INTRODUCTION:

Oral candidiasis also called as oral thrush, is a commonest oral fungal infection, which affects mucous membranes of the mouth^1,2. Candida albicans in the main causative agent of this disease, a highly versatile commensal microorganism³ and apart from these it may also be caused by C. tropicalis, C. glabrata, and some other species of Candida as well⁴.

In recent decades, this invasive fungal infection has been increasing significantly, contributing to high incidences and mortality in immunosuppressed patients⁵. Current antifungal agents have several limitations, including that only a small number of classes of antifungals are available, certain of which have severe toxicity and high cost.

Moreover, the emergence of drug resistance is a new limitation to successful patient outcomes. Drug resistance in candidiasis is exacerbating the threat this disease is posing to human beings. Therefore, the development of antifungals with novel targets is an essential strategy for the efficient management of C. albicans infections^6,7.

From the literature survey it has been clear that, the new triazol antifungal voriconazole is a potent antifungal drug as compared to fluconazole in the treatment of esophageal candidiasis and it was also suggested that this new azole derivative may be a useful alternative for fluconazole-resistant Candida strains⁸. As it is a new drug molecule so, there is little strains resistant to voriconazole. In one of our study we had combined voriconazole with beauvericin to synergize the effect of voriconazole and combat the multi drug resistance of C. albicans⁹. Beauvericin is a cyclic hexadepsipeptide which belongs to the enniatin antibiotic family^10,11. It is normally a mycotoxin, produced from another fungus Beaveria bassiana^12,13. Numerous studies proved the anti-fungal potential of beauvericin against C. albicans ^14-16. Besides this, one study proved that the MIC of voriconazole (0.5μg/ml) was dramatically reduced by 128^th times (0.0039μg/ml) following combination with beauvericin⁹.

Furthermore, the efficacy of the treatment was enhanced by incorporation of both drugs in mucoadhesive niosomal nanogel. Niosomes are vesicular drug delivery system, which can entrap both hydrophilic and lipophilic drugs. The mucoadhesive gel provides the advantages of localized drug delivery over an extended period of time.

In the present study we developed and optimized a muco-adhesive niosomal nanogel for the co-delivery of voriconazole and beauvericin to treat the oral candidiasis effectively.

MATERIALS AND METHODS:

Materials:

Voriconazole was received as a gift sample from Acros Organics, Belgium and beauvericin was received as a gift sample from Medchem express, USA. Cholesterol, chitosan and span 60 were purchased from Yarrow Chem Products, Mumbai. All reagents were of highest analytical grade.

Preformulation profiling:

Both the drugs were identified by determining absorption maxima by UV spectrophotometer in phosphate buffer of pH 6.8 using Shimadzu-1700 UV-Visible Spectrophotometer and FT-IR analysis¹⁷.

The shake flask method was used to determine the solubility of drugs¹⁸. Four different solvents, including distilled water, ethanol, 0.1 N Hydrochloric acid, and phosphate buffer pH 6.8, were used to asssess the solubility.

The drug's compatibility with excipients was established by FT-IR analysis. All samples were completely mixed with excipients at a ratio of 1:1, and were kept in closed vials at 40°C and 75% RH for 21 days before the test. To rule out potential physical and chemical incompatibilities, the spectra of pure drugs and mixes of drugs with excipients were compared with the standards ¹⁹.

Preparation of niosomes:

Niosomes were prepared by the hydration of lipid thin films. In a round bottom flask, 0.01% w/v each of voriconazole and beauvericin was dissolved in 5ml of ethanol. Cholesterol and span 60 were dissolved in 5ml of chloroform and placed in a separate beaker²⁰. Both the solutions were mixed and the organic solvent from both the solutions was evaporated until a fully dry film was formed at the bottom of round bottom flask²¹. Then, phosphate buffer pH 6.8 was used to hydrate this dry film.

Incorporation of niosomes in to mucoadhesive nanogel:

The gel base was consist of HPMC (8.7% w/w), poloxamer P407 (0.4% w/w), and chitosan (0.12% w/w). HPMC and P407 were dissolved in deionized water and chitosan was dissolved in sodium acetate-acetic acid buffer pH 4.2. Then, both solutions were mixed at room temperature with proper stirring. The resulted mixture will acted as gel base^22,23.

