Limits...
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

hCol1 protects against cartilage loss in mid to late stage murine PTOA.Panel (A) presents an array of representative 40x Safranin O/Fast Green stained sagittal sections from the medial compartment of sham and MLI joints 12 weeks post-injury under various treatment conditions (control = vehicle, LD = 3.8mg hCol1/day, HD = 38mg hCol1/day). Joint structures are labeled (F = femur, M = meniscus, T = tibia) and the tidemarks are denoted with a yellow dashed line in the zoomed images. Black scale bars depict 100μm. Cartilage architecture was evaluated using the Osteomeasure System to determine the tibial uncalcified cartilage area (B), the tibial calcified cartilage (C), the number of chondrocytes in the tibial uncalcified cartilage (D), and the number (E) and percentage (F) of Safranin-O positive (SafO+) chondrocytes in the tibial uncalcified cartilage. OARSI scoring of the sections analyzed by histomorphometry was also performed (G). For histomorphometry and cell counting, symbols (○) represent the average measurement made from 3 sections/joint. For OARSI Scoring, symbols (○) represent the average score for each joint based on scoring of 3 sections/joint by four observers. Bars in all graphs represent the average for each experimental group (± SEM, N = 6). Significant differences between experimental groups in the histomorphometry data (B-F) were identified via one-way ANOVA with a Tukey’s multiple comparisons post-test (*p<0.05, **p<0.01, ***p<0.001 compared to Control Sham; xp<0.05, xxp<0.01 compared to Control MLI, N = 6). Significant differences between experimental groups in the OARSI data (G) were identified via a Kruskal-Wallis Test with a Dunn’s multiple comparisons post-test (*p<0.05, **p<0.01 compared to Control Sham).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5383229&req=5

pone.0174705.g004: hCol1 protects against cartilage loss in mid to late stage murine PTOA.Panel (A) presents an array of representative 40x Safranin O/Fast Green stained sagittal sections from the medial compartment of sham and MLI joints 12 weeks post-injury under various treatment conditions (control = vehicle, LD = 3.8mg hCol1/day, HD = 38mg hCol1/day). Joint structures are labeled (F = femur, M = meniscus, T = tibia) and the tidemarks are denoted with a yellow dashed line in the zoomed images. Black scale bars depict 100μm. Cartilage architecture was evaluated using the Osteomeasure System to determine the tibial uncalcified cartilage area (B), the tibial calcified cartilage (C), the number of chondrocytes in the tibial uncalcified cartilage (D), and the number (E) and percentage (F) of Safranin-O positive (SafO+) chondrocytes in the tibial uncalcified cartilage. OARSI scoring of the sections analyzed by histomorphometry was also performed (G). For histomorphometry and cell counting, symbols (○) represent the average measurement made from 3 sections/joint. For OARSI Scoring, symbols (○) represent the average score for each joint based on scoring of 3 sections/joint by four observers. Bars in all graphs represent the average for each experimental group (± SEM, N = 6). Significant differences between experimental groups in the histomorphometry data (B-F) were identified via one-way ANOVA with a Tukey’s multiple comparisons post-test (*p<0.05, **p<0.01, ***p<0.001 compared to Control Sham; xp<0.05, xxp<0.01 compared to Control MLI, N = 6). Significant differences between experimental groups in the OARSI data (G) were identified via a Kruskal-Wallis Test with a Dunn’s multiple comparisons post-test (*p<0.05, **p<0.01 compared to Control Sham).

Mentions: A similar set of manual histomorphometric analyses were performed in Safranin O/Fast Green-stained sagittal joint sections harvested 12 weeks post-MLI. At this time point, hCol1 appeared to provide more significant protection than at 3 weeks, with representative sections depicting apparent preservation of articular cartilage area and cellularity (Fig 4A). Use of Osteomeasure again facilitated quantification of key cartilage structural elements, with clear hCol1 dose-dependent preservation of uncalcified tibial cartilage following MLI (Fig 4B). Complimenting this effect was a protection against MLI-induced expansion of the tibial calcified cartilage zone, with joints from HD hCol1-treated mice showing a significant reduction in calcified cartilage area (Fig 4C). Regarding cellularity, MLI-induced nearly complete loss of chondrocytes in the uncalcified tibial cartilage by 12 weeks, with hCol1 dose dependently protecting against this effect (Fig 4D). Remarkably, chondrocyte number in the HD hCol1 group was not significantly different from sham operated control group, indicating substantial efficacy of the supplement in preserving cartilage cellularity. Regarding the number and percentage of Safranin O-positive chondrocyte lacunae in the tibial uncalcified cartilage, the nearly complete loss of cells producing proteoglycan matrix 12 weeks post-MLI was fully rescued in mice supplemented with hCol1 (Fig 4E and 4F). In fact, suggestive of a chondroregenerative effect, both LD and HD hCol1 groups displayed both a greater number and greater percentage of Safranin O-stained lacunae in tibia uncalcified cartilage compared to Control Sham joints (Fig 4E and 4F). While, as expected, the OARSI score was significantly increased 12 weeks post-MLI, hCol1 supplemented mice only showed modest improvement of the score, with the LD group trending toward significance (Fig 4G). As discussed for analytics performed on joints at the 3 week time point, it should be noted that there were no significant differences between experimental groups in uncalcified and calcified cartilage area on the femoral condyles, and in the number of hypertrophic chondrocytes in any zone of the cartilage on the femoral condyles and tibial plateaus (data not shown).


