Implantable compositions used for cartilage defect repair in postoperative rehabilitation.

Abstract

The present invention relates to an implantable composition comprising a piezoelectric polymer loaded with magnetic microparticles or magnetic nanoparticles as its main component, and further comprising cells derived from stem cells, chondrocyte progenitor cells, or mature chondrocytes. This composition is useful in postoperative regenerative rehabilitation as part of the treatment of joint diseases caused by cartilage defects (e.g., osteoarthritis).

Description

[Technical Field]

[0001] The present invention relates to a composition comprising a piezoelectric polymer loaded with magnetic microparticles or magnetic nanoparticles as a main component, and further comprising cells derived from stem cells, chondrocyte progenitor cells, or mature chondrocytes. This composition is useful in postoperative regenerative rehabilitation as part of the treatment of joint diseases caused by cartilage defects (e.g., osteoarthritis).

[0002] This invention was first disclosed at the 13th Annual Meeting of The Orthobiologic Institute (TOBI), held from June 9-11, 2022, at The Diplomat Beach Resort Hollywood, Curio Collection by Hilton, Hollywood, Florida, United States. [Background technology]

[0003] Musculoskeletal disorders resulting from cartilage defects are conditions that cause severe functional impairment, posing a widespread and increasing health burden that significantly impacts patients, healthcare systems, and associated medical costs. Among the diseases associated with cartilage defects, osteoarthritis is one of the most representative. Given the importance and prevalence of osteoarthritis, the following description will primarily refer to this disease, but the present invention is generally applicable to all diseases associated with cartilage defects.

[0004] Several epidemiological studies have shown that this condition is highly correlated with the aging of the global population and the rising rate of obesity, and its importance is increasing year by year. These studies estimate that approximately 250 million people currently suffer from symptomatic osteoarthritis with limited activity. To date, no effective treatment has been developed, and the only definitive treatment is total joint replacement (TJR). However, several observational studies have reported that the failure rate of TJR remains high in younger, more active patients, those with comorbidities, and those who underwent surgery at low-case centers. In particular, it is noteworthy that up to 20% of artificial joints require revision within 20 years.

[0005] More advanced and promising approaches are based on cartilage repair techniques (RCTs) such as microfractures (MF), autologous chondrocyte transplantation (ACI), and matrix-assisted autologous chondrocyte transplantation (MACI). The rationale is to repair cartilage defects by inducing a pro-regenerative response in situ by stimulating the formation of new functional tissue. However, the success rates of these innovative techniques remain too low. In fact, the main limitation of RCTs lies in the biological inactivity of the matrix used as a cell support. Numerous attempts have been made to overcome this limitation, resulting in a focus on developing pro-regenerative rehabilitation protocols that rely on a clearly defined pro-regenerative response of chondrocytes after mechanical stimulation. Unfortunately, however, due to the lack of techniques to produce effective mechanical stimulation at the cellular level, rehabilitation is not entirely efficient, and beneficial effects in vivo are highly variable and insufficient.

[0006] In reality, regardless of the repair technique employed, restoring overall pre-operative mobility requires long and challenging postoperative rehabilitation. Furthermore, each patient is an individual case requiring a specially developed (ad hoc) individualized rehabilitation protocol. This significant heterogeneity leads to longer recovery periods, potentially up to 25 months.

[0007] Due to these problems, the RCT approach still has a high failure rate, and postoperative outcomes are severely limited, especially in younger patients.

[0008] The treatment of diseases related to cartilage defects remains an unresolved issue that still requires clinical solutions.

[0009] The current technical challenge is to develop a matrix that actively supports cells after implantation and integrates various already developed techniques to fully realize their potential.

[0010] The objective of this invention is to obtain a biocompatible material that functions as an active support in RCT surgery through biofabrication. This type of matrix can enhance the regenerative potential of cells by simultaneously providing in situ physical regenerative stimuli. Notably, this matrix enables the implementation of individualized regenerative regimens at the cellular level through a cost-effective and minimally invasive approach.

[0011] The matrix described herein is intended for use in postoperative regenerative rehabilitation performed as part of the treatment of diseases pathologically related to cartilage defects (e.g., osteoarthritis). [Overview of the project]

[0012] This and other objectives are achieved by the present invention. A first aspect of the present invention relates to the following biocompatible compositions. A biocompatible composition used for implantation in a body site that has undergone surgery for the treatment of a disease characterized by cartilage defects, a) A matrix comprising an intrinsically piezoelectric biocompatible polymer, or a non-intrinsically piezoelectric biocompatible polymer loaded with an inorganic or organic filler that imparts piezoelectricity to a non-intrinsically piezoelectric polymer, b) 1 × 10 5 ~1 × 10 9 Magnetic material with nanoparticles / mL, c) 1 × 10 per 1 mL of composition 5 ~1 × 10 8 A cell comprising a cell selected from the group consisting of mesenchymal stem cells, mature chondrocytes, and chondrocyte progenitor cells, Biocompatible composition.

