Curcumin and Freshwater Clam Extracts Alleviate the Progression of Osteoarthritis by Reducing Synovial Inﬂammation and Allowing Cartilage Regeneration

: Osteoarthritis (OA) is a common degenerative disorder and is accompanied by numer-ous pain symptoms. With increased age, individuals develop a chronic inﬂammatory status, and pro-inﬂammatory cytokines as well as mediators contribute to the progression of OA. However, no desirable remedies have been completely able to inhibit OA progression or safely provide effective symptomatic relief. Natural component extracts or dietary-derived compounds are widely used for anti-inﬂammatory diseases. Curcumin and freshwater clam extract (FCE) have been proven as functional foods that are able to regulate immune systems. This study demonstrated that curcumin and FCE had synergistic effects on alleviating the progression of OA by assuaging inﬂammation and repairing the cartilage within the joints. After consumption of curcumin and FCE, the severity of synovitis was quantiﬁed by the infrapatellar fat pad inﬂammation scoring system and the Osteoarthritis Research Society International (OARSI) scoring system. Signiﬁcant improvement and articular cartilage regeneration were noted. Moreover, once the inﬂammation within the joints was reduced, the animals redistributed their body weight on the OA-induced hindlimb. In summary, curcumin and FCE possess desirable anti-inﬂammatory and repair functions, suggesting their potential as alternative remedies in the management of OA or other inﬂammatory diseases. p > 0.05) and IL-1 β (HCF, p < 0.01; LCF, p > 0.05) expression. These results are consistent with the WBS test, and both indicate that, in the rats that consumed high doses of curcumin and FCE orally, the inﬂammatory response can be successfully reduced in the IFP and the microenvironment within the joint. With the decreased inﬂammatory responses, the curcumin and FCE lessen the pain in rats, leading to improved clinical symptoms. desirable anti-inﬂammatory effects in local tissues, but also the systemic inﬂammatory responses of MIA-induced OA rats signiﬁcantly decrease. Compared with the VC group, inﬂammatory factors MCP-1, IL-1, IL-6, and TNF- the serum of rats were signiﬁcantly reduced when p < 0.01, A similar effect on reducing pro-inﬂammatory cytokines could also be in the LCF group, though it only signiﬁcantly MCP-1 and IL-1 production p < 0.05, 5a,b). These results indicate that curcumin and clam extract could have anti-inﬂammatory effects in the rat OA model.


Introduction
Osteoarthritis (OA) is a complex disease characterized by loss of articular cartilage and failure of the synovial joint [1,2]. Clinical signs of OA include joint swelling, joint pain, limitation of activity, stiffness, and immobility. The chronic and progressive illness ultimately leads to irreversible destruction of articular cartilage and inflammation of the infrapatellar fat pad (IFP) [3]. Even though OA is one of the most common degenerative disorders in ageing society, the exact etiology or pathogenesis has not been well-defined [4,5]. To date, no desirable remedies have been completely able to inhibit OA progression or provide effective symptomatic relief [1,2,4].
Surgical intervention, pain relievers, and complementary medicines are now mainstream medicines for OA patients. Surgical intervention includes partial or total joint replacement (TJR), which can immediately improve severe clinical symptoms [2,6]. Nevertheless, it is rather aggressive and costly [7]. Pain relievers mainly target pain. Specifically, non-steroidal anti-inflammatory drugs (NSAIDs) are usually considered the first-line medical treatment against OA [2,8]. However, there is some question about their safety. Namely, Figure 1. Experimental schema, weight-bearing status (WBS) test results, and body weight change of MIA-induced osteoarthritis rat model. (a) Sprague-Dawley rats were injected with 2.5 mg monoiodoacetic acid (MIA) dissolved in 50 μL sterile 0.9% saline per joint to induce osteoarthritis (OA) at the beginning of the experiment. Saline only was administrated into the right knee articular cavity of rats as a control. Oral gavage of curcumin and freshwater clam extract (FCE) or 100% corn oil for the control group was executed every single day. Body weight and weight-bearing status (WBS) were measured as presented above. (b) WBS test was performed on days 0 (baseline), 1, 4, 6, 8, 13, 15, 17, 21, and 28. WBS test results of the left hindlimb on day 28 showed a significant difference between vehicle control and HCF group (p < 0.05). (c) Body weights of all groups had no significant differences at every measurement timepoint with an upward tendency. All data are presented as mean ± S.D. WBS test and body weight were analyzed with two-way ANOVA, followed by Tukey's post hoc test (HCF, high-dose curcumin and FCE p.o.; LCF, low-dose curcumin and FCE p.o.; n.s., no significant difference; *, p < 0.05).

