Advertisement

The Effect of Ketorolac Tromethamine, Methylprednisolone, and Platelet-Rich Plasma on Human Chondrocyte and Tenocyte Viability

      Purpose

      The purpose of this study was to evaluate the effect on cell viability of the isolated and combined use of allogeneic platelet-rich plasma (PRP) and ketorolac tromethamine on human chondrocytes and tenocytes in a highly controlled in vitro environment.

      Methods

      PRP was produced from 8 subjects. Human chondrocytes (Lonza, Hopkinton, MA) and tenocytes isolated from samples of the long head of the biceps tendons were treated in culture with PRP, ketorolac tromethamine, and methylprednisolone, both alone and in combination. Control samples were treated in media containing 2% or 10% fetal bovine serum (FBS). Cells were exposed for 1 hour. Luminescence assays were obtained to examine cell viability after 24 hours and long-term effects on cell viability after 120 hours. Radioactive thymidine assay was used to measure proliferation after 120 hours.

      Results

      For chondrocytes, cell viability (120 hours) increased significantly with the treatment of PRP alone (43,949 ± 28,104 cells; P < .001) and with the combination of ketorolac tromethamine and PRP (43,276 ± 31,208; P < .001), compared with the 2% FBS group (7,397 ± 470). Cell viability decreased significantly after exposure to methylprednisolone (1,323 ± 776; P < .001) and its combination with PRP (4,381 ± 5,116; p < .001). For tenocytes, cell viability (120 hours) was significantly higher with the treatment of PRP (61,287 ± 23,273; P < .001) and the combined treatment of ketorolac tromethamine and PRP (52,025 ± 17,307; P < .001), compared with the 2% FBS group (23,042 ± 2,973). Cell viability decreased significantly after exposure to methylprednisolone (3,934 ± 1,791; P = .001) and its combination with PRP (5,201 ± 2,834; P = .003), compared with 2% FBS.

      Conclusions

      Tendon and cartilage cells showed increased cell viability after an exposure to allogeneic PRP and ketorolac tromethamine. Exposure to methylprednisolone alone decreased cell viability, and addition of PRP could partially reverse this negative effect.

