Quality by Design-Based Development and In Vitro–In Vivo Evaluation of Baicalin-Loaded Thermoresponsive Nanocomposite Hydrogel for Targeted Immunomodulatory Therapy in Chronic Plaque Psoriasis

Jagadeeswara Rao Batna; Sujith Thomas; Meman Rahil Salim; Sheik Nasar I; Mohit Kumar; Senthil Rajan Dharmalingam; Neelam Pawar; D. Suresh

doi:10.29284/c9xwp940

Authors

Jagadeeswara Rao Batna Lecturer in Pharmacy, Government Polytechnic, Visakhapatnam, Andhra Pradesh, India. 530007
Sujith Thomas Professor cum HOD, Department of Pharmacology, Moulana College of Pharmacy , Perinthalmanna, Kerala.
Meman Rahil Salim Associate Professor, Ismail Mehta College of Pharmacy, Beed Road, Ambad, Jalna, Maharashtra. 431204
Sheik Nasar I Professor, East Point College of Pharmacy, East Point Group of Institutions, Jnana Prabha, East Point Campus, Virgo Nagar, Bidarahalli, Bengaluru, Karnataka. 560049
Mohit Kumar Assistant professor, Teerthanker Mahaveer College of Pharmacy, Teerthanker Mahaveer University, Moradabad, Uttar Pradesh.
Senthil Rajan Dharmalingam Professor & Head Department of Pharmaceutics, Swamy Vivekanandha College of Pharmacy (affiliated to The Tamil Nadu Dr. M.G.R. Medical University, Chennai), Elayampalayam, Tiruchengode, Namakkal, Tamil Nadu, India. 637205
Neelam Pawar Department of Pharmaceutical Sciences, Hargobind Khurana Building, Prem Nagar, Chaudhary Bansilal University, Bhiwani, Haryana. 127031
D. Suresh Professor, KMCT College Of Pharmaceutical Sciences, Kallanthode, Nitc-Po, Kozhikode, Kerala. 673601

DOI:

https://doi.org/10.29284/c9xwp940

Keywords:

Baicalin; Thermoresponsive hydrogel; Nanocomposite; Quality by Design; Chronic plaque psoriasis; Immunomodulatory therapy.

Abstract

Chronic plaque psoriasis is a persistent inflammatory skin disorder characterized by excessive keratinocyte proliferation and immune dysregulation. The present study aimed to develop a Quality by Design (QbD)–based baicalin-loaded thermoresponsive nanocomposite hydrogel for targeted topical immunomodulatory therapy. Baicalin nanoparticles were optimized using a three-factor Box–Behnken design to evaluate the influence of polymer, stabilizer, and nanoclay concentrations on particle size, polydispersity index, and encapsulation efficiency. The optimized formulation exhibited nanoscale particle size (~162 nm), narrow size distribution, and high drug entrapment. These nanoparticles were incorporated into a poloxamer-based thermoresponsive hydrogel that underwent sol–gel transition near skin temperature and demonstrated favourable rheological properties and spreadability. In vitro release studies revealed sustained drug release with reduced burst effect compared to free drug gel. Ex vivo permeation studies demonstrated enhanced dermal deposition and controlled transdermal flux. Cytocompatibility assessment using HaCaT and RAW 264.7 cells confirmed formulation safety. Furthermore, significant suppression of pro-inflammatory cytokines, including TNF-α and IL-6, indicated improved immunomodulatory potential. Overall, the QbD-guided nanocomposite hydrogel system demonstrated enhanced pharmaceutical and biological performance, supporting its potential as a localized therapeutic strategy for chronic plaque psoriasis.

References

1. Adarmanabadi, S. R., Jafari, M., Farrash, S. M. H., & Heidari, M. (2021). Effect of nano clay, nano-graphene oxide and carbon nanotubes on the mechanical and tribological properties of crosslinked epoxy nanocomposite. PLoS One, 16(11), e0259401. https://doi.org/10.1371/journal.pone.0259401

2. Adin, S. N., Gupta, I., Aqil, M., & Mujeeb, M. (2023). Application of QbD based approach in development and validation of RP-HPLC method for simultaneous estimation of methotrexate and baicalin in dual-drug-loaded liposomes. Biomed Chromatogr, 37(4), e5581. https://doi.org/10.1002/bmc.5581

