly structure, and the length is uniformly distributed between 10 μm and tens of micrometers.
Example 2
(1) Taking aluminum sulfate octadecahydrate powder, fully grinding and sieving the powder, and weighing 0.45g of the powder;
(2) weighing calcium sulfate dihydrate powder, fully and uniformly mixing the calcium sulfate dihydrate powder with the weighed aluminum sulfate octadecahydrate, and filling the mixture into a 150ml beaker;
(3) buckling filter paper on a beaker filled with calcium sulfate by using weighing paper, sleeving a rubber band on the beaker, and placing the beaker in a pressure cooker;
(4) controlling the voltage of the pressure cooker to be kept between 0.14 and 0.15MPa, keeping the pressure for 10 hours and then taking out;
(5) drying at 80 deg.C for 12h to obtain alpha-calcium sulfate hemihydrate, and packaging in a sealed bag.
(6) And (3) fully and uniformly mixing the beta-TCP ceramic and the calcium sulfate according to the ratio of 1:7, filling the mixture into a beaker, and repeating the steps 4 and 5.
(7) And filling the powder into a 50-hole plate, sealing the opening in a 60-DEG C oven, hydrating for 24h, and opening and drying for 24h to finally obtain the porous calcium sulfate/beta-TCP biological scaffold.
The process flow diagram of this example is shown in FIG. 1. The SEM appearance diagram of the prepared calcium sulfate/beta-TCP composite ceramic material is shown in figure 3, the particle size of the calcium sulfate/beta-TCP composite ceramic material is between 10 and 50 microns, and the comparison of the SEM appearance diagram 2 of a pure calcium sulfate sample shows that the length-diameter ratio of the whisker is reduced, and the flaky structure on the surface of the powder basically disappears and becomes smooth. The XRD spectrum is shown in FIG. 4, and characteristic peaks and CaSO can be seen4The powder is completely consistent with beta-TCP, and the graph analysis shows that the 2 theta angle of the powder prepared by the experiment is well corresponding to the PDF card of the beta-TCP, and the three strong peaks are respectively at the crystal face positions of (214), (0, 2, 10) and (220), and the relative diffraction intensity is equivalent. HAP diffraction peaks appear on the two crystal faces of (112) and (300), and can be seen by comparing calcium sulfate PDF card with CaSO4The three strong peaks of the two were completely coincident. The invention successfully prepares the calcium sulfate/beta-TCP bracket.
Table 1 shows the cell proliferation ratio (RGR) value of the leaching solution of the prepared calcium sulfate/beta-TCP ceramic composite scaffold material and 293T cells after 24h of coculture in a CCK-8 cytotoxicity test. The cytotoxicity result shows that when the powder concentration is 0.05-0.2g/ml, the cell proliferation degree is between 110 and 160, and the cytotoxicity is 0 grade by combining with the relevant regulation of the national standard GB/T14233.3-2005. Thus, the beta-TCP powder prepared by the experiment has no cytotoxicity.
Example 3
(1) Taking aluminum sulfate octadecahydrate powder, fully grinding and sieving the powder, and weighing 0.45g of the powder;
(2) weighing calcium sulfate dihydrate powder, fully and uniformly mixing the calcium sulfate dihydrate powder with the weighed aluminum sulfate octadecahydrate, and filling the mixture into a 150ml beaker;
(3) buckling filter paper on a beaker filled with calcium sulfate by using weighing paper, sleeving a rubber band on the beaker, and placing the beaker in a pressure cooker;
(4) controlling the voltage of the pressure cooker to be kept between 0.14 and 0.15MPa, keeping the pressure for 10 hours and then taking out;
(5) drying at 80 deg.C for 12h to obtain alpha-calcium sulfate hemihydrate, and packaging in a sealed bag.
(6) And (3) fully and uniformly mixing the beta-TCP ceramic and the calcium sulfate according to the proportion of 1:5, filling the mixture into a beaker, and repeating the steps 4 and 5.
(7) And filling the powder into a 50-hole plate, sealing the opening in a 60-DEG C oven, hydrating for 24h, and opening and drying for 24h to finally obtain the porous calcium sulfate/beta-TCP biological scaffold.
Example 4
(1) Taking aluminum sulfate octadecahydrate powder, fully grinding and sieving the powder, and weighing 0.45g of the powder;
(2) weighing calcium sulfate dihydrate powder, fully and uniformly mixing the calcium sulfate dihydrate powder with the weighed aluminum sulfate octadecahydrate, and filling the mixture into a 150ml beaker;
(3) buckling filter paper on a beaker filled with calcium sulfate by using weighing paper, sleeving a rubber band on the beaker, and placing the beaker in a pressure cooker;
(4) controlling the voltage of the pressure cooker to be kept between 0.14 and 0.15MPa, keeping the pressure for 10 hours and then taking out;
(5) drying at 80 deg.C for 12h to obtain alpha-calcium sulfate hemihydrate, and packaging in a sealed bag.
(6) And (3) fully and uniformly mixing the beta-TCP ceramic and the calcium sulfate according to the ratio of 1:9, filling the mixture into a beaker, and repeating the steps 4 and 5.
(7) And filling the powder into a 50-hole plate, sealing the opening in a 60-DEG C oven, hydrating for 24h, and opening and drying for 24h to finally obtain the porous calcium sulfate/beta-TCP biological scaffold.