[0013] A second aspect of the present invention is a method for treating a joint disease in a subject requiring treatment, comprising the step of using a biocompatible composition described herein. [Brief explanation of the drawing]

[0014] [Figure 1] The electrical signals generated by the material of the present invention under compressive stress are shown, with the cases of no preload (Figure 1a) and with a preload of 5N (Figure 1b). [Figure 2] The results of MRI imaging obtained for one of the scaffolds of the present invention are shown, with coronal sections (Figure 2a) and sagittal sections (Figure 2b) respectively. [Figure 3] The X-ray measurements obtained for one of the scaffolds of the present invention are shown, with Figure 3a showing the results before magnetic excitation and Figure 3b showing the results after magnetic excitation, respectively. [Modes for carrying out the invention]

[0015] In the following explanation, the following terms and abbreviations will be used with the meanings set forth below. "NPs" stands for nanoparticles. "ES" stands for electrospun. "MF" stands for microfracture. "MACI" stands for Matrix-assisted Autologous Chondrocyte Implantation. • "PBS" refers to phosphate-buffered saline. • "GelMA" refers to gelatin methacrylate. • "hACs" refers to human articular chondrocytes. • Unless otherwise specified, percentages of components described below mean weight / volume (w / v)%, representing the gram amount of a component dissolved or dispersed in 100 mL of the composition. • Unless otherwise specified, cell density is expressed in cell / mL, representing the number of cells dispersed in 1 mL of suspension medium.

[0016] The inventors have found that when a piezoelectric biocompatible matrix loaded with magnetic microparticles or magnetic nanoparticles, which has been cellularized by chondrocyte progenitor cells (e.g., perichondroblasts, synoviocytes, etc.), mesenchymal stem cells, or mature chondrocytes, is implanted at the surgical intervention site by MF / MACI and subjected to magnetic irradiation during post-operative regenerative rehabilitation, it can maximize the effect of post-operative regenerative rehabilitation, particularly in the treatment of osteoarthritis. Magnetic nanoparticles embedded in the polymer matrix vibrate in response to an external pulsed magnetic field, causing controlled deformation of the matrix. The piezoelectric polymer generates electrical discharge in response to this deformation, promoting type II collagen production and the differentiation of progenitor cells into a chondroid phenotype (direct effect). Furthermore, the cyclic deformation of the matrix reproduces the therapeutic shear / compression deformation induced by regenerative rehabilitation therapy (indirect effect). This approach enables potent chondrogenesis in situ by an individualized and non-invasive method.

[0017] The first component of the composition of the present invention is or comprises a biocompatible polymer.

[0018] The polymers may have intrinsic piezoelectricity and include, for example, polyvinylidene fluoride (known as PVDF in the polymer field), poly(vinylidene fluoride-trifluoroethylene) (P(VDF-TrFE)), poly-L-lactic acid (PLLA), poly(3-hydroxybutyrate-co-3-hydroxyvaleric acid) (PHBV), cellulose, collagen, polyacrylonitrile (PAN), nylon-11, or mixtures thereof. These polymers may be used dissolved or suspended in a suitable solvent such as water, PBS at pH 7.4, culture medium (cellulose, collagen), tetrahydrofuran, diethyl ether (especially when the polymer is PLLA), dimethylformamide, hexafluoroisopropanol, methyl ethyl ketone (especially in combination with P(VDF-TrFE), particularly in the form of ES fibers), or mixtures thereof. The amount of polymer is typically 10-20% w / v.