The niosomal dispersion was then mixed with gel base until the final weight was 10g.

Box–Behnken Design (BBD) Experiment for optimization of Niosomal gel:

The voriconazole and beauvericin loaded niosomes were optimized by Box-Behnken response surface methodology (Design-Expert^® Software Version 12). The independent variables included span 60 concentration (X1), cholesterol concentration (X2), and the sonication time (X3) at three levels i.e. low, medium, and high for the preparation of fifteen formulations. These responses were studies with respect to these three responses were studies which were particle size (Y1), entrapment efficiency (Y2), and percent cumulative drug release (Y3) as presented in Table 1. Moreover, 3D response surface graphs and contour plots were plotted to depict the effects of the predetermined factors on the measured responses^24-26.

Table 1: Independent variables in Box–Behnken design for the optimization of niosomes

Factor	Independent variables	Units	Level
Factor	Independent variables	Units	Low	Medium	High
A (X₁)	Span 60	% w/v	60	80	100
B (X₂)	Cholesterol concentration	% w/v	10	20	30
C (X₃)	Sonication Time	Sec	2	4	6
S. No.	Response	Goal
1	Y₁(Particle Size)	Minimize
2	Y₂(Entrapment efficiency of voriconazole)	Maximize
3	Y₂(Drug release of voriconazole)	Maximize

The following non-linear quadratic model expression, where Y is the dependent variable, b0 is the arithmetic mean and Y1– Y123 are the regression coefficients of respectable variables, was used to calculate the effect of independent variables on dependent variables at three levels. The interaction between the several parameters is indicated by the factors X1, X2, and X3.

Y=b₀+Y₁X₁+ Y₂X₂+ Y₃X₃+ Y₁Y₂X₁X₂+ Y₁Y₃X₁X₃+ Y₂Y₃X₂X₃+ Y₁²X₁²+ Y₂² X₂²+Y₃²X₃²

Eq. 1

CHARACTERIZATION OF NIOSOMES:

Particle size and zeta potential:

The size and zeta potential of niosomes were determined by the Zeta sizer 2000 (Malvern Instruments) at 25°C and 90° scattering angle. The mean and standard deviation were calculated after each measurement, in triplicate^27-29.

Surface and Shape Analysis by TEM:

The shape and surface characteristics of niosomes were determined by Transmission Electron Microscope (Model H-7500 Hitachi, Japan)^30,31.

Drug Entrapment Efficiency:

The amount of voriconazole and beuvericin entrapped in the niosomes was measured by the gel filtration method. Sephadex G-50 column was utilized to eliminate the free drug that was present in the niosomes and triton X-100 (0.5%v/v) was used for lysing of vesicles. The entrapment efficiency was determined by measuring the drug content using a UV-visible spectrophotometer at a wave length of 255nm for voriconazole and 208 nm for beauvericin^32-34.

In vitro drug release studies:

The in vitro drug release was studied out using Franz diffusion cell using egg membrane. To maintain the appropriate temperature condition, the entire assembly was placed on a magnetic stirrer and swirled at a speed of 100rpm^35,36. The required volume of phosphate buffer (pH 6.8) was placed into the receptor compartment of the diffusion cell. One ml of niosomal dispersion was applied to the egg membrane. To keep the sink conditions, a 1 ml aliquot of the receptor medium was taken at the predetermined time intervals and immediately replaced with an equivalent volume of fresh phosphate buffer (pH 6.8)^37-39. For in-vitro drug release experiments, the sample was spectrophotometrically examined at a maximum wavelength of 255nm for voriconazole and 208nm for beauvericin.

Characterisation of Mucoadhesive Gel:

Physical appearance:

The color, clarity, homogeneity and appearance of the developed niosomal gel was examined visually⁴⁰.

pH study:

The pH of the niosomal gel was determined by dispersing 2.5g of gel in 25ml of distilled water with the help of digital pH meter⁴¹.

Spreadability:

The spreadability of the gel was assessed by applying 0.5g of the gel to a circle that had been previously marked on a glass plate with a diameter of 2cm, and then using a second glass plate. Five minutes were given for a half kilogram of weight to rest on the upper glass plate. After the gel had spread, the circle's diameter was measured⁴².

