Patent Application
20220409775
The present invention relates to an injectable hydrogel composition having the recruitment of endogenous progenitors or stem cells and the induction of vascular differentiation of recruited cells, and more specifically to an injectable hydrogel composition having the recruitment of endogenous progenitors or stem cells and the induction of vascular differentiation of recruited cells, which consists of: a first solution including anionic hyaluronic acid into which a vascular differentiation inducing factor is introduced; and a second solution including a cationic material, wherein a stem cell recruitment factor is further included in the first solution and/or the second solution, and wherein a hydrogel is formed by electrostatic interaction.
With the exception of cornea and cartilage, all tissue engineering substitute materials require vascular neural networks to supply nutrients and oxygen necessary for survival in vivo. Since angiogenesis in artificial tissues can take several weeks to occur spontaneously, the tissue develops necrosis during this time due to a lack of supply of essential nutrients. Therefore, the formation of blood vessels in regenerative medicine is a very important problem in the regeneration of damaged tissues or the development of artificial organs.
Angiogenesis is a process in which endothelial cells of existing blood vessels decompose, migrate, divide and differentiate the extracellular matrix (ECM) to form new capillaries, and it is involved in various physiological and pathological phenomena such as wound repair, embryogenesis, tumor formation, chronic inflammation, obesity and the like. The angiogenic process includes the proliferation and migration of vascular endothelial cells from the vessel wall to surrounding tissues in the direction of stimulation. Subsequently, various proteolytic enzymes are activated such that vascular endothelial cells infiltrate the basement membrane and form loops, and the formed loops are differentiated to form tubes.
In order to induce or promote angiogenesis, many studies have been conducted to induce differentiation of stem cells into vascular endothelial cells, but in fact, it has been mainly reported that angiogenesis of the host is induced by growth factors secreted from stem cells rather than stem cells themselves induce angiogenesis. In addition, it has been reported that among the cells produced by decomposing adipose tissue, a structural vascular fraction (SVF) can be transplanted into animals without culturing to differentiate into vascular endothelial cells. However, the method does not induce proliferation by subculture of adipose stem cells, and thus, the amount of vascular endothelial cells differentiated from adipose stem cells is very small, and in particular, since the proliferation and differentiation rates of differentiated vascular endothelial cells are low, the application thereof is limited.
Among the methods using stein cells, the method of using “self-regeneration” activates the endogenous progenitor cells/stein cells originally present in the patient's own body, thereby regenerating damaged organs and/or tissues to recover of their functions. Endogenous progenitor cells/stem cells can differentiate into different types of cells depending on the different types of cells in contact, the content of extracellular matrix and the microenvironment in which the cells are present, such as cytokines and growth factors.
In order to recruit endogenous progenitor cells/stem cells into the wound site or the tissue damaged site, stem cell recruitment factors can be used, and among them, substance P (SP), which is known as a stem cell recruitment factor, has been reported to be helpful in wound treatment, but since it is used in a simple solution state, it has the disadvantage that it does not stay for a long period of time in the tissue damaged site, and thus, the development of a method that can continuously release substance P from the damaged area is required.
Accordingly, in the present invention, as a result of diligent efforts to develop a substance that can effectively recruit endogenous progenitor cells/stem cells and promote angiogenesis by inducing vascular differentiation at the same time, it was confirmed that when the vascular endothelial growth factor mimic peptide (VP), which is capable of differentiating progenitor/stem cells into vascular cells, was mixed with HA-VP, which was chemically introduced into anionic hyaluronic acid, and a cationic material, a hydrogel was formed by electrostatic interaction, and the present invention was completed by confirming that when the progenitor cell/stem cell recruitment factor (substance P, WKYMVM, SDF1α, G-SCF, MCP-1, etc.) was physically supported and injected in vivo, the stem cell recruitment factor was released from the injected hydrogel such that endogenous progenitor cells/stem cells were recruited in the hydrogel, and the induction of angiogenesis was promoted by differentiation into vascular cells by VP which was chemically introduced into hyaluronic acid.
