Formulation and In Vitro Assessment of Tragacanth Gum-Based Hydrogel Loaded with Artemisia vestita Leaf Extract for Wound Healing
Abstract
:1. Introduction
2. Materials and Methods
- Collection of Plant and Extract Preparation
- Gel Preparation
- Drug–Excipient Compatibility Study
- Preformulation Studies
Physiochemical Characterization
- Physical appearance: The physical appearance of a formulated gel is an essential aspect with regard to its overall quality and user acceptance. Evaluation of color, appearance, and clarity helps detect any abnormalities or changes in the formulation that may affect its effectiveness or esthetics. To evaluate the color, appearance, and clarity of the hydrogel formed by incorporating A. vestita leaf extract (ALE) into tragacanth gum, the following procedures were systematically and reliably performed. Samples were prepared in identical containers to ensure uniformity. Color assessment was performed by trained observers in triplicate. Standardized color charts were used for precise measurement. Assessments were performed under standardized lighting conditions. The same trained observers evaluated the hydrogel thrice to observe visible particulates, uniformity, and texture. Clarity was measured using a transparent grid behind the gel, and a numerical scale (ranging from 1 to 5) with detailed criteria was used for scoring. Turbidity or transmittance was spectrophotometrically measured at 600 nm, with lower turbidity or higher transmittance indicating higher clarity.
- pH: The pH of the A. vestita leaf extract hydrogel (ALEH) was measured thrice using a calibrated Seven Excellence S400 pH meter (Mettler Toledo, India) to ensure accuracy. To determine the pH of the formulations using the calibrated pH meter, an optimal pH range was required (e.g., for skin compatibility). Hence, 1 g of the gel formulation was dispersed in 100 mL of distilled water and allowed to stand for 2 h. The pH of the resultant solution was then measured. Data were presented as the mean ± SEM.
- Spreadability: In total, 1 mL of the prepared ALEH was transferred onto a glass plate using a sterile syringe. A calibrated plate was placed on top of the hydrogel. Weights of increasing mass (25, 50, 100, 200, 300, 400, and 500 g) were then sequentially placed on the plate at 20 s time intervals to enable ALEH to stabilize and spread under the applied weight. After placing each weight, the radii of the ALEH formulation were measured. To ensure the reliability and variability of the results, spreadability was assessed in triplicate at ambient temperature. The area covered by the prepared ALEH was calculated using the following formula:P = πr2,
- Rheological properties: The viscosity of the formulation was assessed using a Brookfield viscometer. Viscosity affects spreadability, product handling, and release kinetics. The digital viscometer was loaded with the hydrogel and placed into the flow jacket of the viscometer. The flow jacket is a component of the apparatus used to regulate temperature or create specific conditions during the measurement process. An adapter was used to measure the viscosity of the formulated gel at a rotation speed of 20 rotations per minute (rpm). The temperature was kept constant at 24.8 °C by circulating water through a thermostated water jacket. Before taking readings, the sample was allowed to settle for 5–6 min. The viscosity of the hydrogels was determined by increasing the shear rate.
- Extrudability: The formulated gel (10 g) was filled into standard caps containing eight collapsible aluminum tubes. They were sealed at the end by crimping. To assess extrudability, the weights of the tubes were recorded. Subsequently, the tubes were securely positioned between two glass slides and firmly clamped in place. A 500 g weight was placed on top of the slides, following which the cap was removed to allow the extrusion of the gel. The gel extruded from the tubes was gathered and weighed. Based on the amount of gel extruded, the percentage of extruded gel was calculated.
- Excellent: more than 90% extrudability;
- Good: more than 80% extrudability;
- Fair: more than 70% extrudability.
- Drug Release Study
- Fickian diffusion or case I transport: n = 0.45;
- Non-Fickian or anomalous transport: 0.45 < n < 0.89;
- Case II transport or relaxation-controlled release: n = 0.89;
- Super case II transport: n > 0.89.
