Department of Pharmacology, Vidhyabharti college of Pharmacy, camp road, Amravati 444601, Maharashtra India
Osteoarthritis (OA) is a degenerative joint disorder characterized by the progressive breakdown of articular cartilage, changes in subchondral bone, and inflammation of synovial structures. Fagonia arabica, a medicinal plant known for its anti-inflammatory and antioxidant properties, has been traditionally used to treat various ailments. This study aimed to evaluate the anti-osteoarthritic potential of the ethanolic extract of Fagonia arabica (EEFA) in a formaldehyde-induced OA rat model. Wistar rats were divided into five groups: control, formaldehyde-induced (0.1 ml), standard (diclofenac sodium 10 mg/kg), and two EEFA treatment groups (200 mg/kg and 400 mg/kg). OA was induced by subplantar administration of formaldehyde, and treatments were administered orally for 4 weeks. Paw diameter, body weight, and swelling ratio were assessed to evaluate the anti-osteoarthritic effects. Phytochemical analysis of EEFA revealed the presence of carbohydrates, glycosides, saponins, phenols, tannins, and flavonoids. EEFA treatment significantly reduced paw diameter and swelling ratio while improving body weight compared to the formaldehyde-induced group. The 400 mg/kg EEFA dose showed efficacy comparable to diclofenac sodium. Histopathological findings corroborated the anti-inflammatory effects of EEFA, with reduced joint damage and inflammation. These results suggest that Fagonia arabica possesses anti-inflammatory and anti-osteoarthritic properties, which may be attributed to its bioactive constituents, such as flavonoids and tannins. Further clinical studies are warranted to validate the therapeutic potential of Fagonia arabica in the management of osteoarthritis.
Osteoarthritis is a degenerative joint disease that occurs when the protective cartilage cushioning the ends of bones wears down over time to pain localized to a joint, regardless of the origin of the pain (which may or may not be due to joint inflammation). Arthritis is a term used to describe inflammation of the joints, which can lead to symptoms such as pain, stiffness, swelling, and reduced range of motion. It is a common condition that affects people of all ages, though it becomes more prevalent with age The condition arises from various causes, including wear and tear of cartilage (as in osteoarthritis), autoimmune reactions (as in rheumatoid arthritis), or the accumulation of uric acid crystals (as in gout). Other factors like infections, genetics, and lifestyle choices can also contribute to its development There are several types of arthritis. The most common forms are osteoarthritis (most commonly seen in weightbearing joints) and rheumatoid arthritis, Osteoarthritis usually occurs as an individual ages and often affects the hips, knees, shoulders, and fingers. Rheumatoid arthritis is an autoimmune disorder that often affects the hands and feet. Other types of arthritis include gout, lupus, and septic arthritis. These are inflammatory based types of rheumatic disease. Osteoarthritis is the most common form of arthritis affecting more than 3.8% of people, while rheumatoid arthritis is the second most common affecting about 0.24% of people. OA commonly affects the knees, hips, hands, and spine, and symptoms tend to worsen over time. Risk factors include aging, joint overuse or injury, obesity, genetics, and being female, particularly after menopause. Osteoarthritis (OA) is a degenerative joint disorder and the most prevalent form of arthritis, particularly affecting the elderly population. Age-related osteoarthritis refers to the form of OA that develops as a natural consequence of aging. It is characterized by the progressive breakdown of articular cartilage, changes in subchondral bone, and inflammation of synovial structures, leading to joint pain, stiffness, and functional limitations. Fagonia is a genus of wild, flowering plants in the caltrop family, Zygophyllaceae, having about 34 species. Fagonia arabica is a tropical herb found in the entire Indian subcontinent and it is commonly known as Dhamasa, Suchi boti, dhamanian kunda, Damoo, Shaukat-e-Albeefa and Shokat-e-albaiza. It is a green herb of about 1 to 3 feet in height often grown on calcareous rocks mostly in Africa, Afghanistan, India and Pakistan. Fagonia species are widely studied due to their antitumor, antioxidant, analgesic, astringen febrifuges and prophactic against small pox agents. They are also known traditionally for treatment of cancer, fever, asthma, toothaches, urinary diseases, stomach problems and kidney diseases. Fagonia Arabica is an ethno-pharmacologically ayurvedic herb known for many medicinally important properties. It has anti-inflammatory, analgesic and antipyretic effect. Different parts of F. arabica are used to treat different ailments like hematological, neurological, endocrine and inflammatory disorders. Its twigs are used as treatment for snake bite and its paste is applied on tumors and swollen portions of neck. F. arabica is traditionally used for treatment of inflammation, open wounds, boils, skin diseases and various allergies. Dhamasa is found to be thrombolytic agent for the treatment of Atherthrombotic disease. Fagonia arabica is used as alcoholic and hydroalcohlic extract for the treatment of microbial infections. Fagonia arabica contains a variety of phytochemicals with potential medicinal properties. Some of the key phytochemicals identified in this plant include flavonoids, saponins, sterols, glycosides, tannins, alkaloids, carbohydrates, and amino acids. These compounds contribute to its antioxidant, antimicrobial, and pharmacological activities, making it a valuable plant in traditional medicine. Due to the Presence of flavonoids, Polyphenols and Tannins. The present study was planned to anti-osteoarthritis potential of Fagonia arabica in formaldehyde induced osteoarthritis in experimental animal.
