Poloxamer-chitosan-based Naringenin nanoformulation used in brain targeting for the treatment of cerebral ischemia
Authors: Md Aftab Alam
Wasim Ahmad
Mohd Amir
Abuzer Ali
Niyaz Ahmad
Rizwan Ahmad
Farhan Jalees Ahmad
Journal: Saudi Journal of Biological Sciences (2020)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: ANOVA., analysis of variance; AUC, area under curve; BA, bioavailability; CLSM, confocal laser scanning microscopy; CS, chitosan
Affiliations: Department of Pharmaceutics, College of Clinical Pharmacy, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia., Department of Pharmaceutical Chemistry, College of Clinical Pharmacy, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia., Department of Natural Products and Alternative Medicine, College of Clinical Pharmacy, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia., Nanomedicine Lab, Department of Pharmaceutics, School of Pharmaceutical Education and Research, Jamia Hamdard, Hamdard Nagar, New Delhi, India., Department of Pharmacy, Mohammad Al-Mana College for Medical Sciences, Safaa, Dammam-34222, Saudi Arabia., Department of Pharmaceutics, School of Medical and Allied Sciences, Galgotias University, Gautam Budh Nagar, Greater Noida 201310, India., College of Pharmacy, Taif University, Taif 21974, Saudi Arabia.

Abstract:  Objective: Here, the aim is to improve the bioavailability of Naringenin (NRG) in brain and to establish the highest remedial benefit from a novel anti-ischemic medicine i.e. NRG. Methods: A novel Naringenin-loaded-nanoemulsion (NE)-(in situ)-gel (i.e. thermoresponsive), was formulated with the help of Poloxamer-407 (20.0% w/v). Chitosan (CS, 0.50% w/v) was used to introduce the mucoadhesive property of NE-(in situ)-gel and finally called as NRG-NE-gel + 0.50%CS. A novel UHPLC-ESI-Q-TOF-MS/MS-method was optimized and used for NRG-NE-gel + 0.50%CS to quantify the Pharmacokinetic-(PK)-parameters in plasma as well as brain and to evaluate the cerebral ischemic parameters after MCAO i.e. locomotor activity, grip strength, antioxidant activity, and quantity the infarction volume in neurons with the safety/toxicity of NRG-NE-gel + 0.50%CS after i.n. administration in the rats. Results: The mucoadhesive potency and gelling temperature of NRG-NE-gel + 0.50%CS were observed 6245.38 dynes/cm2 and 28.3 ± 1.0 °C, respectively. Poloxamer-407 based free micelles size was observed 98.31 ± 1.17 nm with PDI (0.386 ± 0.021). The pH and viscosity of NRG-NE-gel + 0.50%CS were found to be 6.0 ± 0.20 and 2447 ± 24cp (at 35.0 ± 1.0 °C temperature), respectively. An elution time and m/z NRG were observed 1.78 min and 270.97/150.96 with 1.22 min and m/z of 301.01/150.98 for Quercetin (IS) respectively. Inter and intra %precision and %accuracy was validated 1.01-3.37% and 95.10-99.30% with a linear dynamic range (1.00 to 2000.00 ng/ml). AUC0-24 of plasma &amp; brain were observed 995.60 ± 24.59 and 5600.99 ± 144.92 (ng min/ml g) in the rats after the intranasal (i.n.) administration of NRG-NE-gel + 0.50%CS. No toxicological response were not found in terms of mortalities, any-change morphologically i.e. in the microstructure of brain as well as nasal mucosa tissues, and also not found any visual signs in terms of inflammatory or necrosis. Conclusion: Intranasally administered NRG-NE-gel + 0.50%CS enhanced the bioavailability of Naringenin in the brain. In the cerebral ischemic rats, significantly improved the neurobehavioral activity (locomotor &amp; grip strength) followed by antioxidant activity as well as infarction volume. Finally, the toxicity studies carried out and established the safe nature of optimized-NRG-NE-gel + 0.50%CS. 

Retracted: No

PubPeer comments:
 Above here:  Representative photo-micrographs showing the TS of ratsu2019 brain ... for **normal control, CS-PCL-NPs (placebo)**, CS-GA-PCL-NPs treated groups, respectively after 14 days , from [Ahmad et al (2019)] . Below: Fig 11:  Representative photomicrographs showing the TS of ratsu2019 brain (A, B, and C) ... for SHAM (Main control group), **NE-gel + 0.50%CS (placebo), NRG-NE-gel + 0.50%CS treated groups**, respectively after 14 days. ![file] , 
 #1 Extending the comparisons, the present 11B *[right]* and 10(d) from [Ahmad et al (2019)] *[middle]* have much in common with *[left]* 10A from [Ahmad et al (2014)] . ![file] , 
 Fig. 3.  CLSM images of Naringenin (NRGu2013NE) [A], mucoadhesive Naringenin (NRG)u2013NEu2013gelu202f+u202f0.50%CS [B]. Are these really two different materials, or the same material that has been assigned different colours? Overlap marked. ![] ,