Protective effects of colchicine against osteoarthritis in rat induced by monosodium iodoacetate

doi:10.21203/rs.3.rs-5174979/v1

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Research Article

Protective effects of colchicine against osteoarthritis in rat induced by monosodium iodoacetate

https://doi.org/10.21203/rs.3.rs-5174979/v1

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Journal Publication

published 31 May, 2025

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Background

Knee osteoarthritis (OA) is a debilitating condition that can severely limit an individual’s mobility and quality of life. This study evaluated the efficacy of colchicine therapy in promoting cartilage healing in a rat model with monosodium iodoacetate (MIA)-induced knee OA. This was assessed through semiquantitative radiographic features as well as histological and biochemical alterations.

Methods

Rats were given an intra-articular injection of MIA on day zero to induce knee OA. After that, 40 Wistar albino female rats were split into 4 groups at random (10 rats/each group): a negative control group, an osteoarthritic control group, an osteoarthritic reference group receiving a meloxicam^®, and an osteoarthritic group receiving an intraperitoneal injection of colchicine. The body weight and knee diameter were recorded once per week. Semiquantitative radiographic imaging and enzyme-linked immunosorbent assay (ELIZA) analysis of serum inflammatory cytokines interleukin-1beta (IL-β) and anti-inflammatory cytokines interleukin-10 (IL-10), were carried out before the end of the trial. Finally, hematoxylin/and eosin stains were used for histological investigation.

Results

Colchicine significantly reduced the osteoarthritic conditions after six weeks of supplementation. We observed decreased joint diameters in response to treatment in OA animals. Colchicine significantly decreased IL-1β (p = 0.000) and increased the IL-10 (p = 0.000) in the serum of osteoarthritic rats in response to treatment in OA rats compared to the OA group with no treatment. Colchicine improved the histological structure of the knee joint and lowered the radiographic scores of osteoarthritic rats.

Conclusion

These results point to the potential benefit of colchicine in treating rats with MIA-induced knee OA by postponing cartilage deterioration and reducing the activity of inflammatory mediators.

knee osteoarthritis

colchicine

Rats

monosodium iodoacetate

One of the main causes of disability in the elderly is osteoarthritis (OA) of the knee. There is currently no approved medication that can decrease the disease's course, despite the disease's high frequency and widespread effects [1]. It has been noted that OA and hyperuricemia can coexist. We observed substantial associations between serum uric acid and the radiographic and histological severity of knee OA in a population of patients without clinical gout [2]. Additionally, we saw a significant relationship between the radiographic severity of osteoarthritis in the knee and serum uric acid and interleukin, which is generated by macrophages via the inflammasome assembly triggered by uric acid crystals [3]. It was postulated that the degeneration of cartilage in OA would promote the deposition of crystals of monosodium urate, a mechanism that aids in the advancement of OA [4]. This understanding of the pathophysiology of OA supports the idea that colchicine, an established gout therapy, may make a compelling case for being a successful OA treatment. Rats can be made to develop arthritis by injecting monosodium iodoacetate (MIA) into the knee joint. It causes meniscus and ligament damage as well as decreased stability. As a result, the mechanical stress affects the metabolism of cartilage and synovial cells, mimicking the symptoms of osteoarthritis [5]. For this reason, medicinal compounds with potential to treat osteoarthritis are usually investigated using the MIA-induced rat model [6]. In order to examine the effects of colchicine in an MIA-induced osteoarthritis rat model, the articular structures of the knee joint were subjected to semiquantitative radiographic characteristics, haematoxylin and eosin (H&E) staining, and analysis of the inflammatory cytokines.

2.1. Type of study:

It was an experimental-based study that was carried out in the department of radiology and physiology. Ethics was approved by the Research Ethics Committee considering the care and use of laboratory animals (No. : Soh-Med-22-07-12) and were performed in compliance with the recommendations of the National Institutes of Health Guide for Care and Use of Laboratory Animals (publication no. 85 − 23, revised 1985).

