*3.3. Effect of Independent Variables (A, B, C, D) on Entrapment Efficiency (R2)*

EE of DC-BMs was found between 50.23% (F1) and 93.11% (F16). The formulation (F1) prepared with composition cholesterol (A, 0.1%), lipid (B, 0.5%), surfactant (C, 0.5%), and bile salt (D, 1%) showed minimum EE. The maximum EE was shown by formulation (F19) having composition cholesterol (A, 0.5%), lipid (B, 1.5%), surfactant (C, 0.5%), and bile salt (D, 1%). There was a significant (*p* < 0.01) variation in EE was found due to variation in ratio of independent variables. The polynomial Equation (6) and 3D response surface plot (Figure 2) showed the effect of independent variables on EE. The increase in cholesterol (A) leads to enhancement in EE of DC. This effect was found due to deposition of CHO between free spaces of lipid bilayers, which reduces the flexibility, weakens the drug mobility, and reduces diffusion for DC from BMs [34,35]. The second factor lipid (B) also plays an important role on EE. The increase in lipid concentration lead to increase in EE due to enhancement in lipid viscosity. This prevents leaching of DC from lipid bilayer due to increase in hydrophobicity and longer alkyl chain length [36]. However, surfactant (C) showed a positive effect on EE of DC in BMs. The increase in surfactant concentration led to reduction in interfacial tension and increase in viscosity protects leakage of DC from BMs. The fourth variable bile salt (D, SD%) also showed a positive effect on EE. The increases in bile salt led to an increase in EE. It showed a lesser effect than surfactant. It also had surfactant-like properties—it reduced the interfacial tension and then drug easily assimilated into lipid bilayer due to enhanced solubility and flexibility [37].

EE (%, R<sup>2</sup> ) = 76.55 + 12.17 A + 8.054 B + 6.73 C + 4.46 D − 5.25 AB − 0.40 AC − 1.25AD − 0.55 BC − 4.78BD − 3.5 CD (6)

> The second order design model (2 F1) was found to be the best fit model for EE. The model F-value 36.53 implies that model was significantly fitted (*p* < 0.001). The lack of fit was found to be non-significant (*p* = 0.0942), and indicates model is well fitted. The regression coefficient of best fit model was found to be 0.958 and it indicates lesser variation between actual and predicted value (Table 2). The polynomial equation (Equation (2)) and ANOVA of best fitted model showed coded terms, i.e., A, B, C, D, AB, BD, CD, are significant (*p* < 0.05) which means these factors had a significant effect on EE of DC in BMs (Table 3).

#### *3.4. Optimized Formulation (DC-BMs-opt)*

The formulation (DC-BMs-opt) was selected from point prediction of software. The composition CHO (0.3% *w*/*v*), lipid (1% *w*/*v*), surfactant (0.5% *w*/*v*), and bile salt (1% *w*/*v*)

depicted an experimental vesicle size of 270.21 ± 3.76 nm and EE of 79.01 ± 2.54%. The software showed a predicted value of vesicle size of 274.36 nm and EE of 76.56%. There was non-significant variation in result observed between experimental and predicted value. The closeness in result revealed that model is valid and reproducible.

#### *3.5. Vesicle Evaluation*

The size of prepared DC-BMs (F1-F27) was found between 169.34 nm and 380.14 nm. The optimized composition (DC-BMs-opt) showed VS of 270.21 ± 3.76 nm (Figure 3A). PDI was found to be (0.26 ± 0.03) and revealed homogeneity of BMs-opt [38]. The zeta potential of DC-BMs-opt was of high negative (−36.34 mV) indicating that formulation was highly stable and in disaggregated form. The surface morphology exhibited spherical shape vesicles with a smooth surface without any aggregation (Figure 3B). *Pharmaceutics* **2022**, *14*, x FOR PEER REVIEW 10 of 16

**Figure 3.** Vesicle size (**A**), and (**B**) TEM image of optimized diclofenac bilosomes (DC-BMs-opt marked with arrow). **Figure 3.** Vesicle size (**A**), and (**B**) TEM image of optimized diclofenac bilosomes (DC-BMs-opt marked with arrow).