[0019] Alternatively, the biocompatible polymer does not need to possess piezoelectric properties itself; in that case, it is loaded with piezoelectric inorganic filler microparticles or nanoparticles. In this case, the polymer can be of any type, but naturally derived polymers such as animal gelatin are preferred. These polymers can be used as hydrogels consisting of a polymer phase swollen with water, PBS, or culture medium. When the matrix is ​​a hydrogel, the amount of polymer is about 2-30% w / v of the total composition. A typical aqueous phase used for hydrogel preparation is phosphate buffer with a pH of 6-10. Non-piezoelectric polymers must be loaded with one or more piezoelectric inorganic fillers. Examples of inorganic fillers include barium titanate (BaTiO3) and lead zirconate titanate (Pb[Zr x Ti 1-x Examples include ¹O3 (0 ≤ x ≤ 1), zinc stanate (ZnSnO3), lithium niobate (LiNbO3), and sodium niobate (NaNbO3). These inorganic compounds are in the form of nanoparticles with a diameter (or maximum axial length) of 1 to 100 nm, or microparticles with a diameter (or maximum axial length) of 101 nm to 100 μm. Inorganic fillers may be present in the composition in an amount of 4 to 40% w / v.

[0020] The second component of the composition of the present invention is a magnetic material, which is dispersed in a matrix in the form of nanoparticles (NPs).

[0021] Many classes of magnetic materials are known (e.g., composite ceramic magnetic materials, single-molecule magnets (SMM), organic magnets of composition V(TCNE)₂·1 / 2CH₂Cl₂, wherein TCNE represents tetracyanoethylene), but preferred for the purposes of the present invention are magnetic metals, magnetic metal oxides, or magnetic metal alloys. Examples of magnetic metals include iron, nickel and cobalt. Examples of magnetic oxides include oxides of these metals, such as ferric oxide (Fe₂O₃), magnetite (Fe₃O₄), or mixed oxides containing these metals. Examples of magnetic alloys include steel, the alloy known as AlNiCo (comprising iron, nickel, cobalt and aluminum as main components), the alloy known as TiCoNiAl (comprising iron, nickel, cobalt, aluminum and titanium as main components), or alloys based on rare earth elements such as samarium-cobalt, neodymium-iron-boron. For the purposes of the present invention, it is also possible to use a mixture of NPs of different magnetic materials.

[0022] Magnetic NPs may be present in the composition in an amount of 1×10 5 to 1×10 9 NPs / mL.

[0023] The composition of the present invention further contains 1×10 5 to 1×10 8 cells per mL, selected from the group consisting of mesenchymal stem cells, chondrocyte progenitor cells, and mature chondrocytes. In a preferred embodiment, the present invention contains 2×10 6 to 2×10 7 cells per mL, selected from the group consisting of mesenchymal stem cells, mature chondrocytes, and chondrocyte progenitor cells.

[0024] In addition to the essential components mentioned above, the compositions of the present invention may also contain one or more additional components selected from, for example, antioxidants (e.g., ascorbic acid), preservatives (e.g., ethyl alcohol, benzyl alcohol, sodium benzoate, etc.), or bioactive agents (growth factors, drugs, antibiotics, antivirals, enzymes, vitamins, etc.).

[0025] A second aspect of the present invention is a method for treating a joint disease in a subject requiring treatment, comprising the step of using a biocompatible composition described herein.

[0026] Subjects (patients) requiring treatment with this joint disease treatment method may have cartilage defects or diseases that are etiologically related to cartilage defects. In particular, the disease related to cartilage defects may be osteoarthritis. [Examples]

[0027] The present invention is further illustrated by the following embodiments.

[0028] (Example 1: Gelatin hydrogel doped with BaTiO3 as a piezoelectric inductive agent) This example relates to the production of a scaffold based on porcine gelatin loaded with a piezoelectric filler consisting of BaTiO3 in nanopowder form. To this end, a 5% w / v gelatin solution was prepared by dissolving 500 mg of gelatin (Sigma-Aldrich, USA) in 10 mL of a 0.15% w / v genipine (Sigma-Aldrich, USA) solution in pH 7.4, 1 M phosphate-buffered saline (PBS) under magnetic stirring at 600 rpm at 60°C. Next, an amount of BaTiO3 nanopowder to 40% w / v was gradually added to the mixture and uniformly dispersed (the remainder of the composition, when considered as 100%, was the PBS solution). Genipine acts as a crosslinking agent to prevent the gelatin scaffold from dissolving after immersion in water, PBS, or culture medium. Magnetic stirring was stopped when complete dissolution was achieved, and the gelatin / genipine solution was then maintained at 60°C to allow crosslinking to proceed. The endpoint of the reaction was determined by tracking the change in the solution's color from pale yellow to medium blue or dark blue as the carboxyl group of genipine reacted with the amino group of gelatin. In this example, after the initial pale blue color appeared, the solution was held at 60°C for 15 minutes. Multiple disc-shaped scaffolds (φ=2.5cm, h=0.5cm) were prepared.