2.2. Animals:

In this investigation, forty nonpregnant female Wistar rats were used. The rats weighed between 180 and 250 grammes on average when they were about 3 months old. They were provided with unlimited access to food and tap water. The rats were kept in 20 x 32 x 20 cm polycarbonate cages in groups of five.The housing conditions included a normal light-dark cycle and a temperature of 23 ± 2°C in a room equipped with air conditioning at the Experimental Animals General Services Centre. Prior to the commencement of the study, the rats were kept under observation for a period of 10 days to ensure the absence of any infections. Throughout the study, no rats were lost, and at the end of the designated time point, all rats were euthanised using an overdose of inhaled isoflurane.

2.3. Reagents

Monosodium iodoacetate (MIA) was acquired from Sigma-Aldrich, located in St. Louis, MO, USA. The Egyptian manufacturer, El Nasr Pharmaceutical Chemicals Co., provided the colchicine. Meloxicam® was purchased from Global Pharmaceutical Industries Co., also located in Egypt. For the measurement of serum IL-1β, the Rat IL-Iβ ELISA Kit was obtained from ELK Biotechnology Co., Ltd., which is based in China. Additionally, the Rat IL10 (Interleukin 10) ELISA Kit (ELK) was acquired from Biotechnology Co., Ltd., also based in China, for the measurement of serum IL-10.

2.4. Induction of the OA model

To construct the OA model on the first day, a single injection of 0.3 mg of MIA mixed with 50 µL of sterile saline was administered directly into the joint, as was previously reported [7]. Following the administration of anaesthesia, the patellar ligament was exposed through a surgical incision made in the middle of both knees. Every rat was placed on its back, with its leg bent at the knee joint at a 90° angle. Using a 26.5-gauge 0.5-inch needle, MIA was gently injected into the inner side of the knee ligaments in both knees, taking care to avoid the cruciate ligaments. After the injection, the animals were observed every day by qualified individuals who were not informed of the specifics of the treatment in order to evaluate any knee joint swelling or dysfunction.

2.5. Animal Grouping and Experimental Design

Following the accommodation period, four groups consisting of ten Wistar rats each were randomly chosen: the MIA-induced OA group, which involved injecting isotonic sterile saline containing MIA once intra-articularly into the right knee joint; the MIA + meloxicam® group, which received daily subcutaneous injections of meloxicam® (0.2 mg/kg) as an anti-inflammatory protocol [8]; and the MIA + colchicine group, which received treatment with colchicine following the MIA injection. Colchicine was administered intraperitoneally once a day (0.5 mg/kg) for 4 weeks [9]. Every other day, animals were watched to measure body weights and knee diameters. At the end of the experiment, serum was used for the determination of inflammatory biomarkers (IL-10 and IL-1β). Radiographic imaging and histological examination of the rats' knees were also performed. In Fig. 1, a flowchart illustrating the study design is displayed.

2.6. Radiographic Assessment.

Radiographs of the right joints were performed using X-ray equipment (Fuji Medical Systems Corporation, Japan) equipped with a digital detector (FDR D-EVO wireless digital radiography system, Fuji, Japan) on days 42 post-OA induction and 28 post-colchicine therapy. After thorough disinfection, the knees were placed on the tape. The digital radiographic images of each affected knee included two projections of craniocaudal and lateral views in the same manner by the same radiographer. The following exposure parameters were employed to get the radiographs: 10 mA tube current, 10–30 kV tube voltage, 1 m object-focus distance, and 10–100 ms exposure time. One radiologist with fifteen years of experience independently evaluated the images, and the image processing was based on the sets of image-processing parameters provided by the manufacturers. Prior to the official assessment of image quality, a radiologist with experience evaluated the sets of images acquired from multiple rats using different processing parameters. The objective was to optimise the brightness, contrast resolution, and detailed enhancement of the images while standardising the parameters. Following the identification of the ideal settings, postprocessing was carried out, and the image data from the radiography using a flat-panel detector was sent to a shared workstation (Easy-Vision, Philips Medical Systems, Hamburg, Germany). A 19-inch screen computer was used to perform the measurements. Based on the Kellgren and Lawrence (KL) score, which divides radiographic lesions into five phases of escalating severity [10], the knees were categorized. It correlated with an index that considered joint space in addition to osteophytes. The measurements were done by a single author (MI, with 15 years of radiology experience). This KL score included five grades: no, minimal, mild, moderate, and advanced arthritis, as represented in Table 1.

