Effect of SL-1010 (sodium hyaluronate with high molecular weight) on experimental osteoarthritis induced by intra-articularly applied papain in rabbits]

Sodium hyaluronate (HA) with a molecular weight of approximately 600,000-1,200,000 is reportedly effective against osteoarthritis (OA). However, since HA with higher molecular weight is expected to be more effective against OA, we investigated the effects of HA (SL-1010) newly produced by fermentation with a molecular weight of 1,800,000-2,100,000 on the experimental OA induced by intraarticular injection of papain, into the knee joint of the rabbit, in comparison with those of HA with a molecular weight of about 950,000 (HA-95). When 0.4, 0.8, and 1.6% papain (0.5ml) was injected into the knee joint of the animal twice with a 3-day interval, there were dose-dependent degenerative changes and a decrease in sulfated glycosaminoglycan (S-GAG) in the articular cartilage with slight synovial inflammatory changes 6 weeks after the final injection of papain. In this OA model, intraarticular application of SL-1010 slightly reduced the degeneration of articular cartilage, compared with the injections of HA-95 or saline (control). SL-1010 also caused a significant recovery in the S-GAG level which was decreased in the cartilage of the OA model, compared with the control. In addition, SL-1010 inhibited the release of 35S-GAG from the cartilage obtained from normal and OA model joints. These results suggest that SL-1010 is effective in inhibiting the degeneration of cartilage in the OA model, probably due to the recovery of the S-GAG level by reducing the release of S-GAG from the cartilage.     

Intraarticular drug delivery in osteoarthritis

Osteoarthritis (OA) is a primarily non-inflammatory, degenerative joint disease characterized by progressive loss of articular cartilage, subchondral bone sclerosis, osteophyte formation, changes in the synovial membrane, and an increased volume of synovial fluid with reduced viscosity and hence changed lubrication properties. As OA is the most common type of arthritis and a leading cause of disability, there is a largely unmet medical need for disease-modifying and symptomatic treatment. Due to the localized nature of the disease, intraarticular (IA) drug injection is an attractive treatment approach for OA. The various glucocorticoid and hyaluronic acid (HA) formulations, which are currently available on the market for IA treatment, provide only short-term pain relief or/and often do not provide adequate pain relief. The available oral drugs for symptomatic treatment also have shortcomings, most notably side effects. Therefore, there is still a large unmet need for novel OA drugs, which provide effective long-term pain relief and/or have disease-modifying properties. To achieve long-term drug exposure, different established formulations such as suspensions and hydrogels, and also novel approaches such as lipid based formulations and nano- or microparticles are currently in development. The development of novel drugs in combination with new formulations for IA treatment of OA, represents a promising approach in this challenging area of research.     

Intra-articular Injection of Urinary Bladder Matrix Reduces Osteoarthritis Development

Micronized porcine urinary bladder matrix (UBM) is an extracellular matrix biomaterial that has immunomodulatory and pro-regenerative properties. The objective of this study was to assess the ability of UBM to alter disease progression in a mouse model of post-traumatic osteoarthritis (OA). Ten-week-old wild-type C57BL/6 male mice underwent anterior cruciate ligament transection (ACLT) to induce OA. Two weeks after ACLT, UBM (50 mg/mL) or saline was injected into the mouse joint. At 4 and 8 weeks post-ACLT, cartilage integrity was assessed using OARSI scoring of histology, pain was evaluated, and joints were harvested for quantitative RT-PCR analysis of cartilage-specific and inflammatory gene expression. UBM-treated animals showed improved cartilage integrity at 4 and 8 weeks and reduced pain at 4 weeks compared to saline-injected mice. Animals injected with UBM expressed higher levels of genes encoding structural cartilage proteins, such as collagen2α1 and aggrecan, as well as anti-inflammatory cytokines, including interleukins 10 and 4. UBM decreased cartilage degeneration in the murine ACLT model of OA, which may be due to reduced inflammation in the joint and maintenance of high expression levels of proteoglycans.     

mTOR: A Potential Therapeutic Target in Osteoarthritis?

