Fast-Dissolving Protein Nanofibrous Membrane for Dual Drug Oral Delivery
Round 1
Reviewer 1 Report
1. The manuscript presents a novel drug delivery platform that could be used for elder patients and kids. This drug delivery platform shows novelty and have the potential of a rapid release of either hydrophobic or hydrophilic drugs. It could even be used to the release of both types of drugs. This work is well designed and shows novelty and the scientific soundness; therefore, I consider this manuscript it is suitable for publication in this journal.
We thank reviewer for the effort reviewing our paper and the positive feedback.
2. The authors just need to correct some typos and certain mistakes such as changes in the letter size that are all around the manuscript.
We gratefully thank for the comment. According to the comment, we have carefully checked and revised the mistakes in the revised manuscript.
The word “vitamin B12” has been revised as “vitamin B12”. (on page 2, line 57)
The word “(Figure 2d, e)” has been revised as “(Figure 2d, 2e)”. (on page 5, line 172)
The phrase “may be” has been revised as “might be”. (on page 5, line 179)
The word “(Figure 3d, e)” has been revised as “(Figure 3d, 3e)”. (on page 5, line 199)
Reviewer 2 Report
1. The authors prepared the gelatin membrane using trifluoroethanol (TFEA). TFEA is highly toxic and has a boiling point of 78°C, higher than the drying temperature at 37°C. How did the authors confirm that the resulting gelatin membrane contained no TFEA?
We gratefully thank for reviewer’s comment. We quite agree with the comment that TFEA has a relatively higher boiling point of 78°C than the treated temperature 37°C. Actually, in this study, two strategies are implemented to ensure the complete remove of TFEA. 1) As it is well investigated, during the electrospinning process, nanofiber formation is accompanied by solvent evaporation. When the nanofiber is ejected from the spinning nozzle and further flied to the collector at a very fast rate, and fast movement of nanofiber favors for quick evaporation of TFEA. In addition, during the electrospinning process, the nanofiber becomes thinner and thinner due to the electrical repulsion. More solvent molecules can be diffused from the interior of the nanofiber to the surface of the nanofiber, which also benefits for the solvent evaporation. 2) In order to ensure the complete remove of the solvent TFEA, we put the nanofiber membrane into the 37°C oven for a relatively long time 24 hours. As the nanofiber membrane is highly porous and the nanofiber possesses large specific surface area, the residual solvent in the nanofiber can be easily and completely removed.
In Results and discussion, the sentences “In this study, two strategies were implemented to ensure the complete remove of TFEA. 1) As it was well investigated, during the electrospinning process, nanofiber formation was accompanied by solvent evaporation. When the nanofiber was ejected from the spinning nozzle and further flied to the collector at a very fast rate, and fast movement of nanofiber favored for quick evaporation of TFEA. In addition, during the electrospinning process, the nanofiber became thinner and thinner due to the electrical repulsion. More solvent molecules could be diffused from the interior of the nanofiber to the surface of the nanofiber, which also benefited for the solvent evaporation. 2) In order to ensure the complete remove of the solvent TFEA, we put the nanofiber membrane into the 37°C oven for a relatively long time 24 hours. As the nanofiber membrane was highly porous and the nanofiber possessed large specific surface area, the residual solvent in the nanofiber could be easily and completely removed.” have been added in the revised manuscript. (on page 4, line 150-161)
2. The dissolving speed should depend on the membrane thickness. Did the authors determine the membrane thickness?
We quite agree with the comment. According to the comment, we have carefully measured the thickness of the pristine Gel membrane, Gel/Rhb membrane, Gel/Flu membrane, and bilayered membrane, and they were 46, 44, 45, and 88 μm, respectively.
In Materials and methods, the sentence “Thickness of the membrane was measured by a thickness gauge.” has been added. (on page 3, line 131; on page 4, line 166; on page 5, line 195-196; on page 7, line 230)
3. The dissolution processes are qualitative in all Figures for the dissociation process. The authors need to quantitatively and kinetically analyze the release of drugs from the gelatin membrane, for instance, by determining a release speed using time-concourse in a UV spectrum.
According to the comment, we have quantitatively investigated the cumulative release profile of drugs from the gelatin membrane by using a UV-Vis spectrophotometer. The new results coincide with the qualitative results. The new results have been added in the revised manuscript (Figure S1 and S2). (on page 5, line 182-184; on page 6, line 209-210)
4. What advantage does the dual drugs membrane have in oral administration over the simultaneous gelatin membranes loading the hydrophilic and hydrophobic drugs? Concurrent using the membranes and separately loading them may be helpful because the proportion of hydrophilic and hydrophobic drugs depends on the patients.
