Tissue gel with kidney tissue endocrine recovery function and preparation method thereof
By mixing renal cortical tissue into a hydrogel matrix material to prepare tissue gel, the problems of ectopic renal tissue reconstruction and endocrine function recovery were solved. Stable colonization and functional recovery of renal tissue in the spleen were achieved, and endocrine function was restored in mice with renal injury.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-30
- Publication Date
- 2026-03-13
AI Technical Summary
There is currently no effective method to use the spleen as a host organ to achieve ectopic reconstruction of kidney tissue and restoration of endocrine function.
A tissue gel is provided, which is prepared by mixing renal cortical tissue into a hydrogel matrix material. The preparation method includes pretreatment and mixing of the renal cortical tissue, and is used to achieve stable colonization of renal tissue and restoration of endocrine function in the spleen.
It achieved good colonization and vascularization of kidney tissue in the spleen, restored the renal tubule and glomerular-like structure, and had renal endocrine functions, such as the secretion of erythropoietin (EPO), renin, and active vitamin D3, thus restoring normal endocrine function in mice with kidney injury.
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Figure CN121648159A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biomedical and regenerative medicine technology, specifically, it relates to a tissue gel that restores the endocrine function of kidney tissue and its preparation method. Background Technology
[0002] In addition to their excretory functions, the kidneys also play a crucial endocrine role, including secreting erythropoietin (EPO), renin, and activated vitamin D3 (1,25(OH)2D3). Traditional kidney transplantation primarily restores excretory function, with limited research on the reconstruction of endocrine function. The liver and spleen, due to their rich blood supply and stable microenvironment, are considered excellent biological "culture dishes" for xenografted plants. Existing studies have shown that the spleen can support the survival and functional maintenance of various xenograft tissues (pancreatic islets, liver tissue, etc.).
[0003] However, there is currently no systematic research on methods for using the spleen as a host organ to achieve ectopic reconstruction of kidney tissue and restoration of endocrine function. Summary of the Invention
[0004] To address the challenge of restoring endocrine function after kidney injury, this invention provides a tissue gel that restores the endocrine function of kidney tissue and its preparation method. By leveraging the advantages of spleen blood supply and immune regulation, it enables stable colonization of kidney tissue within the spleen and restoration of endocrine function.
[0005] Therefore, the present invention provides the following technical solution.
[0006] The first aspect of the present invention provides a tissue gel that restores the endocrine function of renal tissue, the tissue gel being mixed with renal cortical tissue in a hydrogel matrix material; the mass ratio of the hydrogel matrix material to the renal cortical tissue is (1-10):1.
[0007] In a preferred embodiment of the present invention, the hydrogel matrix material is selected from one or a mixture of two or more of sodium alginate, gelatin, hyaluronic acid, collagen, fibroin, polyethylene glycol, and polylactic acid-glycolic acid copolymer.
[0008] A second aspect of the present invention provides a method for preparing a tissue gel as described above that restores the endocrine function of renal tissue, the method comprising the following steps: S1: Isolate the renal cortex tissue from the kidney; S2: The renal cortical tissue is pretreated and then mixed with a hydrogel matrix material to obtain a tissue gel.
[0009] In a preferred embodiment of the present invention, in step S1, the kidney is derived from experimental animals commonly used in biological experiments, such as mice, rats, guinea pigs, rabbits, cats, dogs, pigs, monkeys, etc.
[0010] In a preferred embodiment of the present invention, step S2, the preprocessing process includes: S21: Cut the renal cortex into tissue blocks; S22: Rinse the tissue block and then treat it with collagenase I.
[0011] In a preferred embodiment of the present invention, in step S21, the size of the tissue block is 0.5-2 mm.
[0012] In a preferred embodiment of the present invention, in step S22, the rinsing is performed using a PBS buffer containing penicillin and streptomycin.
[0013] In a preferred embodiment of the present invention, in step S22, the mass concentration of collagenase I is 0.1%.
[0014] In a preferred embodiment of the present invention, the processing time in step S22 is 2-10 min.