Table 1

Kellgren-Lawrence Grading System for OA
Grades	Radiographic findings
0	No radiographic features of OA are present
1	Doubtful JSN and possible osteophyte lipping
2	Definite osteophytes and possible JSN on anteroposterior weight-bearing radiograph
3	Multiple osteophytes, definite JSN, sclerosis, and possible bony deformity
4	Large osteophytes, marked JSN, severe sclerosis, and definite bony deformity
JSN, joint space narrowing; OA, osteoarthritis

2.7. Assessment of body weight (gm) and knee diameter (mm):

Body weight

body weight was measured before induction (week 0), then measured every week until the end of the experiment.

Knee diameter

right knee diameter was measured before induction (day 0) and after induction (day 1), then measured every week in the experiment using a manual calliper until the end of the experiment.

2.8. Assessment of serum levels of interleukin (IL-1b and IL-10).

On day 42, blood samples were taken from all animal groups (control and other experimental groups) by deep ether inhalation in order to evaluate the levels of cytokines. Heparinised capillary tubes in dry, clean screw-capped tubes were used to draw fresh blood samples from the rats' neck veins. The samples were then allowed to coagulate for 30 minutes at room temperature and separated by centrifugation at 3000 rp.m. for ten minutes. The clear, pure serum was transferred into dry, sterile Eppendorf tubes using automated micropipettes. Aliquots of serum were kept at -20°C until the levels of IL-1b and IL-10 were measured. The thawed serum aliquots were used using a commercially available rat ELISA kit from ELK Biotechnology Co., Ltd. to quantitatively estimate the serum IL-1b and IL-10 concentrations. Every step was completed in accordance with the manufacturer's guidelines.

2.9. Tissue sampling and histological analysis of the joints

Rats from both the experimental and control groups were slaughtered and their knee joints dissected to obtain tissue samples at the conclusion of the experimentation period, which occurred 42 days following the adjuvant injection. The soft tissues surrounding the right and left knee joints were removed, and the joints themselves were dissected. For 48 hours, the knee joints were fixed in 10% paraformaldehyde. The joints were imbedded in a paraffin block after being decalcified for three weeks in 10% ethylenediaminetetraacetic acid (EDTA). The materials underwent light microscopy examination or were cut into 5-um slices and stained with hematoxylin, eosin, and periodic acid Schiff (PAS) staining. The histopathology of OA is graded using the modified Mankin grading system (Table 2). This technique was initially created to evaluate the cartilage in the hips of OA patients. Since then, it has also been used to assess cartilage regeneration, healing, and degeneration in a variety of OA animal models. Four factors are evaluated by the MANKIN system: tidemark integrity, cellularity, Safranin O staining, and cartilage structure. There are subcategories for each parameter, and the quantity of the scores for each yields a total score that varies from 0 (normal) to 14 (the most severe OA) [11].

Table 2

The modified Mankin score of microscopic observation of OA articular cartilage.
Category	Subcategory	Score
Structures	Normal	0
	Surface irregularities	1
	Pannus and surface irregularities	2
	Clefts to transitional zone	3
	Clefts to radial zone	4
	Clefts to calcified zone	5
	Complete disorganization	6
Cells	Normal	0
	Diffuse hypercellularity	1
	Cloning	2
	Hypocellularity	3
Safranin-O staining	Normal	0
	Slight reduction	1
	Moderate reduction	2
	Severe reduction	3
	No dye noted	4
Tidemarks	Intact	0
Tidemarks	Damaged	1
Mankin score is the sum of structure, cells, Safranin-O staining and tidemark integrity.