Osteoarthritis (OA) is a chronic degenerative joint disease characterized by the progressive loss of articular cartilage, remodeling of the subchondral bone, and synovial inflammation. Mammalian target of rapamycin (mTOR) is a serine/threonine protein kinase that controls critical cellular processes such as growth, proliferation, and protein synthesis. Recent studies suggest that mTOR plays a vital role in cartilage growth and development and in altering the articular cartilage homeostasis as well as contributing to the process of cartilage degeneration associated with OA. Both pharmacological inhibition and genetic deletion of mTOR have been shown to reduce the severity of OA in preclinical mouse models. In this review article, we discuss the roles of mTOR in cartilage development, in maintaining articular cartilage homeostasis, and its potential as an OA therapeutic target.     

膝关节液中透明质酸和Ⅱ型胶原羧基端端肽含量与滑膜炎关系的实验研究

目的探讨关节液中透明质酸(HA)和Ⅱ型胶原羧基端端肽(CTX-Ⅱ)含量与骨关节炎(OA)滑膜炎的关系。方法采用酶联免疫吸附试验对膝OA67例患者(OA组)、膝关节其他疾病77例患者(非OA组)的关节液中HA和CTX-Ⅱ含量进行检测,并在关节镜下利用Ayral滑膜炎评分法对滑膜炎程度进行评价。结果OA组关节液中HA含量明显低于非OA组(P<0.05);OA组关节液中CTX-Ⅱ含量高于非OA组(P<0.05);OA组关节液中HA含量与滑膜炎程度呈负相关,CTX-Ⅱ含量与滑膜炎程度呈正相关(P<0.05)。结论关节液中HA和CTX-Ⅱ含量是可反映OA滑膜炎的标志物。     

降钙素对膝骨关节炎保护作用的动物实验研究

目的通过观察降钙素（Calcitonin,CT）对动物膝骨关节炎（OA）软骨的影响,探讨降钙素对OA的治疗作用。方法6个月龄新西兰大白兔18只,随机分为3组,每组各6只,正常空白对照组（A组）：非手术,正常喂养;模型对照组（B组）：行双膝关节部分软骨缺损手术;CT组（C组）：行双膝关节部分软骨缺损手术,术后第2周开始,A、B组不予任何处理;C组按15U/kg皮下注射CT,1次/2天;共给药12周;术后12周处死动物。切取胫骨平台关节软骨大体观察,HE及甲苯胺蓝染色,检测血清、关节液。结果A组动物模型关节软骨正常;B组动物模型关节软骨缺损部分修复;C组动物模型关节软骨缺损完全修复。与B组比较,C组关节软骨形态学上明显改善,血清及关节液中SOD明显升高（P〈0.01）。结论降钙素能够明显降低兔膝关节骨关节炎软骨的退变,通过抑制关节软骨炎症,加强关节软骨下骨的吸收应力的能力来保护关节软骨。     

鹿瓜多肽对膝骨关节炎保护作用的动物实验研究

目的通过观察鹿瓜多肽对动物膝骨关节炎（OA）软骨的影响,探讨鹿瓜多肽对骨关节炎的治疗作用。方法6个月龄新西兰大白兔18只,采用随机数字表法分为3组,每组各6只,正常空白对照组：非手术,正常喂养;模型对照组：行双膝关节部分软骨缺损手术;观察组：行双膝关节部分软骨缺损手术,术后第2周开始,正常空白对照组、模型对照组不予任何处理;观察组按0．6ml／kg腹腔注射鹿瓜多肽,1次／d,术后12周处死动物。切取胫骨平台关节软骨大体观察,苏木素-伊红及甲苯胺蓝染色,检测血清、关节液。结果正常空白对照组动物模型关节软骨正常;模型对照组动物模型关节软骨缺损部分修复;观察组动物模型关节软骨缺损完全修复。观察组关节软骨形态学上明显改善,血清及关节液中超氧化物歧化酶（superox—idedismutase,SOD）分别为（117．31±11．49）mol／L、（76．28±19．11）mol／L,明显高于模型对照组[（102．37±11．54）mol／L,（57．58±17．31）mol／L]（P〈0．01）。结论鹿瓜多肽能够降低兔膝关节骨关节炎软骨的退变,通过抑制关节软骨炎症,加强关节软骨下骨的吸收应力的能力来保护关节软骨。     

