Formulation and Development of Bioadhesive Oral Films Containing Usnea barbata (L.) F.H.Wigg Dry Ethanol Extract (F-UBE-HPC) with Antimicrobial and Anticancer Properties for Potential Use in Oral Cancer Complementary Therapy
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. The Development of UBE-Loaded Bioadhesive Oral Films
2.3. Physico-Chemical Characterization of the UBE-Loaded Bioadhesive Oral Films
2.3.1. SEM Analysis
2.3.2. Atomic Force Microscopy (AFM)
2.3.3. Fourier Transform Infrared Spectroscopy (FTIR)
2.3.4. Powder X-ray Diffractometry
2.3.5. Thermogravimetric Analysis
2.4. Pharmacotechnical Properties of the UBE-Loaded Bioadhesive Oral Films
2.4.1. Weight Uniformity
2.4.2. Thickness
2.4.3. Folding Endurance
2.4.4. Tensile Strength and Elongation Ability
2.4.5. Moisture Content
2.4.6. Surface pH
2.4.7. In Vitro Disintegration Time
2.4.8. Swelling Ratio
2.4.9. Ex Vivo Bioadhesion Time
2.5. Antimicrobial Activity
2.5.1. Inoculum Preparation
2.5.2. Samples and Standards
2.5.3. Microdilution Method
2.5.4. Reading and Interpreting
2.6. Cytotoxic Activity of UBE-Loaded Bioadhesive Oral Films on A. salina Larvae
2.6.1. Fluorescent Microscopy
2.6.2. Data Processing
2.7. In Vitro Analysis of the Biological Effects of UBE-Loaded Bioadhesive Oral Films on Human Blood Cell Cultures and CLS-354 Tumor Cell Line
2.7.1. Equipment
2.7.2. Data Processing
2.7.3. Human Blood Cells Cultures
2.7.4. CLS-354 Cell Line, Cell Culture
2.7.5. Samples and Control Solutions
2.7.6. Annexin V-FITC Apoptosis Assay
2.7.7. Evaluation of Caspase 3/7 Activity
2.7.8. Evaluation of Nuclear Condensation and Lysosomal Activity
2.7.9. Evaluation of Total ROS Activity
2.7.10. Cell Cycle Analysis
2.7.11. Evaluation of Cell Proliferation
2.8. Data Analysis
3. Results
3.1. Development of the Bioadhesive Oral Films
3.2. Physico-Chemical Characterization of the Bioadhesive Films
3.2.1. SEM Analysis
3.2.2. AFM Analysis
3.2.3. FTIR Analysis
3.2.4. XRD Analysis
3.2.5. TG Analysis
3.3. Pharmacotechnical Properties of the Bioadhesive Oral Films
3.4. Antimicrobial Activity
3.5. Cytotoxic Activity on A. salina Larvae
3.6. In Vitro Analysis of the Biological Effects of UBE-Loaded Mucoadhesive Oral Films on Human Blood Cell Cultures and CLS-354 Cancer Cell Line
3.6.1. Caspase 3/7 Activity
3.6.2. Nuclear Condensation and Lysosomal Activity
3.6.3. ROS Levels
3.6.4. Cell Cycle Analysis
3.6.5. Apoptosis
3.6.6. Cell Proliferation
3.6.7. Principal Component Analysis
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Ingredients | Quantity (g) | |
---|---|---|
F-UBE-HPC | R | |
UBE | 0.30 | - |
Ethyl alcohol 96% (v/v) | 10.00 | 10.00 |
PEG 400 | 5.00 | 5.00 |
HPC 20% water dispersion (w/w) | 84.70 | 85.00 |
Film | 1st Step (%) | 2nd Step (%) | 3rd Step (%) |
