Transdermal enhancement regulation with different electroporation parameters and different drug concentrations. The relationship between the different frequency, waveform, and intensity of electroporation and the amount of SH that permeated in the in vitro transdermal experiments mice and miniature pig skins were studied under the fixed drug concentration. To study the amount of SH that permeated at different drug concentrations, in vitro transdermal experiments mice and miniature pig skins were performed using fixed parameters of electroporation. Transdermal permeation experiments were done in vitro.
The skin was then dehydrated using gradient alcohol and xylene, and then embedded in paraffin according to standard processes. The histological properties of the skin were analyzed using hematoxylin and eosin HE staining and were observed with a Leica DM Microscopy. The conditions were as follows: nm excitation wavelength, nm emission wavelength, 0 to fluorescence intensity. These tests did not change the original treatment plan of patients. All patients in the tests were diagnosed with arthritic diseases RA or OA according to a detailed medical history for knee pain, a comprehensive physical exam, and imaging studies.
In patients with bilateral arthritis, only the knee that showed more advanced disease was considered. The protocol was approved by Guangzhou Hospital of Integrated Traditional Chinese and Western Medicine Ethical Committee, and informed consent was obtained from all patients. All programs were performed in conformity to the ethical standards of the institutional research committee and in accordance with the Helsinki Declaration and its latter amendments.
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Evaluating the therapeutic efficacy and detecting the concentration of SH in the patient SF in clinical tests. For evaluating clinical efficacy, the test group had nine patients, and the positive control group had six patients. The therapeutic program in the test group was transdermal treatment of SH solution with optimized electroporation combined with standard therapy eg, celecoxib.
Meanwhile, the therapeutic program in the positive control group was only the standard therapy eg, celecoxib. Clinical efficacy parameters were assessed after 8 days. The clinical improvement rate was calculated according to Eq 3. After vortexing to allow homogenization, the mixture was absorbed by two pieces of cotton. The two pieces of cotton containing the liquid mixture were placed in two electrodes, and the two electrodes were subsequently placed on one knee of the patients.
Clinical transdermal tests were done using the liquid mixture with electroporation for 30 minutes.
The electroporation conditions were as follows: 3 KHz, exponential waveform, and intensity For detecting the concentration of SH in the patient SF, the test group had nine patients and the control group had six patients. The clinical transdermal drug delivery in the test group was done using the liquid mixture with electroporation for 30 minutes. The clinical transdermal drug delivery in the control group was done using the liquid mixture without electroporation for 30 minutes.
After transdermal administration, the patient SF samples were collected by the hospital staff using the SF collection equipment. The transdermal treatment method using SH solution with electroporation was done as described earlier. After vortexing to allow homogenization, the mixture was centrifuged at 19, g for 30 min. After vortexing to homogenize, the mixture was centrifuged at 19, g for 30 min. The data were recorded, and the system was controlled using the MassHunter software version B. The conditions were as follows: column, Zorbax SB-C18, 3.
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Collision energy was set at 26 for SH and 18 for IS. The low limit of quantitative analysis was 0. The flux enhancement ratio ER induced by electroporation was computed according to Eq 2. One-way analysis of variance was used to evaluate statistical differences. Taken together, the data obtained for the SH in samples was in accordance with the standard substance. Effect of electroporation parameters and drug concentration on SH penetration in mice and miniature pig skins.
The results demonstrated that the total Q n of 3 KHz was 6. The ER of 3 KHz was 3. Taken together, 3 KHz was found to be the optimal pulse frequency that had the most significant enhancement-promoting effect on SH penetration. The situation above was exactly opposite to our primary hypothesis that 6 KHz might induce the strongest transdermal enhancement. The reason for this phenomenon might be that 6 KHz produced too many pulses, and 1. Thereby, Q n induced by 6 KHz and 1. To summarize, electroporation did enhance the SH penetration in mice skins, and 3 KHz was the optimal pulse frequency.
