Injection Of Monosodium Iodoacetate Research Articles

Umbilical cord derived mesenchymal stem cells attenuate rodent Osteoarthritis progression via preserving articular cartilage superficial layer cells and inhibiting synovitis

Purpose: Osteoarthritis (OA) is one of the most common joint diseases, which is characterized by pain and joint dysfunction. Inhibition of synovial inflammation and promotion of cartilage regeneration is considered to effectively attenuate the manifestation. Mesenchymal stem cells showed tissue regeneration effects mainly through tissue replacement effects, inflammation and immune response modulation, and initiation of endogenous repair. Umbilical cord derived MSCs (UCMSCs) are becoming increasingly popular as it is highly proliferative and could be noninvasively harvested compared with other MSCs. Therefore, we aim to test the effects of UCMSCs on rodent inflammatory arthritis induced by Monosodium iodoacetate (MIA) and posttraumatic OA (PTOA) models induced by DMM surgery. Methods: Intra-articular injection of 0.3 mg Monosodium iodoacetate (MIA) in 50 μL saline was injected into the left knee of the 200g weight male SD rat to induce rat knee inflammatory arthritis, and saline as veh group on contralateral right knees. 2.5× 105 UCMSCs or chondrocytes (CHO) in 50 μL normal saline were then given on day1, day 7, and day 14 respectively two weeks after MIA injection. Joints were harvested 4 weeks after MIA injection for paraffin section and then Safranine-O, H&E and immunohistochemistry staining were performed to observe cartilage erosion and synovium. In addition, PTOA mouse model was established on 3 months old C57 male mice by DMM (destablization of medial meniscus) surgery. 2.5× 105 UCMSCs were injected in to the DMM knees during the surgery, and joints were harvested 3 months post surgery for histology. For invitro studies, migration ability of SFCs by UCMSCs were accessed by transwell assay. Furthermore, catabolism change of MIA induced SFCs by UCMSCs is performed by real-rime PCR of Col-X and BCL-2 genes. CCK-8 assay was done to check proliferation ability of SFCs by UCMSCs conditioned media. Results: First, we performed Safranine-O staining and found delayed OA progression by UCMSCs treatment, compared with vehicle (Fig. 1 A). Further Mankin Score confirmed a significant decrease in UCMSCs treatment (4±0.45 VS 12.5±0.34 in vehicle, P<0.0001) (Fig. 1 B). Based on histology, we quantified the cartilage thickness, and found a 125% increase in UCMSCs treatment compared with vehicle (Fig. 1 C). The ratio of uncalcified zone to total thickness shows similar result (Fig. 1 D). To understand the molecular mechanism, we performed immunohistochemistry and found that UCMSCs treatment had more type 2 collagen staining, less ADAMTS-5 and less MMP-13 staining compared with vehicle group (Fig. 2 A C D). In addition, increased hypertrophic chondrocytes, another cartilage catabolism marker, by MIA injection is attenuated by UCMSCs (Fig. 2 E). Those data suggested that UCMSCs are capable of switching the catabolism to anabolism of chondrocytes during OA. Interestingly, we found that UCMSCs treatment moderately reserve the loss of SFCs after MIA treatment (Fig. 3 A B), possibly due to elevated proliferation (Fig. 3 C), decreased catabolism gene expression: Col-X and BCL-2 gene expression (Fig. 3 D E). by UCMSCs conditioned media. Interestingly, we observed that SFCs may accumulate in cartilage erosion area during the process OA (Fig 4 A), we performed a transwell assay and revealed that that SFCs were more likely to migrate to injured chondrocytes by serum starvation compared to normal chondrocytes (Fig 4 B D). Moreover, UCMSCs conditioned media could promote the migration of SFCs compared to vehicle (Fig 4C E). Combining those data, we hypothesized that migration of SFCs is crucial to initiate cartilage repair and UCMSCs could effectively promote this capacity, which may attribute to its rescue effects on OA progression. As synovitis is also a major manifestation of OA and main cause of pain, we performed H&E staining on synovium and revealed that MIA induced obvious inflammatory cells infiltration and obvious hyperplasia (Fig. 5A). On the contrary, UCMSCs treatment could greatly diminish these effects and significantly bring the synovitis score down (1.60 vs vehicle 5.33, P=0.0002, Fig 5 A B). To further identify the identity of those infiltrated inflammatory cells in the synovium, we did IHC staining of CD68 and CD4. Interestingly, both CD4+ Th cells and CD68+ macrophages were elevated by MIA, and obviously attenuated by multiple UCMSCs treatment (Fig. 5 C D). As MIA induced arthritis is an inflammatory arthritis model, so we introduced another typical OA model by DMM surgery, and we also found that UCMSCs could effectively attenuate cartilage erosion (Fig 6 A) and thus delay OA progression by significantly bringing down the Mankin Score (5.75±0.37 VS 12.20±0.84, N=9, P<0.0001, Fig 6 B). Further H&E staining also showed preserved SFCs numbers by UCMSCs treatment after DMM surgery (2.57±0.98 VS 0.83±0.75, N=9, P<0.05, Fig 6 C D) Conclusions: This study highlights the alleviating effects on OA progression by UCMSCs on two arthritis models. Furthermore, the effects are possibly via inhibited synovial inflammation and preserved SFCs on the articular cartilage. However, detailed celluar and molecular mechanisms are needed in future studies.