3. Alexa, R. L., Iovu, H., Trica, B., Zaharia, C., Serafim, A., Alexandrescu, E., Radu, I. C., Vlasceanu, G., Preda, S., Ninciuleanu, C. M., & Ianchis, R. (2021). Assessment of Naturally Sourced Mineral Clays for the 3D Printing of Biopolymer-Based Nanocomposite Inks. Nanomaterials (Basel), 11(3). https://doi.org/10.3390/nano11030703

4. Alorabi, A. Q., Hassan, M. S., Alam, M. M., Zabin, S. A., Alsenani, N. I., & Baghdadi, N. E. (2021). Natural Clay as a Low-Cost Adsorbent for Crystal Violet Dye Removal and Antimicrobial Activity. Nanomaterials (Basel), 11(11). https://doi.org/10.3390/nano11112789

5. Andrade-Guel, M., Cabello-Alvarado, C., Romero-Huitzil, R. L., Rodríguez-Fernández, O. S., Ávila-Orta, C. A., Cadenas-Pliego, G., Medellín-Banda, D. I., Gallardo-Vega, C., & Cepeda-Garza, J. (2021). Nanocomposite PLA/C20A Nanoclay by Ultrasound-Assisted Melt Extrusion for Adsorption of Uremic Toxins and Methylene Blue Dye. Nanomaterials (Basel), 11(10). https://doi.org/10.3390/nano11102477

6. Ashrafudoulla, M., Yun, H., Ashikur Rahman, M., Jung, S. J., Jie-Won Ha, A., Anamul Hasan Chowdhury, M., Shaila, S., Akter, S., Park, S. H., & Ha, S. D. (2024). Prophylactic efficacy of baicalin and carvacrol against Salmonella Typhimurium biofilm on food and food contact surfaces. Food Res Int, 187, 114458. https://doi.org/10.1016/j.foodres.2024.114458

7. Bernasinska-Slomczewska, J., Hikisz, P., Pieniazek, A., & Koceva-Chyla, A. (2024). Baicalin and Baicalein Enhance Cytotoxicity, Proapoptotic Activity, and Genotoxicity of Doxorubicin and Docetaxel in MCF-7 Breast Cancer Cells. Molecules, 29(11). https://doi.org/10.3390/molecules29112503

8. Cao, Z., Ma, N., Shan, M., Wang, S., Du, J., Cheng, J., Sun, P., Sun, N., Jin, L., Fan, K., Yin, W., Li, H., Yin, C., & Sun, Y. (2024). Baicalin Inhibits FIPV Infection In Vitro by Modulating the PI3K-AKT Pathway and Apoptosis Pathway. Int J Mol Sci, 25(18). https://doi.org/10.3390/ijms25189930

9. Chai, J., Hu, J., Wang, T., Bao, X., Luan, J., & Wang, Y. (2024). A Multifunctional Liposome for Synergistic Chemotherapy with Ferroptosis Activation of Triple-Negative Breast Cancer. Mol Pharm, 21(2), 781-790. https://doi.org/10.1021/acs.molpharmaceut.3c00903

10. Chavan, M. J., Wakte, P. S., & Shinde, D. B. (2010). Analgesic and anti-inflammatory activity of Caryophyllene oxide from Annona squamosa L. bark. Phytomedicine, 17(2), 149-151. https://doi.org/https://doi.org/10.1016/j.phymed.2009.05.016

11. Chen, C., Liu, W., Gu, X., Zhang, L., Mao, X., Chen, Z., & Tao, L. (2024). Baicalin-loaded Polydopamine modified ZIF-8 NPs inhibits myocardial ischemia/reperfusion injury in rats. J Biomater Sci Polym Ed, 35(12), 1863-1878. https://doi.org/10.1080/09205063.2024.2358640

12. Chen, J., Chen, F. Y., Lu, C. J., & Yi, S. W. (2024). Baicalein alleviates palmitic acid-induced endothelial cell dysfunction via inhibiting endoplasmic reticulum stress. Clin Hemorheol Microcirc, 88(2), 235-245. https://doi.org/10.3233/ch-242230

13. do Nascimento Silva, H., Rabêlo, S. V., Diniz, T. C., da Silva Oliveira, F. G., de Andrade Teles, R. B., Silva, J. C., e Silva, M. G., Coutinho, H. D. M., de Menezes, I. R. A., & da Silva Almeida, J. R. G. (2017). Antinociceptive and anti-inflammatory activities of ethanolic extract from atemoya (Annona cherimola Mill x Annona squamosa L.). African Journal of Pharmacy and Pharmacology, 11(18), 224-232.