      Clinical Relevance

      Intra-articular injections of pain-modifying or anti-inflammatory drugs are routinely given in orthopaedic practice. Among the many agents available for intra-articular injection, corticosteroids and local anesthetics are the most common in clinical practice. Potential detrimental side effects of intra-articular injections of corticosteroids and local anesthetics have prompted investigation into alternative treatment options such as combinations of PRP and ketorolac tromethamine. In vitro evaluation of their effect on cell viability might build a basis for further translational research and clinical application.
      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Arthroscopy
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Harvey W.F.
        • Hunter D.J.
        The role of analgesics and intra-articular injections in disease management.
        Med Clin North Am. 2009; 93: 201-211
        • MacMahon P.J.
        • Eustace S.J.
        • Kavanagh E.C.
        Injectable corticosteroid and local anesthetic preparations: A review for radiologists.
        Radiology. 2009; 252: 647-661
        • Behrens F.
        • Shepard N.
        • Mitchell N.
        Alterations of rabbit articular cartilage by intra-articular injections of glucocorticoids.
        J Bone Joint Surg Am. 1975; 57: 70-76
        • Papacrhistou G.
        • Anagnostou S.
        • Katsorhis T.
        The effect of intraarticular hydrocortisone injection on the articular cartilage of rabbits.
        Acta Orthop Scand Suppl. 1997; 275: 132-134
        • Nichols A.W.
        Complications associated with the use of corticosteroids in the treatment of athletic injuries.
        Clin J Sport Med. 2005; 15: 370-375
        • Farkas B.
        • Kvell K.
        • Czompoly T.
        • Illes T.
        • Bardos T.
        Increased chondrocyte death after steroid and local anesthetic combination.
        Clin Orthop Relat Res. 2010; 468: 3112-3120
        • Hall M.P.
        • Band P.A.
        • Meislin R.J.
        • Jazrawi L.M.
        • Cardone D.A.
        Platelet-rich plasma: Current concepts and application in sports medicine.
        J Am Acad Orthop Surg. 2009; 17: 602-608
        • Lopez-Vidriero E.
        • Goulding K.A.
        • Simon D.A.
        • Sanchez M.
        • Johnson D.H.
        The use of platelet-rich plasma in arthroscopy and sports medicine: Optimizing the healing environment.
        Arthroscopy. 2010; 26: 269-278
        • van Buul G.M.
        • Koevoet W.L.
        • Kops N.
        • et al.
        Platelet-rich plasma releasate inhibits inflammatory processes in osteoarthritic chondrocytes.
        Am J Sports Med. 2011; 39: 2362-2370
        • Filardo G.
        • Kon E.
        • Buda R.
        • et al.
        Platelet-rich plasma intra-articular knee injections for the treatment of degenerative cartilage lesions and osteoarthritis.
        Knee Surg Sports Traumatol Arthrosc. 2011; 19: 528-535
        • Kon E.
        • Buda R.
        • Filardo G.
        • et al.
        Platelet-rich plasma: Intra-articular knee injections produced favorable results on degenerative cartilage lesions.
        Knee Surg Sports Traumatol Arthrosc. 2010; 18: 472-479
        • Sanchez M.
        • Anitua E.
        • Azofra J.
        • Aguirre J.J.
        • Andia I.
        Intra-articular injection of an autologous preparation rich in growth factors for the treatment of knee OA: A retrospective cohort study.
        Clin Exp Rheumatol. 2008; 26: 910-913
        • Calmet J.
        • Esteve C.
        • Boada S.
        • Gine J.
        Analgesic effect of intra-articular ketorolac in knee arthroscopy: Comparison of morphine and bupivacaine.
        Knee Surg Sports Traumatol Arthrosc. 2004; 12: 552-555
        • Irwin M.G.
        • Cheung K.M.
        • Nicholls J.M.
        • Thompson N.
        Intra-articular injection of ketorolac in the rat knee joint: Effect on articular cartilage and synovium.
        Br J Anaesth. 1998; 80: 837-839
        • Kostamovaara P.A.
        • Hendolin H.
        • Kokki H.
        • Nuutinen L.S.
        Ketorolac, diclofenac and ketoprofen are equally efficacious for pain relief after total hip replacement surgery.
        Br J Anaesth. 1998; 81: 369-372
        • Morley-Forster P.
        • Newton P.T.
        • Cook M.J.
        Ketorolac and indomethacin are equally efficacious for the relief of minor postoperative pain.
        Can J Anaesth. 1993; 40: 1126-1130
        • Lee S.C.
        • Rha D.W.
        • Chang W.H.