Effects of Pomegranate Concentrate Powder: Eucommiae Cortex: Achyranthis Radix 5:4:1 (w/w) Mixed Formula on Monosodium Iodoacetate-Induced Osteoarthritis in Rats

We examined the antiosteoarthritis effect of a mixture of powdered pomegranate concentrate, eucommiae cortex, and achyranthis radix (5:4:1 w/w) (PEA-Mix). After the injection of monosodium iodoacetate (MIA), PEA-Mixs were orally administered. To assess pain-related behaviors, a von Frey filament test and open field test were performed. We also examined the knee thickness, maximum knee extension angle, bone mineral density (BMD), and compressive strength of the knee joint and performed a histopathologic analysis. The number of COX-2, tumor necrosis factor (TNF)-α, poly (ADP-ribose) polymerase (PARP), and 5-bromo-2′-deoxyuridine immunoreactive cells, the prostaglandin E2 (PGE2) and 5-lipoxygenase (LPO) levels, matrix metalloproteinase (MMP) activity, and mRNA levels of chondrogenesis-related genes were analyzed. PEA-Mix significantly inhibited the MIA-induced decrease in the paw-withdrawal threshold and total distance moved, and the MIA-induced increases in the maximum knee extension angle and knee thickness. Also, the MIA-induced loss of the knee joint articular surface region and decrease in the BMD were significantly suppressed by PEA-Mix. The MIA-induced increases in the 5-LPO, PGE2, MMP-2, MMP-9, COX-2, and TNF-α mRNA levels were reduced by PEA-Mix. PEA-Mix increased the MIA-mediated reduction in the SOX-9 and aggrecan mRNA levels. The number of PARP-positive cells was smaller in PEA-Mix-administered rats than in MIA-administered rats, while the number of 5-bromo-2′-deoxyuridine-positive cells was larger. Therefore, PEA-Mix relieved the MIA-induced pain-related behaviors, chondrocyte proliferation, and anti-inflammatory activity.

Prevention or treatment of enzyme treated royal jelly on monosodium-iodoacetate-induced osteoarthritis

Royal jelly (RJ) is a gelatinous substance that bees produce to feed bees and queen bees. It’s frequently sold as a dietary supplement to treat a variety of physical ailments and chronic diseases. While it has long been used in traditional medicine, its applications in Western medicine remain controversial. The inhibitory effect of royal jelly on osteoarthritis was investigated in primary cultured rat cartilage cells and monosodium-iodoacetate (MIA)-induced arthritis rat model 10-hydroxy-2-decenoic acid (10-HAD) is the main fatty acid present in RJ. Among the criteria for RJ quality analysis, 10-HAD content has been proposed as a freshness parameter. We investigated the effect of RJ on the improvement of osteoarthritis on SD rats and they were divided into five groups. In this study, we examined the effect of enzymatic royal jelly (ERJ) administration on osteoarthritis. To determine the antiinflammatory effects of RJ, tumor necrosis factor alpha (TNF-α) and Interleukin-6 (IL-6) expression were measured after lipopolysaccharide (LPS) activation in RAW 264.7 cells. In in vivo animal study, osteoarthritis was induced by intra-articular injection of MIA into knee joints of rats. As a results, ERJ showed that TNF-α and IL-6 levels were decreased by ERJ treatment in a dosedependent manner. In conclusion, ERJ extract was able to inhibit articular cartilage degeneration by preventing extracellular matrix degradation and cartilage cell damage. It was considered that ERJ extract may be a potential therapeutic treatment for degenerative osteoarthritis.

NF-κB Phosphorylation Inhibition Prevents Articular Cartilage Degradation in Osteoarthritis Rats via 2-Aminoquinoline.

BackgroundOsteoarthritis is a chronic degenerative disease of the joints that is common in older people worldwide. The characteristic features of osteoarthritis include cartilage degradation, synovitis, and remodelling of subchondral bone. The present study investigated the effect of 2-aminoquinoline on knee articular cartilage degradation in an osteoarthritis rat model.Material/MethodsThe rat model of osteoarthritis was established in Wistar rats by intra-articular injection of monosodium iodoacetate. The rats were randomly divided into 6 groups of 10 rats each: a normal control group, an untreated group, and 4 (5, 10, 15 and 20 mg/kg) treatment groups. The rats in treatment groups received 5, 10, 15, or 20 mg/kg doses of 2-aminoquinoline on day 2 of monosodium iodoacetate injection.ResultsThe 2-aminoquinoline treatment of monosodium iodoacetate-injected rats markedly decreased weight-bearing asymmetry, inhibited edema formation, and improved paw withdrawal thresholds. The expression of inflammatory cytokines was markedly higher in the osteoarthritis rats. Treatment with 2-aminoquinoline led to a significant reduction in inflammatory cytokine expression in osteoarthritis rats in a dose-dependent manner. In osteoarthritis rats, the expressions of prostaglandin E2 (PGE2), matrix metalloproteinase-13 (MMP-13), and substance P were also higher in comparison to the control group. The 2-aminoquinoline treatment supressed PGE2, MMP-13, and substance P levels in osteoarthritis rats. Moreover, the expression of phosphorylated nuclear factor kappaB (p-NF-κB) was markedly higher in the untreated rats. However, activation of NF-κB was downregulated in the osteoarthritis rats by treatment with 2-aminoquinoline.ConclusionsThe present study demonstrated that 2-aminoquinoline prevents articular cartilage damage in osteoarthritis rats through inhibition of inflammatory factors and downregulation of NF-κB activation, suggesting that 2-aminoquinoline would be effective in treatment of osteoarthritis.

