INTRODUCTION:

Ficus carica L. is a very significant Ficus species. It is frequently called fig^1,2 and is normally deciduous. Vitamins, minerals, carbohydrates, sugars, organic acids, and phenolic compounds have all been found in the dried fruits of F. carica^3-5. Fibre and polyphenols are abundant in both fresh and dried figs^6,7.

Figs are high in phenolic compounds like proanthocyanidins, although red wine and tea, which are also high in phenolic compounds, have lower levels of phenols than figs^8-10.

The medical effects of fig have been utilized for centuries as a metabolic, circulatory, respiratory, antispasmodic, and anti-inflammatory treatment^11-13. F. carica leaves, fruits, and roots are used in traditional medicine to treat gastrointestinal (colic, indigestion, loss of appetite, and diarrhea), respiratory (sore throats, coughs, and bronchial issues), inflammatory, and cardiovascular ailments^14-16.

Iron oxides (IOs) are chemical compounds made up of iron and oxygen. Iron oxide comes in a variety of forms, including iron (II) oxide (wüstite), iron (II, III) oxide (magnetite), and iron (III) oxide (hematite/maghemite). One of the three types of iron oxides is iron (III) oxide, often known as ferric oxide. It is paramagnetic, reddish-brown in color, and is commonly referred to as rust^17-19.

Nanoparticles (NPs) have found widespread use in a range of industries due to their improved features. Because of their large surface area, low toxicity, and simplicity of separation, they are widely used. IONPs smaller than 20 nm, such as maghemite and magnetite, have unique properties^20-24. They're used in biomedical applications to separate and purify cell populations for MRI diagnosis, medicine delivery to a microenvironment, and cellular biology research^24-27. Because of the concentration and exposure time of iron oxide nanoparticles, their toxicity varies. Lower concentrations (10mg/mL) and exposure times (72 hours) allow them to be removed from the body. At large concentrations, they may cause oxidative cellular stress and affect responses such as DNA and gene expression. More research is still needed in this area^28-30.

Plant extracts including green tea leaf, carob leaf, as well as pomegranate leaf have all been found to produce IONPs³¹. The phenolic, as well as nitrogen components, vitamin, carbohydrate, terpenes, and proteins present in the extract, are known as biomaterials. They have reductive, capping, and stabilizing properties. Several variables influence nanoparticle size, shape, and dispersion. The biomaterial percentage, as well as drying temperature, are the most critical. The drying temperature is critical for protecting the biomaterials, whereas the solvents are required for extraction³². We have aimed to synthesize IONPs by using an extract of fig fruit as well as examine the effect of fig extract and IONPs on wound healing in mice.

MATERIALS AND METHODS:

Materials:

Fresh fig fruits are collected from the local garden with permission, whereas ferric chloride hexahydrate and paraffin wax were purchased from Merck industries (Germany).

Preparation of fig fruit extract:

The fruits were washed to remove dust and cut into small pieces. Ten grams of fig pieces were mixed in the blender with 100mL deionized water. Then the mixture was poured intoa beaker and heated at 65°C for 1h. After the mixture was cooled down to room temperature (27°C), it was filtered by using a Whatman No. 1 paper. The filtrate was stored at 4°C.

Preparation of IONPs:

A modified method of the original method reported byDemirezenet al. (2019)³³ was used for synthesizing IONPs from fig fruit extract. 0.14M of ferric chloride hexahydrate solution was prepared by dissolving the 3.8g of ferric chloride in 100mL deionized water, and the solution was stirred with a magnetic stirrer for 15 minutes for optimum dissolving. After that, one volume of the 0.14M FeCl₃ solution was mixed with two volumes of the plant extract. The mixture was stirred with a magnetic bar for 3h (the brown solution was formed). The solution was centrifuged at 1500 x for 10 minutes and the precipitate was dried at 65°C for 12h.

Characterization of IONPs:

The dry powder of IONPs was examined by using a UV-Vis spectrophotometer (Shimadzu 1800, Japan), and X-ray diffraction (XRD, Xpert, Holland). The morphology of IONPs was characterized by using a field emission scanning electron microscope (FESEM; Tescan, France).

In vivo wound healing:

A paraffin cream was prepared from the fig extract and IONPs. The fig fruit was cleaned and cut into small pieces, and then mixed in the blender. The mixture dried to a powder and the powder was grinded. 1g of fig powder or IONPs were mixed with paraffin at 70°C under continuous stirring for 2h.

Three sets of healthy laboratory mice were placed in a cage and pre-conditioned for the experiment for 2 days. Each set was contained 5 mice, and divided according to the treatment as a control set, fig extract set, and IONPs set. The mice's dorsal area was circled, and the area was localized with a 10% lidocaine sprayer to produce a wound in a radius of 1.0cm using a surgical blade, leaving the incision open until redness indicated acute inflammation. The mice were treated daily, as well as the researchers kept track of what happened during the treatment. The diameter of the wound was measured each day for 14 days, and the results were statistically processed by using the ANOVA test.

