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Daily oral consumption of hydrolyzed type 1 collagen is chondroprotective and anti-inflammatory in murine posttraumatic osteoarthritis

View Article: PubMed Central - PubMed

ABSTRACT

Osteoarthritis (OA) is a degenerative joint disease for which there are no disease modifying therapies. Thus, strategies that offer chondroprotective or regenerative capability represent a critical unmet need. Recently, oral consumption of a hydrolyzed type 1 collagen (hCol1) preparation has been reported to reduce pain in human OA and support a positive influence on chondrocyte function. To evaluate the tissue and cellular basis for these effects, we examined the impact of orally administered hCol1 in a model of posttraumatic OA (PTOA). In addition to standard chow, male C57BL/6J mice were provided a daily oral dietary supplement of hCol1 and a meniscal-ligamentous injury was induced on the right knee. At various time points post-injury, hydroxyproline (hProline) assays were performed on blood samples to confirm hCol1 delivery, and joints were harvested for tissue and molecular analyses were performed, including histomorphometry, OARSI and synovial scoring, immunohistochemistry and mRNA expression studies. Confirming ingestion of the supplements, serum hProline levels were elevated in experimental mice administered hCol1. In the hCol1 supplemented mice, chondroprotective effects were observed in injured knee joints, with dose-dependent increases in cartilage area, chondrocyte number and proteoglycan matrix at 3 and 12 weeks post-injury. Preservation of cartilage and increased chondrocyte numbers correlated with reductions in MMP13 protein levels and apoptosis, respectively. Supplemented mice also displayed reduced synovial hyperplasia that paralleled a reduction in Tnf mRNA, suggesting an anti-inflammatory effect. These findings establish that in the context of murine knee PTOA, daily oral consumption of hCol1 is chondroprotective, anti-apoptotic in articular chondrocytes, and anti-inflammatory. While the underlying mechanism driving these effects is yet to be determined, these findings provide the first tissue and cellular level information explaining the already published evidence of symptom relief supported by hCol1 in human knee OA. These results suggest that oral consumption of hCol1 is disease modifying in the context of PTOA.

No MeSH data available.


Related in: MedlinePlus

Effective bolus delivery of hCol1 and experimental timeline.Hazelnut cream was used as a vehicle to deliver daily bolus doses of hCol1 to mice such that a delivery of a 150mg mixture provided a daily bolus dose of either 3.8mg (LD) or 38mg (HD) hCol1 (Control = hazelnut cream alone). Experimental mixtures were placed on autoclavable ceramic tiles (A) and presented to individually house mice (B) at the same time daily. After 5–7 days of training with vehicle alone, mice consumed the full amount presented within 2 minutes. Panel (C) depicts the experimental timeline. Mice were presented hazelnut cream daily in the bolus feeding regimen for a 1 week training period (blue line), and then Control, LD and HD daily supplements were initiated and continued for the remainder of the experiment (green line). After 4 weeks of supplementation, MLI (right knee) and Sham (left knee) surgery was performed (t = 0), followed by tissue harvests at 3 weeks and 12 weeks post-surgery. (D) To confirm successful delivery of hCol1, serum hProline levels were quantified via ELISA. Serum samples collected 1 week before (-1) and 2 weeks after surgery were harvested 3 hours after the mice consumed supplements (left graph). Serum samples collected 3 and 12 weeks after surgery were harvested 1 hour after consumption of the supplements (right graph). Symbols (○) represent the hProline level in the serum of individual mice. Bars represent the average hProline level for each experimental group (± SEM, N = 6). Significant differences between groups were identified via two-way ANOVA with a Tukey’s multiple comparisons post-test (*p<0.05, **p<0.01 compared to Control).
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pone.0174705.g001: Effective bolus delivery of hCol1 and experimental timeline.Hazelnut cream was used as a vehicle to deliver daily bolus doses of hCol1 to mice such that a delivery of a 150mg mixture provided a daily bolus dose of either 3.8mg (LD) or 38mg (HD) hCol1 (Control = hazelnut cream alone). Experimental mixtures were placed on autoclavable ceramic tiles (A) and presented to individually house mice (B) at the same time daily. After 5–7 days of training with vehicle alone, mice consumed the full amount presented within 2 minutes. Panel (C) depicts the experimental timeline. Mice were presented hazelnut cream daily in the bolus feeding regimen for a 1 week training period (blue line), and then Control, LD and HD daily supplements were initiated and continued for the remainder of the experiment (green line). After 4 weeks of supplementation, MLI (right knee) and Sham (left knee) surgery was performed (t = 0), followed by tissue harvests at 3 weeks and 12 weeks post-surgery. (D) To confirm successful delivery of hCol1, serum hProline levels were quantified via ELISA. Serum samples collected 1 week before (-1) and 2 weeks after surgery were harvested 3 hours after the mice consumed supplements (left graph). Serum samples collected 3 and 12 weeks after surgery were harvested 1 hour after consumption of the supplements (right graph). Symbols (○) represent the hProline level in the serum of individual mice. Bars represent the average hProline level for each experimental group (± SEM, N = 6). Significant differences between groups were identified via two-way ANOVA with a Tukey’s multiple comparisons post-test (*p<0.05, **p<0.01 compared to Control).