[0015] In a preferred embodiment of the present invention, in step S2, the hydrogel matrix material is a hydrogel matrix material / physiological saline solution with a mass concentration of 0.5%-5%.
[0016] A third aspect of the invention provides the use of the tissue gel as described above, or the tissue gel prepared according to the preparation method described above, in the preparation of a medicament for treating renal impairment.
[0017] A fourth aspect of the present invention provides a medicament for treating kidney damage, comprising: (i) the tissue gel as described above or the tissue gel prepared according to the preparation method described above; and (ii) Pharmaceutically acceptable carriers or excipients.
[0018] In a preferred embodiment of the present invention, the renal function impairment is renal failure or renal tissue endocrine function impairment.
[0019] In a preferred embodiment of the present invention, the pharmaceutically acceptable carrier or excipient is a calcium chloride solution.
[0020] In a preferred embodiment of the present invention, the drug is administered via injection.
[0021] In a preferred embodiment of the present invention, the injection volume is 10-100 μL and the injection depth is 0.5-2 mm.
[0022] A fifth aspect of the present invention provides a method for preparing a drug as described above, the method comprising the following steps: Simply mix the tissue gel as described above, or the tissue gel prepared according to the method described above, with the calcium chloride solution.
[0023] In a preferred embodiment of the present invention, the concentration of sodium alginate in the drug is 0.1-10 wt%; In a preferred embodiment of the present invention, the concentration of the calcium chloride solution in the drug is 0.1-10 wt%.
[0024] By employing the above technical solution, the present invention has at least the following advantages: The tissue gel of this invention, after transplantation, achieves excellent colonization and vascularization within the spleen, exhibiting typical renal tubular and glomerular-like structures and possessing renal endocrine functions, such as the secretion of erythropoietin (EPO), renin, and active vitamin D3, thereby restoring normal endocrine function in mice with renal injury. This tissue gel enables long-term survival and functional reconstruction of ectopic kidney tissue within the spleen, featuring simple operation, high success rate, and significant clinical translational potential, providing a novel treatment strategy for renal failure and endocrine disorders.
[0025] The above description is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention and to implement it in accordance with the contents of the specification, the preferred embodiments of the present invention are described in detail below. Attached Figure Description
[0026] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.
[0027] Figure 1 The image shows the H&E staining results of a pathological section of the spleen tissue of the recipient mouse after transplantation according to the present invention.
[0028] Figure 2 This is a statistical graph showing the increase in serum EPO levels in recipient mice after transplantation compared to pre-transplantation levels.
[0029] Figure 3This is a statistical graph showing the rate of increase in renin activity in recipient mice after transplantation according to the present invention.
[0030] Figure 4 This is a statistical chart showing the increase in the rate of active vitamin D3 in recipient mice after transplantation according to the present invention. Detailed Implementation
[0031] To make the technical means, creative features, achieved objectives, and effects of this invention readily understandable, the technical solutions in the embodiments of this invention will be clearly and completely described below in conjunction with the embodiments of this invention. Obviously, the described embodiments are merely some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.
[0032] Unless otherwise specified, the percentage content mentioned in this invention refers to mass percentage for solid-liquid mixtures and solid-phase-solid mixtures, and volume percentage for liquid-phase-liquid mixtures.
[0033] Unless otherwise specified, all percentage concentrations mentioned in this invention refer to the final concentration. The final concentration refers to the proportion of the added component in the system after the addition of that component.
[0034] Unless otherwise specified, the temperature parameters in this invention can be either constant temperature processing or processing within a certain temperature range. The constant temperature processing allows temperature fluctuations within the precision range controlled by the instrument.
[0035] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods.
[0036] Unless otherwise specified, the materials and reagents used in the following examples are commercially available.