Enhanced T-type calcium channel 3.2 activity in sensory neurons contributes to neuropathic-like pain of monosodium iodoacetate-induced knee osteoarthritis.

The monosodium iodoacetate knee osteoarthritis model has been widely used for the evaluation of osteoarthritis pain, but the pathogenesis of associated chronic pain is not fully understood. The T-type calcium channel 3.2 (CaV3.2) is abundantly expressed in the primary sensory neurons, in which it regulates neuronal excitability at both the somata and peripheral terminals and facilitates spontaneous neurotransmitter release at the spinal terminals. In this study, we investigated the involvement of primary sensory neuron-CaV3.2 activation in monosodium iodoacetate osteoarthritis pain. Knee joint osteoarthritis pain was induced by intra-articular injection of monosodium iodoacetate (2 mg) in rats, and sensory behavior was evaluated for 35 days. At that time, knee joint structural histology, primary sensory neuron injury, and inflammatory gliosis in lumbar dorsal root ganglia, and spinal dorsal horn were examined. Primary sensory neuron-T-type calcium channel current by patch-clamp recording and CaV3.2 expression by immunohistochemistry and immunoblots were determined. In a subset of animals, pain relief by CaV3.2 inhibition after delivery of CaV3.2 inhibitor TTA-P2 into sciatic nerve was investigated. Knee injection of monosodium iodoacetate resulted in osteoarthritis histopathology, weight-bearing asymmetry, sensory hypersensitivity of the ipsilateral hindpaw, and inflammatory gliosis in the ipsilateral dorsal root ganglia, sciatic nerve, and spinal dorsal horn. Neuronal injury marker ATF-3 was extensively upregulated in primary sensory neurons, suggesting that neuronal damage was beyond merely knee-innervating primary sensory neurons. T-type current in dissociated primary sensory neurons from lumbar dorsal root ganglia of monosodium iodoacetate rats was significantly increased, and CaV3.2 protein levels in the dorsal root ganglia and spinal dorsal horn ipsilateral to monosodium iodoacetate by immunoblots were significantly increased, compared to controls. Perineural application of TTA-P2 into the ipsilateral sciatic nerve alleviated mechanical hypersensitivity and weight-bearing asymmetry in monosodium iodoacetate osteoarthritis rats. Overall, our findings demonstrate an elevated CaV3.2 expression and enhanced function of primary sensory neuron-T channels in the monosodium iodoacetate osteoarthritis pain. Further study is needed to delineate the importance of dysfunctional primary sensory neuron-CaV3.2 in osteoarthritis pain.

Spinal NF-kB upregulation contributes to hyperalgesia in a rat model of advanced osteoarthritis.

Knee osteoarthritis (OA) pain is the most common joint pain. Currently, dysfunction in the central nervous system rather than knee joint degeneration is considered to be the major cause of chronic knee OA pain; however, the underlying mechanism remains unknown. The aim of this study was to explore whether spinal NF-κB plays a critical role in chronic knee OA pain. In this study, we used a model induced by the intra-articular injection of monosodium iodoacetate. Spinal NF-κB and the phosphorylation and activation status of NF-κB p65/RelA (p-p65) were inhibited by the intrathecal injection of the inhibitor pyrrolidine dithiocarbamate in this model. After behavioral assessment, the knee was dissected for histopathology, and the spinal cord was dissected and examined for NF-κB, p-p65, and cytokine expression. Furthermore, the quantity and activity of neurons, astrocytes, and microglial cells and their colocalization with p-p65 in the spinal dorsal horn were investigated. Our findings included the following: (1) histology, the pathological changes in the joints of the knee OA model were basically consistent with knee OA patients; (2) the protein and transcription levels of NF-κB/p65 and p-p65 increased before day 14, appeared to decrease on day 21 and increased again on day 28, and the tendency of weight bearing was similar; (3) on days 21 and 28, the intrathecal injection of pyrrolidine dithiocarbamate markedly prevented the monosodium iodoacetate-induced reduction in the paw withdrawal threshold; (4) real-time polymerase chain reaction demonstrated that the expression of TNF-α and IL-33 was suppressed in the knee OA model by the intrathecal injection of pyrrolidine dithiocarbamate; and (5) immunofluorescence revealed that astrocytes were activated and that p-p65 was mainly increased in astrocytes. Our findings indicate that the spinal NF-κB/p65 pathway in astrocytes modulates neuroimmunity in rat model of intra-articular monosodium iodoacetate-induced advanced OA.