Viscosity:

The viscosity of the niosomal gel was determined at 22^oC by Brookfield Viscometer⁴².

In-vitro determination of mucoadhesive force of the niosomal gel:

The mucoadhesive force of the niosomal gel was determined by a texture analyzer (TA.XT plus, Stable MicroSystems, UK). Before being placed onto the holding stage, the pig oral mucosa (20x20mm) was equilibrated at 37.0±0.5°C for 15 minutes. The mucoadhesive gel was placed in a beaker. The hydrated disk was attached to the lower end of the probe. The probe holding the tissue attached hydrated disk was then lowered to make contact with the soaked tissue at a predetermined force and held there for a predetermined amount of time. The probe holding the tissue attached hydrated disk was removed from the tissue at a predetermined test speed using the maximum detachment force (Fmax)^{43, 44}.

Stability Studies:

The stability studies were carried out on the optimized formulation F3. The formulation was plced in an airtight container and kept at 25°C and 60% relative humidity (RH) and 40°C and 75% (RH) for 90 days. After 15, 30, 45, 60, and 90 days, samples were checked for the remaining drug content^45,46. Each formulation's initial medication content was assumed to be 100%.

RESULTS AND DISCUSSION:

Preformulation profiling:

The drugs were identified by profiling several techniques including the UV spectroscopy and FTIR spectroscopy. All of the parameters were confirmed to be within acceptable limits and the official compendia specifications. Voriconazole exhibited highest absorbance at 255nm, while beauvericin showed the maximum absorbance at 208nm.

Solubility of drug:

Both the drugs were found to be remarkably soluble in phosphate buffer, propylene glycol, and ethanol and poorly soluble in purified water.

Drug excipient compatibility study:

The results of physical and chemical compatibility study indicate that the APIs are compatible with all the excipients used in the formulations.

Box behnken statistical optimization of niosomes:

The Box-Behnken Design was used for the optimization of niosomes prepared by lipid thin film hydration method. All the fifteen formulations with actual value of all the independent and dependent variables are mentioned in table 2. Along with their binary interactions and polynomial effects, each independent factor was investigated on three different levels. The vesicular size, entrapment efficiency and drug release of voriconazole were examined. The regression analysis and the effect independent variable on dependent variables are shown in Table 3. Equations 2-4 were used to apply quadratic mathematical models to examine the interaction between the independent components and the investigated responses.

Table 2: Box Behnken design with actual value of all the independent and dependent variables

Factor 1	Factor 2	Factor 3	Response 1	Response 2	Response 3
X₁:Span 60 (mg)	X₂:Cholesterol (mg)	X₃:Sonication time (min)	Y₁:Particle size (nm)	Y₂:Entrapment efficiency (%)	Y₃:Drug release at 3 hrs (%)
60	10	4	141	76.43	62.83
80	20	4	138	80.18	66.17
100	20	2	142	89.89	79.98
100	20	6	143	68.73	65.29
80	30	2	146	82.67	72.37
60	30	4	146	72.56	71.09
80	20	4	138	80.18	66.17
80	20	4	138	80.18	66.17
60	20	2	149	76.06	72.63
80	10	2	139	87.12	76.13
80	20	4	138	80.18	66.17
80	30	6	127	76.98	76.49
80	20	4	138	80.18	66.17
100	30	4	162	80.34	68.18
60	20	6	132	65.45	69.37
80	10	6	119	83.18	68.85
100	10	4	153	86.19	69.21

As mentioned in table 3 the p values of the all three responses were less that 0.05, which means the model is significant. Also, the difference between Predicted R² of all the response and Adjusted R² was less than 0.2, hence indicating a reasonable agreement in the study.

Figure1: Contour plots and 3D surface plots of the effects of independent variables on particle size

The contour plots as well as the 3D surface plot shown in Figure 1 and 2, illustrate that with the increase of the concentration of span 60 and cholesterol, the particle size and entrapment also increased. Whereas, the sonication time negatively affect the both particle size. However the entrapment efficiency was found to be increased with respect to sonication time.