An object of the present invention is to provide an injectable hydrogel composition having the recruitment of progenitor cells/stem cells and the induction of vascular differentiation of recruited cells.
In order to achieve the aforementioned object, the present invention provides an injectable hydrogel composition having the recruitment of endogenous progenitors or stem cells and the induction of vascular differentiation of recruited cells, which consists of a first solution including anionic hyaluronic acid into which a vascular differentiation inducing factor is introduced; and a second solution including a cationic material, wherein a stem cell recruitment factor is further included in any one or more of the first solution and the second solution, and wherein when the first solution and the second solution are mixed, a hydrogel is formed by electrostatic interaction.
According to a preferred exemplary embodiment of the present invention, the anionic hyaluronic acid into which the vascular differentiation inducing factor is introduced may be prepared by reacting the vascular differentiation inducing factor and anionic hyaluronic acid in which a carboxylic acid functional group is activated.
According to another preferred exemplary embodiment of the present invention, the cationic material may be at least one selected from the group consisting of chitosan, cationic dextran, polyethyleneimine, polylysine and polyhistidine.
According to still another preferred exemplary embodiment of the present invention, the stem cell recruitment factor may be at least one selected from the group consisting of substance P, WKYMVM, SDF1a, G-SCF and MCP-1.
According to still another preferred exemplary embodiment of the present invention, the ratio of the anionic hyaluronic acid into which the vascular differentiation inducing factor of the first solution is introduced and the cationic material of the second solution may be 3:1 to 1:3.
According to still another preferred exemplary embodiment of the present invention, the storage modulus of the hydrogel formed by mixing the first solution and the second solution may be 10 to 100 Pa.
In addition, the present invention provides an injection for tissue regeneration or an injection for fillers, including the injectable hydrogel composition.
In one aspect, the present invention relates to an injectable hydrogel composition having the recruitment of endogenous progenitors or stem cells and the induction of vascular differentiation of recruited cells, which consists of a first solution including anionic hyaluronic acid into which a vascular differentiation inducing factor is introduced; and a second solution including a cationic material, wherein a stem cell recruitment factor is further included in any one or more of the first solution and the second solution, and wherein when the first solution and the second solution are mixed, a hydrogel is formed by electrostatic interaction.
More specifically, the injectable hydrogel composition consists of a first solution including anionic hyaluronic acid into which a vascular differentiation inducing factor is introduced; and a second solution including a cationic material,
In the present invention, the injectable hydrogel may be formed by the electrostatic interaction of the anionic hyaluronic acid into which a vascular differentiation inducing factor is introduced and the cationic material. Preferably, when the first solution and the second solution are injected into the target site, they are mixed to form a hydrogel, but depending on the purpose, the first solution and the second solution are mixed immediately before injection to form a hydrogel and then injected.
The target site is a place requiring tissue regeneration, such as a damaged organ, a depressed tissue or a wound site, and the injectable hydrogel composition having the recruitment of endogenous progenitor cells or stem cells and the induction of vascular differentiation of the recruited cells according to the present invention has an advantage in that tissue can be regenerated more efficiently, because it recruits endogenous progenitor cells/stem cells with a hydrogel and induces the recruited progenitor cells/stem cells to differentiate into blood vessels.
In the present invention, the stem cells refer to endogenous progenitor cells or endogenous stem cells, and the endogenous progenitor cells or endogenous stem cells refer to cells with self-renewal and pluripotency that are originally present in a specific organ and/or tissue and contribute to the regeneration of the tissue and/or organ, when the corresponding organ and/or tissue is damaged. Specific examples include mesenchymal stem cells, corneal stem cells, myocardial stem cells, neural stem cells and vascular endothelial progenitor cells.