#### *3.6. Thermal Analysis 3.6. Thermal Analysis*

Figure 4 shows DSC spectra of DC, lipid, CHO, Pluronic F127, SC, PM, and DC-BMsopt. DC showed characteristic endothermic peak at 287.5 °C, which corresponds to its melting point [39]. The lipid, cholesterol, Pluronic F127, and SC exhibited a peak at 180°C (Figure 4B), at 150 °C (Figure 4C), at 60 °C (Figure 4D), and 190 °C (Figure 4E), respectively. The physical mixture exhibited endothermic peaks at 56 °C (Pluronic F127), an exothermic peak at 180 °C, and a less intense peak of DC at 287.5 °C (Figure 4F). No characteristic endothermic peak of DC was observed in DC-BMs-opt thermogram. The observation revealed complete encapsulation or solubilization of DC into BMs matrix (Figure 4G). Figure 4 shows DSC spectra of DC, lipid, CHO, Pluronic F127, SC, PM, and DC-BMsopt. DC showed characteristic endothermic peak at 287.5 ◦C, which corresponds to its melting point [39]. The lipid, cholesterol, Pluronic F127, and SC exhibited a peak at 180 ◦C (Figure 4B), at 150 ◦C (Figure 4C), at 60 ◦C (Figure 4D), and 190 ◦C (Figure 4E), respectively. The physical mixture exhibited endothermic peaks at 56 ◦C (Pluronic F127), an exothermic peak at 180 ◦C, and a less intense peak of DC at 287.5 ◦C (Figure 4F). No characteristic endothermic peak of DC was observed in DC-BMs-opt thermogram. The observation revealed complete encapsulation or solubilization of DC into BMs matrix (Figure 4G).

#### *3.7. In Vitro Drug Release*

The release of DC-BMs-opt was analyzed and result was compared with DC-LP and pure DC. The data of release study are shown in Figure 5. DC-BMs-opt exhibited 91.82 ± 4.65% release in 24 h of study. The graph showed biphasic release behavior with an initial fast release and later sustained release. The fast release was due to presence of DC on surface of BM-opt and later the sustained release was found due to release of DC from DC-BMs matrix [16]. There was a significantly (*p* < 0.001) lower DC release achieved from DC-LP (74.54 ± 4.76%) and pure DC (36.32 ± 4.23). The liposomes (DC-LP) showed significantly (*p* < 0.001) higher DC release than the pure DC. The pure DC showed poor release due to poor solubility. The significant high release of DC was achieved from the BMs and LP due to enhanced DC solubility in presence of surfactant. There was also a significant difference in release achieved due to presence of bile salt in BMs. Bile salt showed a synergistic effect with used surfactant and can enhance greater solubility.

**Figure 4.** Thermal analysis of (**A**) diclofenac, (**B**) lipid, (**C**) cholesterol, (**D**) Pluronic F127, (**E**) sodium deoxycholate, (**F**), physical mixture, and (**G**) optimized diclofenac bilosomes (DC-BMs-opt).

**Figure 4.** Thermal analysis of (**A**) diclofenac, (**B**) lipid, (**C**) cholesterol, (**D**) Pluronic F127, (**E**) sodium deoxycholate, (**F**), physical mixture, and (**G**) optimized diclofenac bilosomes (DC-BMs-opt). **Figure 4.** Thermal analysis of (**A**) diclofenac, (**B**) lipid, (**C**) cholesterol, (**D**) Pluronic F127, (**E**) sodium deoxycholate, (**F**), physical mixture, and (**G**) optimized diclofenac bilosomes (DC-BMs-opt). difference in release achieved due to presence of bile salt in BMs. Bile salt showed a synergistic effect with used surfactant and can enhance greater solubility.