Chondrocyte physiopathology and drug efficacy

After a brief exposition on the physiopathology of cartilage, and characteristic features of chondrocytes and proteoglycans (PGs) in osteoarthritis (OA), it is underlined how different molecules of GAGs and aggregated PGs added to the culture media can prevent damage and reduction of GAGs and fibril production in chondrocytes cultured with NSAIDs and corticosteroids. In animal models of OA, the local or general administration of GAGPS reduces the proteinase activity, the level of uronic acid in synovial fluid and the number of inflammatory cells in synovia. In the Pond-Nuki dog, GAGPS improves the cartilage surface. These favourable events can also occur in human OA, where it is, moreover, difficult to monitor the patients. For this purpose, patients must be selected in the first two stages of primary OA, and followed using NMR, the only device able to scan cartilage and subchondral bone, to determine their consistency and thickness, and to provide information on water content.     

机械加载减轻骨关节炎小鼠的病理进展

摘要：目的 探讨机械加载对骨关节炎（OA）模型小鼠早、晚期OA的治疗作用。方法 选用8周龄的C57BL/6 雌性小鼠36只，按随机数字表法分为正常对照组（Control组）、OA组和OA机械加载治疗组（OAL组），每组又分为2 周和8周亚组（n=6）。使用HE和番红O/快绿染色观察机械加载治疗对早、晚期OA小鼠软骨、滑膜和骨赘的治疗作 用。结果 与Control组相比，OA小鼠透明软骨（HC）厚度、全部关节软骨（TAC）厚度均变薄，国际骨关节炎研究学会 （OARSI）评分、滑膜评分、骨赘评分均升高（均P＜0.05）。机械加载治疗后，OAL组HC、TAC厚度均增加，OARSI评 分、滑膜评分、骨赘评分均下降（均P＜0.05）。相对数据表明，与早期OA小鼠相比，晚期小鼠的病理变化更严重，并 且机械加载对晚期OA的治疗效果更好。相关性分析表明，OA早期软骨的退化可能与滑膜炎症有关，而晚期可能与 骨赘形成有关。结论 机械加载可以改善OA小鼠的病理改变，可以作为一种新型的临床治疗手段。     

Osteoarthritis of the knee and hip. Part I: aetiology and pathogenesis as a basis for pharmacotherapy

Objectives Osteoarthritis (OA) of the knee and hip is among the most frequent and debilitating arthritic conditions. Aside from surgical intervention in severe cases, conventional treatment involves relieving painful symptoms with non‐steroidal anti‐inflammatory drugs (NSAIDs), narcotic and non‐narcotic (weak) analgesics and physical therapy. To obtain insight into the extent of pathological changes in hip and knee OA we reviewed current literature on the pathogenesis of this state as a basis for current pharmacotherapy options. Key findings Key features of the pathological joint changes in OA include: cartilage destruction by pro‐inflammatory cytokines, matrix metalloproteinases and prostaglandins, which promote a catabolic environment; subchondral bone remodelling and resorption; hypertrophic differentiation of chondrocytes; neovascularisation of synovial tissue; and focal calcification of joint cartilage. Despite the central involvement of hyaline cartilage in OA pathogenesis, the source of pain likely stems from the richly innervated synovium, subchondral bone and periosteum components of the joint. Tissue damage during joint degeneration generates nociceptive stimuli. The presence of inflammatory mediators, including bradykinin, prostaglandins and leukotrienes, lowers the threshold of the Aδ and C pain fibres, resulting in a heightened response to painful stimuli. Summary It is our opinion that it is important to base and centre the management of OA patients on the severity of patient‐important outcomes, rather than purely an assessment of damage to the joint. The joint damage, as interpreted from radiographs, is not necessarily representative of the symptoms experienced. The management of OA primarily comprises pharmacological therapy, surgical interventions and various non‐pharmacological interventions.     

A novel thermoresponsive hydrogel based on chitosan.

