Intra-articular injection of a nutritive mixture solution protects articular cartilage from osteoarthritic progression induced by anterior cruciate ligament transection in mature rabbits: a randomized controlled trial
© Park et al.; licensee BioMed Central Ltd. 2007
Received: 30 June 2006
Accepted: 26 January 2007
Published: 26 January 2007
Osteoarthritis (OA) is a degenerative disease that disrupts the collagenous matrix of articular cartilage and is difficult to cure because articular cartilage is a nonvascular tissue. Treatment of OA has targeted macromolecular substitutes for cartilage components, such as hyaluronic acid or genetically engineered materials. However, the goal of the present study was to examine whether intra-articular injection of the elementary nutrients restores the matrix of arthritic knee joints in mature animals. A nutritive mixture solution (NMS) was composed of elementary nutrients such as glucose or dextrose, amino acids and ascorbic acid. It was administered five times (at weeks 6, 8, 10, 13 and 16) into the unilateral anterior cruciate ligament transected knee joints of mature New Zealand White rabbits, and the effect of NMS injection was compared with that of normal saline. OA progression was histopathologically evaluated by haematoxylin and eosin staining, by the Mankin grading method and by scanning electron microscopy at week 19. NMS injection decreased progressive erosion of articular cartilage overall compared with injection of normal saline (P < 0.01), and nms joints exhibited no differences relative to normal cartilage that had not undergone transection of the anterior cruciate ligament, as assessed using the mankin grading method. Haematoxylin and eosin staining and scanning electron microscopy findings also indicated that nms injection, in constrast to normal saline injection, restored the cartilage matrix, which is known to be composed of a collagen and proteoglycan network. thus, nms injection is a potent treatment that significantly retards oa progression, which in turn prevents progressive destruction of joints and functional loss in mature animals.
Osteoarthritis (OA) is induced by complex mechanisms such as progressive erosion of articular cartilage, proteoglycan (PG) degradation and disruption of the collagen network, all of which lead to progressive destruction of joints and functional loss . Until recently the only therapies available to patients with OA were short-term relief agents [1–3], oral nutrient supplements [4–6], proliferative or regenerative therapies [7–10] and total surgical replacement of articular cartilage . In Korea, intra-articular injection is becoming increasingly popular because of its convenience and rapid effects. Agents that are commonly administered by injection include analgesics, nonsteroidal anti-inflammatory drugs (NSAIDs), steroids, hyaluronic acid and glucose . Analgesics, NSAIDs and steroids have anti-inflammatory effects. However, analgesics and NSAIDs provide only temporary pain relief [3, 13], and steroids are of limited use because of the resulting symptomatic 'dry' knees [12, 14]. Hyaluronic acid improves only molecular-weight-related short-term viscoelasticity of the joint synovial fluid [15, 16]. Injection of glucose or dextrose is used to manage chronic musculoskeletal pain, soft tissue injuries, and ligament and joint laxity . Although the therapeutic effects of dextrose or glucose are stronger with increased concentration , severe pain caused by inflammatory reactions at the injection site can also occur as a result of increased concentration .
Most patients would prefer treatments that are inexpensive and have long-term efficacy, and are less painful, less invasive and more easily accessible, and with fewer side effects than with existing treatments . Therefore, given the needs of OA patients and the limitations of existing OA treatments, we designed an intra-articular injection material that might confer greater therapeutic benefit in OA and fulfill patients' needs.
This material is a nutritive mixture solution (NMS), and it is formulated to supply nutrients to chondrocytes, which in turn synthesize collagen or proteoglycan (PG) to maintain the matrix network [20, 21]. Collagen fibres, especially type II collagen and PG, hold water to give tensile and compressive stiffness, and cartilage integrity depends on a successful symbiotic relationship between chondrocytes and interstitial matrix [22, 23]. NMS is composed of glucose or dextrose, several amino acids and ascorbic acid. Among the NMS components, glucose or dextrose plays a role in elevating levels of certain growth factors in ligaments after injury  and in serving as an energy substrate for chondrocytes and promoting matrix metabolism . The amino acids that we selected are the substrates for fibril forming collagen or PG in articular cartilage. They include glycine, proline, hydroxyproline, glutamate, alanine, aspartate, serine, glutamine, arginine, lysine and methionine [25, 26]. Cysteine is not a substrate of fibril forming collagen, but it protects cartilage from oxidative damage by acting as a thiol antioxidant . The anti-OA roles that these amino acids play in articular cartilage have been examined in in vitro or in vivo studies [27–38]. Finally, ascorbic acid is required for synthesis of type II collagen and PG as a cofactor in articular cartilage [39, 40].