**Figure 3.** Vesicle size (**A**), and (**B**) TEM image of optimized diclofenac bilosomes (DC-BMs-opt

Figure 4 shows DSC spectra of DC, lipid, CHO, Pluronic F127, SC, PM, and DC-BMsopt. DC showed characteristic endothermic peak at 287.5 °C, which corresponds to its melting point [39]. The lipid, cholesterol, Pluronic F127, and SC exhibited a peak at 180°C (Figure 4B), at 150 °C (Figure 4C), at 60 °C (Figure 4D), and 190 °C (Figure 4E), respectively. The physical mixture exhibited endothermic peaks at 56 °C (Pluronic F127), an exothermic peak at 180 °C, and a less intense peak of DC at 287.5 °C (Figure 4F). No characteristic endothermic peak of DC was observed in DC-BMs-opt thermogram. The observation revealed complete encapsulation or solubilization of DC into BMs matrix (Figure 4G).

marked with arrow).

*3.6. Thermal Analysis*

**Figure 5.** Release study of different treatment groups (pure diclofenac (DC), optimized diclofenac bilosomes (DC-BMs-opt), and diclofenac liposomes (DC-LP)). Study was performed in triplicate and data are shown as mean ± SD. Statistical analysis performed between each group and *p* < 0.05 considered significant. \*\*\* highly significant to pure DC; \*\* significant to pure DC-LP. **Figure 5.** Release study of different treatment groups (pure diclofenac (DC), optimized diclofenac bilosomes (DC-BMs-opt), and diclofenac liposomes (DC-LP)). Study was performed in triplicate and data are shown as mean ± SD. Statistical analysis performed between each group and *p* < 0.05 considered significant. \*\*\* highly significant to pure DC; \*\* significant to pure DC-LP.

The release profile of DC-BMs-opt was fitted into different kinetic models and data showed best fit model as the Korsmeyer–Peppas model (Table 4). The maximum regression value (R2 = 0.9354) confirms best fit. The exponent n-value was 0.58 (0.45 to 0.85) representing non-Fickian mechanism with dual release, i.e., diffusion and swelling release The release profile of DC-BMs-opt was fitted into different kinetic models and data showed best fit model as the Korsmeyer–Peppas model (Table 4). The maximum regression value (R<sup>2</sup> = 0.9354) confirms best fit. The exponent n-value was 0.58 (0.45 to 0.85) repre-

> **Type of Model R2** Zero model 0.7344 First order 0.9257 Higuchi model 0.7744 Korsmeyer–Peppas 0.9354, n = 0.58 Hixon–Crowell model 0.8673

[40].

senting non-Fickian mechanism with dual release, i.e., diffusion and swelling release [40].