14. Dong, J., Cheng, Z., Tan, S., & Zhu, Q. (2021). Clay nanoparticles as pharmaceutical carriers in drug delivery systems. Expert Opin Drug Deliv, 18(6), 695-714. https://doi.org/10.1080/17425247.2021.1862792

15. Haddow, P. J., da Silva, M. A., Kaldybekov, D. B., Dreiss, C. A., Hoffman, E., Hutter, V., Khutoryanskiy, V. V., Kirton, S. B., Mahmoudi, N., McAuley, W. J., & Cook, M. T. (2022). Polymer Architecture Effects on Poly(N,N-Diethyl Acrylamide)-b-Poly(Ethylene Glycol)-b-Poly(N,N-Diethyl Acrylamide) Thermoreversible Gels and Their Evaluation as a Healthcare Material. Macromol Biosci, 22(3), e2100432. https://doi.org/10.1002/mabi.202100432

16. Hemlatha, K., & Satyanarayana, D. (2009). Anti-inflammatory Activity of Annona Squamosa Linn. Biomed. Pharmacol., 2(1).

17. Jumana, S., Hasan, C. M., & Rashid, M. A. (2000). Antibacterial activity and cytotoxicity of Miliusa velutina. Fitoterapia, 71(5), 559-561.

18. Kashif, M. U. R., Sohail, M., Khan, S. A., Minhas, M. U., Mahmood, A., Shah, S. A., & Mohsin, S. (2022). Chitosan/guar gum-based thermoreversible hydrogels loaded with pullulan nanoparticles for enhanced nose-to-brain drug delivery. Int J Biol Macromol, 215, 579-595. https://doi.org/10.1016/j.ijbiomac.2022.06.161

19. Kim, H. P., Son, K. H., Chang, H. W., & Kang, S. S. (2004). Anti-inflammatory plant flavonoids and cellular action mechanisms. Journal of pharmacological sciences, 96(3), 229-245.

20. L, S., U, S. S., & R, Z. (2025). Recent advances in oral in situ gel drug delivery system: a polymeric approach. Drug Dev Ind Pharm, 51(12), 1639-1649. https://doi.org/10.1080/03639045.2025.2559033

21. Mfoafo, K., Omidi, Y., & Omidian, H. (2023). Thermoresponsive mucoadhesive hybrid gels in advanced drug delivery systems. Int J Pharm, 636, 122799. https://doi.org/10.1016/j.ijpharm.2023.122799

22. Pande, S., Vashi, J., & Solanki, A. (2022). QbD-Enabled Systematic Development of Ileo-Colonic Targeted Novel Mucoadhesive Microspheres of Flurbiprofen. Curr Drug Deliv, 19(3), 407-419. https://doi.org/10.2174/1567201818666210708125036

23. Piaserico, S., & Messina, F. (2022). Pharmacological management of severe plaque psoriasis in patients with cardiovascular disease. Expert Opin Pharmacother, 23(7), 853-864. https://doi.org/10.1080/14656566.2022.2060739

24. Pinto, N. C. C., Silva, J. B., Menegati, L. M., Guedes, M., Marques, L. B., Silva, T. P. D., Melo, R. C. N., Souza-Fagundes, E. M., Salvador, M. J., Scio, E., & Fabri, R. L. (2017). Cytotoxicity and bacterial membrane destabilization induced by Annona squamosa L. extracts. An Acad Bras Cienc, 89(3 Suppl), 2053-2073. https://doi.org/10.1590/0001-3765201720150702

25. Purzycka-Bohdan, D., Kisielnicka, A., Zabłotna, M., Nedoszytko, B., Nowicki, R. J., Reich, A., Samotij, D., Szczęch, J., Krasowska, D., Bartosińska, J., Narbutt, J., Lesiak, A., Barasińska, P., Owczarczyk-Saczonek, A., Czerwińska, J., Szepietowski, J. C., Batycka-Baran, A., Czajkowski, R., Górecka-Sokołowska, M., . . . Szczerkowska-Dobosz, A. (2022). Chronic Plaque Psoriasis in Poland: Disease Severity, Prevalence of Comorbidities, and Quality of Life. J Clin Med, 11(5). https://doi.org/10.3390/jcm11051254

