Culture Duration Modulates Collagen Hydrolysate-Induced Tissue Remodeling in Chondrocyte-Seeded Agarose Hydrogels

Abstract

Media supplementation with collagen hydrolysate was hypothesized to increase the collagen content in engineered cartilage. By d28, hydrolysate at 0.5 mg/mL increased type II collagen content and 1 mg/mL increased mechanical properties, total collagen content, and type II collagen content over controls. By d42, however, controls possessed the highest GAG content and compressive Young's modulus. Real-time PCR found that 1 mg/mL increased type II collagen gene expression in d0 constructs, but increased MMP expression with no effect on type II collagen on d28. A 10 mg/mL concentration produced the lowest tissue properties, the lowest type II collagen gene expression on d0, and the highest MMP gene expression on d28. These results indicate that the duration of culture modulates the response of chondrocytes to collagen hydrolysate in 3D culture, transforming the response from positive to negative. Therefore, collagen hydrolysate as a media supplement is not a viable long-term method to improve the collagen content of engineered cartilage tissue.

Improvement of activity-related knee joint discomfort following supplementation of specific collagen peptides

“…Other in vitro experiments showed an increase of protease activity after collagen fragments supplementation (Fichter et al 2006;Jennings et al 2001). There is evidence that the effects of collagen hydrolysate depend on the concentration, the experimental conditions, (Ng et al 2007), and the characteristic of the tested collagen hydrolysates (Schadow et al 2013). However, the clinical relevance of the observed effects needs to be clarified.…”

“…The experiments showed a direct, dose-dependent stimulation of type II collagen and proteoglycans by the addition of collagen peptides. The stimulation of the extracellular matrix (ECM) synthesis is probably caused by specific Hyp-Pro-Gly containing peptides with a molecular size <10 kDa (Ng et al 2007;Oesser and Seifert 2003;Walrand et al 2008;Watanabe-Kamiyama et al 2010). Other in vitro experiments showed an increase of protease activity after collagen fragments supplementation (Fichter et al 2006;Jennings et al 2001).…”

The effect of concentration, thermal history and cell seeding density on the initial mechanical properties of agarose hydrogels

“…Over time, chondrocyte seeded agarose hydrogels have been shown to produce a functional extracellular matrix in free swelling culture (Buschmann et al 1992). For these and other reasons, numerous investigators have utilised agarose hydrogels for cartilage tissue engineering applications specifically to assess the elaboration of pericellular (Quinn et al 2002) and extracellular matrix (Miyata et al 2004;Miyata et al 2005;Mouw et al 2005), chondrogenesis of mesenchymal stem cells (Awad et al 2004;Mauck et al 2006), the interaction of biochemical stimulants (Coleman et al 2007;Ng et al 2007) and as a scaffold material in animal models of cartilage defect repair (Rahfoth et al 1998;Weisser et al 2001).…”

Ефективність І Безпечність Лікування Хворих На остеоартроз Колінних Суглобів Гідролізатом Колагену (FORTIGEL®)

“…Дослідження з ГК припускають, що пептиди можуть бути використані як будівельні блоки для хряща [19,23]. Тим не менше після проведення короткострокових експериментів дані про хондроцити в моношарі не були підтвердже-ні в довгострокових дослідженнях із хондроцитами в 3D-моделях [7,16]. Залишається очевидним, що мож-лива клінічна ефективність ГК спричинена або особ-ливим вмістом амінокислот (відносно велика кількість проліну та гліцину), або впливом пептидів, що всмок-туються через слизову оболонку кишечника.…”