MATERIALS AND METHODS
Collection and Authentication of Plant :
The leaves, and stem of Fagonia arabica were collected from regional farm, Amravati District, Maharashtra, India. The plants were identified and authenticated by Professor L. P. Khalid, Department of Botany, Vidyabharati Mahavidyalaya, Amravati. The leaves and branches were cleaned, dried in shade. The leaves and branches were kept in air tight container for further studies.
Extraction Process
The whole plant including its all parts of Fagonia arabica was processed by washing with clean water, air-drying, pulverizing, and sieving through a 0.3 mm sieve. tool consists of several parts including a heat round bottom flask, Soxhlet extractor, and condenser. The solid coarsely powdered plant (250g) was placed in thimble and placed in an extractor. The bottom end of the extractor was connected to a round bottom flask containing a solvent ( Ethanol 1000ml was chosen as the solvent), and was connected to a reflux condenser. The bottom flask was heated to boil the solvent (Ethanol), the vapor rises through the branch pipe of the extractor, was condensed and drops into the thimble and the solvent (ethanol) was contacted with the solid for extraction. When the solvent (ethanol) surface exceeds the highest point of the siphon, the solvent containing the extract was return back to the round bottom flask. This cycle was repeated until the all the material extracted from the solid plants powder.
The percentage yield of the extract was calculated and the extract was then subjected to different phytochemical tests.
ANIMAL AND HOUSING CONDITION.
Wistar Rats of either male or female sex weighing 200-250 gm was obtained from the animal house of Department of Pharmacology, Vidyabharati College of pharmacy Reg. No: 1504/PO/RE/S/11/CPCSEA, Amravati. All the animal are acclimatized to the animal house prior to use. They are kept in case in animal house with a 12 hr light: 12hr dark cycle at temperature (25°C 1°C) with 50+ 55% of relative humidity. Experiments was performed in accordance with the committee for the purpose of control and supervision of experimental animal (CPCSEA) guideline after the approval of the experimental protocol by the institutional animals ethical committee (IAEC). Animal are fed on pellets and tap water ad libitum. The care and handling of animals in accordance with the internationally accepted standard guidelines of use of animals (CPCSEA).
DRUGS AND CHEMICALS
Diclofenac sodium tablet 10m (Mfg. By Elam Pharma) purchased from Shiv Medical Chaprashipura Amravati, Dist. Amravati,Maharashtra , 444503.
Preparation of Doses
Diclofenac sodium tablet triturated and was diluted to 10 mg/100 ml with distilled water. Two different concentrations ( 200 mg/kg, and 400 mg/kg) of the EEFA were prepared by dissolving the extracts in distilled water. All solutions were freshly prepared at the time of administration to the animals. Extract solution and vehicle (0.9% NaCl) were given orally and inducing drug (Formaldehyde 0.2 %) Subplantar route of Administration & standard drug (Diclofenac sodium) orally
PHYTOCHEMICAL ANALYSIS
The Ethanolic extract of Fagonia arabica undergoes different phytochemical tests for identification of different phytochemical such as Flavonoids, Alkaloids, Tannins, Steroid, etc
EXPERIMENTAL PROCEDURE
Wistar rat weighing 150-200 g will house under standard laboratory condition of light and dark cycle (12:12). Osteoarthritis will induce by 0.1 ml formaldehyde (2%v/v) by sub-planter administration into the left hind paw in all groups ; (except normal control group) for 7 days. The groups were divided into 5 groups (n=6/group) and assigned as control, Inducing , standard and two different test dose groups of ethanolic extract of Fagonia arabica. First test group administered with 200 mg/kg and second test group is administered with 400mg/kg respectively by oral administration for 4 weeks. Arthritis will assess using serum Hb, paw volume, joint diameter, body weight and ESR rate investigation. Formaldehyde induced osteoarthritis in experimental animal model would be adopted for assessing Anti-osteoarthritis activity.