In light of the physiological roles played by each of these elementary nutrients, we investigated the therapeutic effects of intra-articular injection of NMS on osteoarthritic knee joints, as compared with the effect of injection of normal saline (NS). The study was conducted using an experimental model in which OA develops as a result of anterior cruciate ligament transection (ACLT) in New Zealand White Rabbits with closed growth plates.
Materials and methods
Compositions of the NMS
Contents of 100 ml solution
Contents of 0.5 ml NMS
20.0 g (20%)
Amino acids solutionb
20.0 g (20%)
Ascorbic acid solutionb
5.0 g (5%)
Twenty-four mature New Zealand White rabbits (female, age 9 ± 2 months, body weight 3.6 ± 0.2 kg) were examined in this study. Ten rabbits were from the Laboratory Animal Research Center, Samsung Biomedical Research Institute (Samsung Medical Center, Seoul, South Korea) and 14 were from the Laboratory Animal Research Center, ChemOn Institute (Yongin, Gyunggi-do, South Korea). The rabbits were housed individually and had free access to tap water and commercial rabbit diet. The animal experiments were performed in accordance with internationally accredited guidelines, and were approved by each laboratory's Institutional Animal Care and Use Committee.
ACLT surgery for induction of osteoarthritis
Experimental OA in rabbits was induced by ACLT surgery in both groups. The rabbits were anaesthetized with intramuscular injection of ketamine (5 mg/kg) and butorphanol (0.1 mg/kg). After shaving and sterilizing the surgical site, ACLT was performed using a para-medial approach with the skin incision in the left knee medial para-patellar area. To achieve optimal visualization of the anterior cruciate ligament, the patellar bone was displaced laterally and the knee was placed in full flexion. The anterior stability was confirmed by an anterior drawer test . The synovium and the incised skin were sutured, and sterile dressing was applied. Following the surgical procedure, gentamycin (5 mg/kg) was injected intramuscularly into each rabbit once daily for a week.
Experimental protocol for treatment
Histopathological examinations: H&E staining
After the rabbits had been killed, the knee joints of the rabbits were dissected. The medial tibial plateaus and medial femoral condyles of the rabbits were fixed in 10% phosphate-buffered formalin (pH 7.4) with 1% cetylpyridinium (CPC) for 24 hours and decalcified with 20% EDTA. The decalcified specimens were embedded in paraffin and 1 μm thin sections were stained with H&E for light microscopic examination (×100) . The severity of articular cartilage lesions was graded through double-blind observations, using the histological grading method proposed by Mankin and coworkers . The Mankin grading method is a well known and proven method for the histological evaluation of OA cartilage. This method evaluates the severity of erosion and/or fissures of cartilage, disorganization or loss of chondrocytes, and pannus formation. Thus, the method adequately satisfies the criteria for measuring osteoarthritic changes in human and experimental animals [16, 48]. Other parenchymal organs were also examined to investigate possible deleterious effects of the treatment material.
Histopathological examinations: scanning electron microscopy
The extent of fibrillation and abrasion on the cartilage surface was observed in the photographs obtained by SEM (Joel 35CF; Tokyo, Japan; ×6,000). The microsections of cartilage of the medial tibial plateau, which is part of a weight-bearing joint, were washed with normal saline and pre-fixed in 2.5% glutaraldehyde-1/15 M phosphate buffer solution. After serial dehydration with ethanol, the ethanol was replaced with isoamyl acetate, and the samples were completely dried in a dryer. An ionic coater was used for gold deposition, and the coated samples were imaged by SEM .
The histopathological evaluation gradings obtained using the Mankin grading method  were pooled for the normal group (n = 7), the NMS group (n = 7), and the NS group (n = 11). The mean values of the grades were compared among the three groups by one-way analysis of variance and the Tukey HSD test or Kruskal-Wallis test, depending on normality of data (P < 0.05).