Example 5
(1) Taking aluminum sulfate octadecahydrate powder, fully grinding and sieving the powder, and weighing 0.45g of the powder;
(2) weighing calcium sulfate dihydrate powder, fully and uniformly mixing the calcium sulfate dihydrate powder with the weighed aluminum sulfate octadecahydrate, and filling the mixture into a 150ml beaker;
(3) buckling filter paper on a beaker filled with calcium sulfate by using weighing paper, sleeving a rubber band on the beaker, and placing the beaker in a pressure cooker;
(4) controlling the voltage of the pressure cooker to be kept between 0.14 and 0.15MPa, keeping the pressure for 10 hours and then taking out;
(5) drying at 80 deg.C for 12h to obtain alpha-calcium sulfate hemihydrate, and packaging in a sealed bag.
(6) And (3) fully and uniformly mixing the beta-TCP ceramic and the calcium sulfate according to the ratio of 1:11, filling the mixture into a beaker, and repeating the steps 4 and 5.
(7) And filling the powder into a 50-hole plate, sealing the opening in a 60-DEG C oven, hydrating for 24h, and opening and drying for 24h to finally obtain the porous calcium sulfate/beta-TCP biological scaffold.
The graph comparing the degradation results of the composite scaffold materials with different proportions is shown in fig. 5, the abscissa is the degradation days, the weight loss rate is the ordinate, as can be seen from the graph, the first 5-8 days are the weight increasing process, and then the weight losing process gradually occurs, and the analysis reason shows that the weight increasing is due to the deposition of degradation products and SBF active substances, and the weight increasing caused by the deposition products is smaller than the weight loss caused by the degradation of the scaffold after the scaffold is soaked for 5-8 days, so that the degradation curve shows the weight losing situation. After one week, the weight loss rate of the sample begins to increase because the sample itself is degraded, the degradation rate is higher than the deposition rate of the calcium phosphate, and the material begins to lose weight. The scaffold material is soaked in the SBF solution, whether the material has the capacity of inducing the deposition of the apatite or not can be used for judging the biological activity of the material, and if the material has the deposition of the apatite, the biological activity is indicated. Therefore, the five-weight ratio of calcium sulfate/beta-TCP scaffold has good bioactivity according to the analysis.
Table 1 results of cell proliferation experiments on composite ceramic materials (mass ratio of β -TCP ceramic to calcium sulfate 1:7)
Claims (6)
1. CaSO-containing food4And beta-TCP, characterized in that:
the composite ceramic material is made of CaSO4·2H2O and β -TCP ceramic, wherein: the beta-TCP ceramic and CaSO4·2H2The mass ratio of the O is 1: 5-11, and the mass ratio of the beta-tricalcium phosphate to the hydroxyapatite in the beta-TCP ceramic is 7: 3.
2. CaSO-containing composition according to claim 14Preparation method of composite ceramic material of beta-TCPThe method is characterized by comprising the following steps:
(1) taking aluminum sulfate octadecahydrate powder, fully grinding and sieving the powder, and weighing 0.45g of the powder;
(2) weighing calcium sulfate dihydrate powder, fully and uniformly mixing the calcium sulfate dihydrate powder with the weighed aluminum sulfate octadecahydrate, and filling the mixture into a 150ml beaker;
(3) buckling filter paper on a beaker filled with calcium sulfate by using weighing paper, sleeving a rubber band on the beaker, and placing the beaker in a pressure cooker;
(4) controlling the voltage of the pressure cooker to be kept between 0.14 and 0.15MPa, keeping the pressure for 10 hours and then taking out;
(5) drying at 80 deg.C for 12h to obtain alpha-calcium sulfate hemihydrate, and packaging in a sealed bag;
(6) and (3) fully and uniformly mixing the calcium sulfate dihydrate powder and the doping material according to a certain mass ratio, filling the mixture into a beaker, and repeating the steps 4 and 5 for a plurality of times.
(7) Prepared from CaSO4·2H2The powder of the composite ceramic material consisting of O and beta-TCP ceramics.
3. The method of claim 2, wherein:
in the step (1), the aluminum sulfate octadecahydrate powder is fully ground and sieved by a 200-mesh sieve.
4. The production method according to claim 2 or 3, characterized in that:
in the step (6), the beta-TCP ceramic and CaSO4·2H2The mass ratio of O is 1: 5-11.
5. CaSO-containing composition according to claim 14And the application of the composite ceramic material of beta-TCP in constructing tissue engineering artificial bone, which is characterized in that:
(1) filling the powder of the composite ceramic material into a 50-hole plate, sealing the opening in a drying oven at 60 ℃ for hydration for 24 hours, and opening the opening for drying for 24 hours to finally prepare the porous calcium sulfate/beta-TCP biological scaffold;
(2) implanting the porous calcium sulfate/beta-TCP biological scaffold into a human body;
(3) the biological scaffold implanted into a human body has biodegradability and bioabsorbability, and the material can be gradually discharged out of the body through the processes of dissolution and absorption or metabolism, so that the bone defect part is finally replaced by new bone tissues.
6. Use according to claim 5, wherein:
one part of the beta-tricalcium phosphate is discharged out of the body through biochemical reaction, the other part of the beta-tricalcium phosphate participates in the formation of new bones, and the degraded calcium and phosphorus ions can enter the circulatory system of the human body and gradually form new bones.
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