BINARY HETEROJUNCTION NANOSHEET PHOTOSENSITIZER FOR THE PHOTODYNAMIC THERAPY OF RETINOBLASTOMA

Abstract

The invention pertains to a binary heterojunction photosensitizer composed of graphitic carbon nitride (gCN) and black phosphorus (BP) for use in photodynamic therapy (PDT) to treat retinoblastoma. The gCN / BP binary heterojunction nanosheets enhance light absorption and charge carrier separation, activated by near-infrared (NIR) light to generate reactive oxygen species (ROS), inducing apoptosis in cancer cells. The photosensitizer accumulates selectively in tumor tissues, minimizing damage to healthy cells, and addresses BP?s stability in biological environments, offering a durable, non-invasive treatment for retinoblastoma.

Description

21644.92 1 TARIFF DUAL APPROACH TO PHOTODYNAMIC THERAPY OF RETINOBLASTOMA HETEROJOINT NANO-LAYER PHOTOSENSITIZER Technical Field The present invention is in the field of biomedical applications and, specifically, in the area of ​​malnutrition. nano-layer form for the treatment of retinal blastoma tumor Graphite carbon dioxide and black phosphorus (gCN / BP) 10 as photosensitizers including the use of heterojunctive venous insemination in cancer treatment. fotod nam k terap le lg ld r. Previous Tech Retinal blastoma is a tumor that begins in the retina, the light-sensitive tissue at the back of the eye. It is a type of cancer. It is the most common type of childhood eye cancer and the prognosis is... cancer has various aspects, including whether or not it has spread to other parts of the body. It depends on the factors. Among the current treatments for retinal blastoma, the most common is... The options used include cryotherapy (freezing the tumor) and chemotherapy. Additional treatments include thermotherapy, thermo-chemotherapy, radiation, and... Brak therapy is available. It is more serious when other treatment options have been exhausted. In these cases, enucleation (surgical removal of the eye) is the last alternative. After treatment, the focus often shifts to the long-term effects of the disease. The focus shifts to concerns about recurrence and the management of side effects [1]. 25 The graphite is a 2D semiconductor made of polymer KBR with a band gap of 2.7 eV (454 nm). Carbon dioxide (gCN) can be activated and deactivated by visible light, thus making it suitable for biomedical applications. Applications are becoming an attractive CBR candidate. However, the stack gCN has a limited surface area, low light absorption capacity, and low water content. Due to its dispersion, it has low efficiency for photodynamic applications. 21644.92 2 On the other hand, the most stable phosphorus allotrope has a three-dimensional (3D) layered structure. This is due to the extraordinary chemical and electronic properties of black phosphorus (BP). bl msel lg y yen den üzer ne çekm şt r. However, in pure BP, photodynamic therapy photo-generated electron-hole pairs are fast, limiting their applications. It has limitations such as recombination and low stability [2]. 5 Photodetective therapy (PDT), light activation and reacting oxygen species (ROS) a method that uses a photosensitizer to induce the death of cancer cells by producing a photosensitizer treatment. In the previous technique of rejection noblastoma treatment, Photofr n and Verteporf n GB porphyrin-based compounds are commonly used photosensitizers. These 10 The agents accumulate in cancerous tissues and have cytotoxic effects when exposed to light. to treat small tumors in the eye by taking advantage of their production capabilities It is used. However, despite advances in this field, traditional photosensitizers and other therapeutic approaches used for retinal blastoma There are significant limitations. 15 One of the biggest problems with earlier photosensitizers was limited light. Penetration depth is measured in porphyrin-based photosensitizers, which penetrate deep into the tissue. They rely on non-penetrating light sources; this makes their use small, It is limited to superficial tumors. This situation makes deeper tissue treatment necessary for effective treatment. In more advanced cases of retinal blastoma where penetration is necessary, it becomes problematic. This is the result. Consequently, these treatments target tumors located deeper within the eye. It is often ineffective in cases where the tumor or cancer has spread. Another critical point... The problem is the variable effectiveness of current treatments. Traditional photosensitizers... Efficacy depends on the type and concentration of the agent, the light dose, and the specific characteristics of the tumor. 25 It depends on many factors, including these variables. These variables can lead to inconsistent therapeutic outcomes. This leads to standardization of treatment protocols and patient outcomes. This makes predicting their responses difficult. As a result, similar treatment conditions When used, the effectiveness of PDT may vary from patient to patient [3]. System toxicity and side effects also represent significant problems in previous techniques. 30 Traditional photosensitizers induce photosensitivity in healthy tissues. 21644.92 3 (as a cause of photosensitivity), der nn sensitive to light yet gb long period of time This can lead to negative consequences. This situation can affect patients for a long time after treatment. by forcing avoidance of sunlight or bright indoor lighting It can significantly negatively impact quality of life. Furthermore, Retinal neublastoma, other common treatments include systemic chemotherapy; 5 Immunosuppression, nausea, and hair loss are associated with GBC DD side effects. This Side effects can weaken the immune system and increase the risk of infection. and can lead to other complications. The complexity of current PDT systems. Implementation protocols present another challenge. Many existing PDTs... The treatment involves activating photosensitizers by exposing them to multiple light sources or different 10 light sources. It requires specific excitation wavelengths; this increases the complexity of the process. This complexity makes PDT particularly challenging where resources are limited. more for widespread clinical use in settings or when treating pediatric patients This makes it less practical. The need for specialized equipment and expertise... Treatments can limit accessibility and make implementation more difficult. get reb lmekted r [4]. Finally, the biocompatibility of existing photosensitizers in biological environments. There are concerns regarding its stability and reliability. Many previous techniques... The photosensitizer lacks compatibility with healthy cells; this means that 20 It leads to unwanted interactions and potential long-term side effects. Additional However, the stability of these compounds can be impaired in biological environments, and this... It reduces therapeutic efficacy. Poor water solubility and The rapid degradation of photosensitizers in the NV and VO environments further enhances clinical and clinical applications. It restricts. 