1. Paw Diameter :
vernier calliper is used to assess the paw diameter by comparison between induced day and the day after treatment by the two different doses of extract and standard drug by oral administration
2. Body weight :
As induction of the OA in the induced group of the experimental animals resulted into the decreased in the body wight of the induced group.
3. Swelling Ratio
The swelling ratio is generally calculated as the change in volume or mass after swelling divided by the original volume or mass. And it is studied by the comparison study by the common formula is.
Swelling Ratio = DS -- Dd
Dd
Where , Ds = Diameter After FA Induced
Dd = Diameter After Treatment/ Observations
RESULT
1. PHARMACOGNOSTICAL EXAMINATION
Phytochemical Analysis of EEFA
|
Sr. No |
Chemical Constituents |
Results |
|
1 |
Carbohydrate |
+ |
|
2 |
Glycosides |
+ |
|
3 |
Alkaloids |
- |
|
4 |
Phytosterols |
- |
|
5 |
Saponins |
+ |
|
6 |
Phenols |
+ |
|
7 |
Tannins |
+ |
|
8 |
Flavonoids |
+ |
Where (+) indicates Present & (-) indicates Absent
Phytochemical testing carried out to find out the secondary metabolite because secondary metabolic possess biological activity. Phytochemical studies of Fagonia arabica performed for the presence of Carbohydrate, Glycosides, Phytosterols, Alkaloids, Saponins, Tannins, Flavonoids, Protein and Amino acids.
2. PHARMACOLOGICAL EXAMNIATION
Table No.01 : Effect of EEFA on Paw Diameter
|
Day |
Control |
FA 0.1 ml |
DFS 10mg/kg |
EEFA 200mg/kg |
EEFA 400mg/kg |
|
0 |
3.00 ± 0.03 |
3.00 ± 0.03 |
3.00 ±0.03 |
3.00 ± 0.03 |
3.00 ± 0.03 |
|
7 |
3.10 ± 0.03 |
4.50 ± 0.03 |
3.20 ±0.03* |
4.00 ± 0.03* |
3.80 ± 0.03* |
|
14 |
3.30 ± 0.03 |
5.00 ± 0.03 |
3.40 ±0.03* |
4.20 ± 0.03* |
4.00± 0.03* |
|
21 |
3.40± 0.03 |
5.50 ± 0.03 |
3.50 ±0.03* |
4.10 ± 0.03* |
3.90 ± 0.03* |
All Values were expressed as mean ± SEM, (n=6), (*p<0.0001) compared to each group compared with each day on 0,7,14,21, (*p<0.0001) followed by Two-way ANOVA using Tuckey’s test. On day 21, both EEFA-treated groups maintained reduced paw diameters, suggesting sustained anti-arthritic activity. These findings indicate that EEFA effectively inhibits the progression of inflammatory swelling in formaldehyde-induced arthritis. The anti-oedematous effect may be attributed to the presence of bioactive constituents in Fagonia arabica that possess anti-inflammatory, antioxidant, and anti osteo arthritic property.