The Inhibitory Effects of Slow-Releasing Hydrogen Sulfide Donors in the Mechanical Allodynia, Grip Strength Deficits, and Depressive-Like Behaviors Associated with Chronic Osteoarthritis Pain.

Osteoarthritis and its associated comorbidities are important clinical problems that have a negative impact on the quality of life, and its treatment remains unresolved. We investigated whether the systemic administration of slow-releasing hydrogen sulfide (H2S) donors, allyl isothiocyanate (A-ITC) and phenyl isothiocyanate (P-ITC), alleviates chronic osteoarthritis pain and the associated emotional disorders. In C57BL/6 female mice with osteoarthritis pain induced by the intra-articular injection of monosodium iodoacetate, we evaluated the effects of repeated administration of A-ITC and P-ITC on the (i) mechanical allodynia and grip strength deficits; (ii) emotional conducts; and (iii) glial activity and expression of inducible nitric oxide synthase (NOS2), phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt), and antioxidant enzymes (heme oxygenase 1, NAD(P)H:quinone oxidoreductase-1, glutathione S-transferase mu 1 and alpha 1) in the hippocampus. The administration of A-ITC and P-ITC inhibited the mechanical allodynia, the grip strength deficits, and the depressive-like behaviors accompanying osteoarthritis. Both treatments inhibited microglial activation, normalized the upregulation of NOS2 and PI3K/p-Akt, and maintained high levels of antioxidant/detoxificant enzymes in the hippocampus. Data suggest that treatment with low doses of slow-releasing H2S donors might be an interesting strategy for the treatment of nociception, functional disability, and emotional disorders associated with osteoarthritis pain.

Inhibition of miR-203 Ameliorates Osteoarthritis Cartilage Degradation in the Postmenopausal Rat Model: Involvement of Estrogen Receptor α.

miR-203 is known to target estrogen receptor α (ERα) in various cancer cell lines, such as MCF-7. However, whether miR-203 regulates ERα and contributes to the onset and progression of osteoarthritis (OA) is poorly understood. A combined protocol of the bilateral ovariectomy and the intra-articular monosodium iodoacetate injection was applied to establish a postmenopausal OA model in rats. Real-time quantitative polymerase chain reaction was used to detect miR-203 and mRNAs and Western blotting was exploited to quantify the expression levels on the protein level. Enzyme-linked immunosorbent assays were deployed to detect the expression of matrix metalloproteinase-1 (MMP-1), MMP-3, prostaglandin E2 (PGE2), and collagen type II degradation (CTX-II) in serum samples. Dual-luciferase reporter assay was utilized to confirm the direct binding of miR-203 on ERα in postmenopausal OA rats. Expression of miR-203 was elevated; while ERα mRNA and protein were downregulated in postmenopausal OA rats, compared with sham rats. Dual-luciferase reporter assay confirmed miR-203 bound and negatively regulated ERα, resulting in promoted cellular inflammation and cartilage destruction in postmenopausal OA rats. Suppression of miR-203 using a specific inhibitor ameliorated cartilage degradation in postmenopausal OA rats. miR-203 is pivotal in the onset and progression of OA in the postmenopausal rat model, and holds promise for a therapeutic target of OA treatment.

Attenuation of IL-1ß on the use of glucosamine as an adjuvant in meloxicam treatment in rat models with osteoarthritis.

Background Osteoarthritis (OA) is the most prevalent joint disease and a common cause of joint pain, functional loss, and disability. The severity of this disease is always associated with increased levels of proinflammatory cytokines, which play an important role in cartilage damage, synovitis, and other damage to joint tissues. The discovery that many soluble mediators such as cytokines or prostaglandins can increase the production of matrix metalloproteinases by chondrocytes led to the first steps of an inflammatory state. Several studies show that cytokines, such as interleukin 1ß, have a major role in the development of inflammation that occurs in these joints. The use of glucosamine as an adjuvant to meloxicam therapy is expected to inhibit the development of inflammatory OA. Methods The OA model in rat was induced by single injection of intraarticular monosodium iodoacetate (MIA). The development of OA was observed for 21 days. Furthermore, the evaluation of glucosamine potency as an adjuvant of meloxicam therapy for reducing IL-1ß was done by combined treatment at a low dose of meloxicam 1 mg/kg BW with glucosamine at a dose of 125, 250, or 500 mg/kg BW orally for 28 days. Response to hyperalgesia and knee joint diameter was measured on days 0, 7, 14, 21, 28, 35, 42, and 49. IL-1ß levels were measured on day 21 and day 49 after MIA injection. Results MIA injection successfully induced OA as marked by a significant difference in the time of latency to heat stimulus (p < 0.01) and a significant increase in joint diameter (p < 0.01). On day 21, IL-1ß levels showed a significant decrease in MIA injection (p = 0.05). The administration of meloxicam and glucosamine did not induce significant decrease in knee joint diameter (p > 0.10), but was able to significantly increase the latency time to heat stimulus (p < 0.01). IL-1ß levels also showed a significant decrease after administering a combination of glucosamine and meloxicam (p < 0.01). Conclusions Taken together, the use of glucosamine as an adjuvant in meloxicam therapy may be caused by the synergistic mechanism of meloxicam for the attenuation of OA development through systemically reducing IL-1ß.