25 The patent application numbered CN114588279A, included in the previous technical section, concerns specific light. to selectively destroy cancer cells when exposed to wavelengths Development of photosensitizers that produce reactive oxidizing oxidizing oxidizing oxidizing hazards (ROS) Focused photodynamic therapy (PDT) is related to advancements. However, deeper 30 Potential limitations such as limited light penetration that restrict effectiveness on tumors 21644.92 4 There are disadvantages. Off-target effects can still occur, and It can potentially damage healthy tissues. Also, like black phosphorus... Photosensitizers degrade rapidly in biological environments, leading to stability problems. It is possible to survive. In addition, it requires precise timing and coordination. The complex application of PDT and the post-treatment photosensitivity test in patients are also discussed. These are significant challenges. Considering these challenges, better light absorption and deeper tissue penetration are necessary. more advanced and efficient with improved penetration and therapeutic efficacy Photosensitizers require HT. 10 Brief Description of the Invention The primary aim of the invention is to develop photodynamic therapy (PDT) for the treatment of retinal blastoma. The effect of photosensitizers is graphite carbon dioxide (gCN) and black phosphorus (BP) 15 The heterojoint develops a system that absorbs light. by increasing, enabling deeper tissue penetration and killing cancer cells by encouraging the production of reactive oxygen species (ROS) that are essential for inducing radiation It overcomes the limitations of existing photosensitizers. Another aim of the invention is to improve BP-related stability by combining BP with gCN. The goal is to solve these problems by stabilizing the heterojoint BP generated by gCN. In clinical applications, the overall durability and effectiveness of the photosensitizer... It increases. The invention also improves biocompatibility with PDT treatments for retinal blastoma. The aim is to create a gCN / BP Cl heterojunction nanolayer; retinal blastoma. While targeting and destroying cells in an effective way, it also harms healthy retinal cells. It is designed to be selectively designed in a way that will reduce the damage inflicted. In addition, The invention allows the photosensitizer to absorb light more efficiently at larger depths. It enables absorption of light in the NIR spectrum. 21644.92 By improving the gCN / BP heterojunction nanolayer therapy, the eye's ability to see more clearly is enhanced. The lesions do not allow for penetration; this also allows retinal noblastoma to progress further. This makes it possible to treat the stages. Finally, the invention aims to simplify the PDT application. Proposed 5 Photosensitizing, non-invasive and painless, activated with NIR light. It is possible to apply this treatment in clinical settings; this makes it easier to implement the treatment in clinical settings. It provides a faster, less complex solution compared to traditional treatments. It offers. Shapes Figure 1. RB cell viability after BP nanosheet treatment and NIR stimulation. graphs. Figure 1A. BP nano-15 at different concentrations without light irradiation. RB cell viability after treatment of the layers. Figure 1B. BP nanosheet under near-infrared (NIR) laser irradiation. RB cell viability after treatment. Figure 2. BP nanosheet stability and RB cell stability at different incubation times. Potential initiative graphs in viability analysis. 20 Figure 2A. 24-hour analysis of BP nanolayers at different concentrations. RB cell viability after treatment. Figure 2B. BP nanolayers at different concentrations after 48 hours. RB cell viability after treatment. Figure 2C. 25 BP nanolayers with different concentrations for 72 hours. RB cell viability after treatment. Figure 2D. BP nanolayers at different concentrations treated with BP for 96 hours. RB cell viability after treatment. Figure 3. Time-dependent brightness of RB cells incubated with BP nanolayers. Field images; morphological development and clustering behavior over time 30 The white circles represent clustered BP nanolayers. 21644.92 6 Figure 3A. RB cells 0 hours after BP nanolayer exposure. Bright area image. Figure 3B. RB after 24-hour incubation with BP nanolayers. Bright field image of cells. Figure 3C. RB 5 after 48 hours of incubation with BP nanolayers. Bright field image of cells. Figure 3D. RB after 72 hours of incubation with BP nanosheets. Bright field image of cells. Figure 3E. RB after 96 hours of incubation with BP nanolayers. Bright field image of cells. 10 Figure 4. gCN nanosheets under dark and NIR irradiation conditions. Cell viability graphs in RB and RPE-1 cells after treatment. Figure 4A. Different concentrations of gCN nano- under dark conditions. Viability of RB cells after treatment of the layers. Figure 4B. Different concentrations of gCN nano-15 under dark conditions. Viability of RPE-1 cells after treatment of the layers. Figure 4C. Combination treatment of gCN nanosheets and NIR irradiation. then the viability of RB cells. Figure 4D. Combination treatment of gCN nanosheets and NIR irradiation. Then the viability of RPE-1 cells. 20 Figure 5. Treatment of gCN nanosheets under dark and NIR irradiation conditions. I observed the levels of reactive oxygen species (ROS) in RB and RPE-1 cells. graphs. Figure 5A. Treatment with gCN nanolayers at different concentrations. Then, ROS levels in RB cells. 25 Figure 5B. Treatment with gCN nanolayers at different concentrations. then ROS levels in RPE-1 cells. Figure 5C. Combination treatment of gCN nanosheets and NIR irradiation. Then ROS levels in RB cells. Figure 5D. 30 from combined treatment of gCN nanosheets and NIR irradiation. then ROS levels in RPE-1 cells. 21644.92 7 Figure 6. RB and RPE-1 after treatment with gCN / BP heterojunction nanolayer. Graphs for evaluating cellular toxicity in cells. Figure 6A. Different types of gCN / BP heterojunction nanolayer. Viability of RB cells after treatment with concentrations. Figure 6B. Different 5 gCN / BP heterojunction nanolayers. Viability of RPE-1 cells after treatment with concentrations. Figure 6C. gCN / BP Cl heterojunction nanolayer and NIR irradiation. Viability of RB cells after clinical therapy. Figure 6D. gCN / BP Cl heterojunction nanolayer and NIR irradiation. Viability of RPE-1 cells after clinical therapy. 10 Figure 7. gCN / BP Cl heterojunction nanolayer in RB cell culture medium. Bright field images showing its behavior over time. Figure 7A. Post-treatment with gCN / BP heterojunction nanolayer. RB captured in the first stage (first day) and showing early cellular interactions. cells. 15 Figure 7B. 24-hour analysis of gCN / BP heterojunction nanolayer. RB cells after incubation. Figure 7C. gCN / BP Cl heterojunction nanolayer after 72 hours. RB cells after incubation. Figure 8. gCN / BP kl heterojunction nano-20 under dark and NIR irradiation conditions. Reactive oxygen species in RB and RPE-1 cells after treatment of the layer (ROS) level charts. Figure 8A. gCN / BP Cl heterojunction nano- under dark conditions. After treatment with different concentrations of the layer, RB cells showed changes. ROS loves . 25 Figure 8B. gCN / BP Cl heterojunction nano- under dark conditions. RPE-1 after treatment with different concentrations of the layer ROS levels in cells. Figure 8C. gCN / BP Cl heterojunction nanolayer and NIR irradiation. ROS levels in RB cells after combination therapy. 30 21644.92 8 Figure 8D. gCN / BP Cl heterojunction nanolayer and NIR irradiation. ROS levels in RPE-1 cells after combination therapy. Figure 9. gCN / BP heterojunction nanolayer and treatment with NIR irradiation. Then, Western blot analysis of apoptotic pathway markers in RB cells. graphs. 5 Figure 9A. In the control group, only gCN / BP Cl heterojunction nano- after layer treatment, after NIR irradiation alone, and in combination gCN / BP heterojunction nanolayer and RB after NIR treatment Expression levels of Bcl-2 protein in cells. Figure 9B. In the control group, only gCN / BP Cl heterojunction nano-10 after layer treatment, after NIR irradiation alone, and in combination gCN / BP heterojunction nanolayer and RB after NIR treatment Caspase-9 protein expression levels in cells. Figure 9C. In the control group, only gCN / BP Cl heterojunction nano- after layer treatment, only after NIR irradiation and combined 15 gCN / BP heterojunction nanolayer and RB after NIR treatment Expression levels of Bax protein in cells. Figure 10. gCN / BP Cl heterojunction nanolayer and treatment with NIR irradiation. Then, Western blot analysis of apoptotic pathway markers in RPE-1 cells. graphs. 