**Table 4.** Various kinetic release models and their regression value.

#### *3.8. Ex Vivo Permeation Study 3.8. Ex Vivo Permeation Study*

The study of DC-BMs-opt was assessed to compare results with DC-LP and pure DC (Figure 6). The formulation DC-BMs-opt showed significantly (*p* < 0.001) higher permeation (187.59 <sup>±</sup> 9.65 <sup>µ</sup>g/cm<sup>2</sup> ) than DC-LP (119.44 <sup>±</sup> 10.06 <sup>µ</sup>g/cm<sup>2</sup> ) and DC-dispersion (59.52 <sup>±</sup> 7.76 <sup>µ</sup>g/cm<sup>2</sup> ). It also exhibited significant (*p* < 0.05) 3.15-fold (31.26 µg/cm2/h) higher flux than pure DC (9.92 µg/cm2/h) and 1.57-fold higher than DC-LP (19.91 µg/cm2/h). DC-BMs-opt showed the APC of 2.08 <sup>×</sup> <sup>10</sup>−<sup>3</sup> cm/s, which was significantly higher (*p* < 0.05) than pure DC (6.6 <sup>×</sup> <sup>10</sup>−<sup>4</sup> cm/s) and DC-PL (1.33 <sup>×</sup> <sup>10</sup>−<sup>3</sup> cm/s). The pure DC showed lesser permeation due to the poor solubility and not being able to permeate across the biological membrane. The greater amount of DC permeates across the membrane from liposomes due to presence of lipid, cholesterol, and surfactant. The surfactant helps to solubilize drug and due to enhanced solubility, greater effective surface area is available for drug absorption. BMs were prepared with a special component as bile salt which helped to deform the vesicles and also helped to fluidize membrane, possibly because of interaction of phospholipid molecules with membrane layer [41]. Due to this property, it can permeate easily to smaller-sized membrane. The presence of cholesterol helps to extract lipid of membrane and act as a permeation enhancer. The larger amount of drug permeated from BMs. A size of more than 200 nm does not significantly affect permeation of drugs [42]. The study of DC-BMs-opt was assessed to compare results with DC-LP and pure DC (Figure 6). The formulation DC-BMs-opt showed significantly (*p* < 0.001) higher permeation (187.59 ± 9.65 µg/cm2) than DC-LP (119.44 ± 10.06 µg/cm2) and DC-dispersion (59.52 ± 7.76 µg/cm2). It also exhibited significant (*p* < 0.05) 3.15-fold (31.26 µg/cm2/h) higher flux than pure DC (9.92 µg/cm2/h) and 1.57-fold higher than DC-LP (19.91 µg/cm2/h). DC-BMsopt showed the APC of 2.08 × 10−3 cm/s, which was significantly higher (*p* < 0.05) than pure DC (6.6 × 10−4 cm/s) and DC-PL (1.33 × 10−3 cm/s). The pure DC showed lesser permeation due to the poor solubility and not being able to permeate across the biological membrane. The greater amount of DC permeates across the membrane from liposomes due to presence of lipid, cholesterol, and surfactant. The surfactant helps to solubilize drug and due to enhanced solubility, greater effective surface area is available for drug absorption. BMs were prepared with a special component as bile salt which helped to deform the vesicles and also helped to fluidize membrane, possibly because of interaction of phospholipid molecules with membrane layer [41]. Due to this property, it can permeate easily to smaller-sized membrane. The presence of cholesterol helps to extract lipid of membrane and act as a permeation enhancer. The larger amount of drug permeated from BMs. A size of more than 200 nm does not significantly affect permeation of drugs [42].

**Figure 6.** Permeation study of different treatment groups (pure diclofenac (DC), optimized diclofenac bilosomes (DC-BMs-opt), and diclofenac liposomes (DC-LP)). Study was performed in triplicate and result shown as mean ± SD. Statistical analysis performed between each group and *p* < 0.05 considered significant. \*\*\* highly significant to pure DC; ### significant to DC-LP; \*\* significant to pure DC. **Figure 6.** Permeation study of different treatment groups (pure diclofenac (DC), optimized diclofenac bilosomes (DC-BMs-opt), and diclofenac liposomes (DC-LP)). Study was performed in triplicate and result shown as mean ± SD. Statistical analysis performed between each group and *p* < 0.05 considered significant. \*\*\* highly significant to pure DC; ### significant to DC-LP; \*\* significant to pure DC.

The pharmacokinetic study of pure DC, DC-LP, and DC-BMs-opt was conducted and

significant variation in each tested parameter. DC-BMs-opt showed a Cmax value of 2654.76 ± ng/mL and was found to be 2.15-fold higher than pure DC (1232.34 ± ng/mL) and 1.29 fold higher than DC-LP (2054 ± ng/mL). The higher Cmax was achieved due to nano-size of DC-BMs-opt, high permeability, and low first-pass metabolism. The difference was found to be highly significant (*p* < 0.001) compared to pure DC and DC-LP. DC-BMs-opt showed significant (*p* < 0.05) enhancement in AUC0–t (22,340 ng. h/mL) and AUC0–∞ (26,827.92 ng.