Mentions: All handling of mice and in vivo experimental procedures performed in studies reported here were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Rochester (protocol number UCAR-2005-226R). Male C57BL/6J mice were purchased from Jackson Laboratories and were housed individually in micro-isolator cages on a 12 hour light/dark cycle. Male mice were used in this study due to a faster and more temporally predictable progression of degeneration that has been documented in models of PTOA [22]. Mice had ad libitum access to standard chow and fresh water, and were supplemented with hCol1 of bovine origin and a mean molecular weight of 2000 Da (Peptan® B2000, Rousselot) using a method previously described to deliver daily doses of estradiol [23]. Briefly, hCol1 was incorporated into hazelnut cream such that 150mg of the mixture would deliver either low dose (LD, 3.8mg) or high dose (HD, 38mg) hCol1 when completely consumed. The HD is the body weight adjusted mouse equivalent to the 7.4g/day recommended human dose. At the beginning of the experimental time line (Fig 1C), 12 week old mice were presented with an autoclavable ceramic tile loaded with a 150mg aliquot of hazelnut cream vehicle, LD hCol1 or HD hCol1 (Fig 1A). These experimental supplements were provided daily at the same time (in the morning), and once trained, the mice consumed the entire provided amount within 2 minutes (Fig 1B). Modeling the daily consumption regimen suggested for lifelong joint health in humans, mice were fed the supplements daily during the entire experimental protocol until collection of experimental endpoints (Fig 1D).