20 Figure 10A. In the control group, only gCN / BP Cl heterojunction nano- after layer treatment, after NIR irradiation alone, and in combination gCN / BP heterojunction nanolayer and RPE-1 after NIR treatment. BCL-2 protein expression levels in cells. Figure 10B. In the control group, only gCN / BP Cl heterojunction nano-25 after layer treatment, after NIR irradiation alone, and in combination gCN / BP heterojunction nanolayer and RPE-1 after NIR treatment. BAX protein expression levels in cells. Figure 11. gCN / BP Cl heterojunction nanolayer, NIR irradiation and combined ne After therapy, RB cells showed increased cell proliferation and structural integrity for 30 days. immunofluorescence images showing. 21644.92 9 Figure 11A. Clustered nuclei of untreated RB control cells. Morphologically altered DAPI-stained kernels. Figure 11B. K-67 immunofluorescence showing proliferating RB control cells. image. Figure 11C. Image of RB control cells combined with DAPI and K-67; 5 Co-localization of K-67 within the nucleus and active cell It shows proliferation. Figure 11D. gCN / BP Cl heterojunction nanolayer after treatment. RB cells showing disruption of cellular structural integrity (nn DAPI) Painted seeds. 10 Figure 11E. RB after treatment with gCN / BP heterojunction nanolayer. Cells nn K-67 immunofluorescence image. Figure 11F. RB after treatment with gCN / BP heterojunction nanolayer. Compound images showing the nuclear localization of cells nn K-67. DAPI and K-67 image. 15 Figure 11G shows the disruption of cellular structure after NIR irradiation. RB cells showing nn DAPI-stained nuclei. Figure 11H. RB cells nn K-67 after NIR irradiation. Immunofluorescence image. Figure 11I. K-67 20 in the nucleus of RB cells after NIR irradiation. Combined DAPI and K-67 image showing localization. Figure 11J. gCN / BP Cl heterojunction nanolayer and NIR irradiation. after combination therapy, the cellular structure was severely damaged. RB cells showing nn DAPI-stained nuclei. Figure 11K. Combine gCN / BP heterojunction nanolayer and NIR 25 Immunofluorescence image of RB cells after treatment with K-67. Figure 11L. Combine gCN / BP heterojunction nanolayer and NIR. Changes at the nuclear level of RB cells after treatment Combined DAPI and K-67 image showing proliferation behavior. 21644.92 Figure 12. gCN / BP Cl heterojunction nanolayer, NIR irradiation and combined ne Following therapy, RPE-1 cells showed nn cell proliferation behavior. Immunofluorescence images. Figure 12A. Treatment of untreated RPE-1 control cells with DAPI. Painted seeds. 5 Figure 12B. K-67 showing proliferating RPE-1 control cells. Immunofluorescence image. Figure 12C. RPE-1 control cells with K-67 core co- a combined image showing localization and active cell proliferation DAPI and K-67 image. 10 Figure 12D. After treatment with gCN / BP heterojunction nanolayer. RPE-1 cells, nuclei stained with DAPI. Figure 12E. gCN / BP Cl heterojunction nanolayer after treatment. RPE-1 cells nn K-67 immunofluorescence image. Figure 12F. gCN / BP Cl heterojunction nanolayer after treatment 15 RPE-1 cells showed less proliferation compared to the control group. A combined DAPI and K-67 showed no significant change. image. Figure 12G. RPE-1 cells after NIR irradiation with DAPI. Painted seeds. 20 Figure 12H. RPE-1 cells nn K-67 after NIR irradiation. Immunofluorescence image. Figure 12I. Preserved RPE-1 cells after NIR irradiation. Combined DAPI and K-67 image showing prol ferat f akt v tey bel rten b rleşt r lm ş DAPI ve K-67 imaj. Figure 12J. gCN / BP kl heterojunction nanolayer and 25 NIR irradiation. RPE-1 cells stained with DAPI after combination therapy. seeds. Figure 12 shows a 2K gCN / BP heterojunction nanolayer and NIR irradiation. K-67 immunofluorescence of RPE-1 cells after combination therapy image. 30 21644.92 11 Figure 12L. Combination of gCN / BP heterojunction nanolayer and NIR. After treatment, RPE-1 cells were compared to the control group. A combined study showing no significant difference in cell proliferation. DAPI and K-67 image. Figure 13. gCN / BP Cl heterojunction nanolayer, NIR irradiation and combination 5 After therapy, RPE-1 cells showed improved cell integrity and morphological characteristics. immunofluorescence images showing. Figure 13A. Treatment of untreated RPE-1 control cells with DAPI. Painted seeds. Figure 13B. 10 showing the tight junction morphology of RPE-1 control cells. ZO-1 immunofluorescence image. Figure 13C. Conserved cellular morphology of RPE-1 control cells. A combined DAPI and ZO-1 image showing the results. Figure 13D. gCN / BP Cl heterojunction nanolayer after treatment. RPE-1 cells, nuclei stained with DAPI. 15 Figure 13E. gCN / BP Cl heterojunction nanolayer after treatment. RPE-1 cells nn ZO-1 immunofluorescence image. Figure 13F. gCN / BP Cl heterojunction nanolayer after treatment. A composite image showing conserved cellular morphology of RPE-1 cells. DAPI and ZO-1 image. 20 Figure 13G. RPE-1 cells after NIR irradiation with DAPI. Painted seeds. Figure 13H. RPE-1 cells after NIR irradiation nn ZO-1 Immunofluorescence image. Figure 13I. Protected cells 25 after NIR irradiation of RPE-1 cells. A combined DAPI and ZO-1 image showing the morphology. Figure 13J. gCN / BP Cl heterojunction nanolayer and NIR irradiation. RPE-1 cells stained with DAPI after combination therapy. seeds. 21644.92 12 Figure 13. K.gCN / BP Cl heterojunction nanolayer and NIR irradiation. ZO-1 immunofluorescence of RPE-1 cells after combination therapy image. Figure 13L. gCN / BP Cl heterojunction nanolayer and NIR irradiation. After combination therapy, RPE-1 cells were largely preserved. A combined DAPI and ZO-1 image showing the morphology of the epithelium. Figure 14. TEM, HAADF-STEM and TEM analysis of gCN / BP Cl heterojunction nanolayer. Elemental map images. Figure 14A. Low magnification of gCN / BP Cl heterojunction nanolayer. Ratio TEM image. 10 Figure 14B. Higher level of gCN / BP Cl heterojunction nanolayer. High-magnification TEM image. Figure 14C. Layered structure of gCN / BP Cl heterojunction nanosheet. High-resolution TEM (HRTEM) image showing morphology. Figure 14D. gCN / BP kl heterojunction nano-15 confirming the hybrid structure. HAADF-STEM image of the layer. Figure 14E. Carbon (C), nitrogen (N) in gCN / BP Cl heterojunction nanolayer. and elemental map showing the spatial distribution of phosphorus (P). image. Figure 14F. Carbon (C) 20 in gCN / BP Cl heterojunction nanolayer. Elemental map image showing distribution. Figure 14G. Nitrogen (N) in gCN / BP Cl heterojunction nanolayer. Elemental map image showing distribution. Figure 14H. Phosphorus (P) in gCN / BP Cl heterojunction nanolayer. Elemental map showing the distribution. 25 Figure 15. gCN / BP heterojunction nanolayer, X-ray of pure gCN and crystalline BP. X-ray diffraction (XRD) patterns. Figure 16. Optical absorption of gCN / BP heterojunction nanolayer and pure gCN. Features and band gap analysis graphs. Figure 16A. UV–30 treatment of pure gCN and gCN / BP Cl heterojunction nanosheet. vs-NIR d fuz reflection spectra. 21644.92 13 Figure 16B. Optic analysis of pure gCN and gCN / BP Cl heterojunction nanolayer. Tauc plots for band gap energy estimation. Figure 17. Photoluminescence of gCN / BP heterojunction nanolayer and pure gCN. and temporal resolution photoluminescence character st kler graph kler. Figure 17A. Stable 5 of pure gCN and gCN / BP Cl heterojunction nanolayer. Status photoluminescence spectra. Figure 17B. Pure gCN and gCN / BP Cl heterojunction nanolayer. Temporal resolution photoluminescence (TRPL) decay curves. Detailed Explanation 10 The invention is intended for use in photodynamic therapy (PDT) in the treatment of retinal blastoma. Photosensitizers in nano-layer form are found in heterojunctions. From here on, according to the current invention, kl heterojunction nanosheet 15 photosensitizer, gCN / BP kl heterojunction without going beyond the scope of the invention. It can also be called a nano-layer. Photodynamic therapy (PDT) is being considered for the treatment of retinal blastoma. It is a heterojunction nanolayer photosensitizer and contains the following: 20 • Two-dimensional (2D) polymer semiconductor with a band gap of 2.7 eV graph tk carbon n tridile (gCN) and • Low-layer black phosphorus (BP) with a band gap of 1.4 eV; where gCN The heterojunction formed between le BP increases light absorption and 25 Minimizing electron-hole recombination by improving charge-carrier separation. nd rmekted r. gCN is a band of approximately 2.7 eV that allows interaction with visible light. The range of 30 is due to the polymer structure of the polymer. He is known for his determination; this characteristic makes him suitable for PDT gbb yomed kal applications. 