Figure 3 Transdermal enhancement of pulse frequencies, waveforms, and intensity on SH in in vitro mice skins.
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The other fixed parameters of electroporation: sine wave and intensity 4. The other fixed parameters of electroporation: 1. The other fixed parameters of electroporation: sine wave and 1. Different pulse waveforms produced different Q n in mice skins.oxurumigiced.tk
Electrically-assisted transdermal drug delivery.
The total Q n of exponential waveform was The ER of the exponential waveform was 6. That is, exponential pulse produced more intense SH permeation. A previous study showed that exponential waveform was one of the most commonly used pulse waveforms in electroporation. An increase in the electronic pulse duration leads to a decrease in the critical threshold, and so pulse duration of exponential pulse was long. The total Q n and ER at intensity 10 were Therefore, enhanced pulse intensity could enhance skin permeability. For our electroporation equipment, intensity 10 was the maximum intensity that could be given to humans without harm, and for animals it was 4.
Using higher intensity might result in excessive stimulation of the animal and make them feel uncomfortable. The total Q n and ER of intensity 10 was more than those seen at other pulse intensities. As expected, intensity 10 was found to be the optimal pulse intensity for humans. Above all, pulse frequency, waveforms, and intensity could affect the SH penetration in mice skins, and the optimal parameters were 3 KHz, exponential waveform, and intensity 10 for humans.
Transdermal enhancement of electroporation in miniature pig skins by varying the parameters. The total Q n of 3 KHz, exponential waveform, and intensity 10 were 8. Meanwhile, ER for 3 KHz, exponential waveform, and intensity 10 were 8. These results showed that Q n and ER induced by electroporation parameters 3 KHz, exponential waveform, and intensity 10 were higher compared with that induced by other electroporation parameters.
On analysing the overall situation, the results in miniature pig skins were in accordance with that in mice skins, and the optimal parameters were also same in both mice and miniature pig skins, and the optimal electroporation parameters were 3 KHz, exponential waveform, and intensity 10 for humans.
In practical clinical application, we could choose different parameters of electroporation to achieve different kinds of enhancement in transdermal efficiency so as to achieve the purpose of controlled release drugs. Figure 4 Transdermal enhancement of pulse frequencies, waveforms, and intensity on SH in in vitro miniature pig skins. The other fixed parameters of electroporation are as follows: exponential wave and intensity 4. The other fixed parameters of electroporation: 3 KHz and intensity 4.
The other fixed parameters of electroporation: exponential wave and 3 KHz. Effect of different drug concentration of SH on cumulative drug permeation amount in mice and miniature pig skins.
The total Q n of different groups were 0. We also found that Q n increased gradually with the increase of the drug concentration in vitro in mice and miniature pig skin permeation experiments with electroporation Figure 5A1 and 5B1. According to these results, the total Q n of 3. In brief, drug concentration was one of the key factors for electroporation to promote transdermal penetration of SH, and Q n values increased in a drug-dependent manner.
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Figure 5 Relationship between drug concentration and transdermal amount of SH. Notes: The Q n increased as drug concentration rose in the skin taken from the back of mice A1 and in miniature pig abdominal skins B1. Total Q n for different drug concentrations in mice skins A2 and in miniature pig abdominal skins B2.
Transdermal Delivery by Iontophoresis
Electroporation condition: exponential wave, 3 KHz, and intensity 4. Drug penetration routes include intercellular route, intracellular route, and eccrine route eg, sweat glands. As shown above, the Q n of the passive diffusion transdermal experiments without electroporation were all so low that concentration of SH might not reach the targeted therapeutic concentration. Through inducing the change of skin structure, electroporation could enable SH to overcome transdermal defects caused by its hydrophilic property.
All Q n values in the four drug concentrations with electroporation were more than that in control without electroporation, and this was a strong proof that electroporation significantly enhanced the transdermal permeation of SH. The epidermis is the outermost skin layer that consists of that hypodermis, hydrophilic dermis, and epidermis.