Nanomaterials, injectable hydrogels and three dimensional scaffolds

Abstract

The invention relates to nanomaterials, injectable hydrogels and three-dimensional scaffolds capable of carrying drugs or biomolecules, oxygen and metabolites.

Description

[0001] DESCRIPTION

[0002] Nanomaterials, Injectable Hydrogels and Three Dimensional Scaffolds

[0003] Field of Invention

[0004] The present invention relates to nanomaterials, injectable hydrogels and three-dimensional scaffolds that can carry oxygen, drugs or biomolecules, metabolites and can be used in different biomedical fields depending on the properties of the drug or biomolecule they carry.

[0005] State of the Art related to the invention (Prior Art)

[0006] Drug delivery systems or therapeutic strategies are based on biomaterials. Biomaterials are a multidisciplinary field and have become an indispensable element in improving human health and quality of life in the modern era. Today, polymers are the most preferred material in the biomedical field among biomaterial classes. Smart polymers have a very important place in the field of pharmacy. These polymers can be used to carry drugs in a durable, biologically active form in carrier systems and to provide controlled release of drugs specifically in target tissues, organs or cells.

[0007] The biomaterial mentioned in patent document US20210113736A1 includes a hydrogel comprising a number of microparticles suspended in the hydrogel. The microparticles comprise an oxygen carrier encapsulated in a biocompatible hydrophobic material in which the release of oxygen from the oxygen carrier is sustained over a period of four to five weeks. The biomaterial has applications in tissue engineering, osteogenesis, bum and wound treatment, and treatment of heart conditions, and also has antimicrobial properties. This patent document demonstrates the combination of calcium peroxide (CaCh) with polycaprolactone (PCL), a hydrophobic biopolymer, to produce scaffolds that provide continuous oxygen release over long tissue culture periods.

[0008] Patent document KR102083002B1 relates to a hydrogel / hydrocolloid bilayer wet dressing agent comprising probiotics and a method of production thereof and, more particularly, to a hydrogel layer comprising probiotics and a hydrogel layer for wound healing, such as wounds and burns, in which the stability of the probiotics is ensured. The present invention relates to a probiotic-containing double-layer hydrogel / hydrocolloid wet dressing agent comprising a colloidal layer and a method of production thereof. Patent US5578022A describes a portable, self-contained device for topical oxygen application to promote healing of skin wounds. The device is composed of a wound dressing comprising an electrochemical, chemical or thermal means for generating high purity oxygen. The device is capable of regulating the supply of oxygen at various concentrations, pressures and dosages to an area above the wound.

[0009] The current state of the art lacks a drug delivery system that combines functional structures to deliver controlled oxygen and pH responsive drug or biomolecule delivery within the same biomaterial.

[0010] Brief Description and Objectives of the Invention

[0011] The present invention is a single system that combines a drug or biomolecule, oxygen and metabolite transport unit on the same nanoparticle. These nanocomposites can act as a drug delivery system providing continuous oxygen (O2) and pH-sensitive controlled drug or biomolecule, probiotic metabolite co-delivery. Therefore, the direct effect of the toxic drug or biomolecule on healthy cells is reduced and the release of the drug or biomolecule is prolonged.

[0012] In addition, these functional nanocomposites can be combined with organosilica and polymer- encapsulated probiotics to create living injectable hydrogels or wound dressings. In this way, in addition to oxygen and drug release, the generation of the structures that can release pH dependent antimicrobial metabolites produced by probiotics can be possible.

[0013] Thanks to the effect of oxygen, drug and metabolite co-release, it enables oxygen, drug and metabolite therapy, especially in pathological conditions (such as inflammation, cancer, infection) in which hypoxia and acidic pH are active or triggered; it promotes healthy cell viability and proliferation, allowing rapid healing of wounds and damaged tissues; and it can also inhibit inflammation, cancer and infection in hypoxic and acidic conditions.

[0014] Therefore, the release of anticancer and anti-inflammatory drugs and metabolites from these structures (nanomaterials, injectable hydrogels and three-dimensional scaffolds) in an acidic pH (pH 6.0) sensitive manner will reduce the effect of these biomolecules on healthy cell viability (pH 7.4). In addition, prolonged oxygen release would increase healthy cell viability. At the same time, pH-sensitive constructs will release high doses of anticancer and / or antiinflammatory drugs to cells in these tissues due to the local acidic environment of cancer and inflammatory tissues, while releasing less drug molecules at the physiological pH of healthy tissues. Therefore, side-specific and minimally invasive drug delivery will, for example, improve cancer drug therapy, reduce cancer patients' pain and thus help to improve cancer patients' quality of life.