Daily oral consumption of hydrolyzed type 1 collagen is chondroprotective and anti-inflammatory in murine posttraumatic osteoarthritis
hCol1 protects against cartilage loss in mid to late stage murine PTOA.Panel (A) presents an array of representative 40x Safranin O/Fast Green stained sagittal sections from the medial compartment of sham and MLI joints 12 weeks post-injury under various treatment conditions (control = vehicle, LD = 3.8mg hCol1/day, HD = 38mg hCol1/day). Joint structures are labeled (F = femur, M = meniscus, T = tibia) and the tidemarks are denoted with a yellow dashed line in the zoomed images. Black scale bars depict 100μm. Cartilage architecture was evaluated using the Osteomeasure System to determine the tibial uncalcified cartilage area (B), the tibial calcified cartilage (C), the number of chondrocytes in the tibial uncalcified cartilage (D), and the number (E) and percentage (F) of Safranin-O positive (SafO+) chondrocytes in the tibial uncalcified cartilage. OARSI scoring of the sections analyzed by histomorphometry was also performed (G). For histomorphometry and cell counting, symbols (○) represent the average measurement made from 3 sections/joint. For OARSI Scoring, symbols (○) represent the average score for each joint based on scoring of 3 sections/joint by four observers. Bars in all graphs represent the average for each experimental group (± SEM, N = 6). Significant differences between experimental groups in the histomorphometry data (B-F) were identified via one-way ANOVA with a Tukey’s multiple comparisons post-test (*p<0.05, **p<0.01, ***p<0.001 compared to Control Sham; xp<0.05, xxp<0.01 compared to Control MLI, N = 6). Significant differences between experimental groups in the OARSI data (G) were identified via a Kruskal-Wallis Test with a Dunn’s multiple comparisons post-test (*p<0.05, **p<0.01 compared to Control Sham).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5383229&req=5

pone.0174705.g004: hCol1 protects against cartilage loss in mid to late stage murine PTOA.Panel (A) presents an array of representative 40x Safranin O/Fast Green stained sagittal sections from the medial compartment of sham and MLI joints 12 weeks post-injury under various treatment conditions (control = vehicle, LD = 3.8mg hCol1/day, HD = 38mg hCol1/day). Joint structures are labeled (F = femur, M = meniscus, T = tibia) and the tidemarks are denoted with a yellow dashed line in the zoomed images. Black scale bars depict 100μm. Cartilage architecture was evaluated using the Osteomeasure System to determine the tibial uncalcified cartilage area (B), the tibial calcified cartilage (C), the number of chondrocytes in the tibial uncalcified cartilage (D), and the number (E) and percentage (F) of Safranin-O positive (SafO+) chondrocytes in the tibial uncalcified cartilage. OARSI scoring of the sections analyzed by histomorphometry was also performed (G). For histomorphometry and cell counting, symbols (○) represent the average measurement made from 3 sections/joint. For OARSI Scoring, symbols (○) represent the average score for each joint based on scoring of 3 sections/joint by four observers. Bars in all graphs represent the average for each experimental group (± SEM, N = 6). Significant differences between experimental groups in the histomorphometry data (B-F) were identified via one-way ANOVA with a Tukey’s multiple comparisons post-test (*p<0.05, **p<0.01, ***p<0.001 compared to Control Sham; xp<0.05, xxp<0.01 compared to Control MLI, N = 6). Significant differences between experimental groups in the OARSI data (G) were identified via a Kruskal-Wallis Test with a Dunn’s multiple comparisons post-test (*p<0.05, **p<0.01 compared to Control Sham).
Mentions: A similar set of manual histomorphometric analyses were performed in Safranin O/Fast Green-stained sagittal joint sections harvested 12 weeks post-MLI. At this time point, hCol1 appeared to provide more significant protection than at 3 weeks, with representative sections depicting apparent preservation of articular cartilage area and cellularity (Fig 4A). Use of Osteomeasure again facilitated quantification of key cartilage structural elements, with clear hCol1 dose-dependent preservation of uncalcified tibial cartilage following MLI (Fig 4B). Complimenting this effect was a protection against MLI-induced expansion of the tibial calcified cartilage zone, with joints from HD hCol1-treated mice showing a significant reduction in calcified cartilage area (Fig 4C). Regarding cellularity, MLI-induced nearly complete loss of chondrocytes in the uncalcified tibial cartilage by 12 weeks, with hCol1 dose dependently protecting against this effect (Fig 4D). Remarkably, chondrocyte number in the HD hCol1 group was not significantly different from sham operated control group, indicating substantial efficacy of the supplement in preserving cartilage cellularity. Regarding the number and percentage of Safranin O-positive chondrocyte lacunae in the tibial uncalcified cartilage, the nearly complete loss of cells producing proteoglycan matrix 12 weeks post-MLI was fully rescued in mice supplemented with hCol1 (Fig 4E and 4F). In fact, suggestive of a chondroregenerative effect, both LD and HD hCol1 groups displayed both a greater number and greater percentage of Safranin O-stained lacunae in tibia uncalcified cartilage compared to Control Sham joints (Fig 4E and 4F). While, as expected, the OARSI score was significantly increased 12 weeks post-MLI, hCol1 supplemented mice only showed modest improvement of the score, with the LD group trending toward significance (Fig 4G). As discussed for analytics performed on joints at the 3 week time point, it should be noted that there were no significant differences between experimental groups in uncalcified and calcified cartilage area on the femoral condyles, and in the number of hypertrophic chondrocytes in any zone of the cartilage on the femoral condyles and tibial plateaus (data not shown).

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