title: Inflammatory response of cultured rat synoviocytes challenged with synovial fluid from osteoarthritis patients correlates with their radiographic grading: a pilot study.
abstract: The inflammatory nature of synovial fluid (SF) of varying grade osteoarthritis (OA) patients was estimated by measuring pro-inflammatory factors and through a unique cell-challenge experiment. SF samples were collected from six OA and one non-OA patient; spanning Kellgren-Lawrence (KL) grades were analyzed for interlukin-1-beta (IL-1β), nitric oxide (NO) and its derivatives, and glycosaminoglycan (GAG). Levels of IL-1β, NO, and GAG in SF did not correlate with KL grades of the patients studied. In the cell-challenge experiment, cultured rat synoviocyte fibroblasts (RSFs) were challenged by the patient's SFs with and without pre-treatment of IL-1β and lipopolysaccharide (LPS). NO released by the cells was taken as an indicator of inflammation. SFs from KL grades 2 and 3 induced maximum inflammation in cultured RSFs (grade 2 64.61 ± 4.8 and 89.51 ± 5.6 μM/ml after 48 and 72 h, grade 3 58.27 ± 2.7 and 64.22 ± 2.8 μM/ml after 48 and 72 h, respectively). Similar trend was observed in RSF pretreated with either recombinant IL-1β or LPS suggesting that SF from patients KL grades 2 and 3 accumulates more pro-inflammatory factors. IL-1β-pre-treated RSFs challenged by SF for 72 h showed 234.41 ± 17.6 μM/ml increase (patient 3, grade 3), whereas higher NO after LPS pre-treatment was recorded (118.92 ± 6.2 μM/ml; patient 3, grade 3). Interestingly, SFs from grade 1 and non-OA patient could reduce released NO to 27.10 ± 2.2 μM/ml showing potency to alleviate inflammation. These interesting findings, however, need to be confirmed on a wider number of patients, which may offer significant therapeutic application in treatment of OA.
authors.affiliation: Department of Cell and Molecular Biology, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth University, Pune-Satara Road, Pune, Maharashtra, 411 043, India. soumya.koppikar@gmail.com.
authors.forename: Soumya J
authors.initials: SJ
authors.lastname: Koppikar
authors.affiliation: Department of Cell and Molecular Biology, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth University, Pune-Satara Road, Pune, Maharashtra, 411 043, India. pgkulkarni2610@googlemail.com.
authors.forename: Priya G
authors.initials: PG
authors.lastname: Kulkarni
authors.affiliation: Department of Cell and Molecular Biology, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth University, Pune-Satara Road, Pune, Maharashtra, 411 043, India. ingaledhanashri91@gmail.com.
authors.forename: Dhanashri R
authors.initials: DR
authors.lastname: Ingale
authors.affiliation: Department of Kayachikisa, Bharati Ayurvedic Hospital, Bharati Vidyapeeth University, Pune, Maharashtra, India. drshinde249@gmail.com.
authors.forename: Dattatray
authors.initials: D
authors.lastname: Shinde
authors.affiliation: Department of Orthopedics, Bharati Medical Hospital, Bharati Vidyapeeth University, Pune, Maharashtra, India. drnarendrwagh65@gmail.com.
authors.forename: NarendraKumar
authors.initials: N
authors.lastname: Wagh
authors.affiliation: Department of Orthopedics, Bharati Medical Hospital, Bharati Vidyapeeth University, Pune, Maharashtra, India. desh75@gmail.com.
authors.forename: Shantanu
authors.initials: S
authors.lastname: Deshpande
authors.affiliation: Department of Cell and Molecular Biology, Rajiv Gandhi institute of IT & BT, Bharati Vidyapeeth University, Pune, Maharashtra, India. alpanasm@gmail.com.
authors.forename: Alpana S
authors.initials: AS
authors.lastname: Moghe
authors.affiliation: Department of Cell and Molecular Biology, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth University, Pune-Satara Road, Pune, Maharashtra, 411 043, India. pranjekar@gmail.com.
authors.forename: Prabhakar K
authors.initials: PK
authors.lastname: Ranjekar
authors.affiliation: Department of Cell and Molecular Biology, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth University, Pune-Satara Road, Pune, Maharashtra, 411 043, India. aharsulkar@yahoo.com.
authors.forename: Abhay M
authors.initials: AM
authors.lastname: Harsulkar
mesh_terms: D000328:Adult; D000368:Aged; D000818:Animals; D002478:Cells, Cultured; D005260:Female; D006025:Glycosaminoglycans / Q000032:analysis; D006801:Humans; D007249:Inflammation / Q000503:physiopathology*; D053583:Interleukin-1beta / Q000032:analysis; D008297:Male; D008875:Middle Aged; D009569:Nitric Oxide / Q000032:analysis; D010003:Osteoarthritis / Q000000981:diagnostic imaging / Q000503:physiopathology*; D010865:Pilot Projects; D011859:Radiography; D051381:Rats; D017208:Rats, Wistar; D012720:Severity of Illness Index; D013582:Synovial Fluid / Q000737:chemistry / Q000166:cytology* / Q000502:physiology
chemical_list: D006025:Glycosaminoglycans; D053583:Interleukin-1beta; D009569:Nitric Oxide
keywords: Glycosaminoglycan; Interleukin-1-β; Nitric oxide; Osteoarthritis; Synoviocytes-like fibroblast cells