MI and NA were aware of the group allocation at the different stages of the experiment (during the allocation, the conduct of the experiment, the outcome assessment, and the data analysis).

All the values are presented as means ± standard error. Comparisons between different groups were done using one-way ANOVA, followed by Tukey's multiple comparison post hoc tests. A Kruskal-Wallis ANOVA test followed by a Mann-Whitney U test was performed for non-parametric scoring. P < 0.05 was considered significant. SPSS for Windows, version 22.0 (SPSS, Chicago, IL), was used to perform these statistical tests.

4.1. Effect of colchicine on body weight and knee diameter measurements

a) Body weight:

The control group, OA-induced group, and meloxicam® group showed an increase in mean body weight through the experiment from week 0 to week 6. In contrast, the colchicine group shows a decrease in body weight from week 1 to week 3, then there is an increase in body weight till the end of the experiment (see Fig. 2). However, the increase in body weight in the colchicine group was relatively less than in other groups.

b) Knee diameter:

The mean knee diameter does not vary in the control group. As can be seen in Fig. 3, the knee diameters of the OA-induced groups treated with colchicine and meloxicam® increased after induction from day 1 to day 21, at which point only the colchicine group began to improve relative to the meloxicam® group. However, the improvement was gradual and did not reach the normal diameter.

4.2. ELIZA Evaluation

Effect of colchicine on the serum proinflammatory cytokine IL-1β :

The OA-induced (OA) group (16.025 ± 4.158 pg/ml) revealed a significant increase in IL-1b level when compared to the control group (0.62 ± 0.424 pg/ml) (P = 0.000). The two treated groups, the colchicine-treated group (5.984 ± 10.47 pg/ml) and the meloxicam®-treated group (9.811 ± 4.894 pg/ml), revealed significant decreases in IL-1b levels when compared to the OA-induced group (p = 0.000) and (p = 0.025), respectively. As seen in Figs. 4 and 5, there was no discernible difference between the groups receiving colchicine and meloxicam® (p = 0.094).

Effect of colchicine on the serum anti-inflammatory cytokine IL-10

The OA group revealed a significant increase in IL-10 level (9.568 ± 2.987 pg/ml) when compared to the control group (1.815 ± 0.879 pg/ml) (p = 0.000). The two treated groups, the colchicine-treated group (73.957 ± 21.099 pg/ml) and the meloxicam®-treated group (13.601 ± 1.609 pg/ml), revealed a significant increase in IL-10 levels when compared to the OA-induced group (p = 0.000) and (p = 0.008, respectively. The colchicine-treated group revealed a significant increase in IL-10 level when compared with the meloxicam®-treated group (p = 0.000); see Fig. 5.

4.3. Effect of colchicine on histological changes:

The knee joint examination of the control group showed intact tidemarks and a normal histological composition of the articular cartilage, subchondral bone plate, and joint capsule (Fig. 6). Conversely, the group of osteoarthritic rats (MIA group) exhibited localised articular cartilage erosion, a discernible reduction in articular cartilage thickness in certain regions, and a loss of normal chondrocyte zonal distribution in other regions. There was also evidence of articular cartilage fibrillation. In certain areas, the tideline was nonexistent (Fig. 7).

Rats with osteoarthritis who received colchicine treatment showed improvement in the architecture of the knee joint, a regular articular cartilage surface, and focal erosion in some areas. Some areas show a focal absence of the articular cartilage. Additionally, a reduction in joint space width was also observed (Fig. 8). In contrast, the group treated with meloxicam® exhibited a persistent manifestation of OA characterised by reduced staining affinity and fissuring of the articular cartilage, with evidence of trabeculae destruction (Fig. 9). Furthermore, the modified Mankin system's total score was significantly lower in the group treated with colchicine for osteoarthritis (P < 0.05) compared to the control group with OA. This finding confirms the protective effects of colchicine treatment on preventing further cartilage destruction in knee joints affected by OA (Fig. 10).