Measuring Hindlimb Weight-Bearing Distribution
To evaluate the change in weight-bearing capability, hindlimb WBS tests were measured using an incapacitance tester (Linton Instrumentation, Norfolk, UK). Weight distribution was measured between left treated (intra-articular injection of 2.5 mg MIA) and normal right hindlimbs. Rats were carefully placed in a measuring chamber such that each hindlimb rested on a separate force plate. The weight-bearing force exerted by each hindlimb was averaged over 10 s. Each datum is the mean of five readings. The weight distribution ratio onto the treated hindlimb was calculated as (weight on the left hindlimb/weight on the bilateral hindlimb) × 100% [24].

Histopathological Examination for Synovial Inflammation and Cartilage Regeneration
Knee joints were fixed in 10% neutral formalin for 7 days, decalcified in rapid decalcifying solution (Sigma, St. Louis, MO, USA) for 5 days, and then embedded in paraffin wax after being harvested. Deparaffinized slides were sagittally sectioned at 4 μm and stained with Hematoxylin and Eosin (H&E) and with Safranin O/Fast green stain. For the assessment of synovitis after MIA induction, an infrapatellar fat pad (IFP) inflammation scoring system on a scale of 0 to 6 points was used (Table 1) [3,25]. To evaluate the cartilage degeneration, damage, and regeneration, the Osteoarthritis Research Society International (OARSI) score was employed on a scale of 0 to 24 points [26]. rats were injected with 2.5 mg monoiodoacetic acid (MIA) dissolved in 50 µL sterile 0.9% saline per joint to induce osteoarthritis (OA) at the beginning of the experiment. Saline only was administrated into the right knee articular cavity of rats as a control. Oral gavage of curcumin and freshwater clam extract (FCE) or 100% corn oil for the control group was executed every single day. Body weight and weight-bearing status (WBS) were measured as presented above. (b) WBS test was performed on days 0 (baseline), 1, 4, 6, 8, 13, 15, 17, 21, and 28. WBS test results of the left hindlimb on day 28 showed a significant difference between vehicle control and HCF group (p < 0.05). (c) Body weights of all groups had no significant differences at every measurement timepoint with an upward tendency. All data are presented as mean ± S.D. WBS test and body weight were analyzed with two-way ANOVA, followed by Tukey's post hoc test (HCF, high-dose curcumin and FCE p.o.; LCF, low-dose curcumin and FCE p.o.; n.s., no significant difference; *, p < 0.05).

Histopathological Examination for Synovial Inflammation and Cartilage Regeneration
Knee joints were fixed in 10% neutral formalin for 7 days, decalcified in rapid decalcifying solution (Sigma, St. Louis, MO, USA) for 5 days, and then embedded in paraffin wax after being harvested. Deparaffinized slides were sagittally sectioned at 4 µm and stained with Hematoxylin and Eosin (H&E) and with Safranin O/Fast green stain. For the assessment of synovitis after MIA induction, an infrapatellar fat pad (IFP) inflammation scoring system on a scale of 0 to 6 points was used (Table 1) [3,25]. To evaluate the cartilage degeneration, damage, and regeneration, the Osteoarthritis Research Society International (OARSI) score was employed on a scale of 0 to 24 points [26].

Immunohistochemical Staining and Scoring System
The sections which have been prepared as previously described underwent antigen retrieval in boiling citrate buffer (pH = 6.0) for 20 min. To block non-specific endogenous binding, sections were treated with hydrogen peroxide for 60 min; then, the slides were incubated with primary antibody (IL-1β, COX-2, and TNF-α; 1:200 dilution) at room temperature overnight, followed by HRP-conjugated secondary antibody. The expression levels were detected using the TAlink mouse/rabbit polymer detection system (TAHC04D, BioTnA Biotech, Kaohsiung, Taiwan). A semi-quantitative method was used to score cytokine expression in all tissue sections, as previously described [27]. Briefly, Figure 2 shows the cytokines Processes 2021, 9, 931 4 of 13 expressed in sections that were analyzed regarding staining intensity using a score from 0 to 3 (0 = no staining, 1 = weak staining, 2 = moderate staining, 3 = strong staining). The staining intensity was counted in 5 random microscopic fields at 40× objective magnification in each sample independently, and separately scored by two board-certified veterinary pathologists from NTU veterinary hospital, blinded to the experimental history. Table 1. Synovitis score and fibrosis score using the infrapatellar fat pad (IFP) inflammation scoring system (0-6).