[0029] (Example 2: Measurement of the electric field generated from a gelatin / genipin / BaTiO3 scaffold after controlled deformation) The primary objective of this embodiment is to evaluate the inductive piezoelectric properties of the scaffold fabricated in Example 1. One of the scaffolds fabricated in Example 1 was subjected to controlled deformation using an electromechanical test system (Instron 5943, load measurement accuracy: ±0.5% of reading, simultaneous data acquisition option up to 2.5 kHz in load, elongation, and strain channels, speed range 0.05 to 2,500 mm / min (0.002 to 100 in / min), maximum load 1 kN (225 lbf), vertical test space 1,123 mm (44.2 in), INSTRON-USA). The generated electric field was measured using a digital multimeter GBC KDM-120 (GBC, Australia). Specifically, when the scaffold was deformed by 10%, a voltage of 3 mV was read.

[0030] These test results are shown in the graphs in Figures 1a and 1b. In these figures, the dashed line represents the applied strain, and the solid line represents the generated voltage. In particular, Figure 1a shows the voltage generated by a controlled compression cycle using a strain-controlled approach. This type of deformation resulted in a voltage change of 2 mV (Figure 1a).

[0031] Figure 1b shows the voltage change when the same deformation is applied after a preload of 5N. In this case, the voltage change increased to 10mV.

[0032] (Example 3: Gelatin hydrogel loaded with ferromagnetic microparticles that respond to an external magnetic field) A hydrogel crosslinked with genipine, derived from pigskin gelatin, was prepared according to the method described in Example 1. In short, a 5% w / v gelatin solution was prepared by dissolving 500 mg of gelatin (Sigma-Aldrich, USA) in 10 mL of a 0.15% w / v genipine (Sigma-Aldrich, USA) solution in pH 7.4, 1 M phosphate-buffered saline (PBS) under magnetic stirring at 60°C and 600 rpm. Once the polymer / genipine mixture was completely dissolved, a suspension of magnetic microparticles Dynabeads® M-280 Streptavidin (Thermofisher, USA) was added under magnetic stirring. Specifically, 6.7 × 10⁻⁶ 8 500 μL / mL was used from a stock suspension with a beads / mL concentration.

[0033] After obtaining a homogeneous suspension, magnetic stirring was stopped, and the gelatin / genipine solution was maintained at 60°C to allow crosslinking to proceed. After the initial pale blue color appeared, the solution was maintained at 60°C for 15 minutes. Multiple disc-shaped scaffolds (φ=2.5cm, h=0.5cm) were prepared.

[0034] (Example 4: Evaluation of deformation of gelatin hydrogel loaded with ferromagnetic microparticles after application of a controlled external magnetic field)

[0035] To evaluate the responsiveness of the scaffolds fabricated in Example 3, hydrogels consisting of gelatin / genipine loaded with magnetic microparticles were exposed to an external magnetic field controlled using a 0.3T magnetic resonance imaging (MRI) system. Specifically, each scaffold was exposed to a periodic magnetic field, and its diameter was measured in the coronal and sagittal sections.

[0036] Figure 2 shows the results of MRI imaging for one of the scaffolds of the present invention. Figure 2a shows a coronal section, and Figure 2b shows a sagittal section.

[0037] To accurately quantify the deformation of the scaffold, the diameter of each hydrogel was measured by X-ray analysis immediately before and after each MRI cycle.

[0038] Figure 3 shows the X-ray measurement results obtained for one of the fabricated scaffolds. Figure 3a shows that the diameter of the scaffold (in a Petri dish) before excitation was 23.83 mm. Figure 3b shows that the diameter of the scaffold became 24.72 mm after excitation.

[0039] (Example 5: Cellification of cross-linked gelatin scaffolds by human articular chondrocytes (hACs)) Human articular chondrocytes (hACs) were obtained from healthy femoral condyles and tibial plateau cartilage from donors who underwent total knee arthroplasty (TKR).