4.3. Effect of colchicine on radiographic changes:

The KL score was significantly increased by the MIA administration (p < 0.001). However, arthritic rats treated with colchicine presented a lower degree of joint changes according to the KL classification (p < 0.05 and p < 0.01; Fig. 11).

The knees of the MIA-treated group exhibited OA changes, including cartilage deterioration, as demonstrated by the presence of minute marginal osteophytes, a narrow joint space, and bone erosions when compared to those of the normal control rats (Fig. 12). Eight rats (80.0%) with normal and healthy joints (Grade 0) were shown on the MIA-treated group radiographs; one rat (10%) had dubious changes, and another (10%) had mild OA changes (Grades 1 and 2, respectively) (Figs. 13, 14).

The reference group that received meloxicam® injections showed no radiographic changes of OA in one rat (10%), doubtful changes in four (40%), and moderate arthritis in two (20%) of the rats; the rats were graded as grade 0, grade 1, grade 02, and grade 3. The radiographic alterations of the colchicine group were rated as grade 1 and grade 2, respectively, showing no radiographic changes of OA in five rats (50%), dubious changes in four rats (40%), and modest changes in one radiograph (10%), Table 3.

Table 3

Garde of osteoarthritis among experimental groups
Group	Grade osteoarthritis Count, (%)
	0	I	II	III	IV
	No	Minimal	Mild	Moderate	Advanced
Control	8, (80.0%)	1, (10.0%)	1, (10.0%)	0, (.0%)	0, (.0%)
MIA	1, (10.0%)	2, (20.0%)	3, (30.0%)	2, (20.0%)	2, (20.0%)
Colchicine	5, (50.0%)	4, (40.0%)	1, (10.0%)	0, (0%)	0, (.0%)
Meloxicam®	1, (10.0%)	4, (40.0%)	3, (30.0%)	2, (20.0%)	0, (.0%)
Total	15,(37.5%)	11, (27.5%)	8, (20.0%)	4, (10.0%)	2, (5.0%)

Compared to the meloxicam® group, the radiographs collected following therapy demonstrated a significant improvement in the osteoarthritic characteristics in the colchicine group (Fig. 15, 16). Based on Fig. 17, it may be inferred that the meloxicam® group had a statistically significant greater degree of arthritis (p = .02) than the colchicine group. With a mean rank arthritis score of 7.6 for colchicine and 13.4 for meloxicam®, a Kruskal-Wallis H test revealed a statistically significant difference in grade of arthritis between the two forms of treatment (p = .02).

An analysis of variance using one-way ANOVA revealed a statistically significant difference (p = 0.003) between the groups. The degree of arthritis was statistically reduced after taking colchicine compared to meloxicam®, according to a Tukey post hoc test (p = 0.5). Between the colchicine and meloxicam® groups, there was no statistically significant difference (p = 0.1); see Table 4.

Table 4

ANOVA results presenting the effects on control, MIA, Meloxicam® and Colchicine (*, p ≤ 0.05).
Experimental groups (A) (B)		Mean Difference (A-B)	Std. Error	P value
control	MIA	-1.396	.677	.186
	colchicine	.200	.681	.991
	Meloxicam®	.067	.786	1.000
MIA	control	1.396	.677	.186
	colchicine	1.596^*	.387	.001*
	Meloxicam®	1.462	.551	.055
colchicine	control	− .200	.681	.991
	MIA	-1.596^*	.387	.001*
	Meloxicam®	− .133	.556	.995
Meloxicam	control	− .067	.786	1.000
	MIA	-1.462	.551	.055*
	colchicine	.133	.556	.995