Points Histological Features
Cell infiltration at the surface of IFP 0 Normal 1 Cellularity is increased, multinucleated cells present 2 Thickened lining cells, low (<three-fold thickness of the normal synovium) 3 Thickened lining cells, high (>three-fold thickness of the normal synovium)

Points Histological Features
Fibrosis in the body of the IFP 0 No fibrotic lesion 1 Fibrotic lesion in infrapatellar fat pad present, low 2 Fibrotic lesion increased, high 3 Infrapatellar fat pad filled with the fibrotic lesion and fat cells absent 0 Normal 1 Cellularity is increased, multinucleated cells present 2 Thickened lining cells, low (<three-fold thickness of the normal synovium) 3 Thickened lining cells, high (>three-fold thickness of the normal synovium) Points Histological Features Fibrosis in the body of the IFP 0 No fibrotic lesion 1 Fibrotic lesion in infrapatellar fat pad present, low 2 Fibrotic lesion increased, high 3 Infrapatellar fat pad filled with the fibrotic lesion and fat cells absent

Immunohistochemical Staining and Scoring System
The sections which have been prepared as previously described underwent antigen retrieval in boiling citrate buffer (pH = 6.0) for 20 min. To block non-specific endogenous binding, sections were treated with hydrogen peroxide for 60 min; then, the slides were incubated with primary antibody (IL-1β, COX-2, and TNF-α; 1:200 dilution) at room temperature overnight, followed by HRP-conjugated secondary antibody. The expression levels were detected using the TAlink mouse/rabbit polymer detection system (TAHC04D, BioTnA Biotech, Kaohsiung, Taiwan). A semi-quantitative method was used to score cytokine expression in all tissue sections, as previously described [27]. Briefly, Figure 2 shows the cytokines expressed in sections that were analyzed regarding staining intensity using a score from 0 to 3 (0 = no staining, 1 = weak staining, 2 = moderate staining, 3 = strong staining). The staining intensity was counted in 5 random microscopic fields at 40× objective magnification in each sample independently, and separately scored by two board-certified veterinary pathologists from NTU veterinary hospital, blinded to the experimental history.

Statistical Analysis
Data are presented as mean ± standard deviation (S.D.) in this study. Using Graph-Pad Prism 7.0. software, the WBS tests and body weights were analyzed with two-way ANOVA, followed by Tukey's post hoc test. Kruskal-Wallis test followed by post hoc Dunn's tests was used for scoring analysis. p-values less than 0.05 were considered to indicate a significant difference.

High-Dosage Curcumin and FCE Can Effectively Relieve Clinical Symptoms Safely
The main clinical signs of OA are chronic joint pain and limitation of activity. Based on the characteristics of the MIA-induced OA model, measuring hindlimb weight-bearing percentage was considered a universal and practical method to evaluate the development of OA. The WBS test is able to detect joint injury and recovery status from changes in weight distribution between affected and controlled hindlimbs [24]. Intra-patellar injection of MIA led to a painful response in the ipsilateral treated hindlimb, causing the animals to redistribute their body weight to the contralateral limb. For the WBS test, 2.5 mg MIA injection induced a remarkable reduction in ipsilateral weight-bearing on day 6 in all groups ( Table 2). These clinical findings confirmed that our MIA-induced OA rat model presented successfully. Compared with the vehicle control group (39.21% ± 3.59%), the HCF group (47.96% ± 4.33%) exhibited a noticeably higher weight distribution on the MIA-induced joint until the end of the observation period (p < 0.05). On the other hand, the LCF group (43.54% ± 2.08%) showed a slight but not significant increase in weight-bearing capacity until day 28 ( Figure 1b). Furthermore, the body weight of the experimental rats was recorded regularly during the study, and there were no significant differences among all groups (p > 0.05, Figure 1c). As a whole, these results indicated that HCF might safely alleviate the clinical symptoms of OA.