Cartilage protective and anti-analgesic effects of ALM16 on monosodium iodoacetate induced osteoarthritis in rats

BackgroundOsteoarthritis (OA) is an age-related joint disease with characteristics that involve the progressive degradation of articular cartilage and resulting chronic pain. Previously, we reported that Astragalus membranaceus and Lithospermum erythrorhizon showed significant anti-inflammatory and anti-osteoarthritis activities. The objective of this study was to examine the protective effects of ALM16, a new herbal mixture (7:3) of ethanol extracts of A. membranaceus and L. erythrorhizon, against OA in in vitro and in vivo models.MethodsThe levels of matrix metalloproteinase (MMP)-1, −3 and − 13 and glycosaminoglycan (GAG) in interleukin (IL)-1β or ALM16 treated SW1353 cells were determined using an enzyme-linked immunosorbent and quantitative kit, respectively. In vivo, the anti-analgesic and anti-inflammatory activities of ALM16 were assessed via the acetic acid-induced writhing response and in a carrageenan-induced paw edema model in ICR mice, respectively. In addition, the chondroprotective effects of ALM16 were analyzed using a single-intra-articular injection of monosodium iodoacetate (MIA) in the right knee joint of Wister/ST rat. All samples were orally administered daily for 2 weeks starting 1 week after the MIA injection. The paw withdrawal threshold (PWT) in MIA-injected rats was measured by the von Frey test using the up-down method. Histopathological changes of the cartilage in OA rats were analyzed by hematoxylin and eosin (H&E) staining.ResultsALM16 remarkably reduced the GAG degradation and MMP levels in IL-1β treated SW1353 cells. ALM16 markedly decreased the thickness of the paw edema and writhing response in a dose-dependent manner in mice. In the MIA-induced OA rat model, ALM16 significantly reduced the PWT compared to the control group. In particular, from histological observations, ALM16 showed clear improvement of OA lesions, such as the loss of necrotic chondrocytes and cartilage erosion of more than 200 mg/kg b.w., comparable to or better than a positive drug control (JOINS™, 200 mg/kg) in the cartilage of MIA-OA rats.ConclusionsOur results demonstrate that ALM16 has a strong chondroprotective effect against the OA model in vitro and in vivo, likely attributed to its anti-inflammatory activity and inhibition of MMP production.

Effect of Alpinia oxyphylla extract in vitro and in a monosodium iodoacetate-induced osteoarthritis rat model

BackgroundOsteoarthritis (OA) affects the articular cartilage and subchondral bone of synovial joints and induces proinflammatory and anti-inflammatory pathway dysregulation, leading to pain. This study evaluated the anti-inflammatory and antiosteoarthritis effects of Alpinia oxyphylla extract (AOE) in vitro and in vivo. MethodsThe anti-inflammatory effect of AOE was evaluated in vitro in lipopolysaccharide (LPS)-treated RAW264.7 cells. The antiosteoarthritis effect of AOE was investigated in a monosodium iodoacetate (MIA)-induced rat model of OA. Rats were orally administered AOE (150 mg/kg or 300 mg/kg) or the positive control drug indomethacin (1 mg/kg) 3 days before MIA injection and once daily for 21 days thereafter. ResultsAOE significantly decreased the production of nitric oxide (NO, 68.2%), prostaglandin E2 (PGE2, 92.8%), interleukin-1β (IL-1β, 77.2%), interleukin-6 (IL-6, 39.9%), and tumor necrosis factor-alpha (TNF-α, 20.7%) and the activation of extracellular signal-regulated kinase (ERK), Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) in LPS-treated RAW264.7 cells at a dose of 100 µg/ml. In addition, AOE attenuated joint pain, suppressed proinflammatory cytokine and mediator production and inhibited cartilage degradation in the MIA-induced rat OA model. ConclusionThese results demonstrate that AOE exerts potent anti-inflammatory effects and may be a useful therapeutic candidate against OA.

Anti-inflammatory Activity of Nanoemulgel formulated from Ageratum conyzoides (L.) L. and Oldenlandia corymbosa L. Extracts in Rats