[0037] Example 1: Preparation of tissue gel Kidneys were harvested from donor mice (C57BL / 6J strain, purchased from Jiangsu Jicui Pharmaceutical Co., Ltd.). The cortical portion (approximately 70% of the kidney volume) was separated, and the medulla and capsule were removed to obtain renal cortical tissue. The obtained renal cortical tissue was cut into tissue blocks of 0.5-2 mm in size. The obtained tissue blocks were rinsed 3-8 times in PBS containing penicillin and streptomycin, followed by treatment with 0.1% collagenase I for 2-10 minutes to remove the interstitium, resulting in pretreated renal cortical tissue blocks. The pretreated renal cortical tissue blocks were mixed thoroughly with 5 wt% sodium alginate solution to obtain a tissue gel.
[0038] Experimental Example 1: Application of Tissue Gel In this embodiment, mice were used as subjects. The tissue gel was transplanted into the spleen of the recipient mouse. The specific operation method is as follows: 1. Preparation of transplant drugs First, the tissue gel prepared in Example 1 was thoroughly mixed with a 2wt% calcium chloride solution to obtain drug I.
[0039] 2. Construction of a mouse model of renal function impairment The recipient mice used were mice with renal function impairment induced by 5 / 6 nephrectomy. The specific construction method was as follows: Male C57BL / 6J mice aged 8-10 weeks (purchased from Jiangsu Jicui Pharmaceutical Co., Ltd.) underwent partial left nephrectomy under anesthesia and aseptic conditions. The left kidney was exposed through a left lumbar incision, preserving the renal hilar vessels. Approximately two-thirds of the renal parenchyma was removed from the upper and lower poles of the left kidney. After adequate hemostasis, the remaining kidney was repositioned and the incision was sutured. After 5-7 days of recovery, the second step of the surgery was performed: the right kidney was exposed through a right lumbar incision. The right renal artery, vein, and ureter were ligated and transected, and the right kidney was completely removed and the incision sutured. Postoperatively, the animals were kept warm, given fluid replacement, and analgesia, and their general condition was continuously monitored. The model's effectiveness was assessed 2-4 weeks post-transplantation by detecting serum creatinine and blood urea nitrogen levels, as well as renal tissue pathological changes. Generally, blood samples were collected from mice 2-4 weeks after transplantation to measure serum creatinine and blood urea nitrogen levels. Compared to sham-operated controls, successfully modeled mice should show a sustained increase in these indicators, indicating a significant decrease in glomerular filtration function. Simultaneously, residual renal tissue was collected for pathological examination. HE and PAS staining revealed increased glomerular volume, mesangial matrix proliferation, and partial glomerular sclerosis. Masson staining showed increased interstitial collagen deposition and fibrosis, accompanied by varying degrees of inflammatory cell infiltration. If biochemical indicators were abnormally elevated and consistent with the aforementioned typical pathological changes, the 5 / 6 nephrectomy-induced renal function impairment model could be considered successfully established, thus forming a stable chronic renal function impairment phenotype.
[0040] 3. Drug transplantation First, the recipient model mice were divided into two groups: a control group and a KS group, with 5 mice in each group. Then, the mice in each group underwent the following treatments. To avoid significant damage to the kidney tissue during transplantation, the following steps were performed under a microscope: KS group: The spleen was exposed through an incision in the left upper abdomen of recipient mice, and drug I (containing 5-30 tissue blocks) prepared above was slowly injected using a microinjector; Note: The injection volume was 50 μL, and the injection depth was 1 mm. The capsule was kept intact during injection, and the spleen surface was uniformly colored and there was no bleeding after injection; Control group: The spleen was exposed through an incision in the left upper abdomen of recipient mice, and PBS solution was slowly injected using a microinjector; Note: The injection volume was 50 μL, and the injection depth was 1 mm. The spleen capsule was kept intact during injection, and the spleen surface was uniformly colored and there was no bleeding after injection.
[0041] The recipient mice were fed routinely after transplantation. They were given enrofloxacin in drinking water (0.05-0.1 mg / mL, for 3-5 consecutive days) after surgery. They were also given cyclosporine A (10 mg / kg) for immunosuppression for 3-5 days after transplantation to promote graft colonization in the recipients.