In order to physically support the progenitor cell/stem cell recruitment factor (substance P, WKYMVM, SDF1 α, G-SCF, MCP-1, etc.) on the hydrogel, the progenitor cell/stem cell recruitment factor was mixed in each of the HA-VP solution and the chitosan solution, and the HA-VP solution and the chitosan solution including the progenitor/stem cell recruitment formed a hydrogel through electrostatic interaction when injected in vivo. The stem cell recruitment factor was released from the formed hydrogel, and endogenous progenitor cells/stem cells were recruited in the hydrogel, and the endogenous progenitor cells/stem cells recruited in the hydrogel differentiated into vascular cells by chemically introduced VP to promote the induction of angiogenesis.
In the present invention, the vascular differentiation inducing factor is a vascular endothelial growth factor mimic peptide, and may include the amino acid sequence of SEQ ID NO: 1 (KLTWQELYQLKYKGI), but is not limited thereto, and depending on the purpose, angiogenesis promoting factors such as vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and the like, or thrombin may be applied.
In the present invention, the anionic hyaluronic acid into which the vascular differentiation inducing factor is introduced may be prepared by reacting the vascular differentiation inducing factor with anionic hyaluronic acid in which a carboxylic acid functional group is activated.
The molecular weight of the hyaluronic acid is preferably in the range of 500,000 Da to 6,000,000 Da. In this case, if the molecular weight of hyaluronic acid is less than 500,000 Da, the physical properties are too low to form a significant hydrogel, and if it is more than 6,000,000 Da, the viscosity may increase to bring a limitation to application as an injection, and thus, it is not preferable.
In the present invention, the cationic material may be at least one selected from the group consisting of chitosan, cationic dextran, polyethyleneimine, polylysine and polyhistidine.
In the present invention, the stem cell recruitment factor may be at least one selected from the group consisting of substance P, WKYMVM, SDF1 a, G-SCF and MCP-1. Preferably, it may be substance P or a derivative of substance P, but a substance having the endogenous progenitor/stem cell recruitment ability may be used without limitation.
In the present invention, the injectable hydrogel may be formed through electrostatic interaction between anionic hyaluronic acid, into which the vascular differentiation inducing factor is introduced, and a cationic material.
In the present invention, the ratio of the anionic hyaluronic acid, into which the vascular differentiation inducing factor is introduced, and the cationic material (chitosan) in the first solution may be 3:1 to 1:3, and preferably, 2:1 to 1:2.
In the present invention, the storage modulus of the hydrogel formed by mixing the first solution and the second solution may be 10 to 100 Pa.
In a specific exemplary embodiment of the present invention, hyaluronic acid
(HA-VP) into which the vascular endothelial growth factor mimic peptide was introduced was prepared by reacting hyaluronic acid in which a carboxylic acid functional group was activated and the vascular endothelial forming factor mimic peptide (Table 1 and Table 2), and when the chitosan (CH) solution and the HA-VP solution were mixed, it was confirmed that the chitosan and HA-VP formed a hydrogel through electrostatic interaction (
It was confirmed that the hydrogel was prepared through the electrostatic interaction of the cationic polymer such as chitosan (CH), cationic dextran (CD), polyethyleneimine (PEI), polylysine (PL) or poly histidine (PH) and the anionic HA or hyaluronic acid (HA-VP) into which the endothelial growth factor mimic peptide was introduced (
In addition, as a result of confirming whether the hydrogel formation is affected by the chemical introduction of VP and the presence or absence of a stem cell recruitment factor, it was confirmed that the hydrogel was formed through electrostatic attractive regardless of before and after the chemical introduction of VP and whether the progenitor cell/stem cell recruitment factor was mixed (
In another specific exemplary embodiment of the present invention, as a result of confirming the degree of the release of the stem cell recruitment factor and VP in the hydrogel prepared by electrostatic interaction, it was confirmed that the introduction of VP did not affect the characteristics of the hydrogel as a carrier, and when VP was chemically introduced, it had high sustained-release properties compared to the case where VP was simply mixed (
In addition, it was confirmed that the hydrogel of the present invention is non-toxic (