RESULTS AND DISCUSSION:

Characterization of IONPs:

The UV-Vis spectrum of IONPs solution (Figure 1) has shown a maximum absorbance around 400nm, while the fig extract has shown two peaks one around 300nm and the λmax around 450nm. The conformational change of the peaks and the appearance of λmax around 400nm can be attributed to the formation of IONPs. This was in agreement with Guo et al. (2001)³³, and Ali et al. (2017)³⁴.

Figure 1. UV-Vis absorbance spectrum of IONPs solution.

X-ray diffraction analysis was applied to determine the crystalline nature of synthesized IONPs. All the patterns exhibit the characteristic XRD pattern of hematite (Fe₂O₃) nanoparticles (ICDD card no. 33-0664). X-ray diffractograms of synthesized NPs showed characteristic peaks at a diffraction angle of 2 h at 24.125, 33.115 and 35.612 (Figure 2). The crystal average size appeared 67.43nm. In addition, miller indices appeared as (012), (104) and (110), respectively which matched with the structure of rhombohedral α-Fe₂O₃^35,36.

Figure 2. The XRD diffractogram of IONPs.

The FE-SEM image of Fe₂O₃ showed a rough surface with aggregation of particles in different shapes (multiple shapes) with a particle dimensional range of ~40.75nm, as shown in Figure 3. The morphology of obtained IONPs was disagreed with Demirezenet al. (2019)^37,38, despite that the authors have used TEM not FESEM. The authors have reported a particle average below 10nm of IONPs with spherical shape³⁹.

Figure 3. FESEM image of IONPs.

Wound healing:

Table 1 contains the information obtained from the observations of mice post-injury. The significant effect of both fig fruit extract (1.58±0.11cm) and IONPs (1.58±0.08cm) creams were started after 5 days from the treatment compared to control (1.78±0.08cm). Furthermore, the effect of fig fruit extract (0.64±0.17 cm) and IONPs (0.64±0.11cm) was observed to be highly significant (P<0.01) after the 14^th day from injury which has shown complete healing with no sign of inflammation⁴⁰.

Table 1. The effect of Fig extract and IONPs on wound healing in mice.

Healing Period (day)		Control mice	Fig treated mice	IONPs treated Mice	p-value
1^st	Diameter (cm)	2.0±0.0	2.0±0.0	2.0±0.0	-
1^st	Observations	Redness	Redness	Redness	-
3^rd	Diameter (cm)	1.94±0.09	1.88±0.08	1.94±0.06	0.397
3^rd	Observations	Redness	Light Redness	Light redness	0.397
5^th	Diameter (cm)	1.78±0.08	1.58±0.11	1.58±0.08	0.007*
5^th	Observations	Inflamed area	Clot	Clot	0.007*
7^th	Diameter (cm)	1.74±0.11	1.42±0.11	1.34±0.09	0.0001*
7^th	Observations	inflammation	Clogged	Clogged	0.0001*
10^th	Diameter (cm)	1.52±0.08	1.12±0.08	1.14±0.11	0.0001*
10^th	Observations	Clogged	Small clot	Small clot	0.0001*
14^th	Diameter (cm)	1.42±0.15	0.64±0.17	0.64±0.11	0.0001*
14^th	Observations	Clogged	Small clot	Small clot	0.0001*

Mean ± Standard deviation; * Significant at Pequal to or less than 0.05

The fact that the plant has a lot of active substances that interact with the components of ointment may be linked to the fact that the plant contains a lot of active chemicals that interact with the components of ointment to provide the ointment with this efficient therapeutic role. The makeup of the outer envelope that surrounds the open wound is altered by ointment, allowing it to be protected from external forces. The capacity of the ointment to permeate into the skin tissue boosts the skin's tensile strength and raises the epithelium layer's production height and collagen composition around the wound location, increasing the healing process and restoring the mouse skin's natural state³⁸. Furthermore, reports have shown the possibility of using metal oxide NPs in the wound healing line^41-44.

CONCLUSIONS:

The results of the current study have shown that fig fruit extract is a good efficient, low-cost, and eco-friendlymethod for synthesizing IONPs (α-Fe₂O₃). Also, the use of fig fruit extract and IONPs in the wound healing of skin-injured mice have shown fast and safe healing effects starting from day 5. The use of plant extract and NPs as antibacterial and anti-inflammatory agents would provide a more efficient methodology against bacterial resistance.

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Received on 01.07.2022 Modified on 05.10.2022

Research J. Pharm. and Tech 2023; 16(4):1569-1573.

DOI: 10.52711/0974-360X.2023.00256