Injectable thermosetting chitosan hydrogels are attractive systems for drug delivery and tissue engineering that combine biodegradability, biocompatibility and the ability to form in situ gel-like implants. Thermally-induced gelation relies advantageously on biopolymer secondary interactions, avoiding potentially toxic polymerization reactions that may occur with in situ polymerizing formulations. In view of a biomedical use, such formulations have to be sterilizable and storable on extended periods without losing their thermosetting properties. These two key features have been studied in the present paper. Chitosans from two different sources were added with several phosphate-free polyols or polyoses as gelling agents. Despite a reduction in chitosan molecular weight following autoclaving, the hydrogels prepared with autoclaved chitosan showed the desired thermosetting properties. Hence, chitosan steam sterilization combined with aseptic preparation of the hydrogel allows a sterile formulation to be obtained. Whereas thermosetting hydrogels were shown to be unstable when refrigerated, freezing was shown to be conceivable as a storage method. When trehalose or mannitol was used as stabilizing agent, the formulation reconstituted from a lyophilizate displayed thermosetting properties and was still injectable, paving the way to the development of a clinically utilizable, novel chitosan thermosetting hydrogel.     

Structural and biological investigation of chitosan/hyaluronic acid with silanized-hydroxypropyl methylcellulose as an injectable reinforced interpenetrating network hydrogel for cartilage tissue engineering

Cartilage damage continues to pose a threat to humans, but no treatment is currently available to fully restore cartilage function. In this study, a new class of composite hydrogels derived from water-soluble chitosan (CS)/hyaluronic acid (HA) and silanized-hydroxypropyl methylcellulose (Si-HPMC) (CS/HA/Si-HPMC) has been synthesized and tested as injectable hydrogels for cartilage tissue engineering when combined without the addition of a chemical crosslinking agent. Mechanical studies of CS/HA and CS/HA/Si-HPMC hydrogels showed that as Si-HPMC content increased, swelling rate and rheological properties were higher, compressive strength decreased and degradation was faster. Our results demonstrate that the CS and HA-based hydrogel scaffolds, especially the ones with 3.0% (w/v) Si-HPMC and 2.5/4.0% (w/v) CS/HA, have suitable physical performance and bioactive properties, thus provide a potential opportunity to be used for cartilage tissue engineering. In vitro studies of CS/HA and CS/HA/Si-HPMC hydrogels encapsulated in chondrocytes have shown that the proper amount of Si-HPMC increases the proliferation and deposition of the cartilage extracellular matrix. The regeneration rate of the CS/HA/Si-HPMC (3%) hydrogel reached about 79.5% at 21 days for long retention periods, indicating relatively good in vivo bone regeneration. These CS/HA/Si-HPMC hydrogels are promising candidates for tissue compatibility injectable scaffolds. The data provide proof of the principle that the resulting hydrogel has an excellent ability to repair joint cartilage using a tissue-engineered approach.

RESEARCH HIGHLIGHTS

• An injectable hydrogel based on CS/HA/Si-HPMC composites was developed.
• The CS/HA/Si-HPMC hydrogel displays the tunable rheological with mechanical properties.
• The CS/HA/Si-HPMC hydrogel is highly porous with high swelling and degradation ratio.
• Increasing concentration of Si-HPMC promote an organized network in CS/HA/Si-HPMC hydrogels.
• Injectable CS/HA/Si-HPMC hydrogels have a high potential for cartilage tissue engineering.

     

Controlled release of celecoxib inhibits inflammation,bone cysts and osteophyte formation in a preclinical model of osteoarthritis