We quite agree with the comment. As a new formation, dual-loaded nanofibrous membrane is suitable for children and elderly patients with dysphagia. Such membrane has lots of unique advantages. Drug combination therapy by the dual-drug loaded oral instant membrane can enhance efficacy compared to the mono-therapy approach. In addition, the two drugs loading into different layers can avoid interference during the preparation process, which can maximize the retention of bioactivity of the two drugs. Dose of each drug can be easily tuned according to the needs of patients. Besides, not only the combination of hydrophilic and hydrophobic drugs, two hydrophilic drugs or two hydrophobic drugs can be also encapsulated into the membrane depending on the patients. Thus, this study provides a paradigm for the fabrication of dual-drug loaded oral instant membrane, which shows great potential for patients with dysphagia.
In Results and discussion, the sentences “As a new formation, dual-loaded nanofibrous membrane was suitable for children and elderly patients with dysphagia. Such membrane had lots of unique advantages. Drug combination therapy by the dual-drug loaded oral instant membrane could enhance efficacy compared to the mono-therapy approach. In addition, the two drugs loading into different layers could avoid interference during the preparation process, which could maximize the retention of bioactivity of the two drugs. Dose of each drug could be easily tuned according to the needs of patients. Besides, not only the combination of hydrophilic and hydrophobic drugs, two hydrophilic drugs or two hydrophobic drugs could be also encapsulated into the membrane depending on the patients. Thus, this study provided a paradigm for the fabrication of dual-drug loaded oral instant membrane, which showed great potential for patients with dysphagia.” have been added in the revised manuscript. (on page 8, line 270-280)
Reviewer 3 Report
1. The manuscript presents interesting and good results regarding the production of dual drug-loaded with bilayered gelatin oral instant membranes through sequential electrospinning method. The manuscript is succint and well structured.
We thank reviewer for the effort reviewing our paper and the positive feedback.
2. In the introduction section the authors should describe better the used gelatin.
We gratefully thank for reviewer’s comment. The sentences “Gelatin is rich in beneficial amino acids, especially glycine and proline, which can promote wound healing and prevent wrinkles. Besides, previous studies also show that gelatin can accelerate the elasticity of connective tissue and increase cartilage density. It can also help to repair intestinal wall damage and rebuild the protective mucosa of the intestine. The glycine in gelatin has unique anti-inflammatory effect.” have been added in the revised manuscript. (on page 2, line 70-75), Besides, the new reference has also been added. (on page 10, line 386-392)
- Al-Nimry S, Dayah AA, Hasan I, Daghmash R. Cosmetic, Biomedical and Pharmaceutical Applications of Fish Gelatin/Hydrolysates. Mar Drugs 2021,19(3):145.
- Shiao WC, Wu TC, Kuo CH, Tsai YH, Tsai ML, Hong YH, Huang CY. Physicochemical and Antioxidant Properties of Gelatin and Gelatin Hydrolysates Obtained from Extrusion-Pretreated Fish (Oreochromis sp.) Scales. Mar Drugs 2021,19(5):275.
- Ayaz F, Demir D, Bölgen N. Differential anti-inflammatory properties of chitosan-based cryogel scaffolds depending on chitosan/gelatin ratio. Artif Cells Nanomed Biotechnol 2021,49(1):682-690.
3. There are some studies that report similar results. Please revise and compare the results.
E. Guo J, Wang T, Yan Z, Ji D, Li J, Pan H. Preparation and evaluation of dual drug-loaded nanofiber membranes based on coaxial electrostatic spinning technology. Int J Pharm. 2022 Nov 16:122410. doi: 10.1016/j.ijpharm.2022.122410. Epub ahead of print. PMID: 36402289.
According to the comment, we compare the reported work with our study. Although the two works both use the electrospinning method and both load two drugs, the application scenarios and the research contents are different. The reported work focuses on fabrication of nanofibrous membrane loading with two bioactive drugs for promoting skin wound healing; while our study aims to investigate the dual-drug loaded oral instant mucosal administration.
In Results and discussion, the sentence “Previous study had shown that combination therapy by using dual-drug loaded nanofibrous membrane could enhance therapeutic efficiency.” has been added in the revised manuscript. (on page 7, line 251-253) Besides, the new reference has also been added. (on page 10, line 407-408)
- Guo J, Wang T, Yan Z, Ji D, Li J, Pan H. Preparation and evaluation of dual drug-loaded nanofiber membranes based on coaxial electrostatic spinning technology. Int J Pharm 2022, (629)122410.
4. Section 2.3 should be more briefly described and based on similar methods.
Have corrected.
5. Were there any statistical analyses performed? Were the experiments effectuated in duplicate or triplicates?
Yes, experiments are run in triplicate per sample.
Round 2
Reviewer 3 Report
The authors improved the manuscript according to the requirements.