Histopathological results: H&E staining
Histopathologic evaluation of H&E stained cartilage by the Mankin grading method
Normal (n = 7)
NMS (n = 7)
NS (n = 11)
Medial tibial plateau
Cell in tangential zone
Cell in transitional and radial zone
Sum of scores
Medial femoral condyle
Cell in tangential zone
Cell in transitional and radial zone
Sum of scores
Histopathological results: scanning electron microscopy
Our experimental material, NMS, is a nutritive mixture solution that is designed to upregulate chondrocytes' regenerative potential to synthesize a collagen and PG network. The components of NMS are solutions of dextrose or glucose, amino acids and ascorbic acid. These elementary nutrients are substrates that can be delivered into the articular cartilage via the synovial route, which is a major nutrient transport pathway for ligaments and menisci of the articular joint . Articular cartilage has extremely small pores (estimated at 50 Å) in the superficial zone, and so only low-molecular-weight compounds (<20 kDa) in synovial fluid may diffuse into the tissue . All of the components of NMS can move freely through the tissue because they are not heavy molecular compounds. Moreover, articular chondrocytes have special transporter systems for glucose and ascorbic acid . Glucose is delivered to the chondrocytes via synovial microcirculation and taken up by glucose uptake (GLUT) proteins. The intracellular glucose pool is used for glycolysis and extracellular matrix macromolecules . The supply of glucose for anaerobic metabolism is essential to the survival and proliferation of chondrocytes and for the maintenance of matrix integrity. Therefore, impaired glucose uptake would compromise chondrocyte function, and potentially result in an imbalance in cartilage matrix synthesis and degradation, leading to OA . Ascorbic acid is transported into chondrocytes by the sodium-dependent vitamin C transporter (SVCT2), and has been shown to upregulate the expression of type II collagen and aggrecan . Ascorbic acid also plays an important role in chondrocyte proliferation and protection from oxidative stress .
In the case of amino acids, transporter systems in cartilage chondrocytes have not yet been identified, but glycine, proline, glutamine and glutamate transporters in chondrocytes have recently been investigated [53, 54]. Amino acids are expected not only to control chondrocyte gene expression  but also to synthesize collagen by chondrocytes . Therefore, amino acids in NMS were selected to provide substrates for fibril forming collagen and PG, based on their prevalence in the triple-helical structure of collagen, and depending on their specific biochemical and physiologic characteristics [21, 25]. Some amino acids' abilities to maintain cartilage integrity have already been revealed. For example, hydroxyproline stabilizes the collagen fibres to hold water  and glutamate prevents cartilage calcification [37, 38]. Glutamine protects articular chondrocytes from heat stress and nitric oxide induced apoptosis , it regulates collagen gene expression in cultured human fibroblasts , and it also increases collagen gene transcription . Arginine and lysine increase insulin-like growth factor-1 production and collagen synthesis . Lysine also slows the loss of collagen and PG from disrupted articular cartilage surfaces . Methionine stimulates synthesis and deposits of PG in articular cartilage [30, 34], and cysteine activates a signalling pathway in articular chondrocytes  and protects chondrocytes and cartilage from oxidative damage and degenerative processes such as OA . Therefore, sufficient nutrients from the metabolically active synovium reach the chondrocytes, presumably by diffusion through the cartilage matrix via the synovial fluid and various transporter systems. Finally, all of the components of NMS cooperate with each other in promoting chondrocyte activities to regenerate a collagen and PG network, and in preventing degenerative changes in articular cartilage. This is the great benefit of intra-articular injection of NMS, and one that existing OA treatments can not provide.
Existing OA treatments, such as intra-articular injections of either glucose or dextrose solution (5–25%) alone, are expected to yield osmotic changes and production of precursors for extracellular matrix macromolecules in the articular cartilage . Hypertonic solution is known to generate a better therapeutic effect, but it causes more discomfort from an inflammatory reaction [19, 57, 58]. The osmotic change in the knee joint cavity induced by 10% hypertonic dextrose, which we used in this study, is as follows: [296 (synovial fluid) + 505 (10% dextrose)]/2 = 400.5 mOsm. This osmolarity of 400 mOsm is of excellent therapeutic value, because it exerts a strong influence on proliferation of cells such as chondrocytes, osteocytes and fibroblasts . It also influences protein synthesis and amino acid (proline) transport without any cellular toxicity , and produces less pain than 20% dextrose solution does . Oral administration of glucosamine or chondroitin sulphate, which is a component of PG, plays a role as a symptomatic slow-acting drug in degenerative OA, but its effect is slow, small, or temporary [5, 60]. Intra-articular injection of hyaluronic acid plays a role in cartilage as a lubricant that lessens the frictional resistance of the cartilage , but it only generates temporary or placebo effects . Repeated injections of hyaluronic acid may deteriorate chondrocytes' PG biosynthetic ability , because hyaluronic acid is not a substrate for PG but a terminal material. Chondrocyte proliferation therapies, such as arthroscopic abrasion of the articular surface , osteotomy , transplantation of chondrocytes  or soft-tissue grafts [10, 64, 65], injections of growth factors [9, 66] and autologous blood , are also administered into the articular cartilage to stimulate proliferation of chondrocytes and repair cartilage matrix. However, these procedures are too expensive for general use, or they require long-term follow up because of the potential risk for cancer  or haemorrhagic arthritis .