Figure No.1 : Effect of EEFA on the Paw diameter
2. Body weight
Table No. 2 : Effect of EEFA on Body weight
|
Day |
Control |
FA 0.1ml
|
DFS 10mg/kg |
EEFA 200mg/kg |
EEFA 400mg/kg |
|
0 |
180.33 ± 0.4 |
179.33 ± 0.4 |
180.16 ± 0.4 |
179.66 ± 0.4 |
179.33 ± 0.4 |
|
7 |
189.50 ± 0.4 |
174.50 ± 0.3 |
188.50 ± 0.4* |
181.50 ± 0.4* |
183.50 ± 0.4* |
|
14 |
196.50 ± 0.4 |
171.50 ± 0.4 |
195.50 ± 0.4* |
179.50 ± 0.4* |
182.50 ± 0.4* |
|
21 |
203.50 ± 0.4 |
168.50 ± 0.4 |
202.50 ± 0.4* |
177.50 ± 0.4* |
180.50 ± 0.4* |
All Values were expressed as mean ± SEM, (n=6), (*p<0.0001) compared to each group compared with each day on 0,7,14,21, (*p<0.0001) followed by Two-way ANOVA using Tuckey’s test. In the present study, a consistent trend was observed across all groups over the 21-day experimental period. In the formaldehyde (0.1 mL)-induced arthritic group, there was a progressive decline in body weight from day 0 to day 21. , rats treated with the ethanolic extract of Fagonia arabica (EEFA) at both 200 mg/kg and 400 mg/kg demonstrated a dose-dependent protective effect on body weight. Specifically, body weight in the formaldehyde group differed significantly from all treatment groups at each time point, confirming the protective and therapeutic role of EEFA.
Figure No. 2 : Effect of EEFA on Body weight
3. Sweeling ratio :
Table No. 3 : Effect of EEFA on Sweeling ratio
|
Day |
Control |
FA ( 0.1ml )
|
DFS 10mg/kg |
EEFA 200mg/kg |
EEFA 400mg/kg |
|
0 |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
0.00 ± 0.00 |
|
7 |
4.16 ± 0.03 |
52.50 ± 0.76 |
22.50 ± 0.76* |
37.50 ± 0.76* |
32.50 ± 0.78* |
|
14 |
3.16 ± 0.03 |
57.50 ± 0.76 |
16.00 ± 0.57* |
32.50 ± 0.57* |
27.00 ± 0.57* |
|
21 |
3.16 ± 0.03 |
54.50 ± 0.76 |
11.33 ± 0.42* |
30.00 ± 0.57* |
22.50 ± 0.76* |
All Values were expressed as mean ± SEM, (n=6), (*p<0.0001) compared to each group compared with each day on 0,7,14,21, (*p<0.0001) followed by Two-way ANOVA using Tuckey’s test.
The swelling ratio (%) serves as a direct index of the anti-inflammatory efficacy of test substances, In the current study, the formaldehyde (0.1 mL) group exhibited a marked increase in paw swelling, with swelling ratios peaking at days 7 and 14, remaining significantly elevated until day 21. The swelling ratio in the EEFA groups was significantly lower than in the untreated arthritic group, with the 400 mg/kg dose approaching the standard drug in efficacy.
Figure No. 3 : Effect of EEFA on swelling ratio
DISCUSSION
To investigate the Anti-osteo arthritic potential of EEFA on Formaldehyde-induced osteo arthritis ESR (Erythrocyte Sedimentation Rate), C-reactive protein test, White blood cells count, Haemoglobin Count, Paw diameter, body weight, sweeling ratio are evaluated Formaldehyde-induced arthritis (FIA) is a widely used experimental model to study inflammatory arthritis and evaluate potential anti-arthritic agents. It involves the injection of formaldehyde into the joint or paw of rodents, leading to a biphasic inflammatory response that mimics aspects of human arthritis Formaldehyde induces the release of pro-inflammatory mediators such as histamine, serotonin, bradykinin, substance P, and prostaglandins These mediators cause vasodilation and increased vascular permeability, leading to edema and pain at the injection site the NSAID’s drugs is Diclofenac sodium is a widely used nonsteroidal anti-inflammatory drug (NSAID) that effectively manages pain and inflammation associated with various forms of arthritis, including osteoarthritis and rheumatoid arthritis.[86] Its therapeutic effects are primarily attributed to its multifaceted mechanisms of action, which extend beyond traditional COX inhibition The diverse mechanisms of action of diclofenac not only enhance its effectiveness in treating arthritis but also may offer advantages over other NSAIDs, particularly in terms of pain relief and anti-inflammatory effects. However, these benefits must be weighed against potential side effects, especially with long-term use. In the overall research of the evaluation of the anti-osteoarthritic potential of ethanolic extract of fagonia arabica is used as the test drug in the this study and sodium diclofenac is used as the standard drug in this study to evaluate the comparative its EEFA 200mg/kg and 400mg/kg action on the formaldehyde induced rats of either sex male or female, formaldehyde is administered in the left hind of the rats for the induction of the osteoarthritis and by the oral route drugs are administered in the two test dose as 200mg/kg and 400mg/kg to the test animals The ethanolic extract of Fagonia arabica demonstrated significant anti-osteoarthritic effects in the formaldehyde-induced rat model. The observed reduction in paw volume and joint diameter suggests effective anti-inflammatory activity. The improvement in body weight indicates enhanced general health and reduced discomfort. Histopathological findings corroborate the anti-inflammatory effects, with reduced joint damage and inflammation The 400 mg/kg dose of Fagonia arabica extract exhibited efficacy comparable to diclofenac sodium, a standard NSAID Both doses of Fagonia arabica extract (200 mg/kg and 400 mg/kg) demonstrated a significant reduction in paw volume and joint diameter compared to the arthritic control group.