Major hallmarks of osteoarthritis (OA) are cartilage degeneration, inflammation and osteophyte formation. COX-2 inhibitors counteract inflammation-related pain, but their prolonged oral use entails the risk for side effects. Local and prolonged administration in biocompatible and degradable drug delivery biomaterials could offer an efficient and safe treatment for the long-term management of OA symptoms. Therefore, we evaluated the disease-modifying effects and the optimal dose of polyesteramide microspheres delivering the COX-2 inhibitor celecoxib in a rat OA model. Four weeks after OA induction by anterior cruciate ligament transection and partial medial meniscectomy, 8-week-old female rats (n?=?6/group) were injected intra-articular with celecoxib-loaded microspheres at three dosages (0.03, 0.23 or 0.39?mg). Unloaded microspheres served as control. During the 16-week follow-up, static weight bearing and plasma celecoxib concentrations were monitored. Post-mortem, micro-computed tomography and knee joint histology determined progression of synovitis, osteophyte formation, subchondral bone changes, and cartilage integrity. Systemic celecoxib levels were below the detection limit 6?days upon delivery. Systemic and local adverse effects were absent. Local delivery of celecoxib reduced the formation of osteophytes, subchondral sclerosis, bone cysts and calcified loose bodies, and reduced synovial inflammation, while cartilage histology was unaffected. Even though the effects on pain could not be evualated directly in the current model, our results suggest the application of celecoxib-loaded microspheres holds promise as novel, safe and effective treatment for inflammation and pain in OA.     

Anti-arthritic effects of chlorogenic acid in interleukin-1β-induced rabbit chondrocytes and a rabbit osteoarthritis model

Cartilage degradation is one of the pathological changes of osteoarthritis (OA), and accumulating evidence suggests an excess of matrix metalloproteinases (MMPs) plays a role in this cartilage breakdown. Here, we investigated the effects of chlorogenic acid (CGA) on the mRNA and protein expression of MMPs in interleukin (IL)-1β-induced rabbit chondrocytes and evaluated the in vivo effects of CGA in experimental OA induced by anterior cruciate ligament transection (ACLT) in rabbits. Using quantitative real-time PCR and ELISA to investigate the expression levels of MMP-1, MMP-3, MMP-13, and tissue inhibitors of metalloproteinase-1(TIMP-1) in IL-1β-induced rabbit chondrocytes, we showed that CGA inhibits the expression of these MMPs while increasing TIMP-1 expression, at both the mRNA and protein levels. In addition, IL-1β-induced activation of nuclear factor kappa B (NF-κB) and the degradation of inhibitor of κB (IκB)-α were suppressed by CGA. In rabbits, CGA decreased cartilage degradation as assessed by morphological and histological analyses. The down-regulation of MMP-1, MMP-3, and MMP-13 expression and up-regulation of TIMP-1 expression were also detected in CGA-treated cartilage compared with vehicle-treated cartilage, confirming these findings in an in vivo model. Taken together, these findings indicate that CGA may be considered as a possible candidate agent in the treatment of OA.     

HPLC-UV Method Validation for Amobarbital and Pharmaceutical Stability Evaluation When Dispersed in a Hyaluronic Acid Hydrogel: A New Concept for Post-Traumatic Osteoarthritis Prevention

A mitochondrial electron transport chain member complex I inhibitor, amobarbital, can reduce oxidative damage and chondrocyte death, eventually preventing post-traumatic osteoarthritis (PTOA). Viscosupplementation using a crosslinked hyaluronic acid (HA) hydrogel is currently applied clinically for knee OA pain relief. In this work, we utilized the HA hydrogel as a drug delivery vehicle to improve the long-term efficacy of amobarbital. Here we evaluated the pharmaceutic stability of amobarbital when dispersed in a crosslinked HA hydrogel formulated in proportions intended for clinical use. We validated a high-performance liquid chromatography with an ultraviolet detector (HPLC-UV) method following International Conference for Harmonization Q2(R1) guidelines to ensure its suitability for amobarbital detection. The feasibility of this formulation's drug delivery capability was proven by measuring the release, solubility, and drug uniformity. The amobarbital/HA hydrogel showed comparable amobarbital stability in different biological fluids compared to amobarbital solution. In addition, the amobarbital/HA hydrogel imparted significantly greater drug stability when stored at 70°C for 24 hours. In conclusion, we confirmed the pharmaceutical stability of the amobarbital/HA hydrogel in various conditions and biological fluids using a validated HPLC-UV method. This data provides essential evidence in support of the use of this amobarbital/HA formulation in future clinical trials for PTOA treatment.     