Generally, cartilage in mature rabbits does not readily regenerate , and so histological changes after ACLT in rabbit knees, including cartilage hypertrophy, reduced cell density and matrix alterations preceding cartilage fibrillation, lead to progressive degeneration of cartilage . With ageing, the nutritional supply of cartilage diminishes because of degenerative changes of the joint cavity and decreased metabolism. However, if joint cavities were supplied with sufficient nutrients, they might recover from the nutritional deficiency caused by ageing, and OA progression might be inhibited. In this regard, intra-articular injection of NMS has the potential to induce chondrocytes to synthesize a collagen and PG network, which in turn maintains the cartilage matrix and protects against OA progression in the mature rabbit model, whereas NS injection has no such effect.
In summary, 0.5 ml of NMS or NS was intra-articularly administered into the knee joint cavity of mature rabbits for 13 consecutive weeks starting on week 6 after ACLT at 2-week or 3-week intervals, when arthritic changes had begun. It was found that only NMS injection significantly restored the extracellular matrix and inhibited the progression of OA-like changes in articular cartilage that had undergone ACLT. We suggest further comparative studies with other existing OA treatments, because in this study we only examined the effects of NMS on OA progression in comparison with a control (NS) treatment.
This study is the first trial to administer intra-articularly injectable material, not in the form of a macromolecular compound but in the form of a mixture of elementary nutrients, into the osteoarthritic articular cartilage. Each composition of the mixture, NMS, is likely to promote upregulated energy production in chondrocytes and extracellular matrix metabolism in articular cartilage, and to exert antioxidative effects in ageing chondrocytes. Based on the results of this study, NMS injection may be applied to osteoarthritic articular cartilage of adult animals as a very simple and effective treatment without significant adverse effects.
anterior cruciate ligament transection
haematoxylin and eosin
nutritive mixture solution
normal articular cartilage
nonsteroidal anti-inflammatory drugs
scanning electron microscopy.
The authors would like to thank Daehan Pharmaceutical Co. Ltd.(Seoul, South Korea) for supporting experimental materials, and Eugene Kim, the Director of Laboratory Animal Center, ChemOn Institute (Yongin, Gynggi-do, South Korea) for supporting animal handling and care.
- Woessner JF, Howell DS: Joint Cartilage Degradation: Basic and Clinical Aspects. 1993, New York: Marcel DekkerGoogle Scholar
- Brandt KD: Should nonsteroidal anti-inflammatory drugs be used to treat osteoarthritis?. Rheum Dis Clin North Am. 1993, 19: 29-44.PubMedGoogle Scholar
- Creamer P, Hunt M, Dieppe P: Pain mechanisms in osteoarthritis of the knee: effect of intraarticular anesthetic. J Rheumatol. 1996, 23: 1031-1036.PubMedGoogle Scholar
- Shikhman AR, Amiel D, D'Lima D, Hwang SB, Hu C, Xu A, Hashimoto S, Kobayashi K, Sasho T, Lotz MK: Chondroprotective activity of N-acetylglucosamine in rabbits with experimental osteoarthritis. Ann Rheum Dis. 2005, 64: 89-94. 10.1136/ard.2003.019406.PubMed CentralView ArticlePubMedGoogle Scholar
- Bucsi L, Poor G: Efficacy and tolerability of oral chondroitin sulfate as a symptomatic slow-acting drug for osteoarthritis (SYSADOA) in the treatment of knee osteoarthritis. Osteoarthritis Cartilage. 1998, 31-36. 6 Suppl A