        Rapid analgesic onset of intra-articular hyaluronic acid with ketorolac in osteoarthritis of the knee.
        J Back Musculoskelet Rehabil. 2011; 24: 31-38
        • Dogan N.
        • Erdem A.F.
        • Gundogdu C.
        • Kursad H.
        • Kizilkaya M.
        The effects of ketorolac and morphine on articular cartilage and synovium in the rabbit knee joint.
        Can J Physiol Pharmacol. 2004; 82: 502-505
        • Shapiro P.S.
        • Rohde R.S.
        • Froimson M.I.
        • Lash R.H.
        • Postak P.
        • Greenwald A.S.
        The effect of local corticosteroid or ketorolac exposure on histologic and biomechanical properties of rabbit tendon and cartilage.
        Hand (NY). 2007; 2: 165-172
        • Foster T.E.
        • Puskas B.L.
        • Mandelbaum B.R.
        • Gerhardt M.B.
        • Rodeo S.A.
        Platelet-rich plasma: From basic science to clinical applications.
        Am J Sports Med. 2009; 37: 2259-2272
        • Anitua E.
        • Andia I.
        • Sanchez M.
        • et al.
        Autologous preparations rich in growth factors promote proliferation and induce VEGF and HGF production by human tendon cells in culture.
        J Orthop Res. 2005; 23: 281-286
        • de Mos M.
        • van der Windt A.E.
        • Jahr H.
        • et al.
        Can platelet-rich plasma enhance tendon repair? A cell culture study.
        Am J Sports Med. 2008; 36: 1171-1178
        • Baboldashti N.Z.
        • Poulsen R.C.
        • Franklin S.L.
        • Thompson M.S.
        • Hulley P.A.
        Platelet-rich plasma protects tenocytes from adverse side effects of dexamethasone and ciprofloxacin.
        Am J Sports Med. 2011; 39: 1929-1935
        • Wong M.W.
        • Tang Y.Y.
        • Lee S.K.
        • Fu B.S.
        Glucocorticoids suppress proteoglycan production by human tenocytes.
        Acta Orthop. 2005; 76: 927-931
        • Wong M.W.
        • Tang Y.Y.
        • Lee S.K.
        • Fu B.S.
        • Chan B.P.
        • Chan C.K.
        Effect of dexamethasone on cultured human tenocytes and its reversibility by platelet-derived growth factor.
        J Bone Joint Surg Am. 2003; 85: 1914-1920
        • Mazzocca A.D.
        • McCarthy M.B.
        • Chowaniec D.M.
        • et al.
        Platelet-rich plasma differs according to preparation method and human variability.
        J Bone Joint Surg Am. 2012; 94: 308-316
        • Mazzocca A.D.
        • McCarthy M.B.
        • Chowaniec D.M.
        • et al.
        The positive effects of different platelet-rich plasma methods on human muscle, bone, and tendon cells.
        Am J Sports Med. 2012; 40: 1742-1749
        • Haag J.
        • Voigt R.
        • Soeder S.
        • Aigner T.
        Efficient non-viral transfection of primary human adult chondrocytes in a high-throughput format.
        Osteoarthritis Cartilage. 2009; 17: 813-817
        • Pauly S.
        • Klatte F.
        • Strobel C.
        • et al.
        Characterization of tendon cell cultures of the human rotator cuff.
        Eur Cell Mater. 2010; 20: 84-97
        • Yao L.
        • Bestwick C.S.
        • Bestwick L.A.
        • Maffulli N.
        • Aspden R.M.
        Phenotypic drift in human tenocyte culture.
        Tissue Eng. 2006; 12: 1843-1849
        • Mazzocca A.D.
        • McCarthy M.B.
        • Intravia J.
        • et al.
        An in vitro evaluation of the anti-inflammatory effects of platelet-rich plasma, ketorolac, and methylprednisolone.
        Arthroscopy. 2013; 29: 675-683
        • Baker J.F.
        • Byrne D.P.
        • Walsh P.M.
        • Mulhall K.J.
        Human chondrocyte viability after treatment with local anesthetic and/or magnesium: Results from an in vitro study.
        Arthroscopy. 2011; 27: 213-217
        • Maffulli N.
        • Ewen S.W.
        • Waterston S.W.
        • Reaper J.
        • Barrass V.
        Tenocytes from ruptured and tendinopathic achilles tendons produce greater quantities of type III collagen than tenocytes from normal achilles tendons. An in vitro model of human tendon healing.
        Am J Sports Med. 2000; 28: 499-505
        • Ng K.W.
        • Leong D.T.
        • Hutmacher D.W.
        The challenge to measure cell proliferation in two and three dimensions.
        Tissue Eng. 2005; 11: 182-191
        • Ay S.
        • Kucuk D.
        • Gumus C.
        • Kara M.I.
        Distribution and absorption of local anesthetics in inferior alveolar nerve block: Evaluation by magnetic resonance imaging.
        J Oral Maxillofac Surg. 2011; 69: 2722-2730
        • Carofino B.