*3.9. Bioavailability Study*

#### *3.9. Bioavailability Study*

The pharmacokinetic study of pure DC, DC-LP, and DC-BMs-opt was conducted and plasma concentration-time profile is expressed graphically in Figure 7. The result showed significant variation in each tested parameter. DC-BMs-opt showed a Cmax value of 2654.76 ± ng/mL and was found to be 2.15-fold higher than pure DC (1232.34 ± ng/mL) and 1.29-fold higher than DC-LP (2054 ± ng/mL). The higher Cmax was achieved due to nano-size of DC-BMs-opt, high permeability, and low first-pass metabolism. The difference was found to be highly significant (*p* < 0.001) compared to pure DC and DC-LP. DC-BMsopt showed significant (*p* < 0.05) enhancement in AUC0–t (22,340 ng. h/mL) and AUC0–<sup>∞</sup> (26,827.92 ng. h/mL) values. It was about 5.2 and 6.2-fold higher than pure DC (AUC0–t of 4288.48 ng. h/mL and AUC0–<sup>∞</sup> of 4319.12 ng. h/mL) and 1.43 and 1.56-fold higher than DC-LP (AUC0–t of 15,564, AUC0–<sup>∞</sup> of 17,170.09 ng. h/mL). The half-life (t1/2) of DC-LP and DC-BMs-opt was found to be higher (6.62 h and 8.39 h) than pure DC (1.95 h), which revealed that DC-BMs-opt was available for a longer time in circulation. DC-BMs-opt exhibited higher *Tmax* (1 h) than pure DC dispersion (30 min) due to an increase in solubility of DC in BM as well as LP. The elimination rate constant (Ke) for DC-BMs-opt was found to be significantly (*p* < 0.05) lower (0.08 h−<sup>1</sup> ) than pure DC (0.22 h−<sup>1</sup> ) and DC-LP (0.1 h−<sup>1</sup> ) due to slow and prolonged drug release. The relative bioavailability of DC-BMs-opt showed 5.2-fold enhancement compared to pure DC and 1.43-fold higher compared to DC-LP. The higher bioavailability DC in DC-BMs-opt is due to increased DC solubility, longer circulation, lower first-pass metabolism, and higher uptake of BM by Peyer's patch of M-cell of intestine [43]. *Pharmaceutics* **2022**, *14*, x FOR PEER REVIEW 13 of 16 h/mL) values. It was about 5.2 and 6.2-fold higher than pure DC (AUC0–t of 4288.48 ng. h/mL and AUC0–∞ of 4319.12 ng. h/mL) and 1.43 and 1.56-fold higher than DC-LP (AUC0– t of 15,564, AUC0–∞ of 17,170.09 ng. h/mL). The half-life (t1/2) of DC-LP and DC-BMs-opt was found to be higher (6.62 h and 8.39 h) than pure DC (1.95 h), which revealed that DC-BMs-opt was available for a longer time in circulation. DC-BMs-opt exhibited higher *Tmax* (1 h) than pure DC dispersion (30 min) due to an increase in solubility of DC in BM as well as LP. The elimination rate constant (Ke) for DC-BMs-opt was found to be significantly (*p*  < 0.05) lower (0.08 h−1) than pure DC (0.22 h−1) and DC-LP (0.1 h−1) due to slow and prolonged drug release. The relative bioavailability of DC-BMs-opt showed 5.2-fold enhancement compared to pure DC and 1.43-fold higher compared to DC-LP. The higher bioavailability DC in DC-BMs-opt is due to increased DC solubility, longer circulation, lower first-pass metabolism, and higher uptake of BM by Peyer's patch of M-cell of intestine [43].