- Najm WI, Reinsch S, Hoehler F, Tobis JS, Harvey PW: S-adenosyl methionine (SAMe) versus celecoxib for the treatment of osteoarthritis symptoms: a double-blind cross-over trial. [ISRCTN36233495]. BMC Musculoskelet Disord. 2004, 5: 6-10.1186/1471-2474-5-6.PubMed CentralView ArticlePubMedGoogle Scholar
- Levy AS, Lohnes J, Sculley S, LeCroy M, Garrett W: Chondral delamination of the knee in soccer players. Am J Sports Med. 1996, 24: 634-639.View ArticlePubMedGoogle Scholar
- Noguchi T, Oka M, Fujino M, Neo M, Yamamuro T: Repair of osteochondral defects with grafts of cultured chondrocytes. Comparison of allografts and isografts. Clin Orthop Relat Res. 1994, 302: 251-258.PubMedGoogle Scholar
- Buckwalter JA, Lohmander S: Operative treatment of osteoarthrosis. Current practice and future development. J Bone Joint Surg Am. 1994, 76: 1405-1418.PubMedGoogle Scholar
- Bobic V: Arthroscopic osteochondral autograft transplantation in anterior cruciate ligament reconstruction: a preliminary clinical study. Knee Surg Sports Traumatol Arthrosc. 1996, 3: 262-264. 10.1007/BF01466630.View ArticlePubMedGoogle Scholar
- Waikakul S, Vanadurongwan V, Bintachitt P: The effects of patellar resurfacing in total knee arthroplasty on position sense: a prospective randomized study. J Med Assoc Thai. 2000, 83: 975-982.PubMedGoogle Scholar
- Uthman I, Raynauld JP, Haraoui B: Intra-articular therapy in osteoarthritis. Postgrad Med J. 2003, 79: 449-453. 10.1136/pmj.79.934.449.PubMed CentralView ArticlePubMedGoogle Scholar
- Papathanassiou NP: Intra-articular use of tenoxicam in degenerative osteoarthritis of the knee joint. J Int Med Res. 1994, 22: 332-337.PubMedGoogle Scholar
- Raynauld JP: Clinical trials: impact of intraarticular steroid injections on the progression of knee osteoarthritis. Osteoarthritis Cartilage. 1999, 7: 348-349. 10.1053/joca.1998.0193.View ArticlePubMedGoogle Scholar
- Mandell BF, Lipani J: Refractory osteoarthritis. Differential diagnosis and therapy. Rheum Dis Clin North Am. 1995, 21: 163-178.PubMedGoogle Scholar
- Yoshimi T, Kikuchi T, Obara T, Yamaguchi T, Sakakibara Y, Itoh H, Iwata H, Miura T: Effects of high-molecular-weight sodium hyaluronate on experimental osteoarthrosis induced by the resection of rabbit anterior cruciate ligament. Clin Orthop Relat Res. 1994, 298: 296-304.PubMedGoogle Scholar
- Hauser GS: Answers to common questions about prolotherapy. Prolo Your Pain Away. Edited by: Hauser RA, Hauser MA, Pottinger K. 1998, Oak Park, IL: Beulah Land Press, 47-66. 1Google Scholar
- Peyron JG: Intraarticular hyaluronan injections in the treatment of osteoarthritis: state-of-the-art review. J Rheumatol Suppl. 1993, 39: 10-15.PubMedGoogle Scholar
- Reeves KD, Hassanein K: Randomized prospective double-blind placebo-controlled study of dextrose prolotherapy for knee osteoarthritis with or without ACL laxity. Altern Ther Health Med. 2000, 6: 68-74.PubMedGoogle Scholar
- Windhaber RA, Wilkins RJ, Meredith D: Functional characterisation of glucose transport in bovine articular chondrocytes. Pflugers Arch. 2003, 446: 572-577. 10.1007/s00424-003-1080-5.View ArticlePubMedGoogle Scholar
- Bhattacharjee A, Bansal M: Collagen structure: the Madras triple helix and the current scenario. IUBMB Life. 2005, 57: 161-172.View ArticlePubMedGoogle Scholar
- Buckwalter JA, Mankin HJ: Articular cartilage: tissue design and chondrocyte-matrix interactions. Instr Course Lect. 1998, 47: 477-486.PubMedGoogle Scholar
- Quinn TM, Morel V: Microstructural modeling of collagen network mechanics and interactions with the proteoglycan gel in articular cartilage. Biomech Model Mechanobiol. 2007, 6: 73-82. 10.1007/s10237-006-0036-z.View ArticlePubMedGoogle Scholar
- Liu YK, Tipton CM, Matthes RD, Bedford TG, Maynard JA, Walmer HC: An in situ study of the influence of a sclerosing solution in rabbit medial collateral ligaments and its junction strength. Connect Tissue Res. 1983, 11: 95-102.View ArticlePubMedGoogle Scholar