Cell-based therapy for management of osteoarthritis

Osteoarthritis(OA)is a most common form of degenerative joint disease,primarily characterized by the degradation of articular cartilage,subchondral sclerosis and inflammation of the synovial membrane.Mesenchymal stem cells(MSCs),a multipotent adult stem cell population,can be isolated from many connective tissue lineages,including those of the diarthrodial joint.Joint-resident MSCs or MSC-like progenitor cells contribute to the maintenance of healthy microenvironment or to the response to trauma.The onset of degenerative changes in the joint related to abnormal condition or depletion of these endogenous MSCs and native host hyaline cartilage cells,leading to limited selfrepair potential of the joint and advance of the degradation.To date,no acknowledged medical treatment strategies,including non-operative and classical surgical techniques,are efficient in restoring normal anatomy and function of hyaline cartilage in OA.This highlights an urgent need for better celled-based therapeutic strategies that supplement these functional cel s exogenously to recover the tissue homeostasis and repair in joint cavity via chondrogenic and anti-in fl ammatory functions.In this review we focus on the role of native MSCs in healthy or OA joint and recent progress in cel-based researches utilizing culture-expanded chondrocytes,pluripotent stem cel s,or MSCs from different sources for treating OA.     

Effects of diclofenac, aceclofenac and meloxicam on the metabolism of proteoglycans and hyaluronan in osteoarthritic human cartilage

1. Since nonsteroidal anti-inflammatory drugs (NSAIDs) may impair the ability of the chondrocyte to repair its damaged extracellular matrix, we explored the changes in the metabolism of newly synthesized proteoglycan and hyaluronan (HA) molecules produced by aceclofenac, diclofenac and meloxicam in human osteoarthritic (OA) cartilage. 2. Explants were sampled from the medial femoral condyle and were classified by use of the Mankin's histological-histochemical grading system. Cartilage specimens exhibited moderate (M) OA in 20 subjects and had severe (S) OA in 20. 3. Cartilage explants were pulsed with [-3H]-glucosamine and chased in the absence or in the presence of 0.3 - 3 microg ml(-1) of either aceclofenac, diclofenac or meloxicam. After papain digestion, the labelled chondroitin sulphate ([-3H]-proteoglycans) and [-3H]-HA molecules present in the tissue and media were purified by anion-exchange chromatography. 4. In cartilage with MOA and SOA, the metabolic balance of proteoglycan and HA was unaffected by diclofenac. In contrast, and in a dose-dependent manner, aceclofenac and meloxicam both increased the synthesis of proteoglycans and HA in explants with MOA and SOA; these two NSAIDs also reduced significantly the net loss of [-3H]-proteoglycans and [-3H]-HA molecules from cartilage explants. 5. The data obtained in short-term in vitro cultures indicate that, at the concentrations found in synovial fluid, aceclofenac and meloxicam may exert a favourable effect on the overall metabolism of proteoglycans and HA in cartilage with MOA and SOA.     

Salubrinal通过调节破骨细胞发育改善骨关节炎 胫骨平台不均匀沉降

摘要：目的 观察Salubrinal对早期骨关节炎（OA）小鼠胫骨平台不均匀沉降的治疗作用。方法 30只小鼠随机 分为对照（Sham）组、OA组和OA+Salubrinal（OA+Sal）组。采用手术切除小鼠膝关节内侧半月板建立OA模型,Sham 组只切开关节内侧皮肤;OA+Sal组小鼠皮下注射Salubrinal （1 mg/kg）2周,Sham组及OA组给予等量的生理盐水。采 用番红O和抗酒石酸酸性磷酸酶（TRAP）染色,观察胫骨平台内侧和外侧的组织形态学改变以及破骨细胞活性变化。 使用骨髓细胞的破骨细胞形成、迁移、黏附实验检测破骨细胞发育情况。结果 与Sham组相比,OA组破骨细胞发育 显著激活,软骨下骨的破骨细胞活性也明显增高;关节软骨OARSI评分、钙化软骨比例（CC/TAC）均显著升高（P＜ 0.05）;胫骨平台内侧软骨下骨的骨面积分数（B.Ar/T.Ar）明显增加（P＜0.05）,但外侧B.Ar/T.Ar、软骨下骨板（SBP）厚 度均显著降低（P＜0.05）。与OA组相比,OA+Sal组破骨细胞发育及软骨下骨破骨细胞活性被抑制,OARSI评分、CC/ TAC 均有所降低（P＜0.05）;而且,内外侧胫骨平台软骨下骨 B.Ar/T.Ar 和 SBP 厚度的不均匀改变均明显恢复（P＜ 0.05）。结论 Salubrinal通过调控破骨细胞发育对早期OA胫骨平台的骨重建和不均匀沉降有明显的改善作用。     