**Figure 7.** Bioavailability activity of the different treatment groups (pure diclofenac (DC), optimized diclofenac bilosomes (DC-BMs-opt), and diclofenac liposomes (DC-LP)). Study performed with six rats (n = 6) in each group and results shown as mean ± SD. Statistical analysis performed between each group and *p* < 0.05 considered significant. \*\*\* highly significant to pure DC; ### significant to DC- LP; \* significant to pure DC and DC-LP. **Figure 7.** Bioavailability activity of the different treatment groups (pure diclofenac (DC), optimized diclofenac bilosomes (DC-BMs-opt), and diclofenac liposomes (DC-LP)). Study performed with six rats (n = 6) in each group and results shown as mean ± SD. Statistical analysis performed between each group and *p* < 0.05 considered significant. \*\*\* highly significant to pure DC; ### significant to DC- LP; \* significant to pure DC and DC-LP.

#### *3.10. Pharmacodynamic Study 3.10. Pharmacodynamic Study*

The anti-inflammatory activity of pure DC, DC-LP, and DC-BMs-opt was evaluated in carrageenan-induced model and results are expressed graphically in Figure 8. The disease control groups showed about 100% swelling. The pure DC, DC-LP, and DC-BMs-opt exhibited a significant effect in lowering paw edema. The pure DC, DC-LP, and DC-BMsopt showed 26.23 ± 7.83%, 28.43 ± 5.67%, and 31.26 ± 6.13% reduction in paw edema after The anti-inflammatory activity of pure DC, DC-LP, and DC-BMs-opt was evaluated in carrageenan-induced model and results are expressed graphically in Figure 8. The disease control groups showed about 100% swelling. The pure DC, DC-LP, and DC-BMs-opt exhibited a significant effect in lowering paw edema. The pure DC, DC-LP, and DC-BMs-

2 h carrageenan injection, respectively. The pure DC-treated group showed maximum effect at 2 h, whereas DC-LP and DC-BMs-opt treated group showed a maximum effect up

maximum reduction was found to be 8.65 ± 3.87%, 23.76 ± 5.92%, and 64.76 ± 11.12% from pure DC, DC-LP, and DC-BMs-opt. At all-time points, a significant effect was observed from tested groups in comparison to disease control. DC-BMs-opt also exhibited a significant (*p* < 0.05) reduction in swelling than DC-LP. This high reduction in swelling was

opt showed 26.23 ± 7.83%, 28.43 ± 5.67%, and 31.26 ± 6.13% reduction in paw edema after 2 h carrageenan injection, respectively. The pure DC-treated group showed maximum effect at 2 h, whereas DC-LP and DC-BMs-opt treated group showed a maximum effect up to 6 h and 9 h, respectively. There was a highly significant (*p* < 0.001) effect observed from DC-LP and DC-BMs-opt at 3 h, 6 h, 9 h, and 12 h in comparison to pure DC. At 12 h, maximum reduction was found to be 8.65 ± 3.87%, 23.76 ± 5.92%, and 64.76 ± 11.12% from pure DC, DC-LP, and DC-BMs-opt. At all-time points, a significant effect was observed from tested groups in comparison to disease control. DC-BMs-opt also exhibited a significant (*p* < 0.05) reduction in swelling than DC-LP. This high reduction in swelling was achieved due to high penetration capacity of DC-BMs-opt through intestinal mucosa. The nano-sized vesicle having a greater effective surface area, high circulation time, greater solubility and flexibility in presence of surfactant and bile salt led to greater absorption. Therefore, findings revealed that BMs may increase solubility and circulation of drugs which directly increases anti-inflammatory effect. *Pharmaceutics* **2022**, *14*, x FOR PEER REVIEW 14 of 16 achieved due to high penetration capacity of DC-BMs-opt through intestinal mucosa. The nano-sized vesicle having a greater effective surface area, high circulation time, greater solubility and flexibility in presence of surfactant and bile salt led to greater absorption. Therefore, findings revealed that BMs may increase solubility and circulation of drugs which directly increases anti-inflammatory effect.