- Persikov AV, Ramshaw JA, Kirkpatrick A, Brodsky B: Amino acid propensities for the collagen triple-helix. Biochemistry. 2000, 39: 14960-14967. 10.1021/bi001560d.View ArticlePubMedGoogle Scholar
- Mankin HJ, Mow VC, Buckwalter JA, Iannotti JP, Ratclffe A: Articular cartilage structure, composition, and function. Orthopaedic Basic Science: Biology and Biomechanics of the Musculoskeletal System. Edited by: Buckwalter JA, Eidhorn TA, Simon SR. 2000, Rosemont, IL: American Academy of Orthopaedic Surgeons, 443-470. 2Google Scholar
- Li WQ, Dehnade F, Zafarullah M: Thiol antioxidant, N-acetylcysteine, activates extracellular signal-regulated kinase signaling pathway in articular chondrocytes. Biochem Biophys Res Commun. 2000, 275: 789-794. 10.1006/bbrc.2000.3385.View ArticlePubMedGoogle Scholar
- Tonomura H, Takahashi KA, Mazda O, Arai Y, Inoue A, Terauchi R, Shin-Ya M, Kishida T, Imanishi J, Kubo T: Glutamine protects articular chondrocytes from heat stress and NO-induced apoptosis with HSP70 expression. Osteoarthritis Cartilage. 2006, 14: 545-553. 10.1016/j.joca.2005.12.008.View ArticlePubMedGoogle Scholar
- Chevalley T, Rizzoli R, Manen D, Caverzasio J, Bonjour JP: Arginine increases insulin-like growth factor-I production and collagen synthesis in osteoblast-like cells. Bone. 1998, 23: 103-109. 10.1016/S8756-3282(98)00081-7.View ArticlePubMedGoogle Scholar
- Harmand MF, Vilamitjana J, Maloche E, Duphil R, Ducassou D: Effects of S-adenosylmethionine on human articular chondrocyte differentiation. An in vitro study. Am J Med. 1987, 83: 48-54. 10.1016/0002-9343(87)90851-5.View ArticlePubMedGoogle Scholar
- Bellon G, Chaqour B, Wegrowski Y, Monboisse JC, Borel JP: Glutamine increases collagen gene transcription in cultured human fibroblasts. Biochim Biophys Acta. 1995, 1268: 311-323. 10.1016/0167-4889(95)00093-8.View ArticlePubMedGoogle Scholar
- Hering TM, Kollar J, Huynh TD, Varelas JB, Sandell LJ: Modulation of extracellular matrix gene expression in bovine high-density chondrocyte cultures by ascorbic acid and enzymatic resuspension. Arch Biochem Biophys. 1994, 314: 90-98. 10.1006/abbi.1994.1415.View ArticlePubMedGoogle Scholar
- Kawahara K, Nishi Y, Nakamura S, Uchiyama S, Nishiuchi Y, Nakazawa T, Ohkubo T, Kobayashi Y: Effect of hydration on the stability of the collagen-like triple-helical structure of [4(R)-hydroxyprolyl-4(R)-hydroxyprolylglycine]10. Biochemistry. 2005, 44: 15812-15822. 10.1021/bi051619m.View ArticlePubMedGoogle Scholar
- Malemud CJ, Papay RS: Stimulation of cyclic AMP in chondrocyte cultures: effects on sulfated-proteoglycan synthesis. FEBS Lett. 1984, 167: 343-351. 10.1016/0014-5793(84)80154-4.View ArticlePubMedGoogle Scholar
- Beckman KB, Ames BN: Oxidative decay of DNA. J Biol Chem. 1997, 272: 19633-19636. 10.1074/jbc.272.32.19633.View ArticlePubMedGoogle Scholar
- Kahn A, Pottenger LA, Albertini JG, Taitz AD, Thonar EJ: Chemical stabilization of cartilage matrix. J Surg Res. 1994, 56: 302-308. 10.1006/jsre.1994.1047.View ArticlePubMedGoogle Scholar
- Yagami K, Suh JY, Enomoto-Iwamoto M, Koyama E, Abrams WR, Shapiro IM, Pacifici M, Iwamoto M: Matrix GLA protein is a developmental regulator of chondrocyte mineralization and, when constitutively expressed, blocks endochondral and intramembranous ossification in the limb. J Cell Biol. 1999, 147: 1097-1108. 10.1083/jcb.147.5.1097.PubMed CentralView ArticlePubMedGoogle Scholar
- Wang L, Hinoi E, Takemori A, Takarada T, Yoneda Y: Abolition of chondral mineralization by group III metabotropic glutamate receptors expressed in rodent cartilage. Br J Pharmacol. 2005, 146: 732-743. 10.1038/sj.bjp.0706358.PubMed CentralView ArticlePubMedGoogle Scholar