Formulation and evaluation of pH sensitive hydrogel of camptothecin with enhanced solubility by using β-cyclodextrin

The use of a novel injectable biocompatible and biodegradable camptothecin (CPT) formulation for controlled intra-tumoral release of CPT is described. The drug delivery vehicle is an autogelling pH sensitive formulation, which is based on the natural biopolymer chitosan. The formulation was prepared by Crosslinking methods. The formulations were characterized by Fourier transform infrared spectroscopy, X-ray diffraction, gelation time and viscosities were investigated for controlled release hydrogel formulation. The formulation, containing homogeneously dispersed CPT, was studied by MTT assay on tumor cell MCF-7. The effectiveness of treatment was measured in terms of percentage control tumor growth inhibition (TGI). The hydrogel formulation of CPT showed good release profile with polymer [chitosan (C)/glyceryl monooleate (GMO)/β-cyclodextrin (β-CD)] compare to without polymer. The gelation pH of the formulation PF, PF4, PF6, PF10 and PF12 were found to be 6.58, 7.42, 7.20, 6.98 and 7.30 respectively and calculated values of viscosity in centipoises of the formulation PF, PF4, PF6, PF10 and PF12 were 3,413.3, 5,843.1, 6,948.8, 5,212.6 and 6,972.6 respectively at 25 °C. It was found that cumulative percentage drug release for formulations prepared PF, PF4, PF6, PF10 and PF12 were released 48.48, 76.92, 83.72, 78.97 and 59.23 % respectively. The formulation PF6 showed maximum cumulative percentage drug release which has 3 % w/v chitosan, 3 % w/v GMO and 1.5 % w/v β-CD. The TGI was found that for formulations CPT, PF, PF4, PF6, PF10 and PF12 were 10.2 ± 1.22, 10.4 ± 0.85, 14.92 ± 1.06, 16.42 ± 1.11, 13.58 ± 1.21 and 11.71 ± 1.14 % respectively. The formulation PF6 showed maximum TGI in comparison to other formulation. The system formulated with CPT was found to be stable and the release profiles of a formulation with chitosan, GMO and β-CD showed all most effective release kinetics. These findings show chitosan/GMO/β-CD hydrogel to be a safe, effective, homogeneous, injectable and stable formulation for delivery of CPT and this approach represents an attractive technology platform for the delivery of other clinically important hydrophobic drugs.     

Increased expression levels of apolipoprotein J/clusterin during primary osteoarthritis

Osteoarthritis (OA) is a slowly progressive degenerative joint disease that is associated with joint space narrowing, osteophyte formation and subchondral sclerosis. Despite extensive effort actual breakthroughs in the field of genetic or biochemical biomarkers of OA are limited. As secretory apolipoprotein J/clusterin (sCLU) has been implicated in both inflammatory and apoptotic molecular processes which contribute to the OA phenotype, the sCLU concentration in human serum and synovial fluid during advanced primary knee and hip OA was analysed. Elevated sCLU protein levels were shown in these two biological fluids. sCLU mRNA expression was also studied in normal cartilage and in advanced primary knee and hip OA samples. A significant up-regulation of sCLU mRNA expression (~25-fold) was found in samples collected from the tibial bone that was osteotomized during total knee arthroplasty in patients with primary knee OA, as compared to healthy tissue samples collected from the femoral head of macroscopically normal cartilage during the surgical treatment of subcapital fractures. By studying sCLU mRNA expression levels in samples collected during total hip arthroplasty in patients with advanced primary hip OA, an additional up-regulation of the sCLU mRNA expression (~4-fold), as compared to advanced primary knee OA, was found. Taken together, these observations indicate that the sCLU protein or mRNA expression level may be of a significant diagnostic and/or prognostic value during OA progression.