        • Chowaniec D.M.
        • McCarthy M.B.
        • et al.
        Corticosteroids and local anesthetics decrease positive effects of platelet-rich plasma: An in vitro study on human tendon cells.
        Arthroscopy. 2012; 28: 711-719
        • Dang A.B.
        • McCarthy M.B.
        • Chowaniec D.M.
        • Mazzocca A.D.
        Effects of adding epinephrine to arthroscopic irrigation fluid on cultured chondrocyte survival in vitro.
        Arthroscopy. 2011; 27: 1118-1122
        • Burkhardt H.
        • Schwingel M.
        • Menninger H.
        • Macartney H.W.
        • Tschesche H.
        Oxygen radicals as effectors of cartilage destruction. Direct degradative effect on matrix components and indirect action via activation of latent collagenase from polymorphonuclear leukocytes.
        Arthritis Rheum. 1986; 29: 379-387
        • Sharma P.
        • Maffulli N.
        Tendon injury and tendinopathy: Healing and repair.
        J Bone Joint Surg Am. 2005; 87: 187-202
        • Lee S.Y.
        • Suh J.K.
        • Choi J.H.
        • Jeon W.J.
        • Cheong M.A.
        Effect of ketorolac and diclofenac on the impairment of endothelium-dependent relaxation induced by reactive oxygen species in rabbit abdominal aorta.
        Korean J Anesthesiol. 2010; 59: 196-202
        • Mei-Dan O.
        • Carmont M.R.
        • Laver L.
        • Mann G.
        • Maffulli N.
        • Nyska M.
        Platelet-rich plasma or hyaluronate in the management of osteochondral lesions of the talus.
        Am J Sports Med. 2012; 40: 534-541
        • Alsousou J.
        • Thompson M.
        • Hulley P.
        • Noble A.
        • Willett K.
        The biology of platelet-rich plasma and its application in trauma and orthopaedic surgery: A review of the literature.
        J Bone Joint Surg Br. 2009; 91: 987-996
        • Dhillon M.
        • Patel S.
        • Bali K.
        Platelet-rich plasma intra-articular knee injections for the treatment of degenerative cartilage lesions and osteoarthritis.
        Knee Surg Sports Traumatol Arthrosc. 2011; 19 (863-864; author reply 855-856)
        • Sanchez M.
        • Anitua E.
        • Orive G.
        • Mujika I.
        • Andia I.
        Platelet-rich therapies in the treatment of orthopaedic sport injuries.
        Sports Med. 2009; 39: 345-354
        • Cheng J.
        • Abdi S.
        Complications of joint, tendon, and muscle injections.
        Tech Reg Anesth Pain Manag. 2007; 11: 141-147
        • Sendzik J.
        • Shakibaei M.
        • Schafer-Korting M.
        • Lode H.
        • Stahlmann R.
        Synergistic effects of dexamethasone and quinolones on human-derived tendon cells.
        Int J Antimicrob Agents. 2009; 35: 366-374
        • Scutt N.
        • Rolf C.G.
        • Scutt A.
        Glucocorticoids inhibit tenocyte proliferation and tendon progenitor cell recruitment.
        J Orthop Res. 2006; 24: 173-182
        • Oikarinen A.I.
        • Vuorio E.I.
        • Zaragoza E.J.
        • Palotie A.
        • Chu M.L.
        • Uitto J.
        Modulation of collagen metabolism by glucocorticoids. Receptor-mediated effects of dexamethasone on collagen biosynthesis in chick embryo fibroblasts and chondrocytes.
        Biochem Pharmacol. 1988; 37: 1451-1462
        • Davis D.
        • Cyriac M.
        • Ge D.
        • You Z.
        • Savoie F.H.
        In vitro cytotoxic effects of benzalkonium chloride in corticosteroid injection suspension.
        J Bone Joint Surg Am. 2010; 92: 129-137
        • Grishko V.
        • Xu M.
        • Wilson G.
        • Pearsall A.W.
        Apoptosis and mitochondrial dysfunction in human chondrocytes following exposure to lidocaine, bupivacaine, and ropivacaine.
        J Bone Joint Surg Am. 2010; 92: 609-618
        • Maffulli N.
        • Longo U.G.
        • Denaro V.
        Novel approaches for the management of tendinopathy.
        J Bone Joint Surg Am. 2010; 92: 2604-2613
        • Coombes B.K.
        • Bisset L.
        • Vicenzino B.
        Efficacy and safety of corticosteroid injections and other injections for management of tendinopathy: A systematic review of randomised controlled trials.
        Lancet. 2010; 376: 1751-1767
        • Axelsson K.
        • Gupta A.
        • Johanzon E.
        • et al.
        Intraarticular administration of ketorolac, morphine, and ropivacaine combined with intraarticular patient-controlled regional analgesia for pain relief after shoulder surgery: A randomized, double-blind study.
        Anesth Analg. 2008; 106 (table of contents): 328-333