Although knee OA is quite common and has a significant global impact, there is yet no approved medicine to slow down the disease's course [11]. Nonetheless, reports of the coexistence of OA and hyperuricemia have been documented [12]. Significant associations between the amounts of uric acid in synovial fluid and the grade of knee OA as observed on radiographs were discovered in a study including a cohort of knee OA patients who were free of clinical gout [13]. Additionally, elevated uric acid levels and proinflammatory cytokines like IL-1β in the serum have been demonstrated to be strongly correlated with the severity of radiographic knee OA [6]. This interleukin is typically produced by macrophages during gout attacks, triggered by the assembly of inflammasomes induced by uric acid crystals [14]. These findings provide strong evidence for the involvement of uric acid and the innate immune system in the pathology and progression of OA. Therefore, suppression of inflammation has been proposed as a strategy for delaying the progression of OA and slowing down the disease's course. Nonetheless, reports of the coexistence of OA and hyperuricemia have been documented [14]. In a study encompassing a cohort of individuals with knee OA who did not have clinical gout, significant relationships between the levels of uric acid in synovial fluid and the severity of knee OA as shown on radiographs were found [12]. Additionally, it has been shown that there is a high correlation between the severity of radiographic knee OA and elevated levels of uric acid and proinflammatory cytokines such as IL-1β in the serum [15].

We propose that colchicine, which possesses anti-inflammatory properties, has the potential to alleviate or reduce inflammation and cartilage damage that contribute significantly to the development of knee OA. Therefore, colchicine could be a valuable treatment option for knee OA. To test our hypothesis, we induced a knee OA model in rats by injecting MIA into the patellar ligament. Subsequently, we administered colchicine to the knee OA rats and assessed its impact on inflammation and cartilage damage using semiquantitative radiographic features as well as histological and biochemical changes.

The findings of our current investigation revealed that the daily administration of colchicine effectively reduced the knee diameter in rats with MIA. This reduction in knee diameter was accompanied by a noteworthy elevation in the synthesis of biochemical markers, particularly anti-inflammatory cytokines such as IL-10. Additionally, the colchicine-treated rats exhibited significantly diminished tissue damage, as evidenced by histological and radiographic evaluation, in comparison to the rats treated with meloxicam^®.

The current study, to the best of our knowledge, is the first to offer in vivo proof that colchicine can stop or slow the progression of knee OA in the MIA-induced rat model.

Since MIA readily causes OA in rats with clinical characteristics resembling those of OA in people, it is a useful pharmacological agent for inducing OA in rats. As previously reported, all rats in the current investigation acquired OA characteristics following two weeks of MIA administration [7].

The swelling of the knee and the measurement of the diameter of the knee joint are important markers for tracking the advancement of knee OA and assessing the efficacy of an anti-inflammatory medication [1]. In an experiment involving rats, the induction of OA resulted in a notable increase in the diameter of the knee joint, indicating an inflammatory reaction. These findings are consistent with the observations noted by Leung et al., who found a significant increase in knee joint diameters after administering MIA injections [16].

The swelling in the joint can be attributed to the inflammatory exudation and synovial infiltration after the injection of MIA [17]. Microscopic examination confirmed this increase in joint diameter, revealing hyperplasia with fibrosis of the synovial membrane, resorption of the articular cartilage surface, and oedema with inflammatory cell infiltration in the surrounding cartilage, along with congested blood vessels. Our histological studies are consistent with those previously reported by Nwosu et al., who observed abnormal chondrocyte morphology, synovitis, and infiltration of inflammatory cells into the synovium after MIA injection [18].

Colchicine significantly reduced joint diameters as compared to osteoarthritis groups. It has been proposed that colchicine injections are absorbed through peritoneal epithelial cells, allowing for repair of the joint by incorporating the blood supply of the synovial membrane, joint capsule, and tendons. Histological studies also confirmed the impact of colchicine, showing mild hyperplasia in the synovial cell membrane, likely as a result of its anti-inflammatory properties. Therefore, colchicine has successfully attenuated the pronounced hyperplasia observed in osteoarthritic rats.