**Figure 8.** Anti-inflammatory activity of different treatment groups (pure diclofenac (DC), optimized diclofenac bilosomes (DC-BMs-opt), and diclofenac liposomes (DC-LP)) compared with disease control. Study performed with six rats (n = 6) in each group and result shown as mean ± SD. Statistical analysis performed between each group and *p* < 0.05 considered significant. \*\*\* highly significant to diabetic control; ### significant to pure DC and DC- LP; ns, non-significant to pure DC and DC-LP. **Figure 8.** Anti-inflammatory activity of different treatment groups (pure diclofenac (DC), optimized diclofenac bilosomes (DC-BMs-opt), and diclofenac liposomes (DC-LP)) compared with disease control. Study performed with six rats (n = 6) in each group and result shown as mean ± SD. Statistical analysis performed between each group and *p* < 0.05 considered significant. \*\*\* highly significant to diabetic control; ### significant to pure DC and DC- LP; ns, non-significant to pure DC and DC-LP.

#### **4. Conclusions 4. Conclusions**

manuscript.

search grant no (DSR-2021-01-0327).

In the present study, DC-BMs were prepared by solvent evaporation method using sodium deoxycholate as bile salt. The formulations were optimized by Box–Behnken design to select optimum composition. The optimized formulation DC-BMs-opt showed a nano vesicle size and high encapsulation efficiency. The in vitro release and *ex vivo* permeation study showed a prolonged DC release with high permeation flux. The pharmacokinetic and pharmacodynamics study results revealed enhanced bioavailability and anti-inflammatory activity compared to pure DC and DC-LP. Further, prepared formulations need to be evaluated for clinical study. The findings of preclinical data need to be correlated with clinical data for better outcomes. We conclude from our findings that DC-BMs-opt is a promising oral drug delivery for treatment of inflammation. In the present study, DC-BMs were prepared by solvent evaporation method using sodium deoxycholate as bile salt. The formulations were optimized by Box–Behnken design to select optimum composition. The optimized formulation DC-BMs-opt showed a nano vesicle size and high encapsulation efficiency. The in vitro release and ex vivo permeation study showed a prolonged DC release with high permeation flux. The pharmacokinetic and pharmacodynamics study results revealed enhanced bioavailability and anti-inflammatory activity compared to pure DC and DC-LP. Further, prepared formulations need to be evaluated for clinical study. The findings of preclinical data need to be correlated with clinical data for better outcomes. We conclude from our findings that DC-BMs-opt is a promising oral drug delivery for treatment of inflammation.

**Author Contributions:** Conceptualization and methodology, A.Z. and S.S.I.; software and validation, M.Y., S.S.I. and M.K.A.; resources and investigation, N.K.A., O.A.A. and S.A.; data curation, A.A. and A.R.; writing—original draft preparation, A.Z.; writing—review and editing, B.A. and

M.M.G.; funding acquisition, A.Z. All authors have read and agreed to the published version of the

**Funding:** Deanship of Scientific Research at Jouf University for funding this work through re-

**Author Contributions:** Conceptualization and methodology, A.Z. and S.S.I.; software and validation, M.Y., S.S.I. and M.K.A.; resources and investigation, N.K.A., O.A.A. and S.A.; data curation, A.A. and A.R.; writing—original draft preparation, A.Z.; writing—review and editing, B.A. and M.M.G.; visualization and supervision, N.K.A. and O.A.A.; project administration, M.K.A. and M.M.G.; funding acquisition, A.Z. All authors have read and agreed to the published version of the manuscript.

**Funding:** Deanship of Scientific Research at Jouf University for funding this work through research grant no (DSR-2021-01-0327).

**Institutional Review Board Statement:** The animal study protocol was approved by the Institutional Review Board (or Ethics Committee) of Jouf University (Approval Number 04-02-43; Approval Date 28 October 2021).

**Informed Consent Statement:** Not applicable.

**Data Availability Statement:** Not applicable.

**Acknowledgments:** The authors extend their appreciation to the Deanship of Scientific Research at Jouf University for funding this work through research grant no (DSR-2021-01-0327).

**Conflicts of Interest:** The authors declare no conflict of interest.

#### **References**