In still another specific exemplary embodiment of the present invention, as a result of confirming the angiogenesis effect by the hydrogel of the present invention in vitro, compared to the hydrogel (CHHA+VP) prepared by simply mixing the vascular endothelial growth factor mimic peptide with hyaluronic acid, the number of CD31-expressing stem cells was significantly increased in the hydrogel (CHHA-VP) prepared by chemically introducing the vascular endothelial growth factor mimic peptide into hyaluronic acid (
In still another specific exemplary embodiment of the present invention, as a result of confirming the ability to recruit stem cells by the hydrogel of the present invention in vivo, it was confirmed that the mesenchymal stem cells injected into the tail of mice were recruited toward the hydrogel by the stem cell recruitment factor included in the hydrogel (
As a result of confirming whether actual angiogenesis is induced in vivo by the hydrogel of the present invention, it was confirmed that the hydrogels maintained their shape well for 4 weeks, and more blood vessels were observed on the surface of the hydrogel when VP was chemically introduced (
That is, it was confirmed that the injectable hydrogel composition having the recruitment of endogenous progenitor cells or stem cells and the induction of vascular differentiation of the recruited cells according to the present invention releases the stem cell recruitment factor from the injected hydrogel to thereby recruit endogenous progenitor cells/stem cells into the hydrogel, and the induction of angiogenesis was promoted by differentiating into vascular cells by the vascular differentiation inducing factor which was chemically introduced into the hyaluronic acid, and in particular, when the blood vessel differentiation inducing factor was chemically introduced into hyaluronic acid, it was confirmed that it exhibited a high angiogenesis-inducing effect.
Further, in the present invention, since endogenous progenitor cells/stem cells are recruited without injecting progenitor cells/stem cells from the outside, various side effects caused by injection into external cells may be solved. Therefore, since the hydrogel composition of the present invention can be used for tissue regeneration by utilizing endogenous progenitor cells/stem cells in the field of regenerative medicine, it can be applied to tissue improvement, wound treatment, scar improvement, skin tissue improvement, soft tissue connective correction, wrinkle removal or improvement, contour correction, tissue enlargement, breast enlargement or the like.
In another aspect, the present invention relates to an injection for tissue regeneration including the injectable hydrogel composition.
In another aspect, the present invention relates to an injection for fillers including the injectable hydrogel composition.
The injection for fillers is preferably an injection for dermal fillers, and it may be applied to the improvement and treatment of wrinkles, scars or dermal tissue.
In the hydrogel composition of the present invention, it was confirmed that the stem cell recruitment factor was released from the injected hydrogel, and endogenous progenitor cells/stem cells were recruited in the hydrogel, and the induction of angiogenesis was promoted by differentiating into vascular cells by the vascular differentiation inducing factor chemically introduced into hyaluronic acid. In particular, it was confirmed that when the vascular differentiation inducing factor was chemically introduced into hyaluronic acid, a high angiogenesis-inducing effect was observed. Therefore, the hydrogel composition of the present invention has excellent recruitment of endogenous progenitor cells/stem cells and induction of vascular differentiation, and thus, it can be effectively applied to various tissue regenerations and wound treatments in addition to the formation of blood vessels.
Preparation of hyaluronic acid in which vascular endothelial growth factor mimic peptide is introduced
In the present invention, in order to prepare an injectable hydrogel composition having the recruitment of endogenous progenitor cells or stem cells and the induction of vascular differentiation of the recruited cells, first, hyaluronic acid into which a vascular endothelial growth factor mimic peptide was introduced was prepared.
Specifically, an HA solution was prepared by dissolving hyaluronic acid (HA) in distilled water at 10 mg/mL, and then 0.3 mg of 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMTMM, Sigma, USA) was added to 10 mL of the HA solution and stirred for 1 hour to activate the carboxylic acid functional group of hyaluronic acid.
