CN110960733B - A neovascular construct for implantation into subcutaneous tissue of a subject and a method of making the same - Google Patents

A neovascular construct for implantation into subcutaneous tissue of a subject and a method of making the same Download PDF

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CN110960733B
CN110960733B CN201911314139.2A CN201911314139A CN110960733B CN 110960733 B CN110960733 B CN 110960733B CN 201911314139 A CN201911314139 A CN 201911314139A CN 110960733 B CN110960733 B CN 110960733B
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polymeric material
construct
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subcutaneous tissue
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CN110960733A (en
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牛玉清
夏慧敏
张锐忠
付铭
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Guangzhou Women and Childrens Medical Center
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L29/00—Materials for catheters, medical tubing, cannulae, or endoscopes or for coating catheters
    • A61L29/04—Macromolecular materials
    • A61L29/06—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L29/00—Materials for catheters, medical tubing, cannulae, or endoscopes or for coating catheters
    • A61L29/14—Materials characterised by their function or physical properties, e.g. lubricating compositions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L29/00—Materials for catheters, medical tubing, cannulae, or endoscopes or for coating catheters
    • A61L29/14—Materials characterised by their function or physical properties, e.g. lubricating compositions
    • A61L29/148—Materials at least partially resorbable by the body
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces

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Abstract

The invention discloses a neovascular construct for being implanted into subcutaneous tissues of a subject and a preparation method thereof, and relates to the technical field of high polymer materials. The new blood vessel construct is a nano fiber conduit made of the high polymer material in an electrostatic spinning mode, the material is selected from one of levorotatory polylactic acid, dextrorotatory polylactic acid, polylactic-co-glycolic acid and polyglycolic acid, the blood vessel construct made of the high polymer material is transplanted into subcutaneous tissue, and an extracellular matrix space rich in new blood vessels can be rapidly formed under the skin of a subject, so that the new blood vessel construct is used for accommodating islet or islet cells from a donor source to maintain the function for a long time.

Description

A neovascular construct for implantation into subcutaneous tissue of a subject and a method of making the same
Technical Field
The invention relates to the technical field of high polymer materials, in particular to a neovascular construct for being implanted into subcutaneous tissues of a subject and a preparation method thereof.
Background
Type I diabetes, the primary insulin-dependent diabetes mellitus, occurs well in children or adolescents, also occurs at various ages, particularly in menopause, is a disease which is hardly cured, and the deficiency of islet cell function in a patient easily causes ketoacidosis to the patient, so that the patient needs to be treated by insulin injection for life, otherwise, the life is threatened.
The insulin treatment goals for type I diabetes are two, firstly to ensure a good quality of life for the patient (i.e. to avoid as much as possible the occurrence of severe hypoglycemia) and secondly to control the metabolic level in order to prevent diabetic complications. However, long-term insulin injections can cause a number of complications, including dizziness, sweating, and even loss of consciousness or death.
To improve the quality of life of type I diabetic patients, islet or islet cell transplantation may help patients to resolve the problem of insulin deficiency, "Edmonton protocol", by intrahepatic transplantation of a source of donor islets, a preliminary study analysis after transplantation indicating that this approach may achieve glycemic control in patients. However, the longest duration of this method is about 5 years, and intrahepatic transplantation also involves risks of bleeding, inflammation, graft abrasion, and local steatosis at the site of intrahepatic transplantation.
In view of this, the invention is particularly proposed.
Disclosure of Invention
The invention aims to provide application of a biodegradable high molecular material in preparing a neovascular construct for being implanted into subcutaneous tissues of a subject, a preparation method of the neovascular construct for being implanted into the subcutaneous tissues of the subject and the neovascular construct for being implanted into the subcutaneous tissues of the subject.
The invention is realized by the following steps:
in a first aspect, embodiments provide the use of a biodegradable polymeric material in the preparation of a neo-vascular construct for implantation into subcutaneous tissue of a subject, the neo-vascular construct being a nanofiber conduit made by electrospinning the polymeric material, the polymeric material being selected from one of levopolylactic acid, dextropolylactic acid, polylactic-co-glycolic acid, and polyglycolic acid.
In a second aspect, embodiments provide a method of making a neovascular construct for implantation into subcutaneous tissue in a subject, comprising electrospinning a polymeric material selected from one of levopolylactic acid, dextropolylactic acid, a polylactic-co-glycolic acid, and polyglycolic acid to form a nanofiber conduit.