Daily oral consumption of hydrolyzed type 1 collagen is chondroprotective and anti-inflammatory in murine posttraumatic osteoarthritis
Effective bolus delivery of hCol1 and experimental timeline.Hazelnut cream was used as a vehicle to deliver daily bolus doses of hCol1 to mice such that a delivery of a 150mg mixture provided a daily bolus dose of either 3.8mg (LD) or 38mg (HD) hCol1 (Control = hazelnut cream alone). Experimental mixtures were placed on autoclavable ceramic tiles (A) and presented to individually house mice (B) at the same time daily. After 5–7 days of training with vehicle alone, mice consumed the full amount presented within 2 minutes. Panel (C) depicts the experimental timeline. Mice were presented hazelnut cream daily in the bolus feeding regimen for a 1 week training period (blue line), and then Control, LD and HD daily supplements were initiated and continued for the remainder of the experiment (green line). After 4 weeks of supplementation, MLI (right knee) and Sham (left knee) surgery was performed (t = 0), followed by tissue harvests at 3 weeks and 12 weeks post-surgery. (D) To confirm successful delivery of hCol1, serum hProline levels were quantified via ELISA. Serum samples collected 1 week before (-1) and 2 weeks after surgery were harvested 3 hours after the mice consumed supplements (left graph). Serum samples collected 3 and 12 weeks after surgery were harvested 1 hour after consumption of the supplements (right graph). Symbols (○) represent the hProline level in the serum of individual mice. Bars represent the average hProline level for each experimental group (± SEM, N = 6). Significant differences between groups were identified via two-way ANOVA with a Tukey’s multiple comparisons post-test (*p<0.05, **p<0.01 compared to Control).
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pone.0174705.g001: Effective bolus delivery of hCol1 and experimental timeline.Hazelnut cream was used as a vehicle to deliver daily bolus doses of hCol1 to mice such that a delivery of a 150mg mixture provided a daily bolus dose of either 3.8mg (LD) or 38mg (HD) hCol1 (Control = hazelnut cream alone). Experimental mixtures were placed on autoclavable ceramic tiles (A) and presented to individually house mice (B) at the same time daily. After 5–7 days of training with vehicle alone, mice consumed the full amount presented within 2 minutes. Panel (C) depicts the experimental timeline. Mice were presented hazelnut cream daily in the bolus feeding regimen for a 1 week training period (blue line), and then Control, LD and HD daily supplements were initiated and continued for the remainder of the experiment (green line). After 4 weeks of supplementation, MLI (right knee) and Sham (left knee) surgery was performed (t = 0), followed by tissue harvests at 3 weeks and 12 weeks post-surgery. (D) To confirm successful delivery of hCol1, serum hProline levels were quantified via ELISA. Serum samples collected 1 week before (-1) and 2 weeks after surgery were harvested 3 hours after the mice consumed supplements (left graph). Serum samples collected 3 and 12 weeks after surgery were harvested 1 hour after consumption of the supplements (right graph). Symbols (○) represent the hProline level in the serum of individual mice. Bars represent the average hProline level for each experimental group (± SEM, N = 6). Significant differences between groups were identified via two-way ANOVA with a Tukey’s multiple comparisons post-test (*p<0.05, **p<0.01 compared to Control).
Mentions: All handling of mice and in vivo experimental procedures performed in studies reported here were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Rochester (protocol number UCAR-2005-226R). Male C57BL/6J mice were purchased from Jackson Laboratories and were housed individually in micro-isolator cages on a 12 hour light/dark cycle. Male mice were used in this study due to a faster and more temporally predictable progression of degeneration that has been documented in models of PTOA [22]. Mice had ad libitum access to standard chow and fresh water, and were supplemented with hCol1 of bovine origin and a mean molecular weight of 2000 Da (Peptan® B2000, Rousselot) using a method previously described to deliver daily doses of estradiol [23]. Briefly, hCol1 was incorporated into hazelnut cream such that 150mg of the mixture would deliver either low dose (LD, 3.8mg) or high dose (HD, 38mg) hCol1 when completely consumed. The HD is the body weight adjusted mouse equivalent to the 7.4g/day recommended human dose. At the beginning of the experimental time line (Fig 1C), 12 week old mice were presented with an autoclavable ceramic tile loaded with a 150mg aliquot of hazelnut cream vehicle, LD hCol1 or HD hCol1 (Fig 1A). These experimental supplements were provided daily at the same time (in the morning), and once trained, the mice consumed the entire provided amount within 2 minutes (Fig 1B). Modeling the daily consumption regimen suggested for lifelong joint health in humans, mice were fed the supplements daily during the entire experimental protocol until collection of experimental endpoints (Fig 1D).

View Article: PubMed Central - PubMed

ABSTRACT

Osteoarthritis (OA) is a degenerative joint disease for which there are no disease modifying therapies. Thus, strategies that offer chondroprotective or regenerative capability represent a critical unmet need. Recently, oral consumption of a hydrolyzed type 1 collagen (hCol1) preparation has been reported to reduce pain in human OA and support a positive influence on chondrocyte function. To evaluate the tissue and cellular basis for these effects, we examined the impact of orally administered hCol1 in a model of posttraumatic OA (PTOA). In addition to standard chow, male C57BL/6J mice were provided a daily oral dietary supplement of hCol1 and a meniscal-ligamentous injury was induced on the right knee. At various time points post-injury, hydroxyproline (hProline) assays were performed on blood samples to confirm hCol1 delivery, and joints were harvested for tissue and molecular analyses were performed, including histomorphometry, OARSI and synovial scoring, immunohistochemistry and mRNA expression studies. Confirming ingestion of the supplements, serum hProline levels were elevated in experimental mice administered hCol1. In the hCol1 supplemented mice, chondroprotective effects were observed in injured knee joints, with dose-dependent increases in cartilage area, chondrocyte number and proteoglycan matrix at 3 and 12 weeks post-injury. Preservation of cartilage and increased chondrocyte numbers correlated with reductions in MMP13 protein levels and apoptosis, respectively. Supplemented mice also displayed reduced synovial hyperplasia that paralleled a reduction in Tnf mRNA, suggesting an anti-inflammatory effect. These findings establish that in the context of murine knee PTOA, daily oral consumption of hCol1 is chondroprotective, anti-apoptotic in articular chondrocytes, and anti-inflammatory. While the underlying mechanism driving these effects is yet to be determined, these findings provide the first tissue and cellular level information explaining the already published evidence of symptom relief supported by hCol1 in human knee OA. These results suggest that oral consumption of hCol1 is disease modifying in the context of PTOA.

No MeSH data available.


Related in: MedlinePlus