[0015] Descriptions of the Figures Explaining the Invention

[0016] The figures and related explanations necessary for a better understanding of the invention are as follows.

[0017] Figure 1. Oxygen levels of medium containing GA and GA-PMOF under A) hypoxic (H) and B) normoxic (N) conditions.

[0018] Figure 2. Cumulative amount of DOX released (pg / mL) from GA-DOXPMOF and GA-DOXPMOF-PDL at pH 7.4 and pH 6.0.

[0019] Figure 3. Optical density of fibroblast cells on GA, GA-PMOF, GA-DOXPMOF and GA- DOXPMOF -PDL under A) normoxic and B) hypoxic conditions on days 1 and 7 of incubation. (Number of replicates (N)=3; data show significant differences; ANOVA: p<0.05(*), p<0.01(**) and p<0.001(***); and a=significant difference between five groups and b=significant difference between two groups.)

[0020] Figure 4. Optical density of C0I08I8 cells on GA, GA-PMOF, GA-DOXPMOF, GA- DOXPMOF and GA-DOXPMOF -PDL under A) normoxic and B) hypoxic conditions on days 1 and 7 of incubation. (Repetition number (N)=3; data show significant differences; ANOVA: p<0.05(*), p<0.01(**) and p<0.001(***); and a=significant difference between five groups and b=significant difference between two groups.)

[0021] Detailed Description of the Invention

[0022] In this detailed description, the structures contained in the inventive drug delivery system, the properties of these structures and the method of obtaining the drug delivery system are described.

[0023] The invention is a drug delivery system that provides continuous oxygen (O2) and pH dependent drug / active biomolecule and probiotic metabolite co-delivery within the same nanocomposite.

[0024] This transport system includes perfluorocarbon as the Ch-carrying structure, negatively charged periodic mesoporous organosilica (PMO) as the pH-sensitive structure encapsulating hydrophobic and hydrophilic biomolecules (drug or active biomolecule), pH-sensitive polymers that can coat negatively charged PMO surfaces and probiotics that can release pH-sensitive metabolites encapsulated in positively charged periodic mesoporous organosilica (PMO-NH2) and negatively charged polymer.

[0025] The nanocomposite structure, which acts as a drug carrier system and is the subject of the invention, provides oxygen with perfluorocarbon-bonded nanomaterial, drug release with nanomaterials coated with a positively charged polymer, preferably Poly-D-lysine (PDL) and loaded with drugs in its pores, and metabolite release with probiotics encapsulated with organosilica and polymer.

[0026] Perfluorocarbons can dissolve 20 times more oxygen than water. This is explained by Van der Waals interactions between a large number of fluorine and oxygen. Therefore, perfluorocarbons have the ability to dissolve large amounts of oxygen and release it when the oxygen content in the environment decreases. The oxygen delivery mechanism is based on the diffusion of oxygen from concentrated oxygen in perfluorocarbons to a lower hypoxia environment.

[0027] The release of drug molecules and probiotic metabolites from living injectable hydrogels and wound dressings (three-dimensional scaffolds) is pH-dependent and occurs by diffusion into aqueous solution. These prepared pH-sensitive structures will show higher biomolecule dosage release under acidic conditions than under physiological conditions, which will be due to different pH-dependent electrostatic interactions between all charged components of the final structures. For example, for the drug (e.g. anticancer drug: doxorubicin) loaded and positively charged polymer-coated nanomaterial in hydrogel, preferably PDL (1) and organosilica and polymer encapsulated probiotics (OPEP) (2), the charged components and the electrostatic interaction between them are as follows. The term OPEP used here and throughout the detailed description refers to the organosilica and polymer-encapsulated probiotic structure. drug (+) | nanomaterial (-) | PDL (+) | hydrogel (-) (1) probiotics (-) | positively charged nanomaterial (+) | hydrogel (-) (2)

[0028] For example, for the nanomaterial in the hydrogel, at pH 6.0, the negatively charged outer surfaces of the nanomaterial will be protonated, which will cause the nanomaterial to interact less electrostatically with the positively charged drug and PDL, and will cause the PDL to close the pores of the nanomaterial less. This will result in a higher dose of drug release from the pores of the nanomaterial at pH 6.0 compared to pH 7.4. Furthermore, the protonation of the negatively charged hydrogel at pH 6.0 will weaken its interaction with the positively charged PDL-coated nanomaterial, so the drug release from the hydrogel will be higher at pH 6.0 compared to pH 7.4.