21644.92 14 This makes it suitable. However, bulk gCN has a large particle size and limited capacity. Due to its surface area and poor dispersion in water, PDT applications It is not highly effective. The invention involves the conversion of gCN with a single layer of black phosphorus (BP). By combining them, it increases the dispersion and functionality of the photosensitizer. Another 2D semiconductor, the low-layer BP, is highly stable and has a density of 1.4 5 It has a band gap of eV; this makes it critically important for deep tissue treatments. This makes the device sensitive / responsive to near infrared (NIR) light. The wavelength of the NIR light used is 810 nm. However, BP undergoes rapid decay. and has limitations such as the rapid recombination of electron-hole pairs; this da fotod nam k terap applicationdakik etk nl ğ n Sınır Sistemi. gCN le 10 By stabilizing the created heterojoint BP, its stability and effectiveness are increased. It increases. Two heterojunction photosensitizers, gCN and BP, n N-met l-2- p rol don (NMP) liquid phase 15 including ultrasonic dispersion in the solvent. It is prepared using a combination method. Photodetective heterojunction nanolayer (PDT) usage The method for synthesizing the photosensitizer involves the following steps: Synthesis of meso-structured graphite carbon nitride (gCN), . synthesis of phosphate (BP) . s yah phosphorus (BP) is approximately 1.6 la in N-methyl-2-pyrrolidone (NMP). Suspending at a concentration of approximately 1.8 mg / ml kbr and exfoliating with BP To obtain nano-layers, the suspension is kept in a controlled environment of 20–25°C. subjection to final cation at temperature, v. The mass ratio of gCN:BP varied between approximately 4.7:1 and approximately 5.1:1, more first dispersed mesoporous graphitic carbon nitride in NMP The final cation is applied to the BP d spreader, where the addition process takes place. It is carried out under continuous finalization, 30 21644.92 v. Interfacial contact and heterojunction between gCN and BP phases. To stimulate, the final concentration of the combined dispersion is 3.5-4.5 hours. Continuing throughout, v. The resulting gCN / BP composite is approximately 15-20% of the mixed suspension. Isolation by centrifugation at approximately 8500-9500 rpm for 5 minutes Then, to remove the unbound material, the precipitate is mixed with ethanol. washing it once, v. Drying of washed composite material, v. Composite BP loading adjustment, where standard BP load is used. The total mass of the gCN / BP Cl heterojunction nanolayer is approximately 10 It contains 16.5-17.5% BP by weight, thus forming a heterojoint. A photosensitizer is obtained. As a procedure, the percentage of phosphorus (BP) is approximately 1.6 to approximately 1.8 lbs. 15 suspended in N-methyl 1-2-pyrrole donor (NMP) at a concentration of mg / ml kbr BP. For obtaining BP nanolayers, controlled temperatures of 20-25°C were used. It was subjected to final cation at a certain temperature. Then, previously in NMP... dispersed mesoporous graphite carbon nitride (gCN), gCN:BP mass ratio The BP dispersion ratio should be approximately between 4.7:1 and 5.1:1. added; here the addition process was carried out under continuous post-cation. 20 The combined suspension is then placed at the interface between the gCN and BP phases. an additional 3.5-4.5 hours to encourage contact and heterojoint fusion. Ultrasound treatment was applied. After the completion of the fusion procedure, The resulting gCN / BP composite is approximately 8500-9500 over approximately 18-22 minutes. To remove isolated and unbound material by centrifugation at rpm 25 The composite was washed twice with ethanol. The washed composite was then dried. Standard BP loading in the b assembly, gCN / BP Cl heterojunction nanolayer The total mass has been adjusted to approximately 16.5-17.5% by weight, so A heterojunctive photosensitizer was obtained. BP was incorporated into the gCN phase. By varying the amount, heterostructures with alternative BP loading levels can also be obtained. 30 ed lm şt r. 21644.92 16 The method for synthesizing meso-structured graphite carbon dioxide (gCN) involves the following steps: It contains: . guan dnh drochloride, br guan dnh drochloride concentration 5 in ionized water, varying between approximately 0.8 and approximately 1.2 g / ml dissolution, thus obtaining an aqueous precursor solution, The precursor solution, under vigorous stirring, contains guanine hydrochloride. The mass ratio of sl ka varied between approximately 0.37:1 and approximately 0.47:1. kolo dal sl ka ç ne kademel olarak dah l ed lmes , böyle sl kanın sert 10 allowing it to function as a template, ...obtained to ensure proper mixing and precursor filtration. d spers yonun 48-52°C in an oil bath overnight mixing throughout, v. Cooling the mixture to room temperature and recovering the solid content, 15 then the resulting material is ground into a fine powder, v. placing the ground powder into a lidded quartz crucible and The material is subjected to an argon atmosphere for 1.5-2.5 hours, approximately 540- Calculating by heating to 560°C, maintaining this temperature for an additional 1.5-2.5 hours. protected throughout and then the oven automatically cools to ambient temperature 20 left to cool, v. kals ne ed lm ş sarı malzemen n 190-210 ml'l k 3.8-4.2 M ammonium immersion in hydrogen defluoride (NH₄HF₂) solution and approximately Erosion of the SLKA template by mixing for 46-50 hours, v. Collection of the obtained meso-structured gCN by centrifugation, followed by 25 To remove residual fluoride ions and reaction byproducts. repeated washing with distilled pure water and ethanol, v. subsequent composite formation suitable meso-structured graphitic carbon nitrile To obtain it, the final product must be dried under ambient conditions. 21644.92 17 Meso-structured gCN is synthesized using a rigid template-supported strategy. It was prepared by thermal polymerization of hydrogen chloride. In the synthesis of T pkbr, Guan dnh drochloride (GndCl), a GndCl concentration varying between approximately 0.8 and 1.2 g / ml. to obtain an aqueous solution with a certain concentration, ionized water is used The dissolved and obtained solution, the sl ka particles function as a sacrificial template 5 To ensure visibility, GndCl was mixed vigorously with the rock. Colloidal group where the mass ratio varies between approximately 0.37:1 and approximately 0.47:1 Slurry rock (Ludox HS40) was added slowly. The mixture was formed within the slary rock network of the precursor. An oil kept at 48-52°C (321-325 K) to ensure even distribution. It was stirred in the bath overnight. After cooling to room temperature, it solidified. 10 The phase was separated and ground into a fine powder. The resulting material was placed in a lidded container. The quartz was transferred to a crucible and the temperature was raised to 540-560°C (813-) for 1.5-2.5 hours. (833 K) it is taken out, kept at this temperature for another 1.5-2.5 hours and then In a tube furnace where it is left to cool naturally, it remains under an argon atmosphere. ed lm şt r. Calcined yellow powder, 190-210 15, for removing silica skeleton. 