In a third aspect, the embodiments provide a neovascular construct for implantation into the subcutaneous tissue of a subject, made by the method of any one of the preceding embodiments for making a neovascular construct for implantation into the subcutaneous tissue of a subject.
The invention has the following beneficial effects:
the embodiment of the invention provides application of a biodegradable high polymer material in preparing a new blood vessel construct for implanting into subcutaneous tissues of a subject, wherein the blood vessel construct is a nano fiber catheter made of the high polymer material in an electrospinning mode, the high polymer material is selected from one of levorotatory polylactic acid, dextrorotatory polylactic acid, polylactic-co-glycolic acid and polyglycolic acid, and the new blood vessel construct prepared by the high polymer material is implanted into subcutaneous tissues, so that a new blood vessel-rich extracellular matrix space can be rapidly formed under the skin of the subject, and the new blood vessel-rich extracellular matrix space can be used for accommodating islet or islet cells from donor sources for long-term maintenance.
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In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a cross-sectional view of the subcutaneous skin of type I diabetic Balb/c mice and a histochemical staining and immunofluorescence staining graph of the subcutaneous prevascularization space of type I diabetic Balb/c mice after islet cell implantation in the experimental example of the present invention, wherein, A in FIG. 1 is a diagram of the subcutaneous tissue forming a prevascularized collagen-rich transplantation space, and B in FIG. 1 is a diagram of laser confocal microscope;
FIG. 2 is a result of quantitative analysis of a blood glucose tolerance test 60 days after transplantation in the test example of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The embodiment of the invention provides an application of a biodegradable high polymer material in preparing a new blood vessel construct implanted into subcutaneous tissues of a subject, wherein the blood vessel construct is a nano fiber catheter prepared by the high polymer material in an electrospinning mode, and the high polymer material is selected from one of levorotatory polylactic acid, dextrorotatory polylactic acid, polylactic-co-glycolic acid and polyglycolic acid.
In an optional embodiment, the molecular weight of the polymer material is 3000-400000.
In an alternative embodiment, the molecular weight of the polymer material is 29000-48000.
In an alternative embodiment, the poly (lactic-co-glycolic acid) is prepared by mixing (75-50): (25-50) lactic acid and glycolic acid.
In an alternative embodiment, the electrospinning liquid of the nanofiber conduit is prepared by mixing an organic solvent and the high molecular material, wherein the organic solvent is at least one selected from tetrahydrofuran, 1, 2-dichloromethane, trifluoroethanol, chloroform and hexafluoroisopropanol.
In alternative embodiments, the organic solvent comprises at least one of trifluoroethanol, tetrahydrofuran, hexafluoroisopropanol.
In an alternative embodiment, 8 to 12ml of the organic solvent is mixed with 1 to 1.4g of the polymer material.
In an alternative embodiment, the electrospinning flow rate of the electrostatic spinning is 2.8-3.2 mL/h, and the voltage is 17-19 kV.
In an optional embodiment, the electrostatic spinning receiving device is a cylindrical tube with the outer diameter of 1.7-1.9 cm, and the rotating speed of the receiving device is 780-820 rpm.
In an optional embodiment, during electrostatic spinning, the distance between the spray head and the receiving device is 10-14 cm.
On one hand, the nanofiber catheter prepared by adopting the material has degradability and good mechanical property, can meet the requirements of subcutaneous implantation of different individuals and is not easy to deform; on the other hand, the nanofiber catheter has good biocompatibility, can cause quick immune stress reaction of a host, and can avoid or reduce subcutaneous long-term inflammatory reaction.
In an optional embodiment, the outer diameter of the nanofiber conduit is 2.1-5.0 cm, the length is 2-6 cm, and the thickness is 0.5-1000 μm.
The embodiment of the invention also provides a preparation method of the new blood vessel construct used for being implanted into subcutaneous tissues of a subject, which comprises the step of preparing the nano fiber catheter by adopting a high polymer material in an electrostatic spinning mode, wherein the high polymer material is selected from one of levorotatory polylactic acid, dextrorotatory polylactic acid, polylactic-co-glycolic acid and polyglycolic acid.
In an optional embodiment, the molecular weight of the polymer material is 3000-400000.
In an alternative embodiment, the molecular weight of the polymer material is 29000-48000.
In an alternative embodiment, the poly (lactic-co-glycolic acid) is prepared by mixing (75-50): (25-50) lactic acid and glycolic acid.