[0042] The following tests were then performed: (1) Graft survival was assessed 6 weeks post-transplantation. Results are shown in […]. Figure 1 .
[0043] (2) Serum EPO, renin activity, and 1,25(OH)2D3 levels in recipients were measured 6 weeks post-transplantation. The results are shown in […]. Figure 2-4 .
[0044] 4. Results and Analysis like Figure 1 As shown, the H&E staining results of pathological sections of spleen tissue from recipient mice after transplantation are illustrated; Figure 1 It can be seen that after transplantation, the kidney tissue block can continue to survive in the spleen of mice and has typical renal tubule and glomerular structures.
[0045] like Figure 2 As shown, a statistical graph illustrates the increase in serum EPO levels in recipient mice after transplantation compared to before transplantation; Figure 2 It can be seen that the long-term survival rate of transplanted tissues reached 100%, and serum EPO increased by 100% after surgery compared with before surgery.
[0046] like Figure 3 As shown, a statistical graph illustrating the rate of increase in renin activity in recipient mice after transplantation is presented; Figure 3 It can be seen that the renin activity in transplanted mice increased by 70%.
[0047] like Figure 4 As shown, a statistical graph illustrating the increase in the rate of active vitamin D3 in recipient mice after transplantation is presented; Figure 4 It can be seen that the active vitamin D3 increased by 80%.
[0048] The above results demonstrate that the tissue gel and transplantation method of the present invention can achieve good survival and functional recovery of kidney tissue in the spleen of recipient animals in a short period of time.
[0049] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the methods and techniques disclosed above without departing from the scope of the present invention to create equivalent embodiments. Any simple modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.
Claims
1. A tissue gel that restores the endocrine function of kidney tissue, characterized in that, The tissue gel is mixed with renal cortical tissue in a hydrogel matrix material; the mass ratio of the hydrogel matrix material to the renal cortical tissue is (1-10):
1. Preferably, the hydrogel matrix material is selected from one or a mixture of two or more of sodium alginate, gelatin, hyaluronic acid, collagen, fibroin, polyethylene glycol, and polylactic-co-glycolic acid copolymer.
2. The method for preparing the tissue gel with renal endocrine function restoration according to claim 1, characterized in that, The preparation method includes the following steps: S1: Isolate the renal cortex tissue from the kidney; S2: The renal cortical tissue is pretreated and then mixed with a hydrogel matrix material to obtain a tissue gel.
3. The preparation method according to claim 2, characterized in that, In step S2, the preprocessing process includes: S21: Cut the renal cortex into tissue blocks; S22: Rinse the tissue block and then treat it with collagenase I.
4. The preparation method according to claim 3, characterized in that, In step S21, the size of the tissue block is 0.5-2 mm.
5. The preparation method according to claim 3, characterized in that, In step S22, the rinsing is performed using PBS buffer containing penicillin and streptomycin; The mass concentration of collagenase I is 0.1%; The processing time is 2-10 minutes.
6. The use of the tissue gel according to claim 1 or the tissue gel prepared according to any one of claims 2-5 in the preparation of a medicament for treating renal impairment.
7. A drug for treating kidney damage, characterized in that, include: (i) The tissue gel of claim 1 or the tissue gel prepared according to any one of claims 2-5; as well as (ii) Pharmaceutically acceptable carriers or excipients.
8. The medicament according to claim 7, characterized in that, The kidney function impairment refers to kidney failure or kidney tissue endocrine function impairment. The pharmaceutically acceptable carrier or excipient is calcium chloride solution.
9. The drug according to claim 7, characterized in that, The drug is administered via injection. Preferably, the injection volume is 10-100 μL and the injection depth is 0.5-2 mm.
10. A method for preparing the drug according to any one of claims 7-9, characterized in that, The method includes the following steps: The tissue gel described in claim 1 or the tissue gel prepared according to any one of claims 2-5 is thoroughly mixed with calcium chloride solution; Preferably, the concentration of sodium alginate is 0.1-10 wt%; Preferably, the concentration of the calcium chloride solution is 0.1-10 wt%.