[0029] Similarly for OPEP in hydrogel, at pH 6.0 the negatively charged outer surfaces of the probiotics will be protonated, which will cause less electrostatic interaction of the probiotics with the positively charged nanomaterial, resulting in a higher dose of metabolite release from OPEP at pH 6.0 compared to pH 7.4. Moreover, the protonation of the negatively charged hydrogel at pH 6.0 will weaken its interaction with the positively charged OPEP, which will result in higher metabolite release from the aforementioned structures at pH 6.0 compared to pH 7.4.

[0030] Hypoxic cells are dependent on anaerobic glycolysis, which leads to lactic acid accumulation and thus to a decrease in pH. Therefore, around tumor tissue and in inflamed tissues, areas of hypoxia often occur with lower than average pH values. In other words, acidic pH conditions are present in the hypoxic cell environment, tumor tissues, wounds and inflamed areas of tissue. Therefore, the release of anticancer and anti-inflammatory drugs and metabolites from these structures (nanomaterials, injectable hydrogels and three-dimensional scaffolds) in an acidic pH (pH 6.0) sensitive manner will reduce the effect of these biomolecules on healthy cell viability (pH 7.4). In addition, prolonged oxygen release would increase healthy cell viability. At the same time, pH-sensitive constructs will release high doses of anticancer and / or antiinflammatory drugs to cells in these tissues due to the local acidic environment of cancer and inflammatory tissues, while releasing fewer drug molecules at the physiological pH of healthy tissues. Therefore, side-specific and minimally invasive drug delivery will, for example, improve cancer drug therapy, reduce cancer patients' pain and thus help to improve cancer patients' quality of life.

[0031] The nanocomposite structure, which acts as a drug carrier system and is the subject of the invention, provides oxygen with perfluorocarbon bonded nanomaterial, drug release with nanomaterials coated with positively charged polymer and loaded with drugs in their pores, and metabolite release from organosilica and polymer encapsulated probiotics.

[0032] In the preferred embodiment of the invention, PMO, which is used as a negatively charged nanomaterial to load drugs or active biomolecules into its pores, is preferably obtained from a mixture of hexadecyltrimethylammonium bromide (CTAB) and l,2-bis(trimethoxysilyl)ethane (BTME). However, CTAB is then removed from this mixture and a porous PMO structure is obtained. Especially these pores are where the drugs are loaded. The size of the pores is the same everywhere in the PMO as it is determined by the size of the CTAB. The chemical composition and formula of the negatively charged PMO preferably used is (SiO3C2H4)2n(C3He)m.

[0033] In the preferred embodiment of the invention, nanomaterials with drug loaded pores are preferably coated with Poly-D-lysine (PDL). Poly-D-lysine has no specific barrier properties against gases, i.e. it is not a barrier to prevent the release of oxygen. In the context of the present invention, PDL does not cover the entire surface of the PMO 100%, because in such a case, drug release would not be possible. Since PDL is a polymer, it interacts electrostatically with the PMO surface and at the same time, since it is a polymer chain structure, it is a flexible structure, so it does not have a barrier feature that will prevent drug and oxygen release. The results of the studies carried out within the scope of the present invention confirm this.

[0034] In alternative embodiments of the invention, other positively charged polymers can be used instead of PDL. For example, poly cations such as chitosan, polypeptides, polymers bearing amine groups can also be used in the coating of PMO to provide pH-dependent release.

[0035] The invention relates to a probiotic encapsulated with organosilica and polymer (OPEP) in a nanocomposite structure, where the PMO is positively charged and the polymer negatively charged, which is necessary for the release of the metabolite. Probiotics are facultative anaerobic bacteria that can tolerate oxygen and are sensitive to environmental factors such as temperature, pH changes, chemicals, etc. Therefore, probiotics are encapsulated in a pH- sensitive porous organosilica and polymer shell to protect them from environmental factors and thus increase their bioactivity and control the release of their metabolites. For this, a layer-by- layer coating method is used.