In an alternative embodiment, the electrospinning liquid of the nanofiber conduit is prepared by mixing an organic solvent and the high molecular material, wherein the organic solvent is at least one selected from tetrahydrofuran, 1, 2-dichloromethane, trifluoroethanol, chloroform and hexafluoroisopropanol.
In alternative embodiments, the organic solvent comprises at least one of trifluoroethanol, tetrahydrofuran, hexafluoroisopropanol.
In an alternative embodiment, 8 to 12ml of the organic solvent is mixed with 1 to 1.4g of the polymer material.
In an optional embodiment, the electrospinning flow rate of the electrospinning is 2.8-3.2 mL/h, and the voltage is 17-19 kV.
In an optional embodiment, the electrostatic spinning receiving device is a cylindrical tube with the outer diameter of 1.7-1.9 cm, and the rotating speed of the receiving device is 780-820 rpm.
In an optional embodiment, during electrostatic spinning, the distance between the spray head and the receiving device is 10-14 cm.
In addition, the embodiment of the present invention provides a neovascular construct for implantation into the subcutaneous tissue of a subject, which is prepared by the method for preparing a neovascular construct for implantation into the subcutaneous tissue of a subject according to the foregoing embodiment.
In an alternative embodiment, the vascular construct has mechanical properties, a compressive strength of 10 to 500MPa, an elongation at break of 1 to 55%, and a Young's modulus of 200 to 1000 MPa.
In an alternative embodiment, the vascular construct has an outer diameter of 2.1 to 5.0cm, a length of 2 to 6cm and a thickness of 0.5 to 1000 μm. According to this range setting, a space of extracellular matrix-rich capillaries can be created subcutaneously in the implanted patientThe area may be 2cm3～24cm3。
In an optional embodiment, the vascular construct is a nanofiber catheter, the nanofiber catheter is composed of disordered nanofiber arrays, the length of each nanofiber is 5-20 μm, the porosity is 80-95%, and the pore diameter is 1-30 μm.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
A method of making a neovascular construct for implantation into subcutaneous tissue of a subject, comprising:
1.2g of a polymer material (L-polylactic acid, PLLA, molecular weight 48000Da) was weighed and placed in a 30ml glass beaker, and 10ml of an organic solvent (hexafluoroisopropanol) was added to the beaker and sufficiently dissolved to obtain an electrospinning solution.
And (2) moving the electrospinning liquid from the beaker into a special electrostatic spinning storage needle cylinder by using a liquid transfer machine, installing an electrospinning spray head (22G), adjusting the electrospinning flow rate to be 3.0mL/h and the voltage to be 18kV, wherein the distance between the spray head and a receiving device is 12cm, the receiving device is a cylindrical stainless steel pipe with the outer diameter of 1.8cm, the rotating speed of the receiving device is adjusted to be 800rpm, and electrospinning is carried out for 28 hours to obtain the nanofiber catheter. Cutting the nano fiber catheter into corresponding long sections as required, placing the long sections into a sterile culture dish, sealing the culture dish with a sealing film, and sealing with 12kGy Co60Sterilizing by radiation and reserving for later use.
Example 2
This example provides a method of making a neovascular construct for implantation into the subcutaneous tissue of a subject, substantially the same as that provided in example 1, except that the polymeric material and the organic solvent are different as follows:
the high polymer material is a polylactic-co-glycolic acid copolymer obtained by copolymerizing lactic acid and glycolic acid with a molar ratio of 75: 25;
the organic solvent is trifluoroethanol.
Example 3
This example provides a method of making a neovascular construct for implantation into the subcutaneous tissue of a subject, substantially the same as that provided in example 1, except that the polymeric material and the organic solvent are different as follows:
the high molecular material is polyglycolic acid;
the organic solvent is tetrahydrofuran.
Example 4
This example provides a method of making a neovascular construct for implantation into the subcutaneous tissue of a subject, substantially the same as that provided in example 1, except that the polymeric material and the organic solvent are different as follows:
the polymer material is poly (D-lactic acid) (PDLA) with molecular weight of 22000 Da;
the volume ratio of the organic solvent is 3: 1 of hexafluoroisopropanol with trifluoroethanol.
Example 5
This example provides a method of making a neovascular construct for implantation into the subcutaneous tissue of a subject, substantially the same as that provided in example 1, except that the polymeric material and the organic solvent are different as follows:
the high molecular material is a polylactic-co-glycolic acid copolymer obtained by copolymerizing lactic acid and glycolic acid with a molar ratio of 75:25, and the molecular weight is 40000 Da;
the volume ratio of the organic solvent is 3: 1 of hexafluoroisopropanol with trifluoroethanol.