The use of biochemical markers in blood to evaluate the stage of OA illness and forecast clinical outcomes has gained popularity in recent years [19]. Biochemical markers for OA screening are attractive because of the ease of bioassay collection and immunoassay. In recent times, scholars have focused their attention on examining cytokines, both pro- and anti-inflammatory, due to their significance in this domain [20]. The proper metabolism of knee articular cartilage can be upset by a lack of balance among proinflammatory and anti-inflammatory cytokines, which can result in aberrant regeneration, abnormalities, and eventually the loss of the natural architecture of the knee joint [21]. Previous studies examined the association between colchicine and biochemical markers in OA, yet the casual link between colchicine and both pro- and anti-inflammatory cytokines remains ambiguous [6]. In patients with OA in the knee, Srivastava et al. noticed a significant increase in serum levels of cartilage oligomeric matrix protein, while in the colchicine group, the levels stayed steady [22].

In this study, we evaluated the anti-inflammatory effects of colchicine on the knee OA model. Thus, cytokines, both pro-inflammatory (IL-1β) and anti-inflammatory (IL-10), which are important in the pathogenesis of OA, were determined in rat serum samples by ELISA.

According to our findings, the administration of colchicine in rats with MIA-induced OA resulted in a notable increase in the expression of serum levels of anti-inflammatory cytokines (IL-10), in comparison to the group treated with meloxicam^®. Numerous studies have indicated that anti-inflammatory cytokines should be regarded as inhibitors of proinflammatory cytokines rather than possessing direct protective effects on cartilage [22].

Reduced ex vivo whole-blood IL-10 synthesis raises the risk of OA, and 1IL-10 concentrations in serum are lower in OA. Therefore, the higher serum level of IL-10 with the administration of colchicine could reflect a greater systemic anti-inflammatory cytokine status and protect against the catabolic events governed by pro-inflammatory cytokines.

In the present research, we examined the impact of colchicine on the prevention of joint destruction. The KL grading system was utilised to estimate the osteoarthritic knee of each rat through radiographs. Analysis of the knee joint radiographs confirmed the presence of destructive joint changes in all MIA groups while demonstrating the mitigating effects of colchicine on knee joint pathology.

Our radiographic findings, consistent with the studies conducted by Jaleel and Hamdalla et al., demonstrated that chondral injury induced by MIA, along with inflammation, led to the development of osteophytosis, bone sclerosis, and a reduction in joint space, similar to what is observed in human OA [17, 24]. Conversely, the radiography images revealed that the administration of colchicine mitigated the effects of MIA, resulting in reduced tissue swelling, decreased periosteal bone formation, less narrowing of the joint spaces, and increased bone density compared to MIA rats. Furthermore, the treatment with colchicine also led to lower radiographic scores when compared to the rats treated solely with MIA.

A common way of determining the degree of structural improvement or deterioration in the joint space and surrounding joint tissues, knee OA is classified using radiographic KL grading.

Nonetheless, there are a number of conceptual and technological difficulties with utilising radiography to determine the degree of OA. For instance, there is little evidence to establish a clear correlation between structural changes shown on radiography and complaints related to the knee, and limitations in resolution preclude the ability to differentiate between menisci and degeneration of the joint cartilage. As radiographs are not useful for observing or assessing changes in soft tissues, it has been suggested that a long period of follow-up is required to identify significant outcomes [24].

Many researchers have observed histological alterations in the articular cartilage of the tibiofemoral joint following MIA injection, including articular cartilage degradation and subchondral bone necrosis, as well as chondrocyte necrosis [2, 3]. The same histopathological changes were detected in the current study after OA induction in the knee joint.

The histological findings in the present research indicated a significant decrease in cartilage degradation and Mankin's histological score in MIA-induced OA rats treated with colchicine. These results were in line with a reduction in joint degenerative changes observed in the knee joints. Conversely, the groups receiving meloxicam^® treatment exhibited more pronounced joint degeneration.