3.8–4.2 M ammonium hydrogen defluoride (NH₄HF₂) solution immersed in and It was mixed for 46-50 hours. After etching, the product was centrifuged. collected, residual fluoride species and other by-products are removed using distilled water and It has been washed with ethanol. The method for synthesizing Black Phosphorus (BP) involves the following steps: The mass ratio of red phosphorus:Sn:SnI4 is approximately 48:2:1 to approximately 52:2:1. red phosphorus, tin (Sn) and tin(IV) iodide (SnI₄) are exchanged between them. weighed and included in a quartz bulb, where the quartz bulb is 25 approximately 18-22 cm long and approximately 1.5-2 cm in diameter sah pt r, To prevent contamination and oxidation, the bulb's temperature should be high. Evacuation and sealing under vacuum conditions, . sealed bulb horizontally 30 into programmable muffle furnace positioning, 21644.92 18 v. volatile species substituting and mineralizing to support phosphorus transport. To activate and maintain the furnace temperature, it is gradually increased to approximately 883-903 K. raising the temperature and maintaining this temperature for approximately 4-5 hours, v. s yah to promote the nucleation of phosphorus and controlled crystal growth To do this, the oven temperature should be slowly reduced to approximately 750-760 K and 5 Keeping the bulb at this temperature for approximately 1.5-2.5 hours, To maintain stability, the system needs to be cooled to approximately 383-403 K. then allowed to cool naturally to ambient temperature, v. To prevent oxidation, the bulb must be switched on under anhydrous toluene and black phosphorus kr staller nn isolated lmes, 10 v. . zole ed len kr staller n mutlak ethanol çinde approximately 25-35 mina ka için purification by ultrasonic treatment, thus removing residue. mineralizers and unreacted red phosphorus removal and extraction of high-purity black phosphorus (BP). High purity s black phosphorus (BP), adapted from previous studies (3,4) using improved mineral vapor transfer (CVT) method It has been synthesized. In this approach, red phosphorus (RP) is used as an elemental precursor. when used, it facilitates crystal nucleation and transitions between intermediate phases. It is known that tin (Sn) and tin (IV) 20 direct the controlled formation of BP through it. Iodide (SnI₄) was used as a mineralizing agent. For synthesis, RP, Sn and SnI₄ is highly sensitive, with mass ratio ranges from approximately 48:2:1 to approximately 52:2:1. a quartz crystal weighed and measuring 18-22 cm in length and 1-2 cm in diameter It was loaded into the bulb. The bulb was then subjected to oxidation and external contamination. To prevent this, it was completely evacuated and sealed under high vacuum. 25 After being sealed, it is placed horizontally inside the programmable lbr muffle furnace. placed; here all volatile components except Sn are completely To ensure sublimation, the temperature was gradually increased to 883-903 K. and maintained for 4.5-5.5 hours. The furnace temperature was then used for BP core preparation. and was slowly lowered to 748-768 K, the established regime for crystal growth, and 30 It was held for 1.5-2.5 hours. Then the bulb was removed to minimize thermal shock and crystallization. 21644.92 19 To stimulate cooling, it was cooled to 383-403 K, which then spontaneously returned to ambient temperature. The cooling process is complete. The ampoule is opened under anhydrous toluene to prevent oxidation. The resulting BP crystals were collected. Residual mineralizers and reactions... To remove the g rmem ş RP, the crystals were soaked in absolute ethanol for 25-35 minutes. Ultrasound treatment was applied throughout and it is suitable for further use, 5 Purified BP has been obtained. The use of photosensitizers in phototherapy is a key approach. It includes the following steps: ...to ensure that the photosensitizer is applied to the targeted tumor tissues. application within a given environment for the purpose of The photosensitizer is activated using near infrared (NIR) light from the target area. exposure to light, . The formation of reactive oxygen species (ROS) upon light activation is 15 initiation. The invention relates to the use of a photosensitizing agent (a heterojunction photosensitizer) in ROS. They operate according to the PDT principle, which involves using light to generate radiation. These ROSs, While healthy retinal cells remain undamaged, retinal noblastoma cells show a 20% deterioration. It triggers apoptosis (programmed cell death). Two heterojunctions Photosensitizer, compared to visible light, penetrates biological tissues more deeply. It is activated by penetrating NIR light. This allows the eye to see deeper. treatment of retinoblastoma tumors located in these regions It is recognized. When activated with photosensitizing NIR light, heterojoint tumor 25 It produces ROS efficiently in the region. Heterojunction, electron-hole By minimizing recombination, cancer cells are effectively destroyed. ROS, which is of critical importance for the production process, increases production efficiency. Furthermore, kl heteroeklem photosensitizing tumor tissues selectively accumulate br c r and reduces the risk of collateral damage to healthy retinal cells. This discovery is based on 30 key principles. The advantage is the increased stability and biocompatibility of the photosensitizer. 21644.92 BP, when used alone, tends to degrade rapidly in biological environments. However, when combined with gCN, stability increases, improving efficacy during treatment. It is developing. It is a heterojunction composed of carbon, nitrogen, and phosphorus. Photosensitizers are biocompatible and non-toxic, and may elicit an immune response or It minimizes the risk of side effects. The treatment is non-invasive; because cli 5 Heterojoint photosensitizers are administered intravenously, targeting the tumor. It is performed by cutting and activating with NIR light. This procedure is painless and takes 30 minutes. The process continues; the photosensitizer is eventually broken down and eliminated from the body. Healthy control cell line RPE-1 cells (Retinal Pigment Epstein-Barr Cells) 10 and using retinal blastoma (RB) cells, a type of cancer cell line, nv tro Experiments were carried out. RPE-1 cells were treated with 10% Bovine Calf Serum (BCF). pen sln and streptom sec supplement Dulbecco's Modified Eagle Five places le Bes n Mixture F-12 (DMEM-F12) is a controlled mixture at 37 °C containing 5% CO2. Cultured under these conditions. Similarly, RB cells, 20% BCF, penicillin 15 and streptomin-containing RPMI 1640 cells were grown in a culture medium. This cell culture To prepare the nanolayers to be used in the experiments; 3200 μg / ml BP, gCN and gCN / BP Cl heterojunction nanolayer dissolved in ddH2O and heated at 17 °C for 24 hours. It was exposed to the final cation for hours while protected from light. The final cation... Subsequently, the dispersion was centrifuged at 2000 rpm for 15 minutes at 4 °C and nano-20 The supernatant containing the layers was transferred to a new tube for use. 100 μL of the stock solution for the cell culture experiment is mixed with 900 μL of cell culture medium. Diluted RPE-1 and RB cells were then added to 96-well plates. to be applied in concentrations of 320 μg / ml, 160 μg / ml, 80 μg / ml, 40 μg / ml and 20 μg / ml. Series dilutions were performed to obtain varying concentrations. 