Curcumin and FCE Simultaneously Alleviated Local and Systemic Inflammatory Responses
To evaluate the status of synovitis, the IFP of MIA-induced OA rats was histologically assessed after a knee joint fixation process. In the vehicle control (VC) and the LCF groups, the thickness of lining cells at the surface of the IFP highly increased. Meanwhile, fibrotic lesions were observed and occupied the majority of the IFP (Figure 3a). In short, these pathological findings concurred with previous findings that MIA-induced OA rats simulated the natural progression of OA disease in vivo. The IFP inflammation scoring system was used to assess cell infiltration at the surface of the IFP and fibrosis in the body of the IFP. The inflammation scores were significantly higher in the VC and LCF groups than in the HCF group (p < 0.01) (Figure 3b). Inflammatory cytokines which play vital roles in OA progressions, such as COX-2, TNF-α, and IL-1β, are also analyzed by IHC within the IFP. The strong staining signal of COX-2, TNF-α, and IL-1β can be observed in the VC group, indicating a progressive inflammatory response in the OA joints. In contrast, there is a significantly reduced inflammation after the curcumin and FCE consumption (Figure 4a,b). Interestingly, low-and high-dosage intake of curcumin and FCE can reduce COX-2 (p < 0.01); however, only HCF can effectively suppress TNF-α (HCF, p < 0.01; LCF, p > 0.05) and IL-1β (HCF, p < 0.01; LCF, p > 0.05) expression. These results are consistent with the WBS test, and both indicate that, in the rats that consumed high doses of curcumin and FCE orally, the inflammatory response can be successfully reduced in the IFP and the microenvironment within the joint. With the decreased inflammatory responses, the curcumin and FCE lessen the pain in rats, leading to improved clinical symptoms.
(p < 0.01); however, only HCF can effectively suppress TNF-α (HCF, p < 0.01; LCF, p > 0.05) and IL-1β (HCF, p < 0.01; LCF, p > 0.05) expression. These results are consistent with the WBS test, and both indicate that, in the rats that consumed high doses of curcumin and FCE orally, the inflammatory response can be successfully reduced in the IFP and the microenvironment within the joint. With the decreased inflammatory responses, the curcumin and FCE lessen the pain in rats, leading to improved clinical symptoms.
Curcumin and clam extract not only exhibit desirable anti-inflammatory effects in local tissues, but also the systemic inflammatory responses of MIA-induced OA rats significantly decrease. Compared with the VC group, inflammatory factors including MCP-1, IL-1, IL-6, and TNF-α in the serum of rats were significantly reduced when treated with HCF (p < 0.01, Figure 5). A similar effect on reducing pro-inflammatory cytokines could also be observed in the LCF group, though it only significantly lowered MCP-1 and IL-1 production (p < 0.05, Figure 5a,b). These results indicate that curcumin and clam extract could have anti-inflammatory effects in the rat OA model.    Curcumin and clam extract not only exhibit desirable anti-inflammatory effects in local tissues, but also the systemic inflammatory responses of MIA-induced OA rats significantly decrease. Compared with the VC group, inflammatory factors including MCP-1, IL-1, IL-6, and TNF-α in the serum of rats were significantly reduced when treated with HCF (p < 0.01, Figure 5). A similar effect on reducing pro-inflammatory cytokines could also be observed in the LCF group, though it only significantly lowered MCP-1 and IL-1 production (p < 0.05, Figure 5a,b). These results indicate that curcumin and clam extract could have anti-inflammatory effects in the rat OA model.

Curcumin and FCE Can Significantly Promote Cartilage Regeneration in High and Low Doses
In consideration of standardized and reliable histopathological OA assessment, we employed the OARSI scoring system to fully examine cartilage damage and regeneration in the patellofemoral joint. Briefly, 6 grades (0-6) and 4 stages (0-4) for depth progression of the cartilage and extent of joint involvement were applied as a semi-quantitative method of OA score (score = grade × stage; range: 0-24). No cartilage degeneration or chondrocyte deaths were observed in the saline-injected right hindlimb. In the left hindlimb from the vehicle control group, however, chondrocyte death, erosion, and severe mineralization of hyaline cartilage were observed (Figure 6a), with an OARSI score of 21.0 ± 4.24 (Figure 6b, p < 0.01 vs. HCF group; p < 0.05 vs. LCF group). Though mild matrix loss and decrease in cartilage thickness occurred, curcumin and FCE repaired joint damage and promoted cartilage regeneration in OA rats, as indicated by OARSI scores of 4.3 ± 1.25 in the HCF group and 4.0 ± 2.83 in the LCF group (Figure 6b, p > 0.05). These results indicated that the severity of OA significantly decreased, and that cartilage would gradually regenerate with continuous consumption of curcumin and FCE.

