Determination of hydroxyproline in tissues and the evaluation of the collagen content of the tissues

Ignat’eva, N. Yu.; Данилов, Н. А.; Averkiev, S. V.; Обрезкова, М. В.; Лунин, В. В.; Sobol, Emil N.

doi:10.1134/s106193480701011x

Cited by 232 publications

(161 citation statements)

References 12 publications

Supporting

Mentioning

151

Contrasting

Unclassified

Order By: Relevance

“…Total collagen content was determined by measuring the hydroxyproline content. Samples were hydrolysed at 110°C for 18hrs in concentrated HCL (38%) and assayed using a chloramine-T assay (Kafienah and Sims 2004) with a hydroxyproline-to-collagen ratio of 1:7.69 (Ignat'eva et al 2007). …”

Section: Quantitative Biochemical Analysismentioning

confidence: 99%

Expansion in the presence of FGF-2 enhances the functional development of cartilaginous tissues engineered using infrapatellar fat pad derived MSCs

Buckley

Kelly

2012

Journal of the Mechanical Behavior of Biomedical Materials

View full text Add to dashboard Cite

A bstractMSCs from non-cartilaginous knee joint tissues such as the infrapatellar fat pad (IFP) and synovium possess significant chondrogenic potential and provide a readily available and clinically feasible source of chondroprogenitor cells. Fibroblast growth factor-2 (FGF-2) has been shown to be a potent mitotic stimulator during ex vivo expansion of MSCs, as well as regulating their subsequent differentiation potential.The objective of this study was to investigate the longer term effects of FGF-2 expansion on the functional development of cartilaginous tissues engineered using MSCs derived from the IFP. IFP MSCs were isolated and expanded to passage 2 in a standard media formulation with or without FGF-2 (5ng/ml) supplementation.Expanded cells were encapsulated in agarose hydrogels, maintained in chondrogenic media for 42 days and analysed to determine their mechanical properties and biochemical composition. Culture media, collected at each feed, was also analysed for biochemical constituents.MSCs expanded in the presence of FGF-2 proliferated more rapidly, with higher cell yields and lower population doubling times. FGF-2 expanded MSCs generated the most mechanically functional tissue. Matrix accumulation was dramatically higher after 21 days for FGF-2 expanded MSCs, but decreased between day 21 and 42. By day 42, FGF-2 expanded MSCs had still accumulated ~1.4 fold higher sGAG and ~1.7 fold higher collagen compared to control groups. The total amount of sGAG synthesised (retained in hydrogels and released into the media) was ~ 2.4 fold higher for FGF-2 expanded MSCs, with only ~25% of the total amount generated being retained within the constructs. Further studies are required to investigate whether IFP derived MSCs have a diminished capacity to synthesise other matrix components important in the aggregation, assembly and retention of proteoglycans. In conclusion, expanding MSCs in the presence of FGF-2 rapidly accelerates chondrogenesis in 3D agarose cultures resulting in superior mechanical functionality.

show abstract

Section: Quantitative Biochemical Analysismentioning

confidence: 99%

Expansion in the presence of FGF-2 enhances the functional development of cartilaginous tissues engineered using infrapatellar fat pad derived MSCs

Buckley

Kelly

2012

Journal of the Mechanical Behavior of Biomedical Materials

View full text Add to dashboard Cite

show abstract

“…This assay is not collagen specific and is not entirely reliable in unpurified tissue samples, with results differing depending on assessment methods. 37 …”

Section: Studies Of Collagen Composition In Uterosacral/ Cardinal Ligmentioning

confidence: 99%

Recent studies of genetic dysfunction in pelvic organ prolapse: The role of collagen defects

Lim

Khoo

Wong

et al. 2014

Aust NZ J Obst Gynaeco

View full text Add to dashboard Cite

Gynaecologists are becoming increasingly aware that women with a family history of prolapse are at an increased risk of prolapse refractory to treatment. In the past five years, several genetic mutations have been shown to correlate with increased prolapse susceptibility. These mutations can result in disordered collagen metabolism, which weaken the fascial support of the pelvic organs. This review examines the contemporary evidence regarding the role of collagen in prolapse.

show abstract

“…Proteoglycan content was estimated by quantifying the amount of sulphated glycosaminoglycan (sGAG) using the dimethylmethylene blue dye-binding assay (Blyscan, Biocolor Ltd., Northern Ireland), with a chondroitin sulphate standard. Total collagen content was determined by measuring the hydroxyproline content, using a hydroxyproline-to-collagen ratio of 1:7.69 [48][49]. Briefly, samples were mixed with 38% hydrochloric acid (Sigma-Aldrich) and incubated at 110°C for 18 hours to allow hydrolysis to occur.…”

Section: Biochemical Analysismentioning

confidence: 99%

Engineering osteochondral constructs through spatial regulation of endochondral ossification

et al. 2013

View full text Add to dashboard Cite

Chondrogenically primed bone marrow derived mesenchymal stem cells (MSCs) have been shown to become hypertrophic and undergo endochondral ossification when implanted in vivo. Modulating this endochondral phenotype may be an attractive approach to engineering the osseous phase of an osteochondral implant. The objective of this study was to engineer an osteochondral tissue by promoting endochondral ossification in one layer of a bi-layered construct and stable cartilage in the other. The top-half of bi-layered agarose hydrogels were seeded with culture expanded chondrocytes (termed chondral layer) and the bottom half of the bi-layered agarose hydrogels with MSCs (termed osseous layer). Constructs were cultured in a chondrogenic medium for 21 days and thereafter were either maintained in a chondrogenic medium, transferred to a hypertrophic medium, or implanted subcutaneously into nude mice. This structured chondrogenic bi-layered co-culture was found to enhance chondrogenesis in the chondral layer, appearing to help re-establish the chondrogenic phenotype that is lost in chondrocytes during monolayer expansion. Furthermore, the bilayered co-culture appeared to suppress hypertrophy and mineralisation in the osseous layer.The addition of hypertrophic factors to the media was found to induce mineralisation of the osseous layer in vitro. A similar result was observed in vivo where endochondral ossification was restricted to the osseous layer of the construct leading to the development of an osteochondral tissue. This novel approach represents a potential new treatment strategy for the repair of osteochondral defects.

show abstract

Determination of hydroxyproline in tissues and the evaluation of the collagen content of the tissues

Cited by 232 publications

References 12 publications

Expansion in the presence of FGF-2 enhances the functional development of cartilaginous tissues engineered using infrapatellar fat pad derived MSCs

Expansion in the presence of FGF-2 enhances the functional development of cartilaginous tissues engineered using infrapatellar fat pad derived MSCs

Recent studies of genetic dysfunction in pelvic organ prolapse: The role of collagen defects

Engineering osteochondral constructs through spatial regulation of endochondral ossification

Contact Info

Product

Resources

About