The expression of nerve growth factor in healthy and inflamed equine chondrocytes analysed by capillary western immunoassay

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Fältnamn	Indikatorer	Metadata
000		04571naa a2200541 4500
001		oai:gup.ub.gu.se/322294
003		SwePub
008		240528s2022 \| \|\|\|\|\|\|\|\|\|\|\|000 \|\|eng\|
009		oai:slubar.slu.se:119810
024	7	a https://gup.ub.gu.se/publication/3222942 URI
024	7	a https://doi.org/10.1016/j.rvsc.2022.08.0152 DOI
024	7	a https://res.slu.se/id/publ/1198102 URI
040		a (SwePub)gud (SwePub)slu
041		a eng
042		9 SwePub
072	7	a ref2 swepub-contenttype
072	7	a art2 swepub-publicationtype
100	1	a Nystrom, S.4 aut
245	1 0	a The expression of nerve growth factor in healthy and inflamed equine chondrocytes analysed by capillary western immunoassay
264	1	b Elsevier BV,c 2022
520		a Nerve Growth Factor (NGF) is a signalling molecule for pain and inflammation. NGF is increased in synovial fluid from osteoarthritic humans and animals, compared to healthy controls. Monoclonal antibody therapy directed against NGF has been approved to treat pain in osteoarthritic dogs but despite many years of trialling, therapy has not been approved for human use. One reason for this is that adverse reactions with rapidly progressing osteoarthritis has occurred in some individuals. More detailed knowledge of NGF expression in joints is needed. In this study, capillary-based Simple Western was used to analyse NGF in cultured equine chondrocytes. Chondrocytes were collected post mortem from three macroscopically healthy intercarpal joints and three intercarpal joints with mild osteoarthritic changes. The chondrocytes were expanded to passage one and seeded in chondrogenic medium to maintain the phenotype. On day four, cells were either stimulated with LPS or kept untreated in medium. All cells were harvested on day five. Wes analysis of lysates did not show mature NGF but two proforms, 40 and 45 kDa, were identified. Results were confirmed with western blot. The same proforms were expressed in chondrocytes from healthy and osteoarthritic joints. Acute inflammation induced by LPS stimulation did not change the forms of expressed NGF. Capillary Simple Western offers a sensitive and sample -sparing alternative to traditional western blot. However, confirmation of peaks is imperative in order to avoid misinterpretation of findings. In addition, in this case the method did not offer the possibility of quantification advertised by the manufacturers.
650	7	a LANTBRUKSVETENSKAPERx Veterinärmedicin0 (SwePub)4032 hsv//swe
650	7	a AGRICULTURAL SCIENCESx Veterinary Science0 (SwePub)4032 hsv//eng
650	7	a LANTBRUKSVETENSKAPERx Veterinärmedicinx Patobiologi0 (SwePub)403022 hsv//swe
650	7	a AGRICULTURAL SCIENCESx Veterinary Sciencex Pathobiology0 (SwePub)403022 hsv//eng
653		a Equine
653		a Chondrocyte
653		a Osteoarthritis
653		a Nerve growth factor
653		a Simple western
653		a synovial-fluid
653		a factor ngf
653		a precursor
653		a beta
653		a prongf
653		a trka
653		a Veterinary Sciences
700	1	a Kendall, Annau Swedish University of Agricultural Sciences,Sveriges lantbruksuniversitet,Inst för biomedicin och veterinär folkhälsovetenskap,Department of Biomedical Science and Veterinary Public Health4 aut0 (Swepub:slu)51201
700	1	a Adepu, S.u Swedish University of Agricultural Sciences,Sveriges lantbruksuniversitet,Inst för biomedicin och veterinär folkhälsovetenskap,Department of Biomedical Science and Veterinary Public Health4 aut
700	1	a Lindahl, Anders,d 1954u Gothenburg University,Göteborgs universitet,Institutionen för biomedicin, avdelningen för laboratoriemedicin,Department of Laboratory Medicine4 aut0 (Swepub:gu)xlandy
700	1	a Skiöldebrand, Evau Swedish University of Agricultural Sciences,Sveriges lantbruksuniversitet,Inst för biomedicin och veterinär folkhälsovetenskap,Department of Biomedical Science and Veterinary Public Health4 aut0 (Swepub:slu)50022
710	2	a Sveriges lantbruksuniversitetb Inst för biomedicin och veterinär folkhälsovetenskap4 org
710	2	a Sveriges lantbruksuniversitet
773	0	t Research in Veterinary Scienced : Elsevier BVg 151, s. 156-163q 151<156-163x 0034-5288x 1532-2661
856	4	u https://pub.epsilon.slu.se/id/eprint/29819/contentsx primaryx Raw objectx freey FULLTEXT
856	4 8	u https://gup.ub.gu.se/publication/322294
856	4 8	u https://doi.org/10.1016/j.rvsc.2022.08.015
856	4 8	u https://res.slu.se/id/publ/119810

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Av författaren/redakt...: Nystrom, S.; Kendall, Anna; Adepu, S.; Lindahl, Anders, ...; Skiöldebrand, Ev ...

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