Curcumin and FCE Can Significantly Promote Cartilage Regeneration in High and Low Doses
In consideration of standardized and reliable histopathological OA assessment, we employed the OARSI scoring system to fully examine cartilage damage and regeneration in the patellofemoral joint. Briefly, 6 grades (0-6) and 4 stages (0-4) for depth progression of the cartilage and extent of joint involvement were applied as a semi-quantitative method of OA score (score = grade × stage; range: 0-24). No cartilage degeneration or chondrocyte deaths were observed in the saline-injected right hindlimb. In the left hindlimb from the vehicle control group, however, chondrocyte death, erosion, and severe mineralization of hyaline cartilage were observed (Figure 6a), with an OARSI score of 21.0 ± 4.24 (Figure 6b, p < 0.01 vs. HCF group; p < 0.05 vs. LCF group). Though mild matrix loss and decrease in cartilage thickness occurred, curcumin and FCE repaired joint damage and promoted cartilage regeneration in OA rats, as indicated by OARSI scores of 4.3 ± 1.25 in the HCF group and 4.0 ± 2.83 in the LCF group (Figure 6b, p > 0.05). These results indicated that the severity of OA significantly decreased, and that cartilage would gradually regenerate with continuous consumption of curcumin and FCE.

Discussion
Our results demonstrated both clinical improvement and histopathological findings in an MIA-induced OA model. Initially, nociception before and after MIA injection was evaluated in this study. The WBS test was applied, and the upward tendency indirectly implied the clinical progress of MIA-induced OA rats. Additionally, to quantify the severity of IFP inflammation within the joints and OA joint histopathological features, our findings were examined with an IFP inflammation scoring system and OARSI scores.
Evidence appeared to show that the combination of curcumin and FCE had a beneficial effect on OA. Notably, a recovery trend was observed in a dose-dependent manner. Curcumin and FCE can reduce the inflammatory response in the synovium. Microscopically, the fibrotic lesions in the body of the IFP and the thickness of lining cells at the surface of the IFP were significantly improved after daily HCF consumption ( Figure 3). Chondrocyte death and mineralization of hyaline cartilage were significantly reduced, and cartilage regeneration was also observed after curcumin and FCE consumption (Figure 6). The regeneration effect may be worthy of attention because NSAIDs fail to slow the progression of OA. In a clinical view, high-dosage curcumin and FCE indeed relieved pain symptoms. After palliation of the severe inflammation in the joints, the rats redistributed their body weight to the MIA-induced limb on day 28 (Figure 1b and Table 2). We suggest the possibility of a synergistic effect on alleviating the OA progression and cartilage regeneration due to inhibition of pro-inflammatory cytokines and mediators.
OA is a whole-joint disease, as the inflammatory environment triggers many immune cells to infiltrate in the tissues [28][29][30][31]. Chronic inflammation and cartilage degeneration are two major problems associated with the progression of OA [5]. Chronic inflammation makes clinical signs more severe, and OA is also characterized by the degradation of articular cartilage and bone matrix [32]. Loss of collagen and the proteoglycan aggrecan leads to the destruction of the cartilage structure and function [4]. Thus, inhibition of aggressive inflammation within joints and reversal of cartilage degradation may largely alleviate the progression of OA patients.
Herein, cytokines and signal pathways related to OA should be briefly described. Interleukin (IL)-1β and TNF-α are considered the most prominent pro-inflammatory cytokines in OA [33,34]. These two cytokines suppress the expression of type II collagen, cleave the articular cartilage matrix, and release proteolytic enzymes to destroy the