10 mL of the HA solution in which the carboxylic acid functional group was activated was added dropwise to a VP solution prepared by dissolving 1.9 mg of the vascular endothelial growth factor mimic peptide (VP, SEQ ID NO: 1: KLTWQELYQLKYKGI) in 1 mL of distilled water, and it was reacted by stirring for 24 hours. The reaction solution was dialyzed for 72 hours and freeze-dried at −80° C. to prepare HA-VP into which VP was introduced.
As shown in Tables 1 and 2 below, the VP introduced into the HA was confirmed through TNBSA analysis (Table 1) and elemental analysis (Table 2).
In the present invention, in order to confirm whether the HA-VP and chitosan prepared in Example 1 above formed a hydrogel through electrostatic interaction, the Zeta potential and rheological properties according to the ratio of chitosan and HA-VP were measured.
First, a CH solution was prepared by dissolving chitosan (CH, sigma, USA) in a 0.1 M aqueous acetic acid solution at a concentration of 20 mg/mL, and HA-VP was dissolved in distilled water at 20 mg/mL to prepare an HA-VP solution.
The CH solution and the HA-VP solution were mixed in proportions and the zeta potential of the hydrogel formed through electrostatic bonding was measured with ELS-Z (Otsuka Electronics, Japan), and the rheological properties were measured with a modular compact rheometer (MCR 102, Anton Paar, Austria). The measurement conditions for rheological properties were that the parallel plate had a diameter of 25 mm, the distance from the bottom surface was 0.3 mm, and the strain was 2% at 25° C. and 1 Hz.
As a result, as shown in
In the present invention, hydrogels were prepared by electrostatic interaction by respectively mixing the cationic polymer chitosan (CH), cationic dextran (CD), polyethyleneimine (PEI), polylysine (PL) or polyhistidine (PH) and anionic HA or hyaluronic acid (HA-VP) introduced with a vascular endothelial growth factor mimic peptide.
First, a CH solution was prepared by dissolving chitosan (CH, sigma, USA) in a 0.1 M aqueous acetic acid solution at a concentration of 20 mg/mL, and CD, PEI, PL, PH, HA and HA-VP were each dissolved in distilled water at 20 mg/mL to prepare a CD solution, a PEI solution, a PL solution, a PH solution, an HA solution and an HA-VP solution.
Then, hydrogels were prepared by mixing the CH solution, CD solution, PEI solution, PL solution or PH solution and HA solution, or the CH solution, CD solution, PEI solution, PL solution or PH solution and HA-VP solution in the same volume.
As a result, as shown in
A CHHA hydrogel or CHHA-VP hydrogel was prepared by mixing the CH solution and the HA solution prepared in Example 3-1 or the CH solution and the HA-VP solution in the same volume. In addition, one of SP, WKYMVM, SDF1a, G-SCF and MCP-1 (Genscript, USA) was dissolved in each of the CH, HA and HA-VP solutions having the same concentration at a concentration of 1 μg/mL, respectively, to prepare solutions, and it was carried out in the same way as forming the hydrogels previously.
As shown in
In order to evaluate the rheological properties of the CHHA and CHHA-VP hydrogels prepared in Example 3 above, the modulus and viscosity were measured using a modular compact rheometer (MCR 102, Anton Paar, Austria). In this case, the used parallel plate had a diameter of 25 mm, an interval from the bottom surface of 0.3 mm, and a strain of 2% at 25° C. and 1 Hz.
As a result, as shown in
In the present invention, it was attempted to confirm the release degree of the stem cell recruitment factor and VP in the hydrogels prepared by electrostatic interaction.
First, an HA+VP solution was prepared by simply mixing VP with the HA solution prepared in Example 3 at a concentration of 320 μg/mL, and SP, which is a stem cell recruitment factor, was mixed with each of the HA solution and HA-VP solution at a concentration of 2 μg/mL to prepare an HA+SP solution and HA−VP+SP solution, respectively.