25 RPE-1 cells 5x10³ cells per well, RB cells se per well Added at a cell density of 2x10⁴ and incubated overnight at 37°C in each sample. ed lm şt r. After incubation, 100 μL of nano-layer dilution was added to each well. The procedure was performed and the treatment lasted 24 hours. Post-treatment cell viability was measured using an MTT (3-30 (4,5-D met l lt azol-2- l)-2,5-D fen l tetrazolium bromide) test using 21644.92 21 It has been evaluated. Another study was conducted to evaluate in vitro PDT. In the experiment, following overnight incubation, 100 μL of culture medium was added. nanolayer solutions were used to modify the process, followed by 38 mW of 810 nm NIR radiation. The cells were exposed to laser irradiation for 30 minutes. After irradiation, the cells... To assess cell viability using the MTT test, incubate for another 24 hours. Experimental groups were: 40 μg / ml and 80 μg / ml, with and without NIR exposure. and 320 μg / ml nanolayer solutions, as well as areas exposed only to light or h included control groups that received no treatment. Before BP was combined with gCN, BP and gCN nanosheets alone were 10 s totoks s te has been tested in terms of. As shown in Figure 1A, the expected n As BP nanolayer concentrations increase, RB cell viability increases. The results are inconsistent after irradiation, as shown in Figure 1B. It has remained there. These unexpected results are shown in the bright area in Figure 3. As also proven with microscopy, BP nanolayers contain 15 cells in the cell culture medium. It is linked to aggregation. Furthermore, as shown in Table 1, BP nanolayers, At 570 nm, the neutral carbon crystal exhibited absorption, affecting the MTT test results. These problems are due to the analytical effects of BP nanolayers, making further experiments difficult. This has shown that it is not suitable. On the other hand, Table 2 shows that gCN nanolayers are 570 It has been shown that there is no intrinsic absorption at nm. However, 20 As shown in Figure 4A-C, light exposure also causes RB cell death. They did not detect it. Absorption (OD570) is an optical measurement at a wavelength of 570 nm. It represents intensity, for example, reflecting the degree of light absorption, and Commonly used biological tests to indicate cell viability or metabolic activity. It is used as. 25 BP nanolayer concentrations (µm / ml) Absorbance (OD570) 320 0.546 0.486 160 0.342 0.307 80 0.239 0.218 40 0.189 0.181 0.167 0.168 21644.92 22 Table 1 gCN nanosheet concentrations (µm / ml) Absorbance (OD570) 320 0.084 0.082 160 0.083 0.083 80 0.083 0.083 40 0.084 0.084 0.089 0.085 Table 2 To address these issues, BP developed a system that significantly improves stability and 5 Heterojunctive photosensitizers that eliminate unwanted signals It has been produced. As shown in Figure 6A, the single heterojoint. Photosensitizer did not affect RB cell viability without NIR stimulation. However, following hair removal therapy applied with nanosheets and NIR, As shown in Figure 6C, RB cell viability was significantly reduced; Figure 6B-10 As shown in D, there are no toxic effects on healthy RPE-1 cells. It has not been observed. Table 3 shows the concentrations of the KL heterojunction photosensitizer at varying levels. It shows the absorption measurement. 15 gCN / BP Cl heterojunction nanolayer concentrations (µm / ml) Absorbance (OD570) 320 0.084 0.087 160 0.094 0.082 80 0.082 0.082 40 0.084 0.083 0.083 0.084 Table 3 Later, the reactant oxygen species, which is the culture mechanism in PDT-induced cell death, The role of ROS (Residual Rotational Antibodies) production in species was evaluated. ROS production decreased in 24 hours after treatment. It was then evaluated and as shown in Figure 8A-C, the RB culture therapy Oxidative stress increases in cells, as shown in Figure 8B-D, RPE-1 21644.92 23 No significant changes were observed in the cells. Apoptotic pathway Western blot The analysis was examined at a more advanced level; as shown in Figure 9, anti-apoptotic A decrease in BCL-2 protein and an increase in Caspase-9 were observed in RB cells. Apoptosis has been confirmed to be active. Interestingly, Bax expression is also present. It has remained unchanged; this means that apoptosis can occur through other pathways. 5 This has led to the belief that it could be directed in a certain way. In terms of cell morphology and proliferation, it involves influencing cell nuclei. To evaluate the activity of 4′,6-damino-2-phen lindol (DAPI) and its tight junctions. Immunofluorescence staining was performed using ZO-1. Figure 13 shows 10 As shown, there was no damage to healthy RPE-1 cells after treatment. No proliferation was observed. Proliferation tests, as shown in Figure 11, during treatment. RB disrupts cell integrity and reduces aggregation; in healthy cells As shown in Figure 12, no significant effect was observed. TEM micrographs reveal the morphology of gCN / BP heterojunction nanolayers. This is revealed. As shown in Figure 14A–C, the observed structures It displays irregular, layer-like areas with varying contrasts, The situation involves the combination of gCN nanolayers and relatively thicker BP flakes. This indicates that it is present. Uniform dispersion and close contact between the two phases, 20 This indicates successful heterojunction formation. The presence of distinct interfacial boundaries. Its presence is critical for promoting charge transfer pathways in photocatalytic applications. It supports the interface interaction between the components that are important. High-angle dark field (HAADF) STEM image, composition tnh br t This further confirms the point. As shown in Figure 14D, image 25 The contrast differences observed throughout are in elemental composition and density. They reflect variations; brighter regions have higher atomic numbers. That's why the darker areas correspond to BP fields, while the darker areas are richer in carbon and nitrogen. zeng n gCN matr sn tems l etmekted r. Net structural integration, BP nano- 30 It shows elemental distribution maps for phosphorus (P), carbon (C) and nitrogen (N). 21644.92 24 This further confirms the successful integration of kb. In Figure 14E–H As shown, the phosphor signal is prioritized in the brighter HAADF areas. Locally, this confirms the presence of BP; however, carbon and nitrogen are absent from gCN. homogeneously distributed throughout the composite in a manner consistent with its structural properties It is dispersed. The superimposed elemental map shows the spatial co-5 of BP and gCN. by exhibiting localization and without significant phase separation. This confirms the formation of an interfacing heterojunction. In general, Microscopic and elemental analyses revealed BP and gCN in a regular heterostructure. This confirms that they have successfully combined, and this is evidenced by visible light irradiation. By improving photocatalytic converter performance, it enhances charge separation and charge transfer. It is expected to make things easier. The XRD pattern of pure mesoporous g-C₃N₄ (gCN) material is shown in Figure 15. Interlayer stacking of conjugated aromatic skeletons with center 2θ ≈ 28° the corresponding (002) plane is assigned a large and dense diffraction pk 15 serg lemekted r. This character st kpk, pol mer k Karbon ntride malzeme ç ntpk which is based on the π–π conjugate planar structuring as previously reported. a layered graphene structure compatible with mesoporous gCN structures is formed This is shown in Figure 15. Also, the XRD of crystalline phosphorus (BP) is shown. The pattern; planes (020), (040) and (060) of the orthorhombic BP crystal structure respectively 20 Three sharp and intense diffraction patterns were indexed at positions 2θ ≈ 17°, 34.5° and 52.5°. pk serg lemekted r. The sharpness and symmetry of these reflections, as previously reported In line with bulk orthorhombic k BP structures, the synthesized BP has a high n-level structure. It verifies the stall and phase purity of the gCN / BP heterojunction nano-. The XRD pattern of the layered h br t material shown in Figure 15 is for both gCN and 25 BP's characteristic diffraction properties are clearly presented by the brin-linked heterostructure. It confirms that it was formed successfully. (002) corresponding to the plane 2θ The main gCN diffraction pk near ≈ 27.4°, in the composite, is due to BP involvement. with a slightly