- Antimicrobial Activity Assay
- Agar Well Diffusion Technique
- Minimum Bactericidal Concentration (MBC) and Minimum Inhibitory Concentration (MIC) Assays
- Cell Culture
- Wound Healing Assay
- Statistical Analysis
3. Results and Discussion
3.1. ALEH Preparation
Physical Assessment of Compatibility
3.2. Physiochemical Characterization
3.2.1. Physical Appearance
3.2.2. pH
3.2.3. Spreadability
3.2.4. Rheological Properties
3.2.5. Extrudability
3.2.6. Optimization
3.3. Drug Release Study
- Antimicrobial Activity
3.4. Wound Healing Activity
4. Conclusions
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Composition | F1 | F2 | F3 | F4 | F5 | F6 | F7 | F8 |
---|---|---|---|---|---|---|---|---|
Aqueous extract (g) | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
Tragacanth gum (g) | 0.25 | 0.5 | 0.75 | 1.0 | 1.25 | 1.5 | 1.75 | 2.0 |
Milli-Q water (mL) | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 |
Methylparaben (mL) | 0.2 | 0.2 | 0.2 | 0.2 | 0.2 | 0.2 | 0.2 | 0.2 |
Propylparaben (mL) | 0.5 | 0.5 | 0.5 | 0.5 | 0.5 | 0.5 | 0.5 | 0.5 |
Propylene glycol (mL) | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
Triethanolamine (mL) | Adjust pH to 6–6.5 |
Batch | Discoloration | Caking | Liquefaction | |||
---|---|---|---|---|---|---|
Initial | After 30 Days | Initial | After 30 Days | Initial | After 30 Days | |
Aqueous extract | No | Consistent | No | Consistent | No | Consistent |
Tragacanth gum | No | Consistent | No | Consistent | No | Consistent |
Aqueous extract + Tragacanth gum | No | Consistent | No | Consistent | No | Consistent |
Formulation Code | pH | Spreadability (g/cm2) | Viscosity (cP) |
---|---|---|---|
F1 | 6.86 ± 0.1 | 15.08 ± 0.085 | 2318 ± 3.78 |
F2 | 6.93 ± 0.15 | 14.75 ± 0.16 | 2540 ± 2.51 |
F3 | 6.83 ± 0.05 | 13.03 ± 0.09 | 2463 ± 5.13 |
F4 | 7.03 ± 0.11 | 15.75 ± 0.19 | 2450 ± 2.08 |
F5 | 7.16 ± 0.05 | 12.84 ± 0.08 | 2637 ± 2.08 |
F6 | 6.90 ± 1.08 | 11.63 ± 0.07 | 2429 ± 3.21 |
F7 | 6.83 ± 0.05 | 12.83 ± 0.11 | 2318 ± 2.08 |
F8 | 6.9 ± 0.10 | 14.31 ± 0.11 | 2411 ± 1.15 |
Time (h) | Cumulative Percentage of Drug Release (25 °C) | |||||||
---|---|---|---|---|---|---|---|---|
F1 | F2 | F3 | F4 | F5 | F6 | F7 | F8 | |
0 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
0.25 | 7.982 ± 0 | 6.390 ± 0 | 12.58 ± 0 | 3.878 ± 0 | 3.291 ± 0 | 13.593 ± 0 | 5.553 ± 0 | 10.578 ± 0 |
0.5 | 12.664 ± 0 | 9.282 ± 0 | 17.362 ± 0 | 11.995 ± 0 | 8.969 ± 0 | 23.831 ± 0 | 10.940 ± 0 | 22.096 ± 0 |
1 | 29.078 ± 0 | 24.540 ± 0 | 25.912 ± 0 | 17.259 ± 0 | 16.349 ± 0 | 31.840 ± 0 | 25.395 ± 0 | 32.333 ± 0 |
2 | 46.067 ± 0 | 32.814 ± 0 | 38.144 ± 0 | 24.633 ± 0 | 21.613 ± 0 | 42.010 ± 0 | 39.883 ± 0 | 40.337 ± 0 |
4 | 60.369 ± 0 | 50.540 ± 0 | 52.285 ± 0 | 33.069 ± 0 | 42.721 ± 0 | 57.061 ± 0 | 53.896 ± 0 | 56.109 ± 0 |