26. Rungjang, A., Meephansan, J., Payungporn, S., Sawaswong, V., Chanchaem, P., Pureesrisak, P., Wongpiyabovorn, J., & Thio, H. B. (2022). Skin Microbiota Profiles from Tape Stripping and Skin Biopsy Samples of Patients with Psoriasis Treated with Narrowband Ultraviolet B. Clin Cosmet Investig Dermatol, 15, 1767-1778. https://doi.org/10.2147/ccid.S374871

27. Scali, E., Dastoli, S., Procopio, A. C., Ricca, D., Mazzei, V., Cinaglia, P., Gulletta, E., Nisticò, S. P., Pellicano, R., Alshuk, N., Fagoonee, S., Luzza, F., & Abenavoli, L. (2022). Evaluation of serum calprotectin as novel biomarker in psoriatic patients: a prospective pilot study. Minerva Med, 113(5), 833-837. https://doi.org/10.23736/s0026-4806.22.08041-7

28. Seddik, A. H., Melzer, N., Tsotra, F., & Ostwald, D. A. (2022). The Societal Economic Impact of Secukinumab in First-Line Treatment of Moderate to Severe Plaque Psoriasis in Germany: An Open-Cohort Simulation. Pharmacoecon Open, 6(2), 265-275. https://doi.org/10.1007/s41669-021-00305-3

29. Shehata, W. A., Shoeib, M., Shoeib, M. M., Shokhba, H., & Shams, A. (2022). Nucleotide binding and oligomerization domain 2 in psoriasis: a clinical and immunohistochemical study. J Immunoassay Immunochem, 43(1), 43-53. https://doi.org/10.1080/15321819.2021.1941095

30. Supachawaroj, N., & Limsitthichaikoon, S. (2024). Lidocaine HCl-Loaded Polyelectrolyte Complex -Poloxamer Thermoresponsive Hydrogel: In Vitro- In Vivo Anesthetic Evaluations for Tooth Socket Wound Delivery. AAPS PharmSciTech, 25(6), 182. https://doi.org/10.1208/s12249-024-02894-8

31. Teng, K., Fu, H., Wu, G., Gong, P., Xie, Y., Zhou, P., Gong, X., & Qu, H. (2023). QbD-Guided Traditional Chinese Medicine Manufacturing Process: Development and Optimization of Fluid-Bed Granulation and Drying Processes for Xiaochaihu Capsules. AAPS PharmSciTech, 24(7), 210. https://doi.org/10.1208/s12249-023-02663-z

32. Vikas, B., Anil, S., & Remani, P. (2019). Cytotoxicity Profiling of Annona Squamosa in Cancer Cell Lines. Asian Pac J Cancer Prev, 20(9), 2831-2840. https://doi.org/10.31557/apjcp.2019.20.9.2831

33. Vilimi, Z., Király, M., Barna Á, T., Pápay, Z. E., Budai, L., Ludányi, K., Kállai-Szabó, N., & Antal, I. (2024). Formulation of Emulgels Containing Clotrimazole for the Treatment of Vaginal Candidiasis. Gels, 10(11). https://doi.org/10.3390/gels10110730

34. Wangusi, B. M., Kanja, L. W., Ole-Mapenay, I. M., & Onyancha, J. M. (2021). Acute Toxicity, Phytochemical Screening, Analgesic, and Anti-Inflammatory Activities of Aqueous and Methanol Root Extracts of Maerua triphylla A. Rich. (Capparaceae). Evid Based Complement Alternat Med, 2021, 3121785. https://doi.org/10.1155/2021/3121785

35. Zeeshan, M., Ain, Q. U., Sunny, A., Raza, F., Mohsin, M., Khan, S., Weigmann, B., & Ali, H. (2023). QbD-based fabrication of transferrin-anchored nanocarriers for targeted drug delivery to macrophages and colon cells for mucosal inflammation healing. Biomater Sci, 11(4), 1373-1397. https://doi.org/10.1039/d2bm01719a