reduced density which can be attributed to lattice perturbation. However, it is protected. Similarly, the BP (020) reflection at approximately 17° is also h br t 30 It has been preserved in its structure, but appears to be enlarged and weakened; this 21644.92 The situation is that BP's previous partial exfoliation into nano-layers and in response The long-range, low-frequency signal indicates a decrease in the current level. This is an important point. As such, both gCN and... BP'n n character st k diffraction p kler nn co-existence, heterostructure's chemical phase transformation mainly through physical interface interaction 5 This proves that it was formed in this way. The single crystal structures within the composite are like this. well-defined protection, suitable for subsequent photon and photocatalytic applications. br gCN / BP kl heterojunction nanolayer formation This confirms it. Optical absorption of pure gCN and gCN / BP composite heterojunction nanolayers. The properties were investigated using UV–Vs–NIR diffuse reflection spectroscopy (DRS). As shown in Figure 16A, pure gCN acts as a polymer semiconductor. has ntr ns k band gap characteristic st kbr feature at approximately 455 nm bel rg nbr absorption edge exhibits. After the participation of BP, gCN / BP 15 KL heterojunction nanolayer emits light throughout the entire visible light region. It exhibits a significant increase in absorption. This is at the absorption edge. The visualization and redshift of the visualization, successful correlation between BP and gCN. h br d zasyona şönmektedir ve gel şörün ışık Toplama kab l yet le This is the conclusion. To further improve the opto-bandwidth energies, 20 The reflection data was transformed using the Kubelka-Munk function and indirectly The Tauc diagrams were analyzed assuming a single electron transition. (Figure 16B) As shown, the band gap energy of pure gCN is estimated to be approximately 2.87 eV. This value is obtained from the literature for mesoporous graphitic carbon nitride. The values ​​are consistent with this. In contrast, the gCN / BP Cl heterojunction nanolayer 25 A narrowed band gap of approximately 2.75 eV was exhibited, indicating that BP participation had an energy advantage. It has been shown that it significantly narrows the band gap. This narrowing of the band gap, interface electron interactions and gCN / BP heterojunction nanolayer It is connected to the formation of intermediate energy levels in the interface. The increasing structure of the hybrid system. Its light-gathering capacity is greater under visible light irradiation than a photo-30. It is expected to stimulate the formation of an excited charge carrier population. 21644.92 26 This situation highlights the photocatalytic and photodynamic implications of light-guided applications. It is extremely advantageous in terms of improving performance. Steady-state of pure gCN and gCN / BP Cl heterojunction nanosheets Photoluminescence (PL) spectra are the radiant spectra of photogenerated charge carriers. It was recorded to evaluate recombination behavior. In Figure 17A. As shown, pure gCN exhibits limited charge separation within a bulk polymer skeleton. an indicator of intense radiative recombination resulting from ver ml l ğ nden It exhibits strong br emission pk. In contrast, gCN / BP kl heterojunction. nanolayer, showing that the recombination process is significantly suppressed, 10 The bar exhibits a damped PL density in the form of a zbr. Emission This significant reduction in density results in more efficient charge carriers within the heterostructure. This indicates the presence of interface electron interactions that encourage differentiation. It is doing so. Time-dependent studies are needed to investigate carrier dynamics at a more advanced level. Photoluminescence (TRPL) spectroscopy was performed. Figure 17B shows 15 As shown, the fluorescence decay curves represent the fast and slow lifetime components. The analysis was performed using a double exponential decay model. Pure Mean carrier lifetime (τ_avg) of gCN, mesoporous carbon nitride systems approximately 7.1 ns, consistent with the values ​​obtained. It has been determined that the formation of a gCN / BP heterojunction nanolayer occurred in 20 years. The average lifetime has increased to approximately 7.8 ns; this indicates an improvement in charge carrier recombination. It has been noticed that the delay in carrier life indicates a change in gCN / BP. Heterojunction nanolayer formation, photo-excited electron-hole pairs nn This clearly demonstrates that it allows for a more effective spatial separation. The transfer of load and suppressed recombination losses in the composite 25 The photocatalytic and photodynamic performance under visible light irradiation is important. It is expected to increase significantly. All statistical analyses were performed using GraphPad Premium 8.0 software. This has been done, and p-values ​​less than 0.05 and 0.01 indicate statistical significance. It is fading. Analysis combined with NIR for retinoblastoma therapy. 21644.92 27 This has confirmed the efficacy and safety of heterojunctive photosensitizers. The results show that BP nanosheets alone cause instability and analytical flexibility. However, the KL heterojoint photosensitizer solves these problems and apoptotic effects on RB cells without damaging healthy retinal cells It has been shown that it has increased significantly. 5 The combination of gCN and BP within a non-ribbed heterojunction structure, NIR By increasing light absorption in the spectrum, the tissue penetration depth is increased. developing and treating tumors located deeper within the tissue and making them more effective. The design also optimizes load-bearing separation, resulting in a higher ROS 10. This results in the production of [method], which in turn destroys the tumor cells. It strengthens [the process]. Furthermore, the invention stabilizes BP and improves biocompatibility. By providing a solution that has fewer side effects, is more durable and patient-friendly. It offers targeted activation of the photosensitizer in tumor cells. By reducing damage to healthy tissues, it increases overall safety and effectiveness. 15 In conclusion, the invention is effective, stable, and offers a unique photodynamic therapy solution for retinal blastoma. This represents a significant step forward by offering a compatible treatment option. The use of heterojunctival photosensitizers causes damage to surrounding healthy tissues. new and powerful for selectively destroying cancer cells while minimizing their presence. br offers a vehicle. 20 References [1] K m, J. W., Jacobsen, B., Zolfaghar , E., Ferrar o, A., Chevez-Barr os, P., Berry, J. L., Lee, D. K., R co, G., Mad , I., Rao, N., Stachelek, K., Wang, L. C., & Gomer, 25 C. (2017). Rabb t model of ocular nd rect photodynam c therapy us ng a ret noblastoma xenograft. Graefe's arch ve for cl n cal and exper mental ophthalmology = Albrecht von Graefes Arch v fur kl n sche und exper mentelle Ophthalmolog e, 255(12), 2363–2373. https: / / do .org / 10.1007 / s00417-017-3805-8 21644.92 28 [2] Gomer, C. J., Ferrar o, A., & Murphree, A. L. (1987). The effect of local zed porphyr n photodynam c therapy on the nduct on of tumour metastas s. Br t sh journal of cancer, 56(1), 27–32. https: / / do .org / 10.1038 / bjc.1987.147 [3] Yamamoto, N., Sery, T. W., Hoober, J. K., W llett, N. P., & L ndsay, D. D. (1994). Effect veness of photofr n II n act vat on of macrophages and n v tro 5 k ll ng of ret noblastoma cells. Photochem stry and photob ology, 60(2), 160–164. https: / / do .org / 10.1111 / j.1751-1097.1994.tb05084.x [4] Walther, J., Schastak, S., Duk c-Stefanov c, S., W edemann, P., Neuhaus, J., & Claudep erre, T. (2014). Eff c ent photodynam c therapy on human ret noblastoma cell l nes. PloS one, 9(1), e87453. https: / / do .org / 10.1371 / journal.pone.0087453 10