Figure 2: Contour plots and 3D surface plots of the effects of independent variables on entrapment efficiency

As illustrated in figure 3, the concentration of span 60 and cholesterol increased, the percent cumulative drug release is decreased. Although, the effect of sonication time is high on percent cumulative drug release and it was found to be decreased.

The formulation F3 having particle size 142 nm, entrapment efficiency (89.89 %), and a drug release at 3 hrs (79.98%) was selected for further studies. This formulation made with 100 mg span 60, 20 mg cholesterol and 2 minutes of sonication time.

Figure 3: Contour plots and 3D surface plots of the effects of sonication time on dependent variables

CHARACTERIZATION OF NIOSOMES:

Particle size analysis by: Particle size analysis was performed by Beckman Coulter and the average size of niosomes was found to be 133.19 nm as mentioned in Figure 4.

Figure 4: Particle size distribution of niosomal dispersions

Zeta potential:

As illustrated in Figure 5, the average zeta potential of optimized formulation (F3) was noted to be -0.29.12 (mV). The highly negatively charged niosomes exhibited excellent stability, where the vesicles tended to be discrete rather than aggregate.

Figure 5: Zeta potential analysis

Surface analysis and shape:

Surface morphology of the niosomes was examined by Transmission Electron Microscope, Hitachi, Japan. The photograph given in figure 6 indicates that the niosomes are identical, smooth, vesicular in form, and free of agglomerations.

Figure 6: TEM image of drug loaded niosomal dispersions

Entrapment Efficiency:

The entrapment efficiency of the formulations (F1-F15) is summarized in Table 4.

Table 4: Entrapment efficiency of drug loaded Niosomes

Batch Code	Entrapment Efficiency (%) ± S.D. of Voriconazole	Entrapment Efficiency (%) ± S.D. of Beauvericin
F1	76.43±0.13	75.87±0.56
F2	80.18±0.29	79.76±0.08
F3	89.89±0.64	92.98±0.35
F4	68.73±0.98	69.08±0.17
F5	82.67±0.56	83.87±0.39
F6	72.56±0.49	74.19±0.62
F7	80.18±0.38	78.14±0.97
F8	81.13±0.47	82.61±0.82
F9	76.06±0.18	74.95±0.79
F10	87.12±0.86	86.25±0.51
F11	80.18±0.91	82.86±0.94
F12	76.98±0.59	74.47±0.38
F13	80.18±0.73	82.90±0.26
F14	80.34±0.09	79.62±0.41
F15	65.45±0.46	64.86±0.83

In vitro drug release study:

The percentage cumulative drug release of optimized formulation is mentioned in table 5 and figure 7. As per the results the voriconazole exhibited a maximum release of 89.52 % whereas the release of beauvericin was found to be 86.37% at 5 hrs.

Table 5: Drug release study of optimized formulation

Time (hrs)	% Cumulative drug release ± S.D. Voriconazole	% Cumulative drug release ± S.D. Beauvericin
0	0	0
0.5	20.76	15.15
1	39.28	31.94
2	62.17	57.83
3	79.98	69.17
4	85.67	81.05
5	89.51	86.37

Figure 7: Drug release profile of optimized formulation (F3)

Stability studies:

The stability studies at 25°C/60% RH showed good stability of all formulations as there was no appreciable change in the drug content. The drug instability was noted at 40±2°C/75 % (RH).

Characterization of Mucoadhesive Gel:

Appearance:

The developed mucoadhesive gel was found to be white translucent with consistent and soft texture.

pH Measurement:

The pH of the drug loaded mucoadhesive gel was found to be within the acceptable limits thus indicating suitability oral mucosa.

Spreadability study of gel:

The spreadability of mucoadhesive gel was found to be 22.94±0.3g.cm/sec which is within the acceptable range.

In-vitro determination of mucoadhesive force of niosomal gel:

The mucoadhesive force of niosomal gel was determined at room temperature. The value of Fmax of all the formulations was found to be in the range of 12.39±1.61 and 23.82±2.15g. The results clearly indicated that the gel formulations have a good mucoadhesion force.