The use of traditional medicine in the treatment of disease has increased globally due to its safety and efficacy. Ageratum conyzoides (L.) L. and Oldenlandia corymbosa L. have been used traditionally as a topical preparation for joint disease in Indonesia. Hence, the study was planned to scientifically study the existing empirical data. A nanoemulgel of combined Ageratum conyzoides (L.) L. (ACE) and Oldenlandia corymbosa L. Extract (OCE) as a new drug focused on plant-based natural products with a good physical characteristic that inhibits inflammatory process in managing osteoarthritis (OA) was formulated. Thirty animals were randomly designated to the 6 groups (n=5) as follows: (1) The normal control group (Normal), (2) negative control groups of Monosodium Iodoacetate (MIA), (3) combination ACE-OCE, (4) single ACE, (5) single OCE, (6) positive control group (Diclofenac). Rats received intraarticular MIA injection dose 3mg/0.05 mL on day zero excluding normal control group. All groups were administered topical preparations allotted to each dose group on day 29. Knee edema profile (every 7 days) and serum cytokine level (on day 57) was evaluated with Enzyme-Linked Immunoabsorbent Assay (ELISA). Till day 42, knee edema profile of all group treatment have been similar with normal control group (p>0.05). Serum cytokines level for some biomarkers, such as S100A8 Protein, Interleukin-1β, Inducible Nitric Oxide Synthase (iNOS), matrix metalloproteinase-13 (MMP-13), a disintegrin and metalloproteinase thrombospondin-like motifs-5 (ADAMTS-5), Collagen Type II and Aggrecan Core Protein were decreased. A significant difference compared with MIA group showed for all group treatment on measurement of S100A8 Protein, IL-1β, and iNOS as a biomarker of inflammatory process (#P<0.05). The developed nanoemulgel ACE-OCE either in single or in combination has good physical characteristic and promising effect of anti-inflammatory activity to enhance MIA induced cartilage damage as potential therapeutic agent for OA and encouraging to conduct further studies.

The Effects of Annatto Tocotrienol Supplementation on Cartilage and Subchondral Bone in an Animal Model of Osteoarthritis Induced by Monosodium Iodoacetate.

Osteoarthritis is a degenerative joint disease which primarily affects the articular cartilage and subchondral bones. Since there is an underlying localized inflammatory component in the pathogenesis of osteoarthritis, compounds like tocotrienol with anti-inflammatory properties may be able to retard its progression. This study aimed to determine the effects of oral tocotrienol supplementation on the articular cartilage and subchondral bone in a rat model of osteoarthritis induced by monosodium iodoacetate (MIA). Thirty male Sprague-Dawley rats (three-month-old) were randomized into five groups. Four groups were induced with osteoarthritis (single injection of MIA at week 0) and another served as the sham group. Three of the four groups with osteoarthritis were supplemented with annatto tocotrienol at 50, 100 and 150 mg/kg/day orally for five weeks. At week 5, all rats were sacrificed, and their tibial-femoral joints were harvested for analysis. The results indicated that the groups which received annatto tocotrienol at 100 and 150 mg/kg/day had lower histological scores and cartilage remodeling markers. Annatto tocotrienol at 150 mg/kg/day significantly lowered the osteocalcin levels and osteoclast surface of subchondral bone. In conclusion, annatto tocotrienol may potentially retard the progression of osteoarthritis. Future studies to confirm its mechanism of joint protection should be performed.

Protective effects of extracorporeal shockwave on rat chondrocytes and temporomandibular joint osteoarthritis; preclinical evaluation with in vivo99mTc-HDP SPECT and ex vivo micro-CT

Extracorporeal shockwave therapy (ESWT) has been shown to have chondroprotective effects on arthritic diseases. We investigated the effects of ESWT on temporomandibular joint osteoarthritis (TMJOA) using rat chondrocytes and TMJOA rat models. Cell viability and expression of pro-inflammatory cytokines, cartilage degradation, and apoptosis markers were measured in control, monosodium iodoacetate (MIA)-treated and ESWT plus MIA-treated chondrocytes invitro, and intra-articular MIA injection (TMJOA) and ESWT on TMJOA rats invivo. Invivo99mTc-hydroxymethylene diphosphonate (HDP) single-photon emission computerized tomography/computerized tomography (SPECT/CT) and ex-vivo micro-CT and histologic examinations were performed in rat models. ESWT plus MIA-treated chondrocytes showed increased cell viability significantly (P=0.007), while decreased genetic expression of pro-inflammatory cytokines [tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6); P<0.001 for each] and cartilage degradation markers [matrix metalloproteinase-3 (MMP3), matrix metalloproteinase-13 (MMP13), and bone morphogenetic protein 7 (BMP7); P<0.001 for each], and number of apoptotic cells (P<0.001) compared to MIA-treated chondrocytes. Changes in cytochrome c and cleaved caspase-3 levels relative to procaspase-3 were decreased over MIA-treated chondrocytes. ESWT on TMJOA rat models was associated with a significant decrease in pro-inflammatory and cartilage degradation markers, as demonstrated by real-time PCR and immunohistochemistry stains (P<0.001 for each). On 99mTc-HDP SPECT/CT, the ESWT group showed a significantly lower uptake ratio compared to the TMJOA group (P=0.008). Micro-CT analysis revealed that the ESWT group showed improved structure and bone quality compared to the TMJOA control group. ESWT was associated with a protective effect on cartilage and subchondral bone structures of TMJOA by reducing inflammation, cartilage degradation, and chondrocyte apoptosis.

Nicotine Attenuates Osteoarthritis Pain and Matrix Metalloproteinase-9 Expression via the α7 Nicotinic Acetylcholine Receptor.