[0036] Positively charged nanoparticles are less toxic to cells than positively charged polymers. Since PMO-based nanoparticles have a large and spherical structure (approximately 200-220 nm in size), they do not easily penetrate the cell membrane and cause cell death like long chain polymers. Therefore, in the preferred embodiment of the invention, PMO-NH2 was preferred as a positively charged PMO to encapsulate probiotics. In addition, PMO are suitable for this study because they are nanomaterials that show useful properties in biomedical uses. The disadvantages of encapsulation with only polymer or silica gel in the literature, such as low mechanical stability or small or large porosity, are intended to be reduced by the use of porous organosilica nanomaterial in combination with polymer. The PMO-NH2 preferably used in the invention can be synthesized to have a porous structure (pore size = 2.5 nm). By optimizing the amount of PMO-NH2, the porosity of the organosilica-polymer shell around the probiotic is ensured. PMO-NH2 forms a porous and mechanically stable structure within the capsule both due to the porosity of its own structure and the gaps between the PMO-NH2 themselves. This structure is further mechanically strengthened by the electrostatic interaction of the negatively charged poly (styrenesulfonate) (PSS) polymer, which is preferably used within the scope of the invention, with the positively charged PMO-NH2 and becomes a structure that is further affected by pH. This is also one of the most important features of layer-by-layer coating. Poly(styrenesulfonate) (PSS) is a biocompatible polymer and interacts well with PMO-NH2. It was therefore chosen for use in the preferred embodiment of the invention. As the probiotic produces lactic acid, the negatively charged PSS will be protonated and its electrostatic interaction with PMO-NH2 will decrease, making the shell of PMO-NH2 and PSS around the probiotic more permeable. And so the amount of nutrients reaching the probiotic will increase and the release of metabolites produced will increase and the amount of metabolites released will increase.

[0037] Instead of the negatively charged PSS polymer, other biocompatible polyanions can be used in alternative embodiments of the invention. For example, cellulose or alginic acid. Furthermore, loading lactose into the pores of PMO-NH2 could also be used as an alternative method, thus providing the necessary nutrients for probiotics directly in the capsule for a certain period of time.

[0038] Injectable hydrogel and three-dimensional scaffold forms are obtained by using gelling agents in addition to the mentioned nanocomposite structure. Of course, photo-initiating cross-linkers are also used in the production of three-dimensional scaffolds with 3D printers. In the preferred embodiment of the invention, gelatin methacryloyl (GelMa) and / or alginate (Alg) are used as gelling agents. The gelling agent should be biocompatible, non-toxic, non-immunogenic, biodegradable, absorb wound fluid and / or keep dry wounds wet, be injectable and have gelforming properties. Due to these complementary properties, GelMa and Alg are used together in the preferred embodiment of the invention. GelMa increases the mechanical stability of the hydrogel and 3D scaffolds. It ensures that the wound dressing is adhesive. Alga increases the injectability of GelMa. GelMa and Alga are biocompatible, biocompatible, non-toxic and immunogenic, biodegradable gelling agents that can absorb the fluid formed in the wound. The hydrogel formed with GelMa and Algae keeps dry wounds moist. Calcium released from alginate causes histamine release and initiates the wound healing process. Alga can remain on the wound surface longer than hydrocolloids. In alternative embodiments of the invention, any gelling agent may be used, but the alternatives to gelling agents that may be used are listed together with their disadvantages (Erfan Rezvani Ghomi et al:) i. Hydrocolloids: Not recommended for infected wounds, they can weaken the skin if not changed at appropriate intervals. ii. Alginate dressings: Since alginate dressings require moisture to function, they should not be used on wounds that are dry and covered with necrotic tissue, otherwise they may leave the wound dehydrated and affect healing negatively. iii. Poly(lactic-cogly colic acid) derivative, poly(vinylalcohol), poly(ethylene glycol) derivative, poly-caprolactone, polyurethane derivative hydrogels: When they absorb wound secretion, they swell until they lose all their adhesive properties. iv. Polyurethane and silicone-based foams: Foams are not suitable for dry and crusted wounds. v. Bioactive wound dressings based on collagen, hyaluronic acid, chitosan: Poor mechanical properties (stability).

[0039] At the same time, the functionality of the nanocomposite structure can be increased by using additional structures in alternative embodiments of the invention. These are; laponite can be added to hydrogels as an additional structure and thus the injectability of the hydrogel is increased and / or calcium peroxide can be added as an additional structure into the structures containing perfluorocarbon and thus the amount of oxygen released is increased and at the same time, long-term release of oxygen is provided by absorbing the released oxygen on the perfluorocarbon.

