Pharmaceutical Chemistry, Rajarambapu College of Pharmacy, Kasegaon- 415404, Sangli. Maharashtra, India.
The methods like green synthesis are advantageous over chemical and physical methods due to eco-friendly and cost-effective synthesis of nanoparticles. Present study was designed for green synthesis of silver (AgNPs) nanoparticles by using Terminalia bellerica plant extract and their pharmacological evaluation. Terminalia bellerica is characterized by powdered microscopical characters and specific chemical tests. Alcoholic extract of Terminalia bellerica were obtained by continuous heat extraction method. A Chemical constituent was Isolated from Terminalia bellerica extract by using TLC and Column chromatography. Plant extract (TB) is mixed with silver nitrate solution respectively, incubated and studied synthesis of nanoparticles by using UV-Vis spectroscopy. Nanoparticles were characterized by FTIR, SEM and XRD. (9) Hair growths potential was determined by using standard protocols. The nanoparticles synthesized were irregular and rectangular in shape having average particle size 10-50 nm. FTIR analysis revealed the possible involvement of phytoconstituents in synthesis of silver nanoparticles. Synthesized silver (TBAgNPs) nanoparticles showed effective enhanced Hair growth activity in comparison to Terminalia bellerica extract. Synthesized silver (TBAgNPs) nanoparticles exhibited dose dependent hair growth potential activity in albino mice. The activity was assessed by studying the follicle count, skin thickness, hair weight. nano-formulate (AgNPs) which will be incorporated into suitable dosage form like fast dissolving film for generation of novel drug delivery system with specific on set of action on epidermis which improved therapeutic efficacy for treatment of hair growth activity. It was concluded that Green synthesized (TBAgNPs) nanoparticles show the enhanced pharmacological activities. (11) It may have potential as a hair growth promoter.
As to keep hair intact is the first and the foremost duty of Sikh. So, that in mammal’s hair plays a vital role in thermal insulation, for social and sexual communication and is protective appendages on the body and considered accessory structure of the integument along with sebaceous glands, sweat glands and nails. Humans are relatively hairless compared other mammals and human hair has no known significance for survival of species. Hair loss is one the dermatological disorder to human race with is common throughout the world and is of great concern for decades. Hair growth is complex and cyclically controlled process that is characterized by finite period of hair fiber production, a brief regression phase and a resting period. The precise mechanism of regulating hair growth cycle was not yet been fully understood. Many of factors such as metabolism, hormones, heredity and side effect of antineoplastics and immunosuppressant drugs, has been negatively affecting on the healthy growth of hair. Terminalia Billerica is a tropical native to Asia including India, China, Sri Lanka, Pakistan, Bhutan, Nepal, Malaysia and Indonesia. In India it is found in Assam, Kerala, Gujarat, Tamil Nadu, Odisha and Andhra Pradesh. (10) This Plant is an important source of medicine with qualities for thousands of years. Baheda plants are the source of many potent and powerful drugs which are medicinally used in various countries. (14) The chemical constituents present in fruit of Baheda plant are gallic acid, galactose, fructose, rhamnose, ?-sitosterol, ellagic acid, galactose, ethyl gallate, chebulagic acid, mannitol and glucose. (4) T. Billerica fruits have about 22.3% (w/w) polyphenolic compounds which include gallic acid and their esters (19.1%), chebulic ellagitannins (0.81%), non-chebulic ellagitannins (1.5%), ellagic acid and derivatives (0.64%), and ellagic glycosides (0.23%). The whole seed contained 12.28 % oil on dry weight basis. Seed oil has been reported to contains oleic (43%), linoleic (29%), palmitic (12%), and stearic acid (16%). (6)
Figure No. 01- Seeds of Terminalia Billerica
Materials and Methods: -
Plant Collection:
The unripe fruits of Terminalia bellerica procured from the local area of Karad. The unripe fruit spiced and dried in the shade, powdered to a coarse consistency and stored in air tight container at room temperature. (15)
Preparation of plant extract:
The dried, powdered fruit was Soxhlet extracted with methanol and aqueous. The extract was weighed after solvent elimination under reduced pressure. The percent yield of the extract was found to be 8% and 10% of the ethanol and aqueous respectively.