[0040] Specifically, tissue samples were collected from three female donors (aged 64, 71, and 82) selected based on clear selection criteria (single-compartment osteoarthritis, no prior knee surgery, and no significant comorbidities). After carefully washing the surgical waste tissue with sterile PBS, it was transferred to sterile plates containing anatomical culture medium (Diss-M: DMEM high glucose (Gibco USA), penicillin / streptomycin / amphotericin 1% v / v (Gibco USA), Fetal Bovine Serum 10% v / v (Gibco USA)). Next, healthy cartilage was carefully excised using a sterile scalpel, taking care not to harvest the underlying subchondral bone, and transferred to a new sterile plate containing Diss-M, then 1 cm 2 The cartilage was shredded into fragments. The shredded cartilage was then collected in a sterile 500 mL conical tube and incubated overnight at 37°C with mechanical stirring at 250 RPM with a 5 mg / mL solution of collagenase A (Worthington, UK) prepared in Diss-M. Finally, the cells were collected, filtered through a 40 μm cell strainer, and counted, resulting in 7.0 × 10⁶ cells. 3 cell / cm 2The cells were seeded in T75 flasks that had been treated for tissue culture. The chondrocytes were cultured in FBS medium for chondrocytes (cFBS-M: DMEM high glucose (Gibco USA), penicillin / streptomycin / amphotericin 1% v / v (Gibco USA), insulin / transferrin / selenium 1% v / v (Thermofisher USA), dexamethasone 0.1 μM (Sigma-Aldrich USA), L-proline 40 μg / mL (Sigma-Aldrich USA), fetal bovine serum 10% v / v (Gibco USA)) until they reached 80% confluence.

[0041] hACs were gently detached by adding 5 mL of TryPLE (Gibco USA) to a T75 flask and holding at 37°C for 10 minutes. Cells were collected by centrifugation at 1200 rpm for 7 minutes (rotor radius = 247 mm), and then hACs from three donors were pooled. The cell pellet was resuspended in 10% w / v GelMA (Cellink, USA) solution in 0.25% w / v lithium phenyl-2,4,6-trimethylbenzoyl phosphinate (LAP) (Cellink, USA) / PBS solution, avoiding bubble formation throughout the process. Finally, the cell suspension was dispensed into a silicone mold and UV irradiation was induced to induce crosslinking of the scaffold by irradiating it for 3 minutes with a UV lamp (LEPRO, USA) equipped with a 395 nm LED. The scaffold was carefully removed from the silicone mold and placed in an untreated 24-well plate. Each scaffold was inoculated with 1 mL of cFBS-M and incubated at 37°C in 5% CO2 for 24 hours. At this point, cFBS-M was replaced with complete cartilage medium (cCM: cFBS-M + TGFβ-3 10 ng / mL, ascorbic acid 50 μg / mL, without FBS).

[0042] (Example 6: Evaluation of viability of hACs cultured in a cross-linked gelatin scaffold) Cells or cell-containing scaffolds were washed twice with PBS to remove culture medium residue, and then stained using the LIVE / DEAD assay (Abcam, UK). This kit contains two fluorescent dyes, calcein AM and ethidium homodimer-1 (EthD-1), which selectively stain live cells (green) and dead cells (red), respectively. Samples were stained with 4 μM calcein and 2 μM EthD-1 (final concentration) in PBS at 37°C and 5% CO2 for 30 minutes. After staining, samples were washed twice with PBS to remove excess dye, and images were obtained using an inverted epifluorescence microscope EVOS M5000 (Thermofisher, USA).

[0043] (Discussion of results) The above experiments confirmed that the scaffold of the present invention, loaded with magnetic microparticles, responds to an external periodic magnetic field. In fact, the diameter of the hydrogel evaluated during MRI stimulation was 19.69 mm (Figures 2a and 2b), which is smaller than the diameter of the same scaffold measured immediately before magnetic stimulation (23.83 mm, Figure 3a). This data suggests that the scaffold of the present invention contracts by 28.6% in response to the magnetic field. Notably, this deformation is reversible, and the scaffold fully recovers to its initial dimensions immediately after deformation, with a diameter of 24.72 mm after relaxation (Figure 3b).

[0044] Even when BaTiO3 was loaded as a piezoelectric filler onto the same material, it showed responsiveness to mechanical compression within a similar deformation range. In particular, the experiments demonstrate that the material of the present invention can respond to compression regimens implemented using a strain-controlled method, generating an electrical effect in the form of a voltage difference. This electrical effect has the ability to stimulate and promote regenerative responses in adjacent tissues. Although not bound by theory, the regenerative signal is provided in situ by cells loaded onto the material itself, enabling personalized treatment for patients.

Claims

1. A biocompatible composition used for implantation in a body site where surgery has been performed for the treatment of a disease characterized by cartilage defects, a) A matrix comprising an intrinsically piezoelectric biocompatible polymer, or a non-intrinsically piezoelectric biocompatible polymer loaded with an inorganic or organic filler that imparts piezoelectricity to a non-intrinsically piezoelectric polymer, b) 1 x 10 ５ ~1 x 10 ９ Magnetic material with nanoparticles / mL, c) 1 × 10 per 1 mL of composition ５ ~1 x 10 ８ A cell comprising a cell selected from the group consisting of mesenchymal stem cells, mature chondrocytes, and chondrocyte progenitor cells, Biocompatible composition.