CONCLUSION:

The study on Voriconazole and Beauvercin loaded mucoadhesive nanogel presented promising and innovative approach to counter oral candidiasis. The study involved the development and optimization of niosomes using the Box-Behnken experimental design and their incorporation in gel. The evaluation of niosomes for vesicular size, surface morphology, entrapment efficiency, drug release, and zeta potential played a pivotal role in establishing the efficiency. The results demonstrated that the optimized niosome formulation (F3) exhibited high entrapment efficiency, 89.89±0.64% for voriconazole and 92.98±0.35% for beauvericin, indicating that a significant amount of the drugs could be effectively delivered to the target site. Moreover, the cumulative drug release of 79.98±1.87% for voriconazole and 69.17±1.98% for beauvericin from the optimized formulation indicated sustained drug release, which could potentially lead to prolonged therapeutic effects, reduction in the frequency of administration and improve patient compliance. The mucoadhesive gel exhibied an optimum mucoadhesion force, ensuring prolonged contact with the affected oral mucosa, which can increase drug retention at the target site and potentially enhanced treatment outcomes.

Summarily, the voriconazole and beauvericin loaded niosomal mucoadhesive gel demonstrated a powerful and synergistic effect against oral candidiasis.

The combination of sustained drug release, improved mucosal adhesion, and formulation stability makes this delivery system a promising candidate for further exploration in clinical trials and potential incorporation into standard clinical practice. As the research continues to evolve, this innovative approach may significantly contribute to the improvement of oral candidiasis management, positively impacting the lives of affected patients worldwide.

CONFLICT OF INTEREST:

None.

ACKNOWLEDGEMENTS:

The authors thankfully acknowledge the support provided by Central Instrument Lab, Panjab University, Chandigarh.

REFERENCES:

1. Mundula T et al. Effect of Probiotics on Oral Candidiasis: A Systematic Review and Meta-Analysis. Nutrients. 2019; Oct 14; 11(10): 2449.

2. Vila T et al. Oral Candidiasis: A Disease of Opportunity. Journal of Fungi. 2020; 6(1):15. https://doi.org/10.3390/jof6010015

3. Zhang LW Fu et al. Efficacy and safety of miconazole for oral candidiasis: a systematic review and meta‐analysis. Oral diseases. 2016; 22(3): 185-95.

4. Alghanem SH et al. Formulation and Characterization of Two Nystatin Lozenge Formulations as a New Pharmaceutical Dosage form in The Syrian Market. Research Journal of Pharmacy and Technology. 2023; Apr 1; 16(4): 1639-43.

5. Taylor M, Raja A. Oral Candidiasis. [Updated 2021 Mar 14]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2021 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK545282/

6. Gheorghe DC et al. Biomaterials for the prevention of oral candidiasis development. Pharmaceutics. 2021; May 27; 13(6): 803.

7. Garcia-Cuesta C et al. Current treatment of oral candidiasis: A literature review. J Clin Exp Dent. 2014; Dec 1; 6(5): e576-82. doi: 10.4317/jced.51798.

8. Song YB et al. Antifungal susceptibility testing with etest for Candida species isolated from patients with oral candidiasis. Annals of Dermatology. 2015; Dec 1; 27(6): 715-20.

9. Rani D et al. Fungicidal combination for the prevention of C. albicans caused oral infections. Indian Patent Publication. 202311027781 A

10. Wu Q et al. A Review on the Synthesis and Bioactivity Aspects of Beauvericin, a Fusarium Mycotoxin. Front Pharmacol. 2018 Nov 20; 9: 1338. doi: 10.3389/fphar.2018.01338.

11. Mallebrera B et al. In vitro mechanisms of Beauvericin toxicity: A review. Food and Chemical Toxicology. 2018; 111: 537-545. https://doi.org/10.1016/j.fct.2017.11.019.

12. Wang Q, Xu L. Beauvericin, a bioactive compound produced by fungi: a short review. Molecules. 2012; Feb 24; 17(3): 2367-77. doi: 10.3390/molecules17032367.

13. Tong Y et al. Beauvericin counteracted multi-drug resistant Candida albicans by blocking ABC transporters. Synth Syst Biotechnol. 2016; Oct 25; 1(3): 158-168. doi: 10.1016/j.synbio.2016.10.001. PMID: 29062940; PMCID: PMC5640798.