Osteoarthritis (OA) is a degenerative joint disease that causes chronic disability among the elderly. Despite recent advances in symptomatic management of OA by pharmacological and surgical approaches, there remains a lack of optimal approaches to manage inflammation in the joints, which causes cartilage degradation and pain. In this study, we investigated the efficacy and underlying mechanisms of nicotine exposure in attenuating joint inflammation, cartilage degradation, and pain in a mouse model of OA. A mouse model of OA was induced by injection of monosodium iodoacetate into the knee joint. Cell culture models were also used to study the efficacy and underlying mechanisms of nicotine treatment in attenuating symptoms of OA. Nicotine treatment reduced mechanical allodynia, cartilage degradation, and the upregulation of matrix metalloproteinase-9 (MMP-9), a hallmark of joint inflammation in OA, in mice treated with monosodium iodoacetate. The effects of nicotine were abolished by the selective α7 nicotinic acetylcholine receptor (nAChR) blocker, methyllycaconitine . In RAW264.7 cells and murine primary bone marrow-derived macrophages, nicotine significantly inhibited MMP-9 production induced by LPS. In addition, nicotine significantly enhanced PI3K/Akt and inhibited NF-κB translocation from the cytosol to the nucleus in an α7-nAChR-dependent manner, suggesting that nicotine acts on α7-nAChRs to inhibit MMP-9 production by macrophages through modulation of the PI3K/Akt-NF-κB pathway. Our results provide novel evidence that nicotine can attenuate joint inflammation and pain in experimental OA via α7-nAChRs. α7-nAChR could thus serve as a highly promising target to manage joint inflammation and pain in OA.

Metformin Augments Anti-Inflammatory and Chondroprotective Properties of Mesenchymal Stem Cells in Experimental Osteoarthritis.

Mesenchymal stem cells (MSCs) can protect against cartilage breakdown in osteoarthritis (OA) via their immunomodulatory capacities. However, the optimization strategy for using MSCs remains challenging. This study's objective was to identify the in vivo effects of metformin-stimulated adipose tissue-derived human MSCs (Ad-hMSCs) in OA. An animal model of OA was established by intra-articular injection of monosodium iodoacetate into rats. OA rats were divided into a control group and two therapy groups (treated with Ad-hMSCs or metformin-stimulated Ad-hMSCs). Limb nociception was assessed by measuring the paw withdrawal latency and threshold. Our data show that metformin increased IL-10 and IDO expression in Ad-hMSCs and decreased high-mobility group box 1 protein, IL-1β, and IL-6 expression. Metformin increased the migration capacity of Ad-hMSCs with upregulation of chemokine expression. In cocultures, metformin-stimulated Ad-hMSCs inhibited the mRNA expression of RUNX2, COL X, VEGF, MMP1, MMP3, and MMP13 in IL-1β-stimulated OA chondrocytes and increased the expression of TIMP1 and TIMP3. The antinociceptive activity and chondroprotective effects were greater in OA rats treated with metformin-stimulated Ad-hMSCs than in those treated with unstimulated Ad-hMSCs. TGF-β expression in subchondral bone of OA joints was attenuated more in OA rats treated with metformin-stimulated Ad-hMSCs. Our findings suggest that metformin offers a promising option for the clinical application of Ad-hMSCs as a cell therapy for OA.

Transcutaneous Electrical Nerve Stimulation Reduces Knee Osteoarthritic Pain by Inhibiting Spinal Glial Cells in Rats

Transcutaneous electrical nerve stimulation (TENS) is commonly used for pain control. However, the effects of TENS on osteoarthritis (OA) pain and potential underlying mechanisms remain unclear. The objective of this study was to investigate the effect of TENS on OA pain treatment and underlying mechanisms related to glial cell inhibition. This was an experimental study. OA was induced by injection of monosodium iodoacetate into the synovial space of the right knee joint of rats. High-frequency (HF) TENS (100 Hz), low-frequency (LF) TENS (4 Hz), or sham TENS was applied to the ipsilateral knee joint for 20 minutes. Paw withdrawal threshold (PWT), weight bearing, and knee bend score (KBS) were measured. Immunohistochemistry for microglia and astrocytes was performed with L3 to L5 spinal segment samples. To investigate the effects of glial inhibition on OA pain, minocycline, l-α-aminoadipate, or artificial cerebrospinal fluid was injected intrathecally, and PWT and KBS were measured. Compared with sham TENS, both HF TENS and LF TENS significantly increased PWT, decreased KBS, and inhibited activated microglia in the L3 to L5 segments but did not decrease the total number of microglia, except in the L4 segment (HF TENS). Astrocyte expression was significantly decreased in the L3 to L5 segments following LF TENS and in the L3 segment following HF TENS. Compared with artificial cerebrospinal fluid, both minocycline and l-α-aminoadipate increased PWT and decreased KBS. These results cannot be generalized to humans. TENS alleviates OA pain in rats by inhibiting activated microglia and reducing astrocyte expression in the spinal cord. Although these results may not be generalizable to chronic pain in patients with OA, within the limitation of the experimental animal model used in the present study, they suggest a possible mechanism and preclinical evidence supporting further experimentation or clinical use of TENS in humans.