Example 6
This example provides a method of making a neovascular construct for implantation into the subcutaneous tissue of a subject, substantially the same as that provided in example 1, except that the polymeric material and the organic solvent are different as follows:
the high polymer material is prepared from the following components in a molar ratio of 50: 50, the molecular weight of the polylactic acid-glycolic acid copolymer obtained by copolymerizing lactic acid and glycolic acid is 30000 Da;
the volume ratio of the organic solvent is 3: 1 of hexafluoroisopropanol with trifluoroethanol.
Example 7
This example provides a method of making a neovascular construct for implantation into the subcutaneous tissue of a subject, substantially the same as that provided in example 1, except that the polymeric material is different as follows:
the high polymer material is prepared from the following components in a molar ratio of 50: the molecular weight of the polylactic acid-glycolic acid copolymer obtained by copolymerizing 50 lactic acid and glycolic acid is 27000 Da.
Example 8
This example provides a method of making a neovascular construct for implantation into the subcutaneous tissue of a subject, substantially the same as that provided in example 1, except that the polymeric material is different as follows:
the high molecular material is a polylactic acid-glycolic acid copolymer obtained by copolymerizing lactic acid and glycolic acid with a molar ratio of 75:25, and the molecular weight is 22000 Da.
Example 9
This example provides a method of making a neovascular construct for implantation into the subcutaneous tissue of a subject, substantially the same as that provided in example 1, except that the polymeric material is different as follows:
the high molecular material is polyglycolic acid obtained by copolymerizing lactic acid and glycolic acid with a molar ratio of 75:25, and the molecular weight is 30000 Da.
Example 10
This example provides a method of making a neovascular construct for implantation into the subcutaneous tissue of a subject, substantially the same as that provided in example 1, except that the polymeric material and the organic solvent are different as follows:
the high molecular material is polylactic acid-glycolic acid copolymer obtained by copolymerizing 75% of lactic acid and 25% of glycolic acid, and the molecular weight is 27000 Da;
the organic solvent is trifluoroethanol.
Example 11
This example provides a method of making a neovascular construct for implantation into the subcutaneous tissue of a subject, substantially the same as that provided in example 1, except that the polymeric material and the organic solvent are different as follows:
the high polymer material is a polylactic acid-glycolic acid copolymer obtained by copolymerizing 75% and 25% glycolic acid, and the molecular weight is 28000 Da;
the volume ratio of the organic solvent is 3: 1 of hexafluoroisopropanol with trifluoroethanol.
Test examples
Three weeks before islet or islet cells transplantation, the nanofiber catheters provided in example 1 were subcutaneously implanted in the abdominal cavity of Balb/c mice, and the control group thereof was non-degradable nylon tubes (6F, GE digital subtraction angiography catheter, Torcon)
Figure BDA0002325356940000091
Advantaged catheater, Bloomington, American) to construct a subcutaneous prevascularized collagen-rich graft space.
After 2 weeks, after the donor islets or islet cells are separated, the subcutaneous nanofiber catheter of the Balb/c mice with type I diabetes is taken out, then the donor islets or islet cells are transplanted into the prevascularized collagen transplanting space by using a blunt syringe, and the gateway is surgically nailed.
And testing the blood sugar of the patient or the experimental animal every 12h for the first three days after transplantation, testing the blood sugar of the patient or the experimental animal every three days after the three days, and periodically carrying out a blood sugar tolerance experiment to evaluate the transplantation effect. And compared with the intrarenal graft group, the nylon catheter graft group and the non-catheter graft group (non-subcutaneous prevascularized graft space graft group), and the results are shown in FIG. 2.
FIG. 1 is a cross-sectional view of the skin of Balb/c mice implanted with a nanofiber catheter subcutaneously and histochemical and immunofluorescence staining patterns of the mice with type I diabetes after islet cells are implanted into the subcutaneous prevascularized collagen-rich space of the Balb/c mice for 100 days.
Wherein, A in figure 1 is a grafting space with rich prevascularized collagen formed by subcutaneous tissues after the nanofiber catheter is implanted under the skin for 2 weeks and the catheter is taken out; in FIG. 1, B is a confocal laser microscopy image, which shows that both insulin and the vascular inner wall marker VWf are positively expressed, indicating that the transplanted islet cells still have the function of releasing insulin.