14. Ramírez-Zavala B et al. A hyperactive form of the zinc cluster transcription factor Stb5 causes YOR1 overexpression and beauvericin resistance in Candida albicans. Antimicrobial Agents and Chemotherapy. 2018; Dec; 62(12): e01655-18.

15. Shekhar-Guturja T et al. Beauvericin Potentiates Azole Activity via Inhibition of Multidrug Efflux, Blocks Candida albicans Morphogenesis, and Is Effluxed via Yor1 and Circuitry Controlled by Zcf29. Antimicrob Agents Chemother. 2016; Nov 21; 60(12): 7468-7480.

16. Niimi M et al. Inhibitor-Resistant Mutants Give Important Insights into Candida albicans ABC Transporter Cdr1 Substrate Specificity and Help Elucidate Efflux Pump Inhibition. Antimicrobial Agents and Chemotherapy. 2022; Jan 18; 66(1): e01748-21.

17. Deshmukh MT, Mohite SK. Preparation and evaluation of mucoadhesive microsphere of fluoxetine HCl. Int J Pharm Sci Res 2017; 8(9): 3776-85.

18. Krishna SR et al. Preformulation, Formulation Development and Drug Release Studies of Dipyridamole Floating microballoons. Internatiomal Journal of Pharmaceutical Sciences and Research. 2020; 11(9): 4637-4647.

19. Rani D et al. Formulation optimization of omeprazole loaded pH sensitive mucoadhesive microspheres forsite specific delivery. Res. J. Chem. Environ. 2022; 26(6).

20. Khatoon M et al. Development and evaluation of optimized thiolated chitosan proniosomal gel containing duloxetine for intranasal delivery. AAPS Pharm Sci Tech. 2019; Oct; 20:1-2.

21. Eid HM et al. Novel chitosan-coated niosomal formulation for improved management of bacterial conjunctivitis: a highly permeable and efficient ocular nanocarrier for azithromycin. Journal of Pharmaceutical Sciences. 2021; Aug 1; 110(8): 3027-36.

22. Uthaiwat P et al. Characteristic evaluation of gel formulation containing niosomes of melatonin or its derivative and mucoadhesive properties using ATR-FTIR spectroscopy. Polymers. 2021; Apr 2; 13(7): 1142.

23. Khalifa AZ, Abdul Rasool BK. Optimized mucoadhesive coated niosomes as a sustained oral delivery system of famotidine. Aaps Pharmscitech. 2017; Nov; 18: 3064-75.

24. Abla KK et al. Application of Box-Behnken Design in the Preparation, Optimization, and In-Vivo Pharmacokinetic Evaluation of Oral Tadalafil-Loaded Niosomal Film. Pharmaceutics. 2023; 15(1): 173. https://doi.org/10.3390/pharmaceutics15010173

25. Chettupalli AK et al. Development and Optimization of Aripiprazole ODT by using box-Behnken Design. Research J. Pharm. and Tech. 2020; 13(12): 6195-6201.

26. Kulkarni P, Rawtani D. Application of box-behnken design in the preparation, optimization, and in vitro evaluation of self-assembly–based tamoxifen-and doxorubicin-loaded and dual drug–loaded niosomes for combinatorial breast cancer treatment. Journal of Pharmaceutical Sciences. 2019; Aug 1; 108(8):2643-53.

27. Seleci DA et al. Niosomes as Nanoparticular Drug Carriers: Fundamentals and Recent Applications. Journal of Nanomaterials. 2016; 1-13.

28. Mehanna MM et al. Tadalafil-Loaded Limonene-Based Orodispersible Tablets: Formulation, in Vitro Characterization and in Vivo Appraisal of Gastroprotective Activity. Int. J. Nanomed. 2020; 15, 10099–10112.

29. Ramadan WM et al. Preparation of Acyclovir Loaded Non ionic Surfactant Vesicles (Niosomes) Using Reverse Phase Evaporation Technique. Research J. Pharm. and Tech. 2009; 2(4): 793-795

30. Karthikeyan D et al. Preparation and In-Vitro Characterization of Diclofenac Sodium Niosomes for Ocular Use. Research J. Pharm. and Tech. 2009; 2(4): 710-713.

