Effect of a solution of hyaluronic acid–chondroitin sulfate–N-acetyl glucosamine on the repair response of cartilage to single-impact load damage

Frances M. D. Henson Department of Veterinary Medicine, University of Cambridge, Cambridge, CB3 0ES, England.

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Alan M. J. Getgood Orthopedics Research Unit, Department of Surgery, Addenbrooke's Hospital, Hills Rd, Cambridge, CB2 0QQ, England.

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David M. Caborn Department of Orthopedic Surgery, College of Medicine, University of Louisville, Louisville, KY 40208.

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C. Wayne McIlwraith Gail Holmes Equine Orthopaedic Research Center, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

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Neil Rushton Orthopedics Research Unit, Department of Surgery, Addenbrooke's Hospital, Hills Rd, Cambridge, CB2 0QQ, England.

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Abstract

Objective—To investigate effects of 1% hyaluronic acid–chondroitin sulfate–N-acetyl glucosamine (HCNAG) on the damage repair response in equine articular cartilage.

Sample—Articular cartilage from 9 clinically normal adult horses.

Procedures—Full-thickness cartilage disks were harvested from the third metacarpal bone. Cartilage was single-impact loaded (SIL) with 0.175 J at 0.7 m/s and cultured in DMEM plus 1 % (vol/vol) HCNAG or fibroblastic growth factor (FGF)-2 (50 ng/mL). Histologic and immunohistochemical techniques were used to identify tissue architecture and apoptotic cells and to immunolocalize type I and II collagen and proliferating nuclear cell antigen (PCNA).

Results—Type II collagen immunoreactivity increased in SIL cartilage, compared with control samples. At days 14 and 28 (day 0 = initiation of culture), control samples had significantly fewer repair cells than did other treatment groups. In control samples and SIL + HCNAG, there was a significant decrease in apoptotic cell number, compared with results for SIL and SIL + FGF-2 samples. At days 14 and 28, there was a significant increase in chondrocytes stained positive for PCNA in the control samples.

Conclusions and Clinical Relevance—1% HCNAG significantly affected apoptotic and repair cell numbers in an SIL damage-repair technique in adult equine articular cartilage. However, HCNAG had no effect on the number of PCNA-positive chondrocytes or on type II collagen immunohistochemical results. The inclusion of 1% HCNAG in lavage solutions administered after arthroscopy may be beneficial to cartilage health by increasing the number of repair cells and decreasing the number of apoptotic cells.

Abstract

Objective—To investigate effects of 1% hyaluronic acid–chondroitin sulfate–N-acetyl glucosamine (HCNAG) on the damage repair response in equine articular cartilage.

Sample—Articular cartilage from 9 clinically normal adult horses.

Procedures—Full-thickness cartilage disks were harvested from the third metacarpal bone. Cartilage was single-impact loaded (SIL) with 0.175 J at 0.7 m/s and cultured in DMEM plus 1 % (vol/vol) HCNAG or fibroblastic growth factor (FGF)-2 (50 ng/mL). Histologic and immunohistochemical techniques were used to identify tissue architecture and apoptotic cells and to immunolocalize type I and II collagen and proliferating nuclear cell antigen (PCNA).

Results—Type II collagen immunoreactivity increased in SIL cartilage, compared with control samples. At days 14 and 28 (day 0 = initiation of culture), control samples had significantly fewer repair cells than did other treatment groups. In control samples and SIL + HCNAG, there was a significant decrease in apoptotic cell number, compared with results for SIL and SIL + FGF-2 samples. At days 14 and 28, there was a significant increase in chondrocytes stained positive for PCNA in the control samples.

Conclusions and Clinical Relevance—1% HCNAG significantly affected apoptotic and repair cell numbers in an SIL damage-repair technique in adult equine articular cartilage. However, HCNAG had no effect on the number of PCNA-positive chondrocytes or on type II collagen immunohistochemical results. The inclusion of 1% HCNAG in lavage solutions administered after arthroscopy may be beneficial to cartilage health by increasing the number of repair cells and decreasing the number of apoptotic cells.

Contributor Notes

Supported by ArthroDynamic Technologies Incorporated.

Presented as an abstract at the International Cartilage Repair Society Meeting, Barcelona, Spain, September 2010.

Address correspondence to Dr. Henson (fmdh1@cam.ac.uk).
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