---|---|---|---|
TG (%) | TG(%)/Tmax (°C) | TG(%)/Tmax (°C) | |
R | 2.4% | 88.4%/357.6 °C | 9.2%/468.3 °C |
F-UBE-HPC | 2.3% | 85.6%/352.3 °C | 12.1%/471.8 °C |
Parameter * | Formulation Code | |
---|---|---|
F-UBE-HPC | R | |
Weight uniformity (mg) | 110 ± 4.77 | 107 ± 5.25 |
Thickness (mm) | 0.093 ± 0.002 | 0.093 ± 0.003 |
Folding endurance value | >300 | >300 |
Tensile strength (kg/mm2) | 2.48 ± 1.58 | 2.57 ± 1.71 |
Elongation % | 63.14 ± 1.94 | 62.75 ± 1.52 |
Moisture content % (w/w) | 6.58 ± 0.44 | 6.23 ± 0.56 |
pH | 7.10 ± 0.02 | 7.07 ± 0.01 |
Disintegration time (seconds) | 118 ± 3.16 | 115 ± 4.19 |
Swelling ratio (% after 6 h) | 289 ± 5.82 | 288 ± 6.13 |
Ex vivo biooadhesion time (minutes) | 98 ± 3.58 | 98 ± 4.17 |
Microdilution | CTR | TRF 10.1 mg/mL | F-UBE-HPC 110 mg/mL | |
---|---|---|---|---|
30 mg/mL | 122 mg/mL | |||
1 | 1.5 | 6.10 | 0.5 | 5.5 |
2 | 0.75 | 4.88 | 0.25 | 2.75 |
3 | 0.375 | 3.904 | 0.125 | 1.375 |
4 | 0.187 | 3.123 | 0.062 | 0.685 |
5 | 0.093 | 2.498 | 0.031 | 0.342 |
6 | 0.046 | 1.998 | 0.015 | 0.171 |
7 | 0.024 | 1599 | 0.007 | 0.085 |
Dil. | S. aureus | P. aeruginosa | ||||||
---|---|---|---|---|---|---|---|---|
CTR F-UBE | F-UBE-HPC | CTR | F-UBE-HPC | |||||
A | B | C | D | E | F | G | H | |
1 | ||||||||
2 | ||||||||
3 | ||||||||
4 | ||||||||
5 | ||||||||
6 | ||||||||
7 | ||||||||
* |
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Popovici, V.; Matei, E.; Cozaru, G.-C.; Bucur, L.; Gîrd, C.E.; Schröder, V.; Ozon, E.A.; Sarbu, I.; Musuc, A.M.; Atkinson, I.; et al. Formulation and Development of Bioadhesive Oral Films Containing Usnea barbata (L.) F.H.Wigg Dry Ethanol Extract (F-UBE-HPC) with Antimicrobial and Anticancer Properties for Potential Use in Oral Cancer Complementary Therapy. Pharmaceutics 2022, 14, 1808. https://doi.org/10.3390/pharmaceutics14091808
Popovici V, Matei E, Cozaru G-C, Bucur L, Gîrd CE, Schröder V, Ozon EA, Sarbu I, Musuc AM, Atkinson I, et al. Formulation and Development of Bioadhesive Oral Films Containing Usnea barbata (L.) F.H.Wigg Dry Ethanol Extract (F-UBE-HPC) with Antimicrobial and Anticancer Properties for Potential Use in Oral Cancer Complementary Therapy. Pharmaceutics. 2022; 14(9):1808. https://doi.org/10.3390/pharmaceutics14091808
Chicago/Turabian StylePopovici, Violeta, Elena Matei, Georgeta-Camelia Cozaru, Laura Bucur, Cerasela Elena Gîrd, Verginica Schröder, Emma Adriana Ozon, Iulian Sarbu, Adina Magdalena Musuc, Irina Atkinson, and et al. 2022. "Formulation and Development of Bioadhesive Oral Films Containing Usnea barbata (L.) F.H.Wigg Dry Ethanol Extract (F-UBE-HPC) with Antimicrobial and Anticancer Properties for Potential Use in Oral Cancer Complementary Therapy" Pharmaceutics 14, no. 9: 1808. https://doi.org/10.3390/pharmaceutics14091808