[0040] Bulu§un tercih edilen uygulamasinda ilag ta§ima sistemi gbrevi gbrecek nanokompozitin igerdigi yapilar a§agida siralanmaktadir: a) Negatively charged periodic mesoporous organosilica (PMO) (PMO) (SiO3C2H4)2n(C3H6)m b) Drug or biomolecule (X) c) Poly-D-lysine (PDL) as a positively charged polymer to coat the negatively charged PMO d) lH,lH,lH,2H,2H,2H-perfluorooctyltriethoxysilane (POTES) as perfluorocarbon structure e) Probiotics (Lactobacillus) encapsulated with positively charged periodic mesoporous organosilica (PMO-NH2) and Poly(styrenesulfonate) (PSS) as a negatively charged polymer (OPEP)

[0041] Bulu§a konu olan ve yukanda tercih edilen yapilan igeren nanokompozit, hidrojel ve tig boyutlu iskelelerin elde edilebilmesi igin uygulanan ybntemin turn basamaklan a§agida anlatilmaktadir:

[0042] (1) Synthesis of PMO: 484.5 mg of hexadecyltrimethylammonium bromide (CTAB) was dissolved in a solution of 90 ml H2O, 33 ml ethanol and 28 wt% ammonia (0.075 g). Then l,2-bis(trimethoxysilyl)ethane (BTME) (1.27 g) and 1H,1H,2H,2H,2H- perfluorooctyltriethoxysilane (POTES) (0.59 g) were added to this mixture and the final mixture was stirred for 48 hours at room temperature. The CTAB mesoporous template is removed by stirring the sample in ethanol (50 ml) containing 36 wt% aqueous hydrochloric acid (HC1) solution (1.5 g) at 50 °C for 6 hours. The resulting solid (PMOF) is recovered by centrifugation, washed several times with ethanol and dried in vacuo at 60 °C.

[0043] (2) Loading of Drug or Biomolecule (X) into PMOF (XPMOF): PMOF (100 mg) is suspended in 1 ml of water and mixed with X (5 mg). This reaction mixture is stirred at room temperature for 1 day. The final productXPMOF is obtained by centrifugation, washed with water and dried at room temperature.

[0044] (3) Coating ofXPMOF with Poly-D-lysine (PDL) (XPMOF-PDL):XPMOF particles (100 mg) were suspended in 1 ml of water, mixed with PDL (0.5 mg ml-1 water). This reaction mixture is stirred overnight at room

[0045] (4) Organosilica-Polymer Encapsulated Probiotics (OPEP): The negatively charged surface of Lactobacillus (xlO51 ml PBS) is first coated with positively charged PMO- NH2 (1 mg). Then, the probiotics are coated with negatively charged poly(styrenesulfonate) (PSS) (1 mg) and PMO-NH2 by electrostatic interactions via an alternating layer-by-layer coating method (e.g. 4 more times) to obtain PMO-NH2 / PSS (organosilica / polymer) coated probiotics. The final product OPEP is obtained by centrifugation, washed with x2 water.

[0046] (5) Preparation of Nanocomposite (NC) Hydrogel Precursors: First, gelatin methacryloyl (GelMa) is synthesised according to previous work (M. Zhu, Y. Wang, G. Ferracci, J. Zheng, N. J. Cho, B. H. Lee, Sci. Rep. 2019, 9, 6863). Then GelMa stock solution (1 g, 10% w / v) was added to phosphate buffered saline (PBS) (10 mL) with eosin Y (0.1 mM) in PBS as photoinitiator, triethanolamine (TEA) as co-initiator (133 pL, 1.3% w / v) and vinylcoprolactane (VC) (0.1 g, 1.0% w / v) as co-monomer and kept at 80°C for 10 min to dissolve GelMA. To obtain hydrogel precursor (GA), the above solution is mixed with alginate (70 mg / mL), which dissolves homogeneously in solution. Or alginate is used alone. Then, to obtain hydrogel precursors containing particles and OPEP (GA-XPMOF-PDL-OPEP or Alg-XPMOF-PDL-OPEP), this GelMA stock solution is mixed withXPMOF-PDL (1.0 mg / mL) and OPEP (1.0 mg / mL) and after sonication for 20 min, alginate (70 mg / mL) is homogeneously mixed into this suspension.