Next, 100 μL of the CH solution and 100 μL of the HA+VP solution were mixed in a vial to prepare 200 μL of a CHHA+VP hydrogel, and 100 μL of the CH solution and 100 μL of the HA-VP solution were mixed in a vial to prepare 200 μL of a CHHA-VP hydrogel.
In addition, 100 μL of the CH solution and 100 μL of HA+SP solution were mixed in a vial to prepare 200 μL of a CHHA+SP hydrogel, and 100 μL of the CH solution and 100 μL of the HA-VP+SP solution were mixed in a vial to prepare 200 μL of a CHHA-VP+SP hydrogel.
3 mL of a physiological saline was placed in each vial and stored in an incubator at 100 rpm and 37° C., and after taking 1 mL of the physiological saline from the vial at a predetermined time, 1 mL of a new physiological saline was added to the vial, and it was carried it out for 28 days to conduct a release experiment.
As a result, as shown in
In addition, as shown in
The presence or absence of toxicity of the hydrogels prepared in the present invention was evaluated.
First, human mesenchymal stem cells (hMSCs) were mixed at a concentration of 1×106 cells/mL in each of the CH, HA, HA-VP solutions prepared in Example 3 above and the HA+VP solution prepared in Example 5 above.
Cytotoxicity evaluation test was performed for the prepared solutions, and in order to form hydrogels, (1) 200 μL of the CH solution and 200 μL of the HA solution were mixed to form 400 μL of a CHHA hydrogel, (2) 200 μL of the CH solution and 200 μL of the HA+VP solution were mixed to form 400 μL of a CHHA+VP hydrogel, and (3) 200 μL of the CH solution and 200 μL of the HA-VP solution were mixed to form 400 μL of a CHHA-VP hydrogel in 24-well plates. As a control group for comparison, 4×105 hMSC cells were added to a 24-well plate. The medium was added by 1 mL and was replaced every 3 days, and cytotoxicity was measured by MTT analysis on days 1, 4 and 7.
As a result, as shown in
In the present invention, the ability of the SP to recruit stem cells was confirmed.
First, human-derived mesenchymal stem cells (hMSCs) were labeled with PKH 26 dye (Sigma, USA) and then dispensed in the upper chamber (8.0 μm pore size) of a 24-well trans well plate (SPL, Korea) to be 5×104 cells. After culturing in the upper chamber for 48 hours using serum-free DMEM (Dubelco's modified eagle medium, Gibco, USA) medium, DMEM containing 1 μg/mL SP and 1% FBS (fetal bovine serum, Gibco, USA) was added to the bottom well, and the medium was replaced every 3 days, and the hMSCs that migrated to the bottom well were observed using a fluorescence microscope (Olympus, Japan). As a control group, DMEM (including 1% FBS) without SP was added to the bottom well and observed.
As a result, as shown in
In order to confirm the angiogenesis-inducing effect of the injectable hydrogel of the present invention, CHHA, CHHA+VP and CHHA-VP hydrogels including human-derived mesenchymal stem cells were prepared in the same manner as in Example 6, respectively, and the medium was exchanged every 3 days and cultured for 4 weeks.
At the 1st 2nd 3rd and 4th weeks of culture, the hydrogel was fixed with formalin, and immunofluorescence analysis was performed to observe the expression of CD31, which is known to be expressed in vascular cells.
As a result, as shown in
After extracting mRNA from the hydrogels of Example 8-1, cDNA was synthesized using the extracted mRNA, and changes in the expressions of the von
Willebrand factor (vWF) gene and the CD31 gene expressed in vascular cells were confirmed through qRT-PCR (quantitative real-time polymerase chain reaction).
As a result, as shown in
Confirmation of recruitment of stem cells by hydrogel in vivo It was confirmed whether the actual stem cells were recruited in vivo by the hydrogel of the present invention.
First, for fluorescence imaging, 1 μg/mL of SP, which is a stem cell recruitment factor, was added to each of an FITC-labeled chitosan (CH) solution and a hyaluronic acid (HA) solution, and a total of 200 μL of the hydrogel by 100 μL, each was injected subcutaneously in nude mice through a dual syringe.