MSC exosomes alleviate osteoarthritis through restoration of matrix homeostasis

Background & Aim The efficacy of mesenchymal stromal cells (MSC) therapies is increasingly attributed to paracrine secretion, particularly exosomes. This study aims to elucidate the mechanisms of action of MSC exosomes in an immunocompetent rat model of temporomandibular joint osteoarthritis (TMJ-OA). Methods, Results & Conclusion Exosomes were isolated from conditioned medium of human MSCs. OA of bilateral TMJs was induced in rats by injection of monosodium iodoacetate. Thereafter, weekly intra-articular injections of exosomes (100µg/50µl) or 50µl of vehicle (PBS) were given over 2, 4 and 8 weeks. Analyses were performed by head withdrawal threshold (HWT) measurement, micro-computed tomography, histology and immunohistochemistry, and Mankin scoring. Gene expression changes were measured by transcriptomic analysis of the condylar cartilage tissue. Cell-based assays were performed to determine the cellular processes and signalling pathways activated by MSC exosomes. Our results showed that exosome-mediated repair of the osteoarthritic TMJs was characterized by early suppression of pain and degeneration with reduced inflammation, followed by sustained proliferation and gradual improvements in matrix expression and subchondral bone architecture, leading to overall joint restoration and regeneration by the end of treatment. By 8 weeks, exosome-treated rats had reduced pain with HWT comparable to that of sham rats, and displayed cartilage and subchondral bone restoration including appropriate cartilage thickness, cellularity, matrix and subchondral bone architecture that closely resembled that of the naive rats. Using an in-vitro chondrocyte model of OA, MSC exosomes increased sulphated glycosaminoglycan synthesis and reduced nitric oxide production in interleukin-1β-treated chondrocytes through exosomal CD73-mediated adenosine activation of AKT and AMPK signalling. These effects were partially abrogated by theophylline, wortmannin or compound C, which inhibited adenosine receptor activation, AKT and AMPK phosphorylation, respectively. Together, our observations suggest that MSC exosomes promote TMJ repair and regeneration in OA through a well-orchestrated mechanism of action that involved multiple cellular processes to restore the matrix and overall joint homeostasis.

The Effect of Mesenchymal Stem Cell Wharton's Jelly on ADAMTS-4 and iNOS Levels in Osteoarthritis Rat Model.

BACKGROUND:Osteoarthritis (OA) is one of the most common diseases among the elderly. OA occurs due to an imbalance between degradation and synthesis in articular joint tissue, causing changes in joint components such as cells, matrices and molecular production. Therefore, knowledge of cartilage-degrading enzymes such as ADAMTS-4 and iNOS is needed.AIM:This study aims to prove the effect of Mesenchymal Stem Cell Wharton Jelly on decreasing ADAMTS-4 levels as cartilage-degrading enzymes and increasing levels of iNOS which showed the immunosuppressive potential of MSC-WJ in cases of osteoarthritis in vivo.MATERIAL AND METHODS:This research is an experimental study with the design of Post-test-Only Control Group Design. The sample consisted of 16 OA rats as a control group and 16 OA rats treated with MSC-WJ as a treatment group. OA induction is done by injection of monosodium iodoacetate (MIA) into the intra-articular right knee. Giving MSC-WJ is done in the third week after MIA induction. The serum ADAMTS-4 and iNOS levels were measured after 3 weeks treated with MSC-WJ using the ELISA method. The statistical test used is an independent t-test. The value of p < 0.05 was said to be statistically significant.RESULT:The result showed that serum ADAMTS-4 levels were lower in the group treated with MSC-WJ than in the control group, but not statistically significant (p > 0.05). Serum iNOS levels were higher in the group treated with MSC-WJ than in the control group (p < 0.05).CONCLUSION:This study concluded that MSC-WJ significantly reduced ADAMTS-4 levels and increased the serum iNOS levels of OA rats.

Condylotomy to Reverse Temporomandibular Joint Osteoarthritis in Rabbits

Osteoarthritis (OA) of the temporomandibular joint (TMJ) is characterized by local tissue degeneration and pain. Treatments have been aimed at reducing symptoms, and only some can interfere with the progression of pathophysiologic changes caused by OA. Condylotomy is an alternative treatment for patients with OA refractory to conventional treatment. The aim of the present animal study was to investigate the influence of condylotomy on TMJ OA in New Zealand rabbits. The 36 rabbits were divided into 3 groups: the osteoarthritis group (OG; induced OA), treatment group (TG; induced OA plus condylotomy), and control group (CG; neither OA nor surgery). OA was induced using intra-articular monosodium iodoacetate injection for 40days. The rabbits in the TG underwent condylotomy and were killed 20, 40, and 60days after treatment. The rabbits in the CG and OG were killed at the same points. The articular joint condition was better in the TG 60days after surgery (P=.032). A direct comparison revealed regression of TMJ OA over time among the treated rabbits (P=.008). Surgical treatment promoted mandibular condylar remodeling in the TG, reversing the tissue degeneration caused by OA. Our findings suggest that condylotomy could be an option for the treatment of OA and prevent damage to TMJ structures. This could be of particular importance for patients without satisfactory responses to more conservative treatment.