- Schwartz ER, Adamy L: Effect of ascorbic acid on arylsulfatase activities and sulfated proteoglycan metabolism in chondrocyte cultures. J Clin Invest. 1977, 60: 96-106.PubMed CentralView ArticlePubMedGoogle Scholar
- Sandell LJ, Daniel JC: Effects of ascorbic acid on collagen mRNA levels in short term chondrocyte cultures. Connect Tissue Res. 1988, 17: 11-22.View ArticlePubMedGoogle Scholar
- Reeves KD, Hassanein KM: Long-term effects of dextrose prolotherapy for anterior cruciate ligament laxity. Altern Ther Health Med. 2003, 9: 58-62.PubMedGoogle Scholar
- Downs JT, Lane CL, Nestor NB, McLellan TJ, Kelly MA, Karam GA, Mezes PS, Pelletier JP, Otterness IG: Analysis of collagenase-cleavage of type II collagen using a neoepitope ELISA. J Immunol Methods. 2001, 247: 25-34. 10.1016/S0022-1759(00)00302-1.View ArticlePubMedGoogle Scholar
- Calliet R: Ligamentous injuries. Knee Pain and Disability. Edited by: Calliet R. 1992, Philadelphia, Seoul: FA. Davis and Yeong Mun, 112-142. 3Google Scholar
- Hellio Le Graverand MP, Vignon E, Otterness IG, Hart DA: Early changes in lapine menisci during osteoarthritis development: Part I: cellular and matrix alterations. Osteoarthritis Cartilage. 2001, 9: 56-64. 10.1053/joca.2000.0350.View ArticlePubMedGoogle Scholar
- Batiste DL, Kirkley A, Laverty S, Thain LM, Spouge AR, Holdsworth DW: Ex vivo characterization of articular cartilage and bone lesions in a rabbit ACL transection model of osteoarthritis using MRI and micro-CT. Osteoarthritis Cartilage. 2004, 12: 986-996. 10.1016/j.joca.2004.08.010.View ArticlePubMedGoogle Scholar
- Kikuchi T, Yamada H, Shimmei M: Effect of high molecular weight hyaluronan on cartilage degeneration in a rabbit model of osteoarthritis. Osteoarthritis Cartilage. 1996, 4: 99-110. 10.1016/S1063-4584(05)80319-X.View ArticlePubMedGoogle Scholar
- Mankin HJ, Dorfman H, Lippiello L, Zarins A: Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips. II. Correlation of morphology with biochemical and metabolic data. J Bone Joint Surg Am. 1971, 53: 523-537.PubMedGoogle Scholar
- Jean YH, Wen ZH, Chang YC, Lee HS, Hsieh SP, Wu CT, Yeh CC, Wong CS: Hyaluronic acid attenuates osteoarthritis development in the anterior cruciate ligament-transected knee: Association with excitatory amino acid release in the joint dialysate. J Orthop Res. 2006, 24: 1052-1061. 10.1002/jor.20123.View ArticlePubMedGoogle Scholar
- Amiel D, Abel MF, Kleiner JB, Lieber RL, Akeson WH: Synovial fluid nutrient delivery in the diathrial joint: an analysis of rabbit knee ligaments. J Orthop Res. 1986, 4: 90-95. 10.1002/jor.1100040111.View ArticlePubMedGoogle Scholar
- Goggs R, Vaughan-Thomas A, Clegg PD, Carter SD, Innes JF, Mobasheri A, Shakibaei M, Schwab W, Bondy CA: Nutraceutical therapies for degenerative joint diseases: a critical review. Crit Rev Food Sci Nutr. 2005, 45: 145-164. 10.1080/10408690590956341.View ArticlePubMedGoogle Scholar
- Mobasheri A, Vannucci SJ, Bondy CA, Carter SD, Innes JF, Arteaga MF, Trujillo E, Ferraz I, Shakibaei M, Martin-Vasallo P: Glucose transport and metabolism in chondrocytes: a key to understanding chondrogenesis, skeletal development and cartilage degradation in osteoarthritis. Histol Histopathol. 2002, 17: 1239-1267.PubMedGoogle Scholar
- Clark AG, Rohrbaugh AL, Otterness I, Kraus VB: The effects of ascorbic acid on cartilage metabolism in guinea pig articular cartilage explants. Matrix Biol. 2002, 21: 175-184. 10.1016/S0945-053X(01)00193-7.View ArticlePubMedGoogle Scholar