Fig. 2 is the results of quantitative glucose tolerance analysis of each group 60 days after islet transplantation.
As can be seen from fig. 2, the quantitative analysis result of the blood glucose tolerance test indicates that the diabetic mice in the prevascularized islet transplantation site group prepared in advance by using the biomaterial catheter have good glucose tolerance compared with the subcutaneous islet cell transplantation group, which suggests that a good blood transport space is required for subcutaneous islet transplantation. Compared with a non-degradable nylon tube group, the diabetic mouse of the nanofiber catheter transplantation group has stronger blood sugar removing capacity and is very close to the intrarenal transplantation group. The subcutaneous prevascularization space created by the nanofiber catheter is suggested to have a blood transport space more suitable for the survival of the islets or islet cells.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (11)

1. Use of a biodegradable polymeric material for the manufacture of a neo-vascular construct for implantation into subcutaneous tissue of a subject, wherein the neo-vascular construct is a nanofiber conduit made by electrospinning the polymeric material, and the polymeric material is selected from one of poly (L-lactic acid), poly (D-lactic acid), poly (lactic-co-glycolic acid), and poly (glycolic acid);
the subject is diabetic, and the outer diameter of the blood vessel construction body is 2.1-5.0 cm.
2. The use of a biodegradable polymeric material according to claim 1, for the preparation of a neo-vascular construct for implantation into the subcutaneous tissue of a subject, wherein the polymeric material has a molecular weight of 3000 to 400000.
3. The use of a biodegradable polymeric material according to claim 2, for the preparation of a neo-vascular construct for implantation into the subcutaneous tissue of a subject, wherein the molecular weight of the polymeric material is from 29000 to 48000.
4. Use of a biodegradable polymeric material according to claim 2, for the preparation of a neo-vascular construct for implantation in the subcutaneous tissue of a subject, wherein the poly (lactic-co-glycolic acid) is prepared from a mixture of poly (lactic-co-glycolic acid) and poly (lactic-co-glycolic acid) in a molar ratio of (75-50): (25-50) lactic acid and glycolic acid.
5. Use of a biodegradable polymeric material according to any one of claims 1 to 4 for the preparation of a neo-vascular construct for implantation into subcutaneous tissue of a subject, wherein the electrospun solution of the nanofiber conduit is prepared by mixing an organic solvent with the polymeric material, wherein the organic solvent is at least one selected from the group consisting of tetrahydrofuran, 1, 2-dichloromethane, trifluoroethanol, chloroform and hexafluoroisopropanol.
6. Use of a biodegradable polymeric material according to claim 5, for the preparation of a neo-vascular construct for implantation into the subcutaneous tissue of a subject, wherein said organic solvent comprises at least one of trifluoroethanol, tetrahydrofuran and hexafluoroisopropanol.
7. The use of a biodegradable polymeric material according to claim 5, for the preparation of a neo-vascular construct for implantation into the subcutaneous tissue of a subject, wherein 8-12 mL of organic solvent is mixed per 1-1.4 g of polymeric material.
8. The use of the biodegradable polymeric material according to any one of claims 1 to 4, for preparing a neovascular construct for implantation into subcutaneous tissue of a subject, wherein the electrospinning flow rate is 2.8 to 3.2mL/h and the voltage is 17 to 19 kV.
9. The use of a biodegradable polymeric material in the preparation of a neovascular construct for implantation into the subcutaneous tissue of a subject according to claim 8, wherein the electrospun receiving means is a cylindrical tube with an outer diameter of 1.7 to 1.9cm and the rotational speed of the receiving means is 780 to 820 rpm.
10. The use of a biodegradable polymeric material in the preparation of a neo-vascular construct for implantation into the subcutaneous tissue of a subject according to claim 8, wherein the distance between the nozzle and the receiving device during electrospinning is 10-14 cm.
11. The use of a biodegradable polymeric material in the preparation of a neo-vascular construct for implantation into the subcutaneous tissue of a subject according to claim 8, wherein the nanofiber conduit has an outer diameter of 2.1 to 5.0cm, a length of 2 to 6cm, and a thickness of 0.5 to 1000 μm.
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US7052829B2 (en) * 2001-03-30 2006-05-30 The Arizona Board Of Regents On Behalf Of The University Of Arizona Prevascularized constructs for implantation to provide blood perfusion
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