6 | 68.155 ± 0 | 61.488 ± 0 | 63.093 ± 0 | 47.778 ± 0 | 52.014 ± 0 | 72.312 ± 0 | 69.515 ± 0 | 75.699 ± 0 |
8 | 80.596 ± 0 | 75.400 ± 0 | 78.201 ± 0 | 61.512 ± 0 | 70.020 ± 0 | 84.246 ± 0 | 77.050 ± 0 | 82.176 ± 0 |
10 | 88.988 ± 0 | 84.709 ± 0 | 86.468 ± 0 | 76.508 ± 0 | 75.468 ± 0 | 87.848 ± 0 | 83.787 ± 0 | 88.752 ± 0 |
12 | 98.440 ± 0 | 93.254 ± 0 | 94.200 ± 0 | 86.587 ± 0 | 84.431 ± 0 | 92.244 ± 0 | 91.636 ± 0 | 96.267 ± 0 |
Time (h) | Cumulative Percentage of Drug Release (F4) | |||
---|---|---|---|---|
pH 4.5 Temperature 37 °C | pH 4.5 Temperature 40 °C | pH 6.8 Temperature 37 °C | pH 6.8 Temperature 40 °C | |
0 | 0.000 | 0.000 | 0.000 | 0.000 |
0.25 | 4.811 | 1.604 | 1.815 | 11.028 |
0.5 | 11.248 | 9.510 | 5.961 | 15.542 |
1 | 18.702 | 14.781 | 15.818 | 22.267 |
2 | 23.784 | 23.031 | 22.083 | 33.138 |
4 | 32.819 | 29.448 | 29.361 | 40.785 |
6 | 42.532 | 34.260 | 43.272 | 50.274 |
8 | 49.760 | 40.792 | 47.879 | 65.198 |
10 | 64.329 | 51.677 | 68.147 | 74.411 |
12 | 74.380 | 54.656 | 81.044 | 79.110 |
Microbes | Zone of Inhibition (mm) | MIC and MBC/MFC (µg/mL) | ||||
---|---|---|---|---|---|---|
25 µg/mL | 50 µg/mL | 100 µg/mL | Standard Drug (Azithromycin/Fluconazole) | MIC | MBC/MFC | |
S. aureus | - | 12.14 ±0.52 | 15.0 ± 0.26 | 20.4 ± 0.22 | 110 | 220 |
E. coli | 6.4 ± 0.84 | 14.6 ± 0.28 | 19.6 ± 0.51 | 22.2 ± 0.31 | 200 | 240 |
B. subtilis | - | 13.46 ± 0.40 | 15.1 ± 0.17 | 17.4 ± 0.23 | 160 | 200 |
S. pyogenes | 12.12 ± 0.9 | 18.1 ±0.70 | 20.3 ± 0.60 | 23.1± 0.20 | 200 | 250 |
A. flavus | 12.3 ± 0.22 | 15.2 ± 0.31 | 17.5 ± 0.21 | 20.3 ± 0.23 | 100 | 260 |
A. niger | - | 10.7 ± 0.42 | 14.8 ± 0.50 | 16.8 ± 0.14 | 100 | 220 |
C. albicans | 10.2 ± 0.36 | 12.1 ± 0.32 | 19.4 ± 0.01 | 22.1 ± 0.24 | 200 | 250 |
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Dogra, S.; Koul, B.; Singh, J.; Mishra, M.; Rabbee, M.F. Formulation and In Vitro Assessment of Tragacanth Gum-Based Hydrogel Loaded with Artemisia vestita Leaf Extract for Wound Healing. Processes 2024, 12, 2750. https://doi.org/10.3390/pr12122750
Dogra S, Koul B, Singh J, Mishra M, Rabbee MF. Formulation and In Vitro Assessment of Tragacanth Gum-Based Hydrogel Loaded with Artemisia vestita Leaf Extract for Wound Healing. Processes. 2024; 12(12):2750. https://doi.org/10.3390/pr12122750
Chicago/Turabian StyleDogra, Shivani, Bhupendra Koul, Joginder Singh, Meerambika Mishra, and Muhammad Fazle Rabbee. 2024. "Formulation and In Vitro Assessment of Tragacanth Gum-Based Hydrogel Loaded with Artemisia vestita Leaf Extract for Wound Healing" Processes 12, no. 12: 2750. https://doi.org/10.3390/pr12122750
APA StyleDogra, S., Koul, B., Singh, J., Mishra, M., & Rabbee, M. F. (2024). Formulation and In Vitro Assessment of Tragacanth Gum-Based Hydrogel Loaded with Artemisia vestita Leaf Extract for Wound Healing. Processes, 12(12), 2750. https://doi.org/10.3390/pr12122750