Claims

21644.92 29 REQUESTS 1. CBR for use in photodynamic therapy (PDT) for the treatment of retinal blastoma. KL heterojunction nanolayer photosensitizers are listed below. It contains: 5 • k-dimensional (2D) polymer kbr semiconductor graphite carbon dioxide (gCN) and • s yah phosphorus (BP).

2. The photosensitizer in claim 1 is g / g, where g / g carbon dioxide (gCN), A two-dimensional (2D) polymer carbon semiconductor with a band gap of 2.7 eV.

3. Claim 1 is the photosensitizer, where s is the band gap of the phosphor (BP). 1.4 eV. 15 4. Like any of the previous systems, it is a photosensitizer, but here... Heterojunctions are reactive when activated with near infrared (NIR) light. It produces ROS (reactive oxygen species), thus inducing apoptosis in neoblastoma cells. It induces. 20 5. Claim 4' is a single GB photosensitizer, where the heterojunction is an 810 nm wavelength. The neck has the ability to absorb near-infrared (NIR) light; this means For effective treatment of retinal blastoma, the injectable material must penetrate deeper into the tissue. It ensures that it is done. 25 6. Photodynamic therapy (PDT) oriented heterojunction nanolayer This is a method for synthesizing the photosensitizer, and it involves the following steps: Synthesis of meso-structured graphite carbon nitride (gCN), 30 . synthesis of phosphate (BP) 21644.92 . s yah phosphorus (BP) is approximately 1.6 la in N-methyl-2-pyrrolidone (NMP). suspended at a CBR concentration of approximately 1.8 mg / ml and For obtaining eksfol BP nanolayers, the suspension should be 20- Subjected to final cation at a controlled temperature of 25°C, v. The mass ratio of gCN:BP is approximately between 4.7:1 and 5.1:

1. The mesoporous graphite that was previously dispersed within NMP has been altered. Addition of carbon dioxide to the BP dispersion subjected to final cation, Here, the insertion process is carried out under continuous post-cation. v. interfacial contact and heterojunction between gCN and BP phases. 10 combined dispersion to encourage unification the final stage should be continued for 3.5-4.5 hours. v. The resulting gCN / BP composite is approximately 15-20% of the mixed suspension. isolating by centrifugation at approximately 8500-9500 rpm for a full minute. ed lmes, then to remove the unbound material. Washing the precipitate with ethanol twice, 15 v. Drying of washed composite material, v. Composite BP loading adjustment, where standard BP load is used. According to the total mass of the gCN / BP heterojunction nanolayer It contains approximately 16.5-17.5% BP by weight, and thus... Heterojoint photosensitizers are obtained. 20 7. The method described in 6 minutes is as follows: here, meso-structured graphite carbon N trioxide. The (gCN) synthesis method involves the following steps: . guan dnh drochloride, br guan dnh drochloride concentration 25 in ionized water, varying between approximately 0.8 and approximately 1.2 g / ml dissolution, thus obtaining an aqueous precursor solution, The precursor solution, under vigorous stirring, contains guanine hydrochloride. The mass ratio of kolo dal sl ka is approximately between 0.37:1 and approximately 0.47:

1. The change in the colloidal structure, thus 30 allowing sl blood to function as a rigid template, 21644.92 31 ...obtained to ensure proper mixing and precursor filtration. d spers yonun 48-52°C in an oil bath overnight mixing throughout, v. Cooling the mixture to room temperature and recovering the solid component, then grinding the resulting material into a fine powder, 5 v. placing the ground powder into a lidded quartz crucible and The material is subjected to an argon atmosphere for 1.5-2.5 hours, approximately 540- Calculating by heating to 560°C, maintaining this temperature for an additional 1.5-2.5 hours. protected throughout and then the oven naturally cools down to ambient temperature. left to cool, 10 v. kals ne ed lm ş sarı malzemen n 190-210 ml'l k 3.8-4.2 M ammonium immersion in hydrogen defluoride (NH₄HF₂) solution and approximately The SLKA template is mixed for 46-50 hours. erosion, v. Collection of the obtained meso-structured gCN by centrifugation, followed by 15 To remove residual fluoride ions and reaction byproducts. repeated washing of the solid with distilled water and ethanol, v. subsequent composite formation suitable meso-structured graphite carbon To obtain the product, the final product must be dried under ambient conditions.

8. The method described in step 6 is for synthesizing S or Phosphorus (BP). It includes the following steps: The mass ratio of red phosphorus:Sn:SnI4 is approximately 48:2:1 to approximately 52:2:

1. red phosphorus, tin (Sn) and tin(IV) iodide (SnI₄) 25 weighing and incorporating into a quartz bulb, here the quartz bulb approximately 18-22 cm long and approximately 1.5-2 cm in diameter sah pt r, To prevent contamination and oxidation, the bulb's temperature should be high. Evacuation and sealing under vacuum conditions, 30 21644.92 32 . sealed bulb horizontally inside programmable muffle furnace positioning, v. volatile species substituting and mineralizing to support phosphorus transport. To activate and maintain the furnace temperature, it is gradually increased to approximately 883-903 K. raising the temperature and maintaining this temperature for approximately 4-5 hours, 5 v. s yah to promote the nucleation of phosphorus and controlled crystal growth to do this, the oven temperature should be slowly lowered to approximately 750-760 K. and keeping the bulb at this temperature for approximately 1.5-2.5 hours, To maintain stability, the system needs to be cooled to approximately 383-403 K. then allowed to cool naturally to ambient temperature, 10 v. To prevent oxidation, the bulb should be switched on under anhydrous toluene and black phosphorus kr staller nn isolated , v. . zole ed len kr staller n mutlak ethanol çinde az 25-35 minka purification by subjecting the patient to ultrasonic treatment throughout, thus removing residue mineralizers and unreacted red phosphorus 15 removal and extraction of high-purity black phosphorus (BP).

9. Use of the GB photosensitizer in phototherapy as described in Claim 1. This is a directional CBR method and includes the following steps: ...the photosensitizer's effect on targeted tumor tissues. implementation within a given environment in order to provide To activate the photosensitizer, the target area must be exposed to near-infrared radiation. Exposure to (NIR) light, . The formation of reactive oxygen species (ROS) upon light activation is 25%. initiation.

10. The method of using the GB photosensitizer in claim 7 is described here. The activation procedure is a 30-minute non-invasive light exposure. It includes and photosensitizer, naturally 30 days after the procedure. It is falling apart.

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