Discussion
Our results demonstrated both clinical improvement and histopathological findings in an MIA-induced OA model. Initially, nociception before and after MIA injection was evaluated in this study. The WBS test was applied, and the upward tendency indirectly implied the clinical progress of MIA-induced OA rats. Additionally, to quantify the severity of IFP inflammation within the joints and OA joint histopathological features, our findings were examined with an IFP inflammation scoring system and OARSI scores.
Evidence appeared to show that the combination of curcumin and FCE had a beneficial effect on OA. Notably, a recovery trend was observed in a dose-dependent manner. Curcumin and FCE can reduce the inflammatory response in the synovium. Microscopically, the fibrotic lesions in the body of the IFP and the thickness of lining cells at the surface of the IFP were significantly improved after daily HCF consumption ( Figure 3). Chondrocyte death and mineralization of hyaline cartilage were significantly reduced, and cartilage regeneration was also observed after curcumin and FCE consumption ( Figure 6). The regeneration effect may be worthy of attention because NSAIDs fail to slow the progression of OA. In a clinical view, high-dosage curcumin and FCE indeed relieved pain symptoms. After palliation of the severe inflammation in the joints, the rats redistributed their body weight to the MIA-induced limb on day 28 (Figure 1b and Table 2). We suggest the possibility of a synergistic effect on alleviating the OA progression and cartilage regeneration due to inhibition of pro-inflammatory cytokines and mediators.
OA is a whole-joint disease, as the inflammatory environment triggers many immune cells to infiltrate in the tissues [28][29][30][31]. Chronic inflammation and cartilage degeneration are two major problems associated with the progression of OA [5]. Chronic inflammation makes clinical signs more severe, and OA is also characterized by the degradation of articular cartilage and bone matrix [32]. Loss of collagen and the proteoglycan aggrecan leads to the destruction of the cartilage structure and function [4]. Thus, inhibition of aggressive inflammation within joints and reversal of cartilage degradation may largely alleviate the progression of OA patients.
Herein, cytokines and signal pathways related to OA should be briefly described. Interleukin (IL)-1β and TNF-α are considered the most prominent pro-inflammatory cytokines in OA [33,34]. These two cytokines suppress the expression of type II collagen, cleave the articular cartilage matrix, and release proteolytic enzymes to destroy the cartilage [34][35][36].
Activation of an inflammatory signal pathway, NF-κB, leads to the upregulation of COX-2, IL-1β, and TNF-α and a chronic inflammatory state within the joints [33,37].
Physiological microenvironments can be modulated or influenced by curcumin and FCE. Curcumin can inhibit the production of inflammatory mediators such as nitric oxide (NO), prostaglandin2 (PGE2), IL-6, and IL-8 stimulated by IL-1β [11,12,38]. Additionally, curcumin prevents IL-1β-induced NF-κB activation and translocation, giving rise to reduced expression of the NF-κB downstream pro-inflammatory gene COX-2 [39]. Through inhibition of the NF-κB pathway, the expression of inflammatory OA targets can be reduced. These targets are IL-1β, TNF-α, COX-2, and matrix metalloproteinases (MMPs). MMPs, especially MMP-13, are the major factors in inflammation and extracellular matrix degradation in OA progression [11]. Furthermore, curcumin exerts upstream control over NF-κB, inhibiting the pathway of toll-like receptor 4 (TLR4), which mediates the inflammatory responses involved in OA pathogenesis [40]. Inhibition of NF-κB also displays a protective function from the accumulation of advanced glycation end-products (AGEs) and upregulation of MMP-13, which can retard OA progression by reducing inflammation and cartilage ageing [41]. FCE has a similar property of inhibitory effects of NF-κB, but it also downregulates mitogen-activated protein kinases (MAPKs), resulting in declines in NO, IL-1β, IL-6, and TNF-α as well [42].
In summary, the present study illustrated the synergistic effects of curcumin and FCE on alleviating the progression of OA by anti-inflammatory function and cartilage regeneration. Curcumin downregulates the activity of COX-2 [18] and targets the NF-κB pathway to inhibit the release of pro-inflammatory cytokine TNF-α [19]. FCE neutralizes inflammation and slows the destruction of articular cartilage by reducing IL-1β and TNFα [23]. The anti-inflammatory function delays the progression of ageing or mineralization of cartilage, thereby allowing the articular cartilage to regenerate. Pain symptoms are ultimately eased by the inhibition of inflammation and regeneration of cartilage. The results suggest that curcumin and FCE have the potential to be an alternative remedy in the management of OA or other inflammatory diseases.