31. Rani D et al. Formulation Development and In-vitro Evaluation of Minoxidil Bearing Glycerosomes. American Journal of Biomedical Research, 2016; 4, 2: 27-37. doi: 10.12691/ajbr-4-2-1

32. Choudhary V et al. Development and evaluation of hydrocortisone-loaded niosomal gel. International Journal of Pharmaceutical Sciences and Research. 2023; 14(3): 1265-1272.

33. Rani D et al. Formulation, Design and Optimization of Glycerosomes for Topical Delivery of Minoxidil. Research Journal of Pharmacy and Technology. 2021; 14(5): 2367-4. Doi: 10.52711/0974-360X.2021.00418

34. Kumar YP et al. Preparation and Evaluation of Diclofenac Niosomes by Various Techniques. Research J. Pharm. and Tech. 2013: 6(10): 1097-1101

35. Chioma ED et al. Formulation and evaluation of etodolac niosomes by modified ether injection technique. Universal Journal of Pharmaceutical Research. 2016; 1(1): 1-6.

36. Abdelkader H et al. Recent advances in non-ionic surfactant vesicles (niosomes): self-assembly, fabrication, characterization, drug delivery applications and limitations. Drug Delivery. 2014 Mar 1; 21(2): 87-100.

37. Srikanth et al. Formulation and Evaluation of Maltodextrin Based Doxorubicin HCl Proniosomes. Research J. Pharm. and Tech. 2019; 12(6): 2776-2780.

38. Sabareesh M et al. Formulation Development, Ex vivo Evaluation and In vivo Antihypertensive study of Losartan Potassium Loaded Nanoproniosomal Gel: A Novel Vesicular Approach for Transdermal Delivery. Research J. Pharm. and Tech 2021; 14(3):1423-1430

39. Chen S et al. Recent advances in non-ionic surfactant vesicles (niosomes): Fabrication, characterization, pharmaceutical and cosmetic applications. European Journal of Pharmaceutics and Biopharmaceutics. 2019; Nov 1; 144: 18-39.

40. Bartelds R, et al. Niosomes, an alternative for liposomal delivery. Plos One. 2018; 13(4): 0194179.

41. Chen J et al. Formulation and evaluation of a topical liposomal gel containing a combination of zedoary turmeric oil and tretinoin for psoriasis activity. J Liposome Res. 2021; Jun; 31(2): 130-144. doi: 10.1080/08982104.2020.1748646.

42. Andleeb M et al. Development, characterization and stability evaluation of topical gel loaded with ethosomes containing Achillea millefolium L. extract. Frontiers in Pharmacology. 2021; Apr 12; 12: 603227.

43. Rençber S et al. Mucoadhesive in situ gel formulation for vaginal delivery of clotrimazole: formulation, preparation, and in vitro/in vivo evaluation. Pharmaceutical Development and Technology. 2017; May 19; 22(4): 551-61.

44. Giri P, Singh I. Development and Evaluation of Mucoadhesive Tablets of Cinnarizine Using Carboxymethylated Guar Gum by Compression Coating Technique. Biointerface Research in Applied Chemistry. 2020; 10(5): 6365 – 6376.

45. Okore VC et al. Formulation and Evaluation of Niosomes. Indian J. Pharm. Sci. 2011; 73(3): 323–328.

46. Mitkari BV et al. Formulation and evaluation of topical liposomal gel for fluconazole. Indian J. Pharm. Educ Res. 2010; Oct 1; 44(4): 324-33.

Received on 24.07.2023 Modified on 13.10.2023

Research J. Pharm. and Tech 2024; 17(6):2549-2555.

DOI: 10.52711/0974-360X.2024.00398

Response	Type of Model	P value	R²	Adjusted R²	Predicted R²	Adequate Precision	Standard deviation	C.V. %
Y₁	Quadratic	0.0001	0.9720	0.9360	0.7520	20.2091	2.56	1.81
Y₂	Quadratic	0.0049	0.9120	0.8990	0.7234	19.5018	2.71	2.03
Y₃	Quadratic	0.0015	0.9415	0.8662	0.695	20.6471	2.83	1.88

Voriconazole and Beauvercin Loaded Niosomal Mucoadhesive Nanogel as an effective Treatment for the Management of Oral Candidiasis

Deepika Rani*, Vinit Kumar Sharma, Ranjit Singh