2. The biocompatible polymer with intrinsic piezoelectricity is selected from the group consisting of polyvinylidene fluoride, poly(vinylidene fluoride-trifluoroethylene), poly-L-lactic acid, poly(3-hydroxybutyrate-co-3-hydroxyvaleric acid), cellulose, collagen, polyacrylonitrile, nylon-11, and mixtures thereof. The biocompatible composition according to claim 1.

3. The intrinsically piezoelectric biocompatible polymer is dissolved or suspended in a solvent selected from water, PBS at pH 7.4, culture medium, tetrahydrofuran, diethyl ether, dimethylformamide, hexafluoroisopropanol, methyl ethyl ketone, and mixtures thereof. The biocompatible composition according to claim 2.

4. The amount of the biocompatible polymer with intrinsic piezoelectricity is 2-30% w / v. A biocompatible composition according to any one of claims 1 to 3.

5. The aforementioned non-intrinsic piezoelectric biocompatible polymer is of natural origin. The biocompatible composition according to claim 1.

6. The biocompatible polymer with non-intrinsic piezoelectricity is animal gelatin. The biocompatible composition according to claim 5.

7. The non-intrinsic piezoelectric biocompatible polymer is a hydrogel containing water, PBS, or a culture medium. The biocompatible composition according to claim 5 or 6.

8. The amount of the polymer is approximately 2-30% w / v of the total composition. The biocompatible composition according to claim 7.

9. The non-intrinsic piezoelectric biocompatible polymer is loaded with piezoelectric inorganic filler microparticles or nanoparticles. A biocompatible composition according to any one of claims 5 to 8.

10. The inorganic filler is selected from barium titanate, lead zirconate titanate, zinc stanate, lithium niobate, sodium niobate, and mixtures thereof. The biocompatible composition according to claim 9.

11. The nanoparticles have a diameter or maximum axial length of 1 nm to 100 nm. The microparticles have a diameter or maximum axial length of 101 nm to 100 μm. The biocompatible composition according to claim 9 or 10.

12. The inorganic filler nanoparticles are present in the composition in an amount of 4 to 40% w / v. A biocompatible composition according to any one of claims 9 to 11.

13. The magnetic material in nanoparticle form is selected from magnetic metals, magnetic metal oxides, magnetic metal alloys, and mixtures thereof. The biocompatible composition according to claim 1.

14. The magnetic metal is selected from iron, nickel, and cobalt. The magnetic metal oxide is an oxide or mixed oxide of iron, nickel, and cobalt. The magnetic metal alloy is selected from steel, AlNiCo group alloys, TiCoNiAl group alloys, or alloys based on rare earth elements. The biocompatible composition according to claim 1.

15. The magnetic metal oxide is selected from ferric oxide or magnetite. The alloy based on the aforementioned rare earth element is selected from samarium-cobalt alloy or neodymium-iron-boron alloy. The biocompatible composition according to claim 1.

16. The magnetic material is present in the composition at a rate of 1 × 10 ６ ~1 x 10 ９ Present in the form of nanoparticles / mL, The biocompatible composition according to claim 13.

17. Cells selected from a group consisting of mesenchymal stem cells, chondrocyte progenitor cells, and mature chondrocytes are used at a rate of 1 × 10⁶ cells per mL. ５ ~1 x 10 ８ Includes one The biocompatible composition according to claim 1.

18. Cells selected from the group consisting of mesenchymal stem cells, chondrocyte progenitor cells, and mature chondrocytes, in an amount of 2×10 per mL ６ to 2×10 ７ cells, The biocompatible composition according to claim 17.

19. Further comprising one or more additional ingredients selected from antioxidants, preservatives, and bioactive agents, The biocompatible composition according to claim 1.

20. The bioactive agent is selected from growth factors, drugs, antibiotics, antivirals, enzymes, vitamins, and mixtures thereof. The biocompatible composition according to claim 19.

21. A method for treating joint diseases in subjects requiring treatment, The process includes a step of using the biocompatible composition described in any one of claims 1 to 20. Treatment methods for joint diseases.

22. The subject requiring treatment suffers from a cartilage defect. The method for treating joint disease according to claim 21.

23. The subject requiring treatment suffers from osteoarthritis. A method for treating joint disease according to claim 21 or 22.

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