[0047] (6) 3D Printing of GA-XPMOF-PDL-OPEP Nanocomposite Hydrogel Scaffolds: The hydrogel precursor (GA-XPMOF-PDL-OPEP) is homogenously mixed with a spatula and the mixed solution is transferred to a syringe. GA-XPMOF-PDL-OPEP is printed on a hexagonal template. A 3 cc syringe barrel and a 0.41 mm needle (LOCTITE Dispense Needle Type:97224) are used. Grid hexagonal structures with dimensions of 10 mm on each side and a height of 1.5 mm are produced with a 3D printer. The speed of the syringe is set to 80 mm / s and 10 mm / s is selected as the injection head speed. Extrusion pressure level and temperature are used as 40 kPa and 25 °C, respectively. Then, the 3D printed GA-XPMOF-PDL-OPEP is photo-crosslinked with visible light (450-550 nm) for 120-180 s using FocalSeal (Genzyme Biosurgical, Cambridge, MA, USA). For ionic cross-linking of alginate, printed constructs are cross-linked with a solution of calcium chloride (CaCh) (100 mM), kept at -80 °C for 24 h and then freeze- dried for 24 h to obtain GA-XPMOF-PDL-OPEP scaffolds.

[0048] In a previous study conducted by the inventors within the scope of the subject matter of the invention, a transport system (GA-DOXPMOF-P) combining the anti-cancer drug Doxorubicin (DOX) and the oxygen transport unit on the same nanoparticle was obtained. GA-DOXPMOF-P provides pH-sensitive sustained release of the anticancer drug for 30 days and sustained O2 release for 15 days. The results (Figure 1-4) show thatDOXPMOF-P reduces cancer cell viability in hypoxic and acidic conditions compared to healthy cells. This study demonstrates the success of oxygen and drug co-distribution. In addition to this study, within the scope of the present invention, oxygen, drug and metabolite co-release is provided by adding periodic mesoporous organosilica (PMO) and probiotic encapsulated with polymer (OPEP) as a structure capable of pH-dependent metabolite release to the nanocomposite structure. Thanks to this co-release effect, it enables oxygen, drug and metabolite therapy especially in pathological conditions (such as inflammation, cancer, infection) where hypoxia and acidic pH are active or triggered, promotes healthy cell viability and proliferation, allows rapid healing of wounds and damaged tissues, and can also inhibit inflammation, cancer and infection in hypoxic and acidic conditions.

[0049] Thanks to the easy synthesis of the main building blocks of the inventive nanocomposite, this drug delivery system can be used as a wound healing or antimicrobial (living) textile material in the textile industry, antimicrobial (living) seats in public transport, antimicrobial in cosmetics, It can be used as anti-inflammatory, wound healing, anti-aging (living) creams, injectable (living) implants as drug delivery systems in biomedicine, (living) wound dressing material, injectable (living) biomaterial for cancer treatment. In addition, this delivery system can also be used in the defence industry as (living) textile and / or wound dressings or injectable hydrogels that can relieve injury-activated pain, stop wound infection and inflammation, or are antimicrobial (living) textile and / or wound dressings or injectable hydrogels that can be applied even by the injured person themself in emergency situations by wounded soldiers or members of law enforcement agencies.

Claims

CLAIMS1. Nanocomposite, characterised by; i. perfluorocarbon as oxygen carrier, ii. negatively charged periodic mesoporous organosilica loaded with a drug or active biomolecule in its pores and coated with a positively charged polymer to allow the release of drug or active biomolecule in a pH sensitive manner, iii. a positively charged periodic mesoporous organosilica and a negatively charged polymer encapsulaed probiotics that can allow pH sensitive release of metabolites.

2. The nanocomposite according to claim 1, characterised in that mentioned perfluorocarbon is lH,lH,2H,2H,2H-perfluorooctyltriethoxysilane (POTES).

3. The nanocomposite according to claim 1, characterised in that mentioned positively charged polymer is Poly -D-ly sine (PDL).

4. The nanocomposite according to claim 1, characterised in that mentioned negatively charged periodic mesoporous organosilica is (SiO3C2H4)2n(C3H6)m.

5. The nanocomposite according to claim 1, characterised in that said negatively charged polymer is poly(styrenesulfonate) (PSS).

6. The nanocomposite according to claim 1, characterised in that mentioned positively charged periodic mesoporous organosilica is PMO-NH2.

7. The nanocomposite according to claim 1, characterised in that mentioned encapsulated probiotic is Lactobacillus.

8. The nanocomposite according to claim 1, characterised in that it comprises a gelling agent.

9. The hydrogel according to claim 8, characterised in that mentioned gelling agent is gelatin methacryloyl and / or alginate.

10. The hydrogel comprising the nanocomposite according to claim 8 or 9.

11. A three-dimensional hydrogel tissue scaffold comprising the nanocomposite according to claim 8 or 9.

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