Phytochemical Screening of TB seed extract:
The various phytochemical screening tests of Terminalia bellerica were performed according to the standard procedure from literature. (16)
Synthesis of Silver Nanoparticle:
Terminalia bellerica extract was used as a reducing agent for the synthesis of silver nanoparticles and silver nitrate (AgNO3) was used as a source of metal (silver) for synthesis of silver nanoparticles (AgNPs) according to the method as described by Abdul-Rehman vigorously mixed in the ratio of (1:1). (5) The mixture was incubated for 24 h at room temperature. The reaction progress observed by naked eye by change in color from yellow to darkish brown and later analyzed by UV-Visible spectroscopy. The obtained nanoparticles (TBAgNPs) were purified through centrifugation at 10,000 rpm for 5 min, washed with water and dried in desiccators for 24 hrs. at 350C. (2)
In vivo Hair growth activity
The mice were divided into three groups of 3 mice each. A 4cm2 area of hair from dorsal portion of all mice was shaved with electric hair clippers. 0.2ml of 1% TBAgNPs extracts in 50% of ethanol were applied to denuded area of the respective groups once a day, a standard group received 2% minoxidil solution (Jay bhaye et al., 2011) and a control group received vehicle treatment. This treatment was continued for 30 days and during this course the hair growth pattern was observed (Jung et al., 2010; Seok-Seon et al., 2002). (1)
Optimization of Formulation
Preparation of casting solutions:
PVOH (Polyvinyl alcohol) and Ethyl cellulose used in casting solutions were prepared by dissolving different weighed quantities polymer ratio such as PVOH (Polyvinyl alcohol) 200 mg and EC (Ethyl Cellulose) 66 mg of polymers, PVOH 200 mg and EC 150 mg of polymers. The obtained nanoparticles (TBAgNPs) 20 mg was dissolved in chloroform and added to the above polymer solution along with polyethylene glycol 300 (1.5 ml), as a plasticizer, and 0.1 ml of DMSO (disodium methyl sulfoxide) as penetration enhancer which was thoroughly mixed to form a homogeneous mixture. The volume was made up to 10 ml with chloroform. Entrapped air bubbles were removed by applying vacuum. (12)
Preparation of Transdermal Patches:
The casting solution (10 ml) was poured into glass molds and dried at room temperature for 24 hrs. for solvent evaporation. The patches were removed by peeling and cut into squares with dimension of 2x2 cm2. These patches were kept in desiccators for 2 days for further drying and wrapped in aluminum foil, packed in self-sealing covers. Transdermal patches were prepared with different polymer ratio, with constant plasticizer concentration and permeation enhancers. (13)
RESULT AND DISCUSSION: -
Determination of Effect of Concentration, pH, and Incubation time on the synthesis of nanoparticles:
The color of the solutions changed from light yellow to pale yellow on increasing crude extract concentration in the absence of silver nitrate solution. But in the presence of silver nitrate solution, with the increase of extract concentration the crude extract color change from yellowish to brown and further to deep brown due to excitation of surface Plasmon vibration, indicating formation in the silver nanoparticles. (7)
Figure No.03: Solution of Terminalia bellerica plant extract (A), Solution of Silver nitrate (B), Solution of synthesized AgNPs (C)
Results of Characterization of Nanoparticles:
Figure No.03: H1 NMR Spectra of Silver nanoparticles (TBAgNPs)
Motic Microscopy Analysis of Silver Nanoparticles (PUAgNPs)
From the primary analysis of Motic microscope images it was suggested that the synthesized silver nanoparticles (TBAgNPs) are rectangular and square in shape and size ranging from 0.040 - 0.210 µm as shown in Figure No.04. (8)
Figure No.04: Motic Microscopy Analysis of Silver Nanoparticles (TBAgNPs)
Scanning Electron Microscope (SEM) analysis of Silver Nanoparticles (TBAgNPs)
SEM micrographs at different magnification levels (Figure No. 05) showed that synthesized silver nanoparticles were in form of rectangular and square shapes in aggregated form. (3)
Figure No. 05: Scanning electron microscope (SEM) images of silver nanoparticles (TBAgNPs)
Preparation of Transdermal Patch of TBAgNPS:
Figure No.06: Formulated Transdermal Patches of TBAgNPs
Hair growth promoting effect of the Terminalia bellerica in Swiss albino mice after 30 days.
(A) Initially shaved skin of mice; (B) Effect of vehicle (Control group); (C) Effect of 2% minoxidil solution; (D). Effect of 1% aqueous extract; (E.) Effect of 1% methanolic extract.
CONCLUSION: -
In conclusion, the effect of TBAgNPs on qualitative hair growth and length was found to be more significant as compared to standard and control group treated animals. The quantitative effect of TBAgNPs definitely promotes hair growth by inducing hair follicles in anlagen phase. Animals treated with TBAgNPs showed better efficacy as compared to control and standard group. The percentage of anlagen induction with TBAgNPs and minoxidil were comparable. On the basis of similarities observed between the minoxidil and the TBAgNPs studies, it is expected that TBAgNPs will have similar hair growth activity as shown by minoxidil. Further research is needed for structural elucidation and identifying the mechanism of action responsible for using TBAgNPs as a potential hair growth promoter.
REFERENCE
Vishwajit S. Patil, Omkar J. Baravkar, Prasad P. Amane, Vyankatesh R. Dharanguttikar*, Development and Industrial Applications of Transdermal Patch: Formulated by Using Silver (AgNPs) Nanoparticles of Terminalia bellerica plant extract, Int. J. of Pharm. Sci., 2024, Vol 2, Issue 8, 2667-2673. https://doi.org/10.5281/zenodo.13237728