- Barker GA, Wilkins RJ, Golding S, Ellory JC: Neutral amino acid transport in bovine articular chondrocytes. J Physiol. 1999, 514: 795-808. 10.1111/j.1469-7793.1999.795ad.x.PubMed CentralView ArticlePubMedGoogle Scholar
- Hinoi E, Wang L, Takemori A, Yoneda Y: Functional expression of particular isoforms of excitatory amino acid transporters by rodent cartilage. Biochem Pharmacol. 2005, 70: 70-81. 10.1016/j.bcp.2005.04.025.View ArticlePubMedGoogle Scholar
- Fafournoux P, Bruhat A, Jousse C: Amino acid regulation of gene expression. Biochem J. 2000, 351: 1-12. 10.1042/0264-6021:3510001.PubMed CentralView ArticlePubMedGoogle Scholar
- Haussinger D: The role of cellular hydration in the regulation of cell function. Biochem J. 1996, 313: 697-710.PubMed CentralView ArticlePubMedGoogle Scholar
- Reeves KD: Prolotherapy: Basic science, clinical studies, and technique. Pain Procedures in Clinical Practice. Edited by: Lennard TA. 2000, Philadelphia: Hanley & Belfus, 172-190. 2Google Scholar
- Reeves KD: Prolotherapy: present and future applications in soft tissue pain and disability. Phys Med Rehab Clin North Am. 1995, 6: 917-926.Google Scholar
- Borghetti P, Della Salda L, De Angelis E, Maltarello MC, Petronini PG, Cabassi E, Marcato PS, Maraldi NM, Borghetti AF: Adaptive cellular response to osmotic stress in pig articular chondrocytes. Tissue Cell. 1995, 27: 173-183. 10.1016/S0040-8166(95)80020-4.View ArticlePubMedGoogle Scholar
- James CB, Uhl TL: A review of articular cartilage pathology and the use of glucosamine sulfate. J Athl Train. 2001, 36: 413-419.PubMed CentralPubMedGoogle Scholar
- Lo GH, LaValley M, McAlindon T, Felson DT: Intra-articular hyaluronic acid in treatment of knee osteoarthritis: a meta-analysis. Jama. 2003, 290: 3115-3121. 10.1001/jama.290.23.3115.View ArticlePubMedGoogle Scholar
- Handley CJ, Lowther DA: Inhibition of proteoglycan biosynthesis by hyaluronic acid in chondrocytes in cell culture. Biochim Biophys Acta. 1976, 444: 69-74.View ArticlePubMedGoogle Scholar
- Weisl H: Intertrochanteric osteotomy for osteoarthritis. A long-term follow-up. J Bone Joint Surg Br. 1980, 62-B: 37-42.PubMedGoogle Scholar
- Beaver RJ, Mahomed M, Backstein D, Davis A, Zukor DJ, Gross AE: Fresh osteochondral allografts for post-traumatic defects in the knee. A survivorship analysis. J Bone Joint Surg Br. 1992, 74: 105-110.PubMedGoogle Scholar
- Paletta GA, Arnoczky SP, Warren RF: The repair of osteochondral defects using an exogenous fibrin clot. An experimental study in dogs. Am J Sports Med. 1992, 20: 725-731.View ArticlePubMedGoogle Scholar
- Isogai N, Morotomi T, Hayakawa S, Munakata H, Tabata Y, Ikada Y, Kamiishi H: Combined chondrocyte-copolymer implantation with slow release of basic fibroblast growth factor for tissue engineering an auricular cartilage construct. J Biomed Mater Res A. 2005, 74: 408-418.View ArticlePubMedGoogle Scholar
- Gilbert MS, Aledort LM, Seremetis S, Needleman B, Oloumi G, Forster A: Long term evaluation of septic arthritis in hemophilic patients. Clin Orthop Relat Res. 1996, 328: 54-59. 10.1097/00003086-199607000-00011.View ArticlePubMedGoogle Scholar
- Chodorowska G, Tomczyk M, Glowacka A: Basic-fibroblast growth factor (b-FGF): its biological role in physiologic and pathologic conditions. Ann Univ Mariae Curie Sklodowska [Med]. 2004, 59: 286-291.Google Scholar
- Caplan AI, Elyaderani M, Mochizuki Y, Wakitani S, Goldberg VM: Principles of cartilage repair and regeneration. Clin Orthop Relat Res. 1997, 342: 254-269. 10.1097/00003086-199709000-00033.View ArticlePubMedGoogle Scholar
- Buckwalter JA: Evaluating methods of restoring cartilaginous articular surfaces. Clin Orthop Relat Res. 1999, S224-S238. 10.1097/00003086-199910001-00022. 367 Suppl
This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.