Previously, there have been various outcomes from clinical trials investigating the impact of colchicine on OA. Some trials have confirmed that colchicine is both effective and safe for treating OA [25]. On the other hand, a different study found that while colchicine lowers inflammation and biomarkers linked to the severity of OA, it did not improve the symptoms of the condition [15]. Our own research adds credence to colchicine's effectiveness in treating OA by showing that it has anti-inflammatory qualities, possibly as a result of elevated serum levels of anti-inflammatory cytokines (IL-10). Furthermore, our research demonstrates that colchicine can enhance cartilage regeneration in vivo. Colchicine has been shown to be safe and effective for the long-term treatment of auto-inflammatory illnesses [39]. This implies that colchicine can potentially help with OA as well, but more research is needed.

Although the study's objectives were met, there were some possible limitations. First off, because this was a novel exploratory study to determine whether colchicine would be helpful in the knee OA model, the sample size was set exceedingly low (n = 5 in each group). It takes a statistically significant sample size to assess the results' dependability. Second, we acknowledge the limitations of radiographic assessment and the possibility that modest knee alterations suggestive of early-stage osteoarthritis may go undetected by basic imaging, irrespective of the grading scheme used. When determining an OA grade, the KL scoring system in particular has come under fire for its propensity to minimise the real importance of joint space narrowing and overemphasise the importance of osteophytes. Third, the administration of colchicine followed a single pattern. Different results can be obtained by varying the amount and frequency of colchicine administration. Fourthly, the study only examined changes that happened up to 42 days following the MIA injection; it is uncertain how well the animal responded to colchicine beyond that point in the chronic phase. Fifth, we are unable to ascertain how colchicine functions as an analgesic because we did not investigate behaviours associated with pain. Further study is required to solve these limits.

This study demonstrates how colchicine modulates inflammation mediators to enhance the radiographic, histologic, and biochemical aspects of MIA-induced knee OA in rats. Colchicine administration is helpful for OA in the knee. By inhibiting the action of inflammatory mediators and postponing cartilage deterioration, it may be able to restore and preserve OA cartilage. These results suggest that colchicine may be a valuable OA treatment.

Osteoarthritis

MIA

Monosodium iodoacetate

ELIZA

Enzyme-linked immunosorbent assay

IL-Iβ

Interleukin-1beta

IL-10

Interleukin-10

Kellgren-Lawrence

kVp

Kilovoltage peak

PACS

Picture Archiving and Communication System

Clinical trial number:

Not applicable

Declarations

Ethics approval and consent to participate

All animal experiments were approved by Research Ethics Committee considering care and use of laboratory animals (No.: Soh-Med-22-07-12)

Consent for publication

Not applicable.

Funding

None.

Author Contribution

N A, HOA, and SE ; Data collection: MI and AM; analysis and interpretation of the results: MI; Draft manuscript preparation. All authors read and approved the final manuscript.

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No competing interests reported.

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published 31 May, 2025

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Protective effects of colchicine against osteoarthritis in rat induced by monosodium iodoacetate

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Journal Publication

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Abstract

Background

Methods

Results

Conclusion

Figures

1. Background

2. Methods

2.1. Type of study:

2.2. Animals:

2.3. Reagents

2.4. Induction of the OA model

2.5. Animal Grouping and Experimental Design

2.6. Radiographic Assessment.

2.7. Assessment of body weight (gm) and knee diameter (mm):

2.8. Assessment of serum levels of interleukin (IL-1b and IL-10).

2.9. Tissue sampling and histological analysis of the joints

3. Statistical analysis

4. Results

4.1. Effect of colchicine on body weight and knee diameter measurements

4.2. ELIZA Evaluation

4.3. Effect of colchicine on histological changes:

4.3. Effect of colchicine on radiographic changes:

5. Discussion

6. Conclusions

Abbreviations

Declarations

Clinical trial number:

Declarations

Funding

Author Contribution

References

Additional Declarations

Status:

Journal Publication

Version 1