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    Biochemical effect evaluation of microbial chondroitin sulphate in experimental knee osteoarthritis model
    (Erenler, Ayse Sebnem; Karabulut, Aysun Bay; Otlu, Onder, 2022) Erenler, Ayse Şebnem; Bay Karabulut, Aysun; Sevimli, Resit; Geçkil, Hikmet; Akpolat, Nusret; Ünver, Tuba; Otlu, Önder
    Osteoarthritis (OA) is the most commonchronic joint disease, primarily due to aging.Chondroitin sulfate (CS) is a glycosaminoglycan(GAG) commonly used to treat osteoarthritis. CS candelay the progression of pathology or reversemorphological changes in joint structure.Traditionally CS is produced from animal sources.However, due to different reasons such ascontamination, ecological risA, and the possibility ofinfectious diseases, the trend towards microbialsources has increased because of its advantages suchas purer, more antiallergic, and lower MolecularWeight (MW) than animal sources. Biochemicalanalysis of Microbial CS (MCS), which is a new andsignificant alternative as a source of CS in the OAhealing process, has not been evaluated in theliterature yet. This study was designed to analyze thebiochemical effects of MCS produced by our teamfrom a microbial source, with an MW value of 269Daltons, on the osteoarthritis healing processcompared to the commercial form. We aim to reachdata that MCS has a higher antioxidant effect thananimalYsourced CS, and in this way, it is a moresuitable production for the treatment ofosteoarthritis. In this study, Anee osteoarthritis wassurgically induced in experimental rabbits; and TGFY1?, CAT, MPO, TOS, and OSI parameters measuredin blood samples before the operation and after thehealing period were analyzed comparatively. Afterthe surgical application, the rabbits were randomlydivided into three groups: control, animalYsourcedCS, and E. coli sourced MCS. The standard rabbitdiet was administered daily to 10 rabbits in Group 1(control), and CS and MCS were applied daily to theother groups as 17 mg/Ag for 12 weeAs. Bloodsamples were taAen from rabbits at the 12th weeAafter surgery, and TGFY1?, CAT, MPO, TOS, andOSI parameters were biochemically evaluated. Thisstudy has confirmed that the antioxidant propertiesof MCS and data on its effectiveness in controllingoxidative stress compared to animalYsourced CS.Based on these results, it can be concluded that MCShas a significant potency of nutraceutical andtherapeutic agents for OA treatment.
  • Küçük Resim Yok
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    Cloning and Expression vgb-kfo Genes in E. coli and Microbial Chondroitin Sulfate Production
    (AMER SCIENTIFIC PUBLISHERS, 2019) Erenler, Ayşe Şebnem; Geçkil, Hikmet; Akpolat, Nusret; Sevimli, Reşit; Ülke, Esra; Aliyeva, Aygün
    This study focused on a new plasmid and new recombinant strain developed for the production of microbial chondroitin sulfate a new and limited area of study the strategies we applied for the production of microbial chondroitin sulfate, and the possible contributions of this study to published research literature. In this study, pETM6-PACF, which carries the genes responsible for capsular chondroitin synthesis [kfA, kfoC, kfoF] was used as the basic plasmid. The Vitreoscilla hemoglobin gene region was transformed into this basic plasmid and the common expression of both gene groups was added to research literature for the first time. This plasmid was transferred to non-pathogenic E. coil (C2987) to produce a completely new chondroitin source specific to this study. Following the transformation by chondroitin synthesis, and the subsequent microbial production of chondroitin by the application of purification protocols, microbial chondroitin sulfate was produced in sulfate form. Consequently, in comparison to published literature, a product with a low molecular weight value of 269 Daltons was developed. This product, which has significant potential drug potency, can be used in many different areas as a novel and unique biomedical product.
  • Küçük Resim
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    Microbial chondroitin sulfate in experimental knee osteoarthritis model investigation of chondroprotective effect
    (Verduci Editore, 2021) Sevimli, Reşit; Erenler, Ayşe Şebnem; Bay Karabulut, Aysun; Akpolat, Nusret; Geçkil, Hikmet
    OBJECTIVE: Chondroitin sulfate (CS) is a glycosaminoglycan with proven anti-inflammatory, anti-apoptotic, anti-oxidant properties. CS increases type II collagen and proteoglycan synthesis in human joint chondrocytes. CS can reduce the production of pro-inflammatory mediators and proteases to improve the anabolic/catabolic balance of the extracellular cartilage matrix (ECM). Due to these characteristics, it is a natural compound that is considered to be Symptomatic Slow-Acting Drugs for Osteoarthritis (SYSADOA). Microbial chondroitin sulfate (MCS) was produced from two different bacterial sources using biotechnological methods by our team. In this study, we aimed to apply microbially produced CS and bovine-derived commercial CS forms to rabbit knees with osteoarthritis experimentally and to evaluate the results. MATERIALS AND METHODS: In this study, a cruciate ligament cutting model was applied to 40 New Zealand rabbits to induce experimental osteoarthritis. Four weeks after the surgical procedure, rabbits were divided into 4 groups as control, animal-derived MCS, E coli-derived MCS and PaJC-derived MCS group. The standard rabbit diet was fed to the control group, and the other groups were additionally fed 17 mg/kg/day CS/MCS for 12 weeks. The rabbits were sacrificed at the 12th week after surgery and the preparations obtained were evaluated histopathologically. RESULTS: As a result, it was observed that regeneration tissue was statistically significant in histopathological cartilage tissue compared to the control group of CS developed from different sources given to rabbits with osteoarthritis. It was determined that among the CS groups produced from different sources, the group with the highest chondroprotective effect was MCS originating from E.coli. CONCLUSIONS: This vegan product (MCS), which we obtained as a result of our study, was produced by our team from a microbial source. According to our analysis, it has the potential to be an effective alternative therapy agent in the treatment of osteoarthritis.