The 400 mg/kg dose showed effects comparable to diclofenac sodium, indicating a dose-dependent response. These findings suggest that Fagonia arabica possesses anti-inflammatory properties that mitigate joint swelling and edema associated with OA.Rats in the arthritic control group exhibited a significant decrease in body weight due to pain and reduced mobility. Treatment with Fagonia arabica extract at both doses resulted in a notable increase in body weight, approaching that of the normal control group. This improvement indicates the potential of Fagonia arabica in enhancing overall health and well-being in arthritic rats. Fagonia arabica is a medicinal herb with established anti-inflammatory activity, known for its ability to reduce inflammation and pain. It's also recognized for its analgesic and antipyretic effects, making it a traditional remedy in Ayurvedic medicine for various ailments. The plant's anti-inflammatory properties are attributed to its chemical composition, which includes flavonoids, triterpenoidal glycosides, and saponins.
Due to the presence of flavonoids and tannins in Fagonia arabica may have potential as anti-antarthritic, anti-inflammatory, anti-oxidant activity. These phytochemicals may inhibit pro inflammatory mediators and may possess anti osteo arthritic activity.
CONCLUSION:
Research on the anti-osteoarthritic action of ethanolic extract of Fagonia arabica in formaldehyde-induced rat models suggests promising results. Studies indicate that the extract contains bioactive compounds such as flavonoids, phenols, saponins, and alkaloids, which contribute to its anti-inflammatory and antioxidant properties, These compounds help reduce joint inflammation, improve haematological parameters, and alleviate osteoarthritic symptoms.
While further clinical validation is needed, the findings support the potential therapeutic role of Fagonia arabica in osteoarthritis management the present experimental findings of pharmacological, radiological, histological and hematological parameters observed from the current investigation, it is concluded that at the doses of 200mg/kg and 400 mg/kg EEFA possesses potentially useful anti- osteoarthritic activity since it gives a positive result in controlling inflammation in formaldehyde induced arthritis model in rats. The high dose of EEFA reflected highly beneficial and treatment of Osteoarthritic disorders.It is concluded from the up taken research that a correlation exists between indigenous knowledge and currently studied anti osteo arthritic activity findings. Fagonia arabica can be used for the extraction of important compounds that have activity.This study can be helpful to revolutionize the pharmaceutical industry. The ethanolic extract of Fagonia arabica has demonstrated significant anti-osteoarthritic effects in formaldehyde-induced rat models. Treatment with the extract resulted in a notable reduction in paw oedema and joint swelling, indicating effective anti-inflammatory activity. Histopathological examinations revealed decreased cartilage degradation and inflammatory cell infiltration, suggesting protective effects on joint tissues. Biochemical analyses showed normalization of various inflammatory markers, further corroborating the therapeutic potential of the extract. The conclusion from studies on the anti-arthritic activity of ethanolic extracts of Fagonia arabica on formaldehyde-induced arthritis in rats is that the extract demonstrates promising anti-inflammatory and anti-arthritic effects. It effectively reduces markers of arthritis, including paw swelling, joint diameter, and arthritis scores. Furthermore, the extract's ability to reduce inflammation and potentially prevent joint damage suggests its potential as a therapeutic agent for osteoarthritis.
REFERENCES
Durgesh Gavande*, Dr. Anjali wankhade, Dr. Vivek Paithankar, Anti-Osteoarthritis Potential of Fagonia Arabica in Formaldehyde Induced Osteoarthritis in Experimental Animal, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 6, 1207-1216. https://doi.org/10.5281/zenodo.15608509
10.5281/zenodo.15608509