Afterwards, 1×106 hMSCs labeled with IR-783 were injected into the tail vein of nude mice, and cell migration was observed through fluorescence imaging. The results are shown in
As a result, as shown in
It was confirmed whether actual angiogenesis was induced in vivo by the hydrogel of the present invention.
First, 1 μg/mL of SP, which is a stem cell recruitment factor, was added to the CH, HA, HA+VP, and HA-VP solutions, respectively, and then, a total of 200 p.1_, of the hydrogel by 100 μL each was injected subcutaneously in nude mice through a dual syringe in the combination of CH and HA, CH and HA+VP, and CH and HA-VP. For comparison, the CH, HA, HA+VP and HA-VP solutions without the recruitment factor were injected in the same way. Afterwards, 1×106 hMSCs labeled with BrdU were injected into the tail vein of nude mice. The hydrogel was extracted at the 1st 2nd 3rd and 4th weeks, and it was observed whether angiogenesis occurred.
As a result, as shown in
Immunofluorescence analysis was performed on BrdU and CD31 to confirm the degrees of hMSCs recruited in the hydrogels extracted in Example 10-1 and angiogenesis. The observation results are shown in
As a result, as shown in
In the case of CD31 stained green, it was confirmed that the number of cells expressing CD31 was increased when VP was chemically introduced, compared to when VP was physically mixed.
Further, in the case of cells stained with BrdU and CD31 at the same time, it was confirmed that it was increased more in the hydrogel in which VP was chemically introduced, and through this, it was confirmed that the differentiation into vascular cells and the promotion of angiogenesis were more effectively induced in the presence of the recruitment factor by VP which was chemically introduced and recruited hMSCs inside the hydrogel than VP which was simply mixed.
The mRNA was extracted from the hydrogels extracted in Example 10-1, and qRT-PCR was performed in the same manner as in Example 8-2.
As a result, as shown in
In the hydrogel composition having the recruitment of endogenous progenitors or stem cells and the induction of vascular differentiation of recruited cells according to the present invention, it was confirmed that the stem cell recruitment factor was released from the injected hydrogel, and endogenous progenitor cells/stem cells were recruited in the hydrogel, and the induction of angiogenesis was promoted by differentiating into vascular cells by the vascular differentiation inducing factor chemically introduced into hyaluronic acid. In particular, it was confirmed that when the vascular differentiation inducing factor was chemically introduced into hyaluronic acid, a high angiogenesis-inducing effect was observed. Therefore, the hydrogel composition of the present invention has excellent recruitment of endogenous progenitor cells/stem cells and induction of vascular differentiation, and thus, since it can be effectively applied to various tissue regenerations and wound treatments in addition to the formation of blood vessels, it has high industrial applicability.
SEQ ID NO: 1 shows the amino acid sequence of the vascular endothelial growth factor mimic peptide, which is a vascular differentiation inducing factor.
SEQ ID NO: 2 shows the amino acid sequence of the substance P (SP) peptide, which is a stem cell recruitment factor.
SEQ ID NO: 3 shows the amino acid sequence of the WKYMVM peptide, which is a stem cell recruitment factor.
SEQ ID NO: 4 shows the nucleotide sequence of the forward primer of vWF.
SEQ ID NO: 5 shows the nucleotide sequence of the reverse primer of vWF.
SEQ ID NO: 6 shows the nucleotide sequence of the forward primer of CD31.
SEQ ID NO: 7 shows the nucleotide sequence of the reverse primer of CD31.
SEQ ID NO: 8 shows the nucleotide sequence of the forward primer of GAPDH.
SEQ ID NO: 9 shows the nucleotide sequence of the reverse primer of GAPDH.
| Number | Date | Country | Kind |
|---|---|---|---|
| 10-2019-0151219 | Nov 2019